Granulocytic stem cells in Friend leukemia. Friend virus induces a leukemia characterized by the proliferation of neoplastic hematopoietic cells believed to be erythroid precursors. In vitro studies were conducted with spleen cells from mice with terminal Firend leukemia in order to determine their capacity for leukocytic differentiation. Spleen cells were obtained from leukemic DBA/2 mice 1 to 2 days before anticipated death and cultured in the presence or absence of colony-stimulating activity (CSA). Growth in liquid culture in dissusion chambers was dependent on CSA and resulted in the generation of normally differentiated granulocytes and macrophages. Colony formation in agar was also dependent on CSA, and the cloning efficiency of leukemic spleen cells was found to be approximately 10 times normal. The colonies formed were composed of leukocytes, which appeared morphologically normal. Total in vitro colony-forming units per leukemic spleen exceeded normal by more than 300-fold, but cells elaborating CSA were decreased. Although it is uncertain whether the stem cells stimulated by CSA are "normal" or leukemic," it is clear that Friend leukemia has profound effects on the proliferation and differentiation of nonerythroid stem cells.


Critical evaluation of lymphocyte functions in urological cancer patients. One hundred three patients with varying stages of urological cancer (bladder, prostate, kidney) were investigated with regard to the following lymphocyte functions. T-cells were assessed numerically (E rosettes), their blastogenic response to phytohemagglutinin (pHA) was determined, and their cytotoxic potential against heterologous target cells in short-term presence of PHA (e.i., PHA-dependent cellular cytotoxocity) was evaluated. Similarly, B cells were numerically assessed (EA rosettes), and their function was evaluated by antibody-dependent cellular cytotoxicity against antibody-coated deterologous target cells. The data on cancer patients, divided on the basis of extent of disease and prior radiation therapy, were compared to those of normal young and age-matched controls. Our investigations emphasize the importance of the following factors: (a) comparison of data with age-matched controls, since several lymphocyte functions appear to change with age; (b) use of multiple controls to compensate for the inherent variability found in certain tests; (c) minimized contamination by nonlymphoid cells in the purified cell preparation; and (d) the influence of certain treatment regimens (radiation, chemotherapy, etc.) on the results. Radiotherapy significantly depressed T-cell number with a depression of PHA blastogenic responses as well as PHA-dependent cellular cytotoxicity. When all of these conditions were taken into account, the urological cancer patients as a group were found to have a lower proportional value of E rosettes (T-cells) and a reduced PHA blastogenic responsiveness. Certain cancer patients displayed an elevated PHA-dependent cellular cytotoxicity as compared to age-matched controls, which may indicate the presence of activated cells in the presence of tumors. With this identification of a group of cancer patients with markedly depressed E rosette values and PHA responsiveness, it will now be possible to follow them clinically in comparison with a group of cancer patients with normal T-cell functions.


Stimulatory effects of 5-lodo-2'-deoxyuridine on number and function of splenic B- and T-cells and of macrophages in C3HeB/FeJ mice. 5-lodo-2'-deoxyuridine (lUdR) significantly increases the total number of spleen cells on the fourth day after administration of 100 mg/kg in normal C3HeB/FeJ male mice. The number of splenic B-cells [cells sensitive to the cytotoxic activity of a rabbit antiserum to mouse F(ab)2, plus complement] increases on the same day. The number of T-cells (cells sensitive to the cytotoxic activity of a rabbit antiserum to mouse brain-associated theta antigen, plus complement) and of macrophages (adherent-phagocytic cells) does not increase. Spleen cells from lUdR-treated mice show heightened responsiveness in vitro to Escherichia coli lipopolysaccharide or concanavalin A on Day 4 and Days 5 to 6, respectively. The rate of clearance of i.v.-injected colloidal carbon is also increased for 3 days after lUdR. Thus, lUdR is able to functionally activate either T- or B-cells or macrophages, but only to increase the number of splenic B-cells.


The ultrastructure of taste and touch receptors of the frog's taste organ. The taste buds from fungiform papillae and the hard palate of frogs were investigated with the scanning and transmission electron microscopes. An immature pre-taste cell and a mature taste cell can be differentiated. Only the mature taste cell exhibits synaptic contact with the afferent taste fibre. Glandular and satellite supporting cells envelop the thin apical processes of the sensory cells. At the base of the taste disc up to 10 Merkel cells form a complex with nerve endings. There are two types of myelinated fibres, large and small. The small fibre innervates the taste cells, the thicker nerve fibre the Merkel cells. The occurrence of two types of receptors explains physiological results.


The fine structure of the perineural endothelium. Fine strands of motor nerves were examined with the electron microscope using thin section as well as freeze-etching techniques. The specimens were taken from frog cutaneous pectoris nerve, rat sciatic nerve, mouse and shrew phrenic nerves and from human skin nerves. The perineural sheath (Henle, Ranvier, Key and Retzius) consists of one to several concentric laminae of endothelial cells; it encases nerve fascicles and eventually individual nerve fibers and terminals. The endothelial cells are extremely thin and fitted togeether smoothly by overlap and dove-tailing of their border zones. The cell contacts are formed by continuous zonulae occludentes, often reinforced by maculae adhaerentes, and in depth they comprise 3-15 strands with an average of 5-6 strands per junction. The membranes of endothelial cells are studded with attachment sites and stomata of plasmalemmal vesicles suggesting a high level of pinocytotic activity. This phenomenon is by no means restricted to the external laminae of the endothelial sheath. Each endothelial lamina is vested with basement membranes on both (epineural and endoneural) sides, and the spaces between laminae contain a few collagen fibers and fibroblasts. Occasionally, punctate tight junctions are seen between laminae. Cytological evidence supports the hypothesis that the perineural endothelium provides a relatively tight and highly selective barrier separating the peripheral nerves from surrounding tissue and its extracellular fluid spaces. This effect is achieved on the one hand by the sealing of pericellular spaces and on the other hand by a membrane controlled transcellular transport mechanism (pinocytosis), both of which are enhanced by their serial arrangement.


[The so-called immunologic classification of non-Hodgkin's malignant lymphoma]. The so-called immunologic classification of malignant lymphomas was used in 100 of suitable cases. Without the simultaneous use of immunocytological methods the classification was  applicable with certain restrictions only, but, in general the entire classification scheme could be used in routine bioptic practice. The so-called immunologic classification permits a more accurate determination of the nature of the neoplasms and a larger flexibility in classifying them considering the possible trend of alterations that may occur during the course of the disease. Some difficulties and points of confusion have been discussed.


[Primary pulmonary histiocytosis X].

[Primary pulmonary histiocytosis X]. The diagnosis of a pulmonary form of histiocytosis X was made on the basis of a lung tissue biopsy in a male aged 60 suffering from progressive dyspnoe of respiratory type and a rentgenological finding of tiny disseminated foci in both lung fields. The bad prognosis of the process related to the fibrous transformation of neoplastic foci rather than to the biological nature of the process as such.


Individual household water supplies as a control measure against Schistosoma mansoni. A study in rural St Lucia. As part of a programme to evaluate single control measures for reducing the transmission of Schistosoma mansoni, household water supplies were installed in 5 rural settlements in the Riche Fond Valley of St Lucia. About 2 000 persons who previously were dependent on rivers and streams are now receiving safe water at their homes. The systems provide useful design data on individual water requirements in rural areas. This experience suggests that future rural water systems can be designed more economically and efficiently by using consumption rates that are closer to the actual requirements and by eliminating water wastage at the taps.


Abnormalities in the microsomal oxidases of the WHO standard reference strain of Musca domestica. Observations made during biochemical and toxicological studies of the housefly, in which the WHO standard reference (SR) strain was used as a standard, indicated that this strain differs from other strains in certain physiological and toxicological aspects. Experiments are described in which microsomal oxidases prepared from the standard reference strain and several other housefly strains, both susceptible and resistant to insecticides, were compared for heat stability, sensitivity to ionic strength, pH optima, and the spectral characteristics of their cytochrome P-450. In a comparison with 8 strains the SR strain microsomes were more stable at high incubation temperatures (40-47.5 degrees C). Compared to two other strains (NAIDM, susceptible, and Isolan-B, resistant) SR was more stable at low pH and more sensitive to high ionic strength. In spectral comparisons with NAIDM and Isolan-B, SR was found to contain microsomal cytochrome P-450 that differed slightly in its carbon monoxide or octylamine binding spectra. The results suggest that this strain may not provide a suitable reference in studies involving the microsomal oxidases. However, these apparent differences should not reduce the value of the SR stain as a reference in insecticide resistance studies.


Field application to malaria studies of the passive haemagglutination (PHA) test with lyophilized cells. Lyophilized cells for the passive (formerly indirect) haemagglutination test for malaria were tested in a small field trial. A comparison was made with results obtained with samples from the same persons by another method in another laboratory (Center for Disease Control, Atlanta, GA, USA). The use of lyophilized cells of standardized reactivity clearly adds to the simplicity and rapidity of the test, and justifies its recommendation for use in field studies.


Single-radial-hemolysis: a new method for the assay of antibody to influenza haemagglutinin. Applications for diagnosis and seroepidemiologic surveillance of influenza. A simple single-radial-haemolysis technique is described that permits the detection and assay of antibody to influenza virus haemagglutinin. The method depends on the passive haemolysis of virus-treated erythrocytes by antihaemagglutinin antibody and complement. Under the "standard" test conditions described antibody to the other surface antigen of the influenza virus (neuraminidase) or to the internal antigens of the virus (nucleoprotein and matrix protein) do not produce haemolysis. Because it requires only small amounts of crude virus antigen and is rapid and simple, the method appears to be of considerable value for large-scale seroepidemiologic studies of new influenza virus variants. Antihaemagglutinin antibody detected by single-radial-haemolysis appears to be relatively strain-specific; the technique may therefore be useful in the antigenic characterization of virus isolates.


Experimental study of the role of inactivated vaccine in two-step vaccination against smallpox. In experiments on rabbits it was found that although administration of inactivated smallpox vaccine did not induce a demonstrable antibody response in the serum it enhanced the immune response to subsequent inoculation with live vaccine. The dose of inactivated vaccine corresponded to 8 x 10(7) PFU before inactivation (by (60)Co gamma-radiation); the dose of live vaccine was 1.2 x 10(5) PFU. When the interval between the two inoculations was 7-days, virus-neutralizing antibody appeared after 5 days and reached levels 2-4 times those obtained with live vaccine alone. With longer intervals (up to 60 days) the enhancement of the immune response was even greater. It seems likely that use of the two-step method may reduce the incidence of post-vaccination encephalitis and clinical studies to determine the optimum conditions for safety and efficacy are at present being undertaken.


Controlled field trial of a typhoid vaccine prepared with a nonmotile mutant of Salmonella typhi Ty2. A controlled field trial was performed in Egypt to evaluate a whole cell typhoid vaccine prepared with a nonmotile mutant of S. typhi Ty2 (TNM1) devoid of flagellar antigen. This vaccine did not elicit an H antibody response, but significant Vi and O agglutinin responses were observed. There were 34 typhoid cases among 21 063 six- to seven-year-old children who received the TNM1 vaccine, and 44 cases among 21 017 children in the control group who received tetanus toxoid. These results suggest that TNM1 vaccine does not provide protection against typhoid fever, and that H antigen may be an important component of an effective vaccine.


Control of Schistosoma mansoni transmission by provision of domestic water supplies. A preliminary report of a study in St Lucia. As an experimental control measure to reduce the transmission of S. mansoni, an individual household water supply was provided in 400 houses in 5 rural settlements of the Riche Fond Valley, St Lucia. This population of about 2 000 had previously been dependent for water on infective streams and rivers. Six other settlements in the valley, all provided with limited piped water from public standpipes, served as the comparison area. After 2 years the incidence, prevalence, and intensity of infection with S. mansoni were significantly lower in the household water supply area, whereas all these indices of infection had increased in the comparison area. An adequate, reliable, and convenient supply of water can reduce the transmission of S. mansoni and should be considered as a control measure in other endemic areas.


Inheritance of resistance to organophosphorus insecticides in Culex tarsalis Coquillet. The mode of inheritance of resistance to parathion, fenitrothion, fenthion, chlorpyr fosmethyl, and parathion-methyl was studied in a strain of C. tarsalis. Resistance in the F(1) hybrid was partially dominant. Backcross experiments indicated that resistance is derived from more than one gene. This conclusion was confirmed by repeated backcrosses of the F(1) to the susceptible strain and selection of the backcross progeny with a discriminating dosage of parathion-methyl.


Left ventricular end-diastolic pressure volume relationships with experimental acute global ischemia. The mechanism of elevation of left ventricular end-diastolic pressure during acute global ischemia was evaluated by examiniation of the relative contributions of a decrease in contractility and an alteration of the pressure-volume relationship. The external circumference (mercury-in-silastic gauge) pressure relationship, as an index of the pressure-volume relationship, was studied in beta adrenergic and ganglionic blocked, open chest dogs on right heart bypass at constant heart rate ane aortic pressure. Ischemia of one and two hours' duration was produced by reducing total coronary blood flow in cannulated left and right coronary arteries until left ventricular end-diastolic pressure rose significantly. At a constant stroke work, left ventricular end-diastolic pressure rose from 5.0 +/- 0.5 to 15.0 +/- 0.5 cm H2O in the experiments of one hour of ischemia, and from 7.0 +/- 1.0 to 17.0 +/- 1.0 cm H2O in experiments of two hours of ischemia. Ischemia was followed by one hour of restoration of coronary blood flow. Ischemia produced a marked depression of ventricular function: stroke work, considered at a left ventricular end-diastolic pressure of 15 cm H2O, decreased from 21.0 +/- 3.0 to 3.5 +/- 0.5 gm-m, and from 15.0 +/- 2.0 to 2.5 +/- 0.5 gm-m, in the experiments of one and two hours, respectively. Neither ischemia nor reflow changed the pressure-volume relationship. Thus, the elevation of left ventricular end-diastolic pressure during ischemia in an otherwise normal canine myocardium is due to a decrease in systolic performance of the heart rather than to an alteration of the pressure-volume relationship.


Enhanced uptake by guinea-pig macrophages of radio-iodinated human aggregated immunoglobulin G in the presence of sera from rheumatoid patients with cutaneous vasculitis. A competitive radiobioassay method for soluble immune complexes in sera has been used to study sera from patients with rheumatoid arthritis. Twenty-eight out of sixty-two sera from selected seropositive rheumatoid patients were found to produce enhanced uptake of radio-iodinated human aggregated IgG by guinea-pig macrophages in contrast with the inhibition of uptake seen with SLE sera. Twenty-two out of the twenty-eight enhancing sera belonged to patients with cutaneous vasculitis. None of the twenty-two sera from seronegative rheumatoid patients possessed enhancing properties and none of these patients had clinical evidence of vasculitis. Three patients with seropositive SLE had sera with enhancing properties and had cutaneous vasculitis. We have shown that the enhancement is probably due to the presence in sera of immune complexes or altered IgG bound to rheumatoid factor. The possible mechanism has been investigated and it appears that there is increased binding of radio-iodinated aggregated IgG to free valencies on rheumatoid factor already bound to immune complexes.


Thyrogastric autoimmune disease. Studies on the cell-mediated immune system and histocompatibility antigens. Cell-mediated immune responses were studied in autoimmune diseases of thyrogastric type, Hashimoto's thyroiditis and autoimmune pernicious anaemia-type gastritis. Specific cell-mediated immunity was investigated by the leucocyte migration inhibition procedure, and general cell-mediated immunity (T-cell performance) was studied by standard in vivo and in vitro tests. In thyrogastric autoimmune diseases inhibition of migration of leucocytes was induced by thyroglobulin and gastric parietal cell microsomes; under conditions of presumably low cellular sensitization, stimulation of migration was observed. There was no depression of general cell-mediated immunity, in contrast to what occurs in systemic lupus erythematosus and related autoimmune diseases. A weak association of autoimmune gastritis with HL-A3 and HL-A7 (P LESS THAN 0.05) lost significance when an appropriate correction was applied; this weakness with HL-A clearly does not explain the strong genetic component in thyroid and gastric autoimmunity.


Correlations among cutaneous reactivity to DNCB, PHA-induced lymphocyte blastogenesis and peripheral blood E rosettes. Comparisons of the results obtained in a study of fifty-two patients with genitourinary malignancies using three assays to monitor the thymus-dependent immune system (delayed cutaneous hypersensitivity to DNCB, PHA-induced lymphocyte blastogenesis, and peripheral blood E rosette-forming lymphocyte counts) yielded statistically significant positive correlations between the DNCB and PHA assays in twenty-one of twenty-eight instances, the DNCB and E rosette assays in thirteen of sixteen instances, the PHA and E rosette assays in twenty-eight of thirty-six instances, and among the DNCB, PHA and E rosette assays in ten of fourteen instances. The results suggest that both PHA-stimulated lymphocyte blastogenesis and peripheral blood E rosette-forming lymphocyte levels provide meaningful in vitro correlates of cell-mediated immunity.


C3 receptor. A marker of a thymus-dependent B-cell subpopulation. Depletion of C3-binding cells from spleen virtually abolishes the T cell-dependent response to sheep erythrocytes as assessed by transfer experiments. It is suggested that a functionally thymus-dependent B-cell subpopulation (B2) can be distinguished by surface receptors for C3.


Effect of azathioprine on in vitro antibody response. Differential effect on B cells involved in thymus-dependent and independent responses. The effect of azathioprine (Az) on the primary in vitro antibody response of mouse spleen cell cultures has been studied. The response towards T cell-dependent antigens is suppressed by low Az concentrations (50% inhibition by 10(-2) mug/ml and 100% suppression by 10(-1) mug/ml). The same pattern is observed when Az addition is delayed until day 2, but the suppression is absent or partial when Az is added on day 3. In the early period (day 0 to day 1) the effect of Az is reversible upon addition of an excess of purine nucleosides. In contrast, the response to a T cell-independent antigen (TNP-T4) is relatively insensitive to Az, since 100-fold higher drug concentrations are required to obtain an inhibition. With the assumption that T helper cells are likely to be highly sensitive to Az, the effect of low Az concentrations on the other two cell populations involved in T cell-dependent responses has been evaluated. Adherent cells appear unaffected. In contrast, the B-cell response is markedly sensitive to Az, as shown by the effect of Az on the response of nude mouse cells to a T cell-dependent antigen in the presence of T-cell products, either specific or non-specific. On the other hand, the B-cell response to mitogens is resistant to az. Thus, Az has a differential effect on B-cell response according to the thymus dependency of the antigen. This may suggest the existence of two pathways for B-cell activation or two different B-cell subpopulations.


Blastogenic response of Toxoplasma-infected mouse spleen cells to T- and B-cell mitogens. In order to differentially test the function of lymphocytes in Toxoplasma gondii-infected mice, the in vitro blastogenic response of spleen cell cultures to non-specific mitogens was studied. Phytohaemagglutinin (PHA) and concanavalin A (Con A) stimulation were used as tests of thymus-dependent lymphocyte (T cell) function and bacterial endotoxin lipopolysaccharide (LPS) was used as a probe of bursal equivalent lymphocyte (B cell) function. For the first 3 weeks following T. gondii infection, the uptake of tritiated thymidine ([3H]TdR) by spleen cells cultured with all three mitogens was markedly reduced in comparison to the uptake in spleen cells from uninfected control mice. Thereafter, the response to LPS returned to normal while stimulation by the T-cell mitogens (PHA and Con A) remained depressed. It is postulated that T. gondii infection either: (1) diluted out T cells in the spleen with unreactive cells; (2) modified T cells in such a way that they were less responsive to mitogens; (3) depleted the peripheral lymphoid tissues of T cells; (4) induced non-specific suppressor cells, which inhibited the T-cell function assays; or (5) activated macrophages which depressed T-cell function non-specifically.


Increased sensitivity of the rosette-forming reaction of human T lymphocytes with sheep erythrocytes afforded by papain treatment of the sheep cells. Human T lymphocytes are known to adhere to sheep erythrocytes. This affinity reaction was found to be consistently increased following treatment of the sheep cells with papain. Reactions with treated erythrocytes were more stable and involved higher percentages of lymphocytes. Using a double marker system of papain-treated sheep erythrocytes together with Fc rosettes as a presumptive B-cell marker, there was an overlap population of only 1% lymphocytes. Sheep erythrocytes treated with papain therefore appear to remain specifically adherent to human T lymphocytes.


T and B lymphocytes in multiple sclerosis. The percentage and total number of E and EAC rosettes, as indicators of T and B lymphocytes respectively, were studied in the blood of subjects with multiple sclerosis (MS) and normals. MS patients in acute exacerbation were found to have a decrease in E rosettes and an increase in EAC rosettes. The relationship of these findings to the pathogenesis of MS is unclear; several possible pathogenetic implications are considered.


Cell-mediated immunity in patients on long-term haemodialysis. Several parameters of cell-mediated immunity were assessed in thirty patients with chronic renal failure treated with long-term haemodialysis. Lymphopenia was uncommon, and only two patients showed diminished numbers of thymus-derived peripheral blood lymphocytes. Skin test energy to three antigens was documented in only one patient. No serious infections with intracellular organisms were noted in any of the patients. Phytohaemagglutinin (PHA) induced DNA synthesis in eleven patients was lower than that shown by normal subjects. This poor responsiveness to PHA was not corrected by haemodialysis, and in only two patients was the response improved by substituting normal human plasma for uraemic plasma in the lymphocyte cultures. Thus with the exception of PHA response, other parameters of cell-mediated immunity were intact in this group of chronic stable haemodialysis patients.


The value of tests for antibodies to DNA in monitoring the clinical course of SLE. A long-term study using the Farr test and the DNA counterimmunoelectrophoretic method. Serial serum samples from fifteen patients with SLE, taken over periods varying from 6 months to 6 years, were tested for DNA binding capacity, DNA electroprecipitins (DNA-EP) and C'3 level to assess the value of these investigations in reflecting clinical disease activity. Patients with renal involvement showed a good correlation between high levels of DNA binding, low serum C'3 and disease activity and typically, their DNA-EP was negative. By contrast, patients without renal involvement in whom vasculitis was prominent, showed a poor correlation of DNA binding capacity to changes in the state of their disease although the DNA-EP test was persistently positive. It was also apparent that both the DNA-BC and C'3 can show marked variation in response to alterations in treatment without accompanying clinical change. Although these serological tests, particularly the DNA binding capacity are of recognized value in the diagnosis of SLE, they serve most usefully as guides to long-term management when they can be related to the clinical pattern of the disease.


Comparative mapping of the local distribution of immunoglobulin-containing cells in ulcerative colitis and Crohn's disease of the colon. The local response pattern of immunoglobulin-containing cells was compared in Crohn's disease and ulcerative colitis by paired immunohistochemistry on specimens of the large bowel wall. In the "Crohn mucosa" with persisting glands the total cell count was on the average raised more than three times compared with controls. The numbers of IgA, IgM and IgG immunocytes were increased 2.0, 4.8 and 28.6 times, respectively. Only 0-2 IgD- and IgE-containing cells were generally found per section. No consistent differences in the mucosal response pattern were revealed when Crohn's disease was compared with ulcerative colitis. The deeper layers of the bowel wall were in both diseases more or less densely infiltrated by immunocytes-IgG cells compromising about 80%. Immunoglobulin-containing cells in the muscularis propria and subserosa were characteristically found in Crohn's disease. There was no indication of a primary defect in the secretory immunoglobulin system which appeared to be normal in areas with intact glands. The pronounced local humoral immune response, particularly that involving IgG, might be of pathogenetic importance by aggravating and perpetuating in the inflammatory bowel disease.


Activation of human peripheral blood lymphocytes by concanavalin A dependence of monocytes. The activation of human peripheral blood lymphocytes or isolated T lymphocytes by concanavalin A (Con A) is hightly potentiated by the presence of autologous, mitomycin C-treated monocytes. The optimal lymphocyte: monocyte ratio within a broad dose range is 1:1 when the incorporation of [14C]thymidine is expressed as total incorporation per culture tube and 1:10 when expressed per lymphocyte. A five-to-ten-fold increase of total DNA synthesis is noted in the presence of 10-90% monocytes. The data may help to explain the wide variations in Con A responsiveness of human peripheral lymphocytes which may be partly related to differences in purification which give rise to cell preparations containing varying amounts of monocytes.


Effect of radiotherapy on lymphocyte cytotoxicity in vitro. The cytotoxic functions of highly purified blood lymphocytes from patients with breast cancer were studied before and after radiotherapy. Addition of PHA or of rabbit antibodies to target cells (chicken erythrocytes) were chosen as two means of inducing lymphocyte cytotoxicity in vitro. The proportion of T and non-T-lymphocytes was determined by means of E and EAC rosette tests. The antibody-induced cytotoxocity of lymphocytes decreased following radiotherapy while that mediated by PHA remained unchanged. There was some reduction in the percentage of EAC rosette-forming cells. These results, as well as our earlier observations, suggest that the decrease in the peripheral blood of the proportion of lymphocytes with receptors for activated complement is responsible for changes in the antibody-mediated lymphocyte cytotoxicity.


In vitro lymphocyte reactivity and T-cell levels in chronic cigarette smokers. Peripheral blood total leucocyte, lymphocyte and thymus-dependent lymphocyte (T cell) levels and in vitro lymphocyte reactivity (LR) to phytohaemagglutinin (PHA) were determined in 153 chronic cigarette smokers and 115 non-smokers ranging in age from 20 to 78 years. Total leucocyte, lymphocyte and T-cell levels were significantly elevated in smokers. There was no correlation with age. LR to PHA was significantly higher in smokers less than 40 years of age or in those with a 20 pack-year or less history of cigarette consumption. Older smokers or those with a history of heavier cigarette consumption did not differ from normals. The results contrast with previous demonstrations of suppression of immune reactivity after exposure to tobacco products. The possible effects of the apparent stimulation of the lymphoid system by chromic cigarette consumption is considered.


Spontaneous autorosette-forming cells in man. A marker for a subset population of T lymphocytes? A subpopulation of human peripheral blood lymphocytes are capable of binding with human (autologous or allogeneic) erythrocytes, forming rosettes. The conditions which lead to autorosette formation are similar to those required for sheep red-cell rosetting. Ageing human erythrocytes are shown to bear less of the determinants involved in the phenomenon than younger ones. Evidence is presented that autorosetting is a T-cell marker. As autorosette-forming cells are very sensitive to the inhibiting effects of ATG they could therefore belong to a T-cell subpopulation.


Hyperimmunoglobulinaemia, T-cell deficiency and plasmacytosis in RFM mice with host versus graft disease induced by the perinatal inoculations (T6XRFM)F1 spleen cells. Host versus graft (HVG) syndrome may be induced in parental strain mice by perinatal inoculations of F1 hybrid spleen cells. The principal manifestations of the disease include thrombocytopaenia, intravascular fibrin deposits, intestinal haemorrhage, hepatic infarcts, lymphosplenomegaly and renal disease. Immune complexes have been shown to be the cause of the renal lesions, and have been implicated as the triggers for disseminated intravascular coagulation. In the present studies of RFM mice perinatally inoculated with (T6 x RFM)F1 spleen cells (RFM/(T6 x RFM)F1 mice), quantitative determinations of serum immunoglobulins (Ig) revealed marked elevations of IgG1, IgG2, IgA and IgM. Electrophoretic analyses revealed the polyclonal pattern which typically follows chronic antigenic stimulation. However, IgG1 levels which reached 29 to 72 times control values suggested disruption of homeostatic mechanisms which control circulating Ig levels. Because antibody responses to histocompatibility antigens were present only occasionally, and then in low titre, it seemed unlikely these antigens were the principal causes of hypergammaglobulinaemia and plasmacytosis. Morphological studies indicated that the elevated levels of Ig seen in end-stage HVG syndrome correlated well with marked plasmacytosis, the third morphological finding in a sequence that included the precocious development of germinal centres and subsequent depletion of thymic-dependent (T) lymphocytes. The fact that spleen cells from RFM/(T6 x RFM)F1 mice were severely impaired in their capacity to cause graft versus host disease in related (T6 x RFM)F1 and unrelated C3H mice provided strong evidence that the HVG reaction resulted in T-cell depletion, rather than specific immunoincompetence.


Thymus dependence of the antibody response to tetanus toxoid in mice. Both the IgM and IgG antibody responses to tetanus toxoid were greatly reduced in adult thymectomized, irradiated, foetal liver reconstituted mice, in contrast with results published for neonatally thymectomized mice. This evidence lends further support to the notion that all IgG responses to protein antigens are thymus dependent.


Complete obstruction of the left main coronary artery associated with congenital pulmonary valvular stenosis. The surgical treatment of the completely obstructed left main coronary artery is aortocoronary saphenous-vein bypass to the left coronary artery system. Without surgical intervention, this lesion has an exceedingly poor prognosis. This report describes a patient with atherosclerotic obstruction of the left main coronary artery and congenital pulmonary valvular stenosis, an unusual combination of lesions not previously documented together. The patient remains asymptomatic 22 months after aortocoronary saphenous-vein bypass and pulmonary valvotomy.


A system-structured medical record for intensive care patient documentation. The problem-oriented approach to the medical record has aroused a long overdue interest in the structuring of the medical case file. Clinical information in the traditional record is source-structured and time-sequenced, whereas the problem-oriented system differs by being a problem-structured record retaining still a chronologic sequence. We have found that in acute illness the multiplicity of interacting pathophysiologic processes makes premature application of the problem-oriented approach cumbersome and unwieldy. The formulation of the problem list at an early stage often led to the reduplication of problems, creating disorder in the clinical picture rather than serving to clarify it. Some used the simple cataloguing of events and data as a substitute for clinical judgment and decision making, focusing more upon style rather than content of the medical record. By using a rigid physiologic system-structured "problem" list and a modification of the SOAP (Subjective Objective, Assessment, Plan) subdivision, we have improved the documentation of our intensive care patients. The summary of the patient's stay in the intensive care unit is structured with active and inactive problems, this summary to be further used as the permanent problem list.


[The quantitative determination of the individual CSF proteins in the diagnosis of inflammatory diseases of the CNS in children (author's transl)]. Albumin, IgG, IgA, IgM, transferrin, and alpha 1-antitrypsin were determined quantitatively in the cerebrospinal fluid (CSF) of 44 healthy children and 37 pediatric patients with central nervous system diseases. Neither IgA nor IgM were found in the CSF of normal children, but they were present in cases of purulent and non-bacterial meningitis. In cases of encephalitis all proteins studied were increased except IgA and IgM, which could not be demonstrated. On the basis of the results it may be concluded that the increase of IgG in the CSF in bacterial and a bacterial meningitis is due to an increased permeability of the blood-CSF barrier. Except in cases with ependymitis, IgG is produced only in small amounts in or near the CSF spaces even in the presence of inflammation of the meninges. The quantitative determination of IgG, IgA, and IgM in case of increased CSF protein content may facilitate the differential diagnosis between purulent and acute viral meningitis and encephalitis.


Adrenaline and the electrogenic sodium pump in Rana catesbeiana sympathetic ganglion cells. The effect of adrenaline on the Na+-pump in bullfrog (Rana catesbeiana) sympathetic ganglion cells was studied by use of electrophysiological methods. The rate of removal of excess Na+ injected into a ganglion cell was increased by adrenaline. The K+-activated hyperpolarization of cell membrane, which might be produced by an electrogenic Na+-pump, was also increased by adrenaline. These results suggested that adrenaline was able to accelerate the Na+-pump, possibly the electrogenic Na+-pump.


[The role of the hypothalamus in the mechanism of positional nystagmus]. The typical positional nystagmus was induced in 30 intact rabbits with injection of 0.1-0.4 ml of 0.03-0.05% dicaine solution into the cerebral cisterna magna. The positional nystagmus resulted, supposedly, from the effect of the anesthetic upon the basal structures of the 3rd ventricle of the brain, upon the hypothalamus in particular. The trigger mechanism of the phenomenon is the displacement of the membraneous ampullae of semicircular canals at the tilting of the head. To prove this, 0.01-0.03 ml of 4% dicaine solution were injected directly into the anterior and posterior hypothalamus through a metallic cannula. When introduced into the anterior hypothalamus the dose of the anesthetic necessary for occurrence of the positional nystagmus is two or three times lesser than when it is injected into the posterior nypothalamus. In two delabyrinthed rabbits we failed to induce the nystagmus in this way.


Differential effects of vasopressin on the water, calcium and lysosomal enzyme contents of mitochondria-rich and lysosome-rich (granular) epithelial cells isolated from bullfrog urinary bladder. Treatment of bullfrog urinary bladder with arginine vasopressin (AVP) elicited a dose-dependent increase in the basal movement of water and sodium across isolated tissues. Epithelial cells from the mucosal surface and incubated with 10 mU AVP/ml for 30 min retained a greater amount of intracellular water and calcium than cells not treated with hormone. The epithelial cells were further separated into two major fractions by density gradient centrifugation; cells damaged during these manipulations were separated from viable cells and discarded. Morphologcal examination of the two respective fractions indicated that they largely consisted of mitochondria-rich (MR) and granular (G) cell types which line the lumen of bullfrog bladder. The calcium content of MR cells averaged 25% greater than that of G-type cells. G cells had a markedly higher content of the characteristic lysosomal hydrolases, acid phsophatase and cathepsin B1, than that found in MR cells. Incubation of G cells with AVP elicited significant increments in water and calcium contents and extracellular release of lysosomal enzymes as compared to untreated cells. Among MR cells treated with AVP, cell calcium declines slightly but no significant increase in water content or extracellular hydrolase activity was detected in comparison  with paired control cells. The physiological significance of acid proteinase release from G cells treated with AVP was evaluated in experiments with intact bladder. Proteinase inhibitors which suppress the activity of cathepsin B1 selectively antagonized the action of hormone on water permeation. The data suggest that alterations in the calcium and lysosomal hydrolase activity associated with G cells exposed to AVP may contribute to the hormone-induced water flow observed across the intact epithelium.


[Influence of imipramine on arousal from winter hibernation]. Imipramine (20 and 50 mg/kg intraperitoneally) is shown to delay the arousal of gophers from hibernation. This effect is similar to the action of serotonin (5 mg/kg) and 5-oxytryptophan (50 mg/kg). In the latter, however, the action on the thermogenesis is most pronounced and, because of this the warming up of the gophers is inhibited, while imipramine acts on the arousal. Tranylcypromine (20 mg/kg) failed to exert any substantial effect on the worming up and arousal of gophers from hibernation and attenuated the action of imipramine.


In vitro studies of inflammatory bowel disease. Surface receptors of the mononuclear cell required to lyse allogeneic colonic epithelial cells. Peripheral blood mononuclear cells from patients with either Crohn's disease or ulcerative colitis (collectively referred to as inflammatory bowel disease) are cytotoxic in vitro for isologous or allogeneic colonic epithelial cells. Utilizing the ability of thymus-derived (T) lymphocytes to bind sheep red blood cells to their surface and the property of bone marrow-derived (B) lymphocytes to display easily detectable surface immunoglobulin determinants, the cytoxicity of these lymphocyte subpopulations was tested. The results indicate that the mononuclear cell required for the lysis of colonic epithelium was not included within the bulk of T or B lymphocytes. Indeed, enrichment for cytotoxicity correlated best with enrichment for a mononuclear cell population lacking classical T or B markers. Additionally, mononuclear cells specifically adhering to plastic petri dishes coated with heat-aggregated immunoglobulin, and thus presumably bearing a surface Fc receptor, were enriched in their cytotoxicity. Alternatively, cells not adhering to aggregated Ig-coated petri dishes were relatively depleted of cytotoxicity. The implications of these findings as they relate to possible interactions between cellular and humoral immune mechanisms as a pathogenic mechanism for the colonic inflammatory process noted in patients with inflammatory bowel disease are discussed.


Isolation and kinetic characteristics of mucosal lymphocytes in Crohn's Disease. Single cell suspensions of lymphocytes have been prepared from human gut mucosa, and short-term culture used to characterize the kinetics of DNA synthesis of mucosal lymphocytes. High rates of DNA synthesis were found in 72-hr cultures of Crohn's disease mucosal lymphocytes, compared with normal subjects. These high rates correlated with the appearance of numerous transformed blast cells in terminated cultures. Lymphocytes from normal and abnormal human gut mucosa were stimulated by phytohemagglutinin-P, except in some patients with Crohn's disease with high rates of "spontaneous" DNA synthesis. The isolation and culture of human gut mucosal lymphocytes by a simple preparative technique will enable the direct study of lymphocyte function in both normal and diseased gut mucosa.


Antibody-dependent cell-mediated cytotoxicity. Comparative ability of murine Peyer's patch and spleen cells to lyse lipopolysaccharide-coated and uncoated erythrocytes. Lymphoid cells from nonsensitized donors can be cytotoxic in vitro for target cells coated with antibody. The antibody can be directed either against antigens which are intrinsic to or experimentally attached to the target cell surface. Killing of target cells by this method has been termed antibody-dependent cell-mediated cytotoxicity (ADCC). In these investigations we compared the ability of Peyer's patch cells and spleen cells from nonsensitized mice to lyse lipoplysaccharide-coated and uncoated target cells by an ADCC mechanism. The results demonstrate that spleen cells, but not Peyer's patch cells, from identical mice can be cytotoxic for lipopolysaccharide-coated chicken erythrocytes (CRBC) in the presence of anti-lipopolysaccharide antibody and uncoated CRBC in the presence of anti-CRBC antibody. Thus, Peyer's patches in mice appear to be deficient in the effector cell type(s) required to kill antibody-coated erythrocyte target cells by an ADCC mechanism as well as the macrophage-like accessory adherent cell type(s) required for the induction of humoral antibody synthesis and cell-mediated cytotoxic allograft reactions in vitro.


Spontaneous bacterial peritonitis induced by intraarterial vasopressin therapy. Two patients developed spontaneous bacterial peritonitis after infusions of vasopressin into the superior mesenteric or gastroduodenal arteries for upper gastrointestinal hemorrhage. The peritonitis in these patients differed from the typical picture in which a single aerobic organism is responsible, by the presence of multiple organisms, some of which were anaerobic. These findings suggest that the arterial vasoconstriction decreased the integrity of the intestinal mucosal barrier and permitted the transmural migration of enteric organisms from the lumen of the bowel into the ascites-filled peritoneal cavity.


[Treatment of Spastic Gait Disorders Using an "Electro-Gait-Stimulator ("Schrittstimulator") (author's transl)]. A short historical outline of electric treatment of spasticity is given. A specially developed management with a "gait-stimulator" is described. Four muscles of each lower extremity being mainly engaged in walk were electrically stimulated in the physiological sequences according to the normal gait. The used electric impulses were of a duration of 0,25 msec and an intensity up to 700 V. Using such a "gain-stimulator" in spastic-paraparetic patients a reduced spasticity has been achieved. Positive effect of this treatment has been mostly pronounced, when the programming of impulses was adjusted to the end of the expected physiological contraction of the corresponding muscles. Physiological and pathological data of the "Silent period" is proposed to be mostly involved. The application of impulses in physiological sequences seems to reactivate normal reflex - mechanisms which are disturbed by supraspinal laesion. The results indicate that the electric impulses activates muscle-sensory - organs and that impulses on these organs produce a pace-making function on the spinal cord, which lessens spasticity.


[Synthesis and biological activity of some cystine peptides from the active center of the basic trypsin inhibitor from bovine organs (Kunitz inhibitor) (author's transl)]. The synthesis of the following 3 model peptides from the active center of the inhibitor molecule is described: (see article). The 3 peptides are split by trypsin with velocities increasing from (I) to (III). The following Ki values were determined: (I) Ki = 2.3 x 10(-2)M; (II) Ki = 4.9 x 10(-3)M; (III) Ki = 2.1 x 10(-3)M. These Ki values differ from the Ki values of the Kunitz inhibitor by a factor 10(11) to 10(12).


Cytogenetic and cell-surface marker studies in two non-Hodgkin's lymphomata of T-cell origin. The chromosomes from 2 cases of lymphoma involving proliferation of abnormal T-lymphocytes are described. The tumours differed histologically and chromosomally, although certain of the structural chromosome changes found are of common occurrence in other, mostly B-cell, lymphomata.


Evidence against T-cell immunoglobulin from radioimmunoassay on serum and cells from bursectomized chickens. A radioimmunoassay with a sensitivity of about 50 pg of chicken immunoglobulin (Ig) was developed and employed for the estimation of Ig in the serum and in extracts of cells from hormonally bursectomized chickens. Chickens, which were judged agammaglobulinaemic by rocket immunoelectrophoresis, had at 12 weeks of age 5-100 ng of Ig/ml of serum. The Ig concentration was decreasing with a half life of 5 days. Thymus and spleen cells from such hypogammaglobulinaemic chickens were extracted with non-ionic detergents, acid urea, or combinations of urea and detergent, and the extracts were analysed for Ig by the inhibition assay. The amount of Ig found in extracts from thymocytes corresponded to less than a hundred molecules of 7S Ig/cell with a lower value of less than five molecules/cell, while extracts of spleen cells contained Ig corresponding to 60-300 molecules of Ig/cell. The inclusion in the media of a number of protease inhibitors did not increase the recovery. Extracts of thymus and spleen cells from untreated chickens contained Ig corresponding to 10-20 X 10(3) and 50-150 X 10(3) molecules of Ig/cell respectively. The results demonstrate the profound hypogammaglobulinaemia of bursectomized chickens and question the concept of an active production of receptor Ig by bursa-independent T cells.


Requirement for T cells in the antibody response of mice to calf skin collagen. The antibody response to triple-helical calf skin collagen is apparently regulated by T cells since nude mice (BALB/c strain) responded to this antigen only after transfer of T cells. Syngeneic transfer of B and T cells was required to prepare thymectomized and irradiated normal C57B1/10 recipients for an anti-collagen response. Reconstitution by B cells alone was unsuccessful. The results are compatible with previous evidence on the H-2 linked genetic control of the antibody response to and the non-repetitious nature of helical antigenic determinants in calf collagen.


Lymphocyte response to T-cell mitogen during experimental gingivitis in humans. This study was conducted to evaluate the dose response relationships of peripheral blood lymphocytes (PBL) by stimulation of phytohemagglutinin (PHA) during the onset of oral inflammation. Eleven dental students underwent a 3-week experimental gingivitis program (Löe et al., 1965). At time zero, weeks 1, 2, and 3, and after 1 week of reinstituted oral hygiene (week 4), the plaque accumulations were evaluated, the degree of gingival inflammation was assessed, and a blood sample was taken. Quadruplicate microcultures each containing 2 x 10(5) PBL in 0.2 ml of tissue culture medium 199 and 10% fetal calf serum were stimulated with five concentrations of PHA (10 to 0.5 mug/ml) and incubated for 78 h at 37 C in 5% CO2. [3H]thymidine was added to each culture for the final 8 h. The cultures were then harvested and counted by liquid scintillation, and stimulation indexes (SI) were determined. At time zero the maximum PBL response occurred at a PHA concentration of 5 mug/ml (SI = 100). During weeks 1, 2, and 3 the location of the maximum PBL response shifted to a lower PHA concentration (1.0 mug/ml) and increased to over SI =400. The phenomenon of shifting peak PHA responses to lower PHA concentrations could be observed after only 1 week of developing gingival inflammation. The PBL response returned to pre-experimental values after 1 week of reinstituted oral hygiene, which resolved the oral inflammation. The findings show that a dose response relationship exists between PHA concentrations and the PBL response. If these dose response changes seen during developing gingival inflammation are ignored, either a decrease, increase, or no change in PBL response can be shown depending upon the PHA concentration evaluated. Owing to the dose-dependent nature of this PBL response, it is advisable to routinely use dose response curves in order to properly evaluate the full responsiveness of PBL to mitogenic substances.


Cellular immunity in Peyer's patches of rats infected with Trichinella spiralis. A rat model of Trichinella spiralis gut infection was used to observe the sequence of developing cellular immunity in Peyer's patches and other lymphoid tissues. Whereas cellular reactivity (lymphocyte blastogenesis) for worm antigens was evident in mesenteric lymph nodes draining the gastrointestinal tract within 3 days after infection, Peyer's patch lymphocytes developed maximal reactivity 2 to 3 weeks later at the same time as the spleen and other lymphoid tissues. Furthermore, the immune reactivity found in Peyer's patches was only transient. Thus, in this parasitic gut infection, the Peyer's patch lymphoid tissue does not appear to be the first site of cellular responsiveness but rather to acquire cellular reactivity only when other lymphoid elements in the infected host have also acquired similar antigen-induced reactivity.


T and CR+ lymphocyte profile in leprosy and the effect of treatment. Thymus-derived lymphocytes (T lymphocytes) and complement receptor-bearing lymphocytes (CR+ lymphocytes) were estimated by using erythrocyte rosettes and erythrocyte-antibody-complement rosettes as markers in untreated lepromatous and untreated tuberculoid patients and in healthy controls. Treated lepromatous cases were also investigated. Ten cases of untreated lepromatous patients were reassessed 6 months or more after therapy commenced. A significant decrease in both percentages and absolute numbers of CR+ cells in the untreated lepromatous leprosy subjects was observed. This decrease showed a return to normal levels after treatment. The percentage of T cells in the untreated lepromatous cases was normal; however, the absolute numbers of T cells and the total lymphocyte count showed a significant decrease. After therapy, the T cell population was unchanged but the total number of lymphocytes increased significantly with treatment. The absolute number of T and CR+ cells was significantly less in the untreated than in the treated lepromatous patients.


Effect of viral and bacterial pneumonias on cell-mediated immunity in humans. Cell-mediated immunity (CMI) was assessed during infection and after convalescence in 12 patients with influenza pneumonia and 10 patients with bacterial pneumonia. The patients with influenza pneumonia had a marked impairment of skin test reactivity, and their lymphocytes showed a diminished response to phytohemagglutinin and streptokinase-streptodornase stimulation in vitro. Suppression of CMI was related to the severity of the pneumonia. Patients with bacterial pneumonia showed as great a suppression of the response to phytohemagglutinin and streptokinase-streptodornase as the patients with viral pneumonia. All parameters of CMI returned to normal in both groups after convalescence. The depression of CMI could not be related to a decrease in the number of thymus-derived lymphocytes or to serum-suppressive factors in these patients.


Quantitation of B and T lymphocytes in guinea pigs with evidence for a release of both cell types from the spleen into the blood. B and T lymphocytes were quantitated in lymphoid organs and in blood of young normal guinea pigs by the use of EAC rosettes and rabbit erythrocyte (RE) rosettes as markers. Special attention was focused on the release of B and T cells from the spleen, estimated from the difference between the content of B and T cells in splenic efferent and afferent blood. The following frequencies of resette-forming cells (RFC) were found. Thymus: 0.3% EAC- and 88% RE-RFC; spleen: 44% EAC- and 46% RE-RFC; lymph nodes: 13% EAC- and 39% RE-RFC; arterial blood: 13% EAC- and 42% RE-RFC, and bone marrow: 2% EAC- and 7% RE-RFC. A fairly large number of cells in the lymph nodes and blood could not be identified by any of the two markers. Some possible explanations for this are discussed. The content of both EAC- and RE-RFC in splenic efferent blood significantly exceeded that in the afferent blood, indicating a release of both B and T cells from the spleen into the blood. The possibility of a release of a third type of mononuclear cell cannot be excluded from the present results.


A case of unusual SLE related syndrome characterized by erythema multiforme, angioneurotic edema, marked hypocomplementemia, and Clq precipitins of the low molecular weight type. Our patient and those of Agnello et al. had identical clinical symptoms such as erythema multiforme, arthralgias and angioneurotic edema and both differed from systemic lupus erthematosus in several important points, i.e., in spite of marked hypocomplementemia the nephropathy is not prominent and it needs high-dose steroids to eliminate the clinical and serological abnormalities. In addition to the features reported by Agnello et al. we found increased viral antibody titers in our patient's sera.


Different surface glycoprotein patterns on human T-, B- and leukemic-lymphocytes. We have labelled the exposed surface glycoproteins of human blood T- and B-lymphocytes and cells from patients with chronic lymphocytic leukemia by the galactose oxidase-tritiated sodium borohydride method. The labelled glycoproteins were separated by polyacrylamide slab gel electrophoresis and visualized by autoradiography. The T- and B-lymphocytes show different and characteristic surface glycoprotein patterns. The surface glycoprotein patterns of the leukemic cells differ from those of normal, non-malignant lymphocytes. A relationship between the altered surface glycoprotein pattern of leukemic cells and the expression of leukemia-associated antigens is discussed.


Non-T-cell resistance against a mouse Moloney lymphoma. We have previously shown that spleen cells from normal mice contain lymphocytes that kill certain in vitro-grown Moloney lymphoma lines in a 51Cr release test. The killing activity shows a marked dependence on the genotype of the donor mouse. In vivo rejection studies with a Moloney lymphoma line in various normal semisyngeneic F1 hybrids showed a clear positive correlation between in vitro natural cytotoxicity and in vivo rejection. Thus mice can be grouped as high- or low-reactive according to their in vitro cytolytic behavior as well as their in vivo rejection potential. A lymphoid cell without detectable T- or B-cell markers is responsible for the in vitro killing effect. The present study also shows that the in vivo growth inhibitory function is T-cell-independent. Lymphoid cells depleted of T- and B-cells and transferred together  with Moloney lymphoma cells into an irradiated syngeneic recipent were highly efficient in delaying tumor growth. Furthermore, syngeneic or semisyngeneic thymecto-mized, irradiated, fetal-liver-reconstituted mice showed if anything an increased in vivo resistance to a Moloney lymphoma compared to control mice. In contrast, tumor cells histoincompatible with regard to the H-2 locus showed the expected preferential growth in the thymectomized animals. We thus conclude that the major in vivo resistance against transplantation of the syngeneic or semisyngeneic Moloney lymphoma used in this study is T-independent.


Association of human alpha1-antitrypsin with anhydrotrypsin. Unlike in the reaction of anhydrotrypsin and soybean trypsin inhibitor, human alpha1-antitrypsin was found not to form a complex with bovine anhydrotrypsin. This was shown using three different methods: electrophoresis, affinity chromatography on an anhydrotrypsin-coupled Sepharose column and equilibrium competitive binding assay. These results indicate the importance of the active site serine of trypsin in formation of a complex with alpha1-antitrypsin.


Adenosine deaminase activity in chronic lymphocytic leukemia. Relationship to B- and T-cell subpopulations. The level, phenotypes, and isozyme distribution of adenosine deaminase (ADA) were determined in lymphocytes from patients with chronic lymphocytic leukemia (CLL). The ADA level in lymphocytes from patients with untreated CLL was consistently lower than in lymphocytes from normal subjects. No significant differences were found in the phenotype or isozyme distribution. In untreated patients, the ADA level was inversely correlated with the lymphocyte count and the percentage of bursa-equivalent (B) cells. After therapy, a diminution in the lymphocyte count was associated with an increase of ADA activity towards normal levels. The ADA levels were slightly higher in the thymus-derived (T) than in the B lymphocytes from normal subjects. The B cells had lower activity than T cells in patients with CLL. They also had a lower activity than the B cells from normal subjects. The ADA level was 2.3-fold higher in T cells from patients with CLL than in normal T cells. The decrease in ADA levels is an anomaly that is reversible and appears to be a reflection of the proliferation of abnormal B cells in this disorder.


Spontaneous cytotoxicity (SCMC) of normal human lymphocytes against a human melanoma cell line: a phenomenon due to a lymphotoxin-like mediator. Spontaneous cell-mediated cytotoxicity (SCMC) of normal human lymphocytes against various allogenic tumor cell lines has recently been identified as a non-T lymphocyte function. In this study evidence is presented that SCMC against a human melanoma cell line (IGR3) involves a nonspecific lymphotoxin-like mediator(s) (LT), which is rapidly produced by lymphocytes that are retained on IgG-anti-IgG columns. LT was shown to inhibit in 48-hr cultures the DNA synthesis of growing IGR3 and HeLa cell monolayers. Furthermore, in short term 51Cr-release assays using IGR3 target cells, LT increased strongly the SCMC of normal allogeneic lymphocytes, although it exhibited little cytotoxicity by itself in this assay. LT was detectable in cell-free supernatants harvested after 6 hr from co-cultures of IGR3 melanoma cells and normal effector lymphocytes, but not in supernatants from melanoma cells alone or lymphocytes alone. Lymphocyte preparations that had been passed through IgG-anti-IgG columns had lost the capacity to generate LT and were poor effectors in SCMC. However, in the presence of LT, a small proportion of null cells, which pass through IgG-anti-IgG columns, was capable of inducing strong SCMC. Absorption of the supernatants, containing LT activity, with an insolubilized rabbit-anti-human IgG antiserum did not remove the mediator, suggesting that it is not an immunoglobulin.


Activation of purified human thymus-derived (T) cells by mitogens. II. Monocyte- macrophage potentiation of mitogen-induced DNA synthesis. Thymus-derived (T) cells from peripheral blood were purified by rosette formation with neuraminidase-treated sheep red blood cells (SRBC) and centifugation on Ficoll-Hypaque. T cells recovered from the pellet were freed of SRBC by treatment with Tris-NH4Cl. T cells purified by this method showed a diminished ability to take up 3H-thymidine (3H-TdR) after mitogen stimulation when compared to the mitogenic response of an equal number of autologous peripheral blood mononuclear lymphocytes (PBL). Autologous monocytes restored the capacity of purified T cells to take up 3H-TdR in the presence of phytohemagglutinin (PHA) or Concanavalin A (Con A). The effect was proportional to the number of monocytes added. Similar restorative effects could be obtained with allogeneic or xenogeneic monocytes. These data suggest that the mitogenic stimulation of human PBL and Con A may reflect the participation of more than one cell type: the T cells and monocyte and that the genetic origin of the monocyte is not critical for augmentation of the mitogenic activation of human T cells.


The distribution of Ia antigens on the surfaces of lymphocytes. The distribution of Ia antigens was studied on murine spleen lymphocytes by an ultrastructural technique employing deep freeze-etched replicas. Ia antigens were labeled on cells from appropriate congenic and recombinant strains of mice by incubating the cells with FITC-conjugated anti-Iak antibody, followed by ferritin-coupled Fab anti-FITC. Ia antigens were detected predominantly on immunoglobulin (Ig)-bearing B lymphocytes. Antigens coded for by the entire Ik region were present on the surfaces of 95% of the positive cells (from B10.BR mice) in densely packed microclusters. Ia specificities coded for by the I-A and I-C subregions (on 4R and B10.HTT mice) exhibited a more variable pattern, with 30 to 35% of the labeled cells having sparsely distributed Ia antigens in relatively discrete microclusters. Binding of anti-Iak antibody at 37 degrees C led to patch formation but not to capping. Modulation of surface Ig left Ia antigens diffusely distributed on the cell surface, indicating that these two membrane proteins are independent molecules.


In vitro generation of secondary cell-mediated cytotoxic response against a syngeneic Gross virus-induced lymphoma in rats. Cell-mediated cytotoxic responses against a syngeneic Gross virus-induced lymphoma, (C58NT)D, in W/Fu rats were generated in vitro by using mixed lymphocyte-tumor cell cultures. The source of responding cells was either spleens from normal rats or spleens from rats carrying or having rejected (C58NT)D tumors. Mitomycin C-treated (C58NT)D tumor cells were used as stimulating cells. The secondary anti-tumor cytotoxic response occurred more rapidly and reached higher levels than the primary response, and it was antigen specific. T cells, but nor B cells or macrophages, were essential for both the induction and the effector phases of the secondary anti-tumor responses. These data suggest that specific memory T cells persist for long periods of time in the lymphoid organs of (C58NT)D immune rats, which can rapidly become cytotoxic upon re-exposure to antigen.


In vitro response of subpopulations of human tonsil lymphocytes. I. Cellular collaboration in the proliferative response to PHA and Con A. Human tonsil lymphocytes have been separated into three subpopulations of cells: purified B cells and two subsets of purified T cells (F1 and F2). B cells were obtained by rosetting with neuraminidase treated SRBC. F1 and F2 were separated by filtration on a nylon wool column using different speeds of elution. Purified B cells contained less than 5% T cells, the T cells preparations contained less than 5% B cells for F1 and 10 to 15% for F2, respectively. A significant contamination in cells not identified by any B or T marker was observed in purified B cells and in F1. Adherent cells enhanced the response of each lymphochte population to PHA and Con A. This explained the paradoxically low responsiveness of the purified T cells. Purified B cells did not respond to these mitogens in different culture conditions. However, a small B cell response was observed when they were cultured in the presence of mitomycin-treated T cells. Striking was the enhancing effect of B cells on the T cell response to PHA and Con A. This enhancing effect was observed even when B cells were treated with mitomycin or depleted in adherent cells. The comparison of the F1 and F2 response suggested that they contained distinct types of T cells.


In vitro response of subpopulations of human lymphocytes. II. DNA synthesis induced by anti-immunoglobulin antibodies. A significant and constant increase in DNA synthesis was observed in human lymphocytes cultured in the presence of purified anti-immunoglobulin antibodies specific for human IgG, IgA, and IgM. This has been found in cultures of lymphocytes isolated from blood, tonsils, spleen, and lymph nodes. The optimal culture conditions for blood and tonsil lymphocytes were determined. As a rule 6-day cultures containing 2 x 10(6) cells/ml and 100 mug/ml of antibody yielded the highest 3H-thymidine uptake. Purified T cell cultures could not be stimulated, whereas a low response could be observed in most of the purified B cell cultures. Optimal culture conditions were the same for the B and total tonsil lymphocytes. However, when the purified B cells were totally depleted of T cells, no response was observed. A T and  B cell synergy has been demonstrated by supplementing B cell cultures with purified T cells, whether treated or not with mitomycin. These experiments indicated a permissive and potentiating effect of T cells on the B cell response. Cultures containing mitomycin-treated B cells and purified T cells (mB + T) could be stimulated by a-Ig, thus indicating a T cell proliferation. In keeping with this finding was the observation of an increased response of total lymphocytes supplemented with T cells but not with B cells. Adherent cells are necessary for an optimal response to a-Ig; they enhanced the B cell proliferation observed in (Tm + B) cultures and suppressed the response of T cells in (T + Bm) cultures.


The role of lymphotoxin in target cell destruction induced by mitogen-activated human lymphocytes in vitro. II. The correlation of temperature and trypsin-sensitive phases of lymphotoxin-induced and lymphocyte-mediated cytotoxicity. The in vitro destruction of phytohemagglutinin (PHA) coated Beta L cells by non-immune human lymphocytes was resolved into two distinct phases--lymphocyte dependent and lymphocyte independent. The initial or lymphocyte-dependent phase occurred within the first 2 hr and proceeded equally well at 34 and 37 degrees C. The amount of lymphotoxin (LT) secreted by PHA-activated human lymphocytes in vitro to PHA stimulation was the same at 34 and 37 degrees C. Antiserum and complement inactivation of the aggressor lymphocytes at various intervals revealed that target cell lysis was lymphocyte independent. However, the latter phase was temperature dependent, i.e., proceeding at the permissive temperature of 37 degrees C, but inhibited at the restrictive temperature of 34 degrees C. Further experiments revealed that LT-induced destruction had the same temperature sensitivity as target cell cytolysis occurring during the lymphocyte-independent step. Trypsin treatment of target cells during an early period of the lymphocyte-independent phase protected the target cell from subsequent death, indicating the aggressor lymphocyte has deposited a cytotoxic effector material on its surface. These results suggest the lymphocyte-dependent stage involves the processes required for the induction of LT synthesis and secretion. The actual cytolysis occurring during the lymphocyte-independent stage may be caused by LT or LT-like material(s) deposited on the target cell surface by the mitogen-activated human lymphocyte.


Rosétte formation between human b lymphocytes and zymosan-C3 complexes using different complement sources. Sera from five animal species were studied as complement source in the preparation of zymosan-C3 complexes used to detect human B lymphocytes. There was no significant difference in the determination of the percentage of ZC rosette forming cells due to differences in complement source. Human B lymphocytes could not discriminate C3 from different species studied. Every one of these sources of complement may be used interchangeably without altering the percentage of B cells detected by this method.


Induction of specific anti-guinea pig T cell sera in rabbits. In an attempt to increase the specificity of antisera raised in rabbits against strain 2 guinea pig thymocytes and brain, the rabbits were screened for titres of natural antibodies to thymocytes and other lymphocytes. Although unimmunized rabbits commonly had moderate titres of cytotoxic antibodies to guinea pig thymocytes, occasional animals had low titres to thymocytes and moderate titres to bone marrow cells. Intravenous immunization of this latter group of rabbits with thymocytes led to the production of high titred anti-thymocyte sera which were easily made specific for thymus-derived lymphocytes (T cells) by absorption with L2 C lymphoma, a bone marrow-derived lymphoma of strain 2 guinea pigs. Sera raised against guinea pig brain in complete Freund's adjuvant which had high titres of antibodies to both thymocytes and bone marrow cells could be made specific for T cells only with great difficulty. The cytotoxic activity of the anti-T cell serum could be absorbed by strain 2 thymocytes and brain homogenates, while high dilutions of this serum inhibited the formation of spontaneous rosettes between guinea pig lymphoid cells and normal rabbit erythrocytes.


75SeL-selenomethionine: a new isotopic marker for lymphocyte localisation studies. L-selenomethionine (SeM), an analogue of methionine containing Se, is incorporated by mouse and rat lymphocytes in vitro: labelled with 75Se (75SeM) it emits gamma-rays distinguishable in energy from those of 125I and 51Cr. The usefulness of 75SeM for lymphocyte localisation studies was investigated. Both B and T cells incorporated 75SeM. Reutilization of the isotope in the mouse after injection of killed cells or allogeneic cells into immunized recipients was 1-2% of the injected dose. The immunocompetence of labelled mouse T and B cells was assessed in irradiated recipients, no effect of 75SeM incorporation being found. The distribution of labelled lymphocytes from various tissues in syngeneic recipients was found to be comparable to that previously obtained with 51Cr.


Mechanism of inhibition of adoptive transfer of tuberculin sensitivity in acute uremia. Cell-mediated immunity is reduced in uremia, but the mechanism of this inhibition is poorly understood. To determine the site of inhibition a study of adoptive transfer of cell-mediated immunity was undertaken. Strain 13 guinea pigs were made uremic by ureteral ligation. Lymph node cells were isolated and transferred in the following three ways: Group 1: tuberculin sensitive nonuremic donors to nonimmune nonuremic recipients; Group 2: tuberculin sensitive nonuremic donors to nonimmune uremic recipients; Group 3: tuberculin sensitive uremic donors to nonimmune nonuremic recipients. There was no difference in the skin test responses of the transfer recipients in Groups 1 and 3, even though the uremic donors in Group 3 had lost their skin test reactivity. Transfer of tuberculin sensitivity was not successful in Group 2. These results indicate that the uremic state inhibits previously established skin test reactivity but does not prevent sensitized T lymphocytes from transferring cellular immunity.


Otosclerotic obliteration of oval window niche: an analysis of the results of surgery. 1. The massive otosclerotic focus, obliterating the oval window niche, has a relatively high case incidence of 11-2 per cent in South Australia. The three classes of obliterated footplate are defined. 2. Basic data of 109 consecutive surgical cases of obliterative otosclerosis are given. A brief description of operative techniques--vein graft and polyethylene tube in nine, and piston technique in 100--is given and the difficulties and complications arising at or after surgery are discussed. 3. Post-operative follow-up of patients with yearly audiometric assessment has been achieved in 105/109 (= 96-33%) at four years, and in 94/109 (= 86-26%) at five years. 4. The results of surgery are presented by many and varied methods including bar diagrams, post-operative hearing gains, post-operative bone-air gaps, speech discrimination studies and standard statistical analysis techniques. 5. The vein graft technique (nine cases) is very much inferior to the piston technique. Vein graft cases had a 33% incidence of profound sensori-neural loss due to bony reclosure of the window. 6. The piston technique (100 cases) gave hearing gains of 20 decibels or more in 91%, 30 decibels or more in 71%, and 40 decibels or more in 41% of cases at five years post-operatively. The bone-air gap at five years post-operatively was diminished to 20 decibels or less in 88%, to 15 decibels or less in 85%, to 10 decibels or less in 77%, and complete closure or over-closure occurred in 52% of the patients. 7. Data analysis has established there is no change with time of the post-operative mean bone-air gap 500--2,000 Hz. over a period of at least five years, and thus that the piston operation gives a persisting and stable hearing result. 8. In predicting the effect of the piston operation only a rough guide can be obtained from other variables. In particular bone hardness, mucosal characteristics, sex and piston diameter appear to be irrelevant. Patients with a large bone conduction reading or a large air-bone gap appear to do slightly worse than patients with smaller values for these variables. There are no grounds for excluding a patient from operation on account of age, or the finding of active otosclerosis (soft bone and thickened vascular muco-periosteum). 9. From the data of piston operations, the estimated mean bone-air gap for the five-year examination period was 4-14 db. with a standard deviation of 12-5 db. 10. The piston operation is highly recommended as a safe and suitable method of alleviating, in the long-term, the conductive hearing loss of patients with obliterative otosclerosis of the oval window.


Histopathological findings in the inner ear caused by measles. An otopathological analysis of three cases of viral labyrinthitis was performed. Six temporal bones cut in serial sections were available for this study. According to the degree of degenerative changes in various parts of the inner ear two types of morphologically distinct labyrinthitis after measles are presented: the first one with the port of entrance through the internal auditory meatus and characterized by, first and most significantly, changes in the spiral ganglion cells; and the second, as described previously by Lindsay, with the port of entrance of the virus in the inner ear through the stria vascularis, with degenerative changes in various structures within the endolymphatic duct.


Very high tension with very little ATP breakdown by active skeletal muscle. The tension developed and the chemical change occurring during stretching at velocities up to 2l0 sec-1 and during isometric contraction of dinitrofluorobenzene-treated and untreated frog sartorius muscle have been measured. At all velocities of stretching the tension was greater than during shortening at the same velocity and the mean rate of chemical change was less than or equal to the rates found in similar studies of shortening. For velocities 0.18, 0.33 and 0.66l0 sec-1, the mean rate of chemical change was significantly less (P less than 0.05) than during shortening at the same velocity. At the highest velocity of stretching the tension at l0 was about 0.8 P0 and the mean rate of chemical change was about the same as that for isometric contraction. At low velocity (0.1 l0 sec-1) the tension was as high as 1.4 P0 and the mean rate of chemical change during the stretches at the three slowest velocities was significantly less (P less than 0.05) than during isometric contraction; the lowest mean rate of chemical change amounted to only about 25% of that for isometric contraction. The fact that the chemical change was so low suggests that during stretching crossbridges can form links and develop tension without breaking down ATP.


Mechanical stresses and morphological patterns in amphibian embryos. 1. Embryos of Rana temporaria have been dissected and shape alterations of different parts of the embryo, taking place within 1 h of separation, have been studied. Two categories of deformation have been revealed. 2. The first category comprises those deformations which take place immediately after separation. They are insensitive to cooling, cyanide and Cytochalasin B treatment. These deformations, which consist of a shortening of initially elongated cells, are considered to be the passive relaxations of previously established elastic tensil stresses. 3. Deformations of the second category proceed more slowly. They are inhibited by cooling, cyanide and Cytochalasin B treatment, are accompanied by elongation and migration of cells and occasionally lead to rather complex morphodifferentiations of isolated fragments. These processes are considered to be the result of the active work of intracellular contractile systems, either pre-existing or induced de novo. 4. By analysing the arrangement of the passive deformations we have constructed maps of mechanical stresses in embryos from late blastula up to the early tail-bud stage. At several embryonic stages drastic transformations of the stress pattern occur, these transformations being separated by periods during which the pattern of stress distribution remains topologically constant. 5. A correlation between the arrangement of stress lines and the presumptive morphological pattern of the embryo is pointed out. 6. Some possible relations between tensile tissue stresses and active mechanochemical processes are discussed.


The effect of ablation of the olfactory pits on the development of the habenular nuclei in Rana pipiens. The habenular nuclei in the diencephalon of the frog, Rana pipiens, are asymmetrical structures: two discrete cell groups develop on the left side (as medial and lateral nuclei), while a single nucleus is formed on the right side. Experimental animals were subjected to bilateral removal of the olfactory pits at an early embryonic stage, and were maintained with normal control animals until metamorphosis was complete. The length, relative volume and cell number for each of the three nuclei were determined in the control and experimental animals at regular intervals during larval development. In the control animals, the left medial nucleus developed similarly to the right nucleus spatially and temporally; however, the left lateral nucleus was significantly different in its development in the three parameters measured. In the experimental animals the left medial and the right habenular nuclei were alone affected by the removal of the olfactory pits. The results provide experimental evidence that the right and left medial, but not the left lateral, habenular nuclei are centers receiving afferent olfactory fibers.


Binding of components of the properdin system to cultured human lymphoblastoid cells and B lymphocytes. Immunofluorescence studies showed that properdin (P) and factor B bind to C3-C3b receptor bearing human lymphoblastoid cells (Raji, Daudi) and B type human peripheral lymphocytes (HPL). P bound to Raji cells first incubated with normal human serum (NHS). EDTA, but not EGTA, halted the binding of P to cells incubated with NHS. However, fixation of P to Raji cells, after incubation with NHS first reacted with inulin, was independent of Ca++ and -g++ ions. Fixation of P to Raji cells depended on the presence of C3 or C3b and occurred in the absence of factor D and factor B. Binding of P to B type HPL was detectable only after incubation of these cells with NHS first reacted with inulin; under these conditions binding of P to Raji cells was also greatly enhanced. With both Raji cells and HPL, factor B was detectable on cell surfaces only after incubation of these cells with NHS first reacted with activators of the P system. Binding of factor B to cells required the presence of C3b and binding or stabilization of cell bound factor B necessitated the presence of activated P. P and factor B were detectable only on cultured cells having C3-C3b receptors. However, incubation of NHS with all lymphoblastoid cell lines studied resulted in activation of P and cleavage of factor B. Binding of P and factor B to cells may follow one of three sequences; (a) activated P in fluid phase combines with C3, factor D, and factor B, and the whole complex fixes to cellular C3-C3b receptors via its C3 moiety; (b) C3b generated in fluid phase combines with P, C3, factor D, and factor B and binds to C3-C3b receptors; or (c) C3 or C3b first binds onto the C3-C3b receptors and thereafter interacts with P, factor D, And factor B. Binding of components of the P system to cells or other particles may relate to such biological phenomena as lysis, phagocytosis, proliferation, attraction of other cell types, and alteration of responsiveness to external stimuli.


Effects of anti-Ia sera on mitogenic responses. II. Differential expression of the Ia marker on phytohemagglutinin and concanavalin A-reactive T cells. Genes mapping in the I region of the H-2 complex control a system of lymphocyte alloantigens (Ia) which are expressed on a subpopulation of T cells and on most B cells. Specific anti-Ia serum in the presence of rabbit complement removed the splenic T-cell subpopulation responsive to Con-A, but did not affect the response to phytohemagglutinin (PHA) or Leucoagglutinin. Antibodies specific for Ia, H-2K, or H-2D membrane antigens were used without complement to pretreat spleen cells. These antibody pretreated cells responded normally to Con-A and PHA.


H-2 compatibility requirement for virus-specific T-cell-mediated cytolysis. The H-2K structure involved is coded by a single cistron defined by H-2Kb mutant mice. Lymphocytic choriomeningitis or vaccinia virus-immune spleen cells of H-2 mutant mice carrying a point mutation in the K region (B6 H-2ba, B6 H-2bf) cannot lyse infected wild-type H-2Kb targets and vice versa. Yet, cytotoxic T cells specific for infected H-2Kba or H-2Kbf targets are generated during virus infections as shown by cold target competition experiments. The critical structure for the apparent restriction by the K or D regions of the H-2 gene complex of cytolytic interactions between T cells and virus-infected target cells are therefore each coded, at least as shown for the K region, by a single cistron. This finding is most readily accommodated within the altered self concept (postulating that T cells are specific for virus-modified self structures) but cannot exclude the possibility of a physiological interaction mechanism being responsible for the apparent H-2 restriction of virus-specific cytotoxic T cells.


T-lymphocyte-enriched murine peritoneal exudate cells. II. Genetic control of antigen-induced T-lymphocyte proliferation. The recent introduction of a reliable, T-lymphocyte proliferation assay, which utilizes thioglycollate-induced, nylon wool column-passed, peritoneal exudate lymphocytes from immune mice (PETLES), allowed us to investigate the genetic control of murine immune responses at the T-lymphocyte level. Examination of the blast cells generated in this population 5 days after stimulation with antigen, revealed that 85% of the cells bore the Thy 1 antigen on their surface, whereas only 5% bore immunoglobulin. Thus, the assay can be considered to measure almost exclusively T-lymphocyte function. This assay was used to examine the T-lymphocyte proliferative responses to seven different antigens: poly(Glu60Ala30Tyr10), poly(Glu58Lys38Tyr4), poly-(Tyr,Glu)-poly-D,L-Ala--poly-Lys, poly-(Phe,Glu)-poly-D,L-Ala--poly-Lys, staphylococcal nuclease, lactate dehydrogenase H4, and the BALB/c IgA myeloma protein, TEPC-15. PETLES from a large number of different inbred mouse strains, including H-2 congenic resistant lines and H-2 recombinants, were studied. The strains could be classified as high responders, low responders, or nonresponders to a particular antigen as judged by the magnitude of the T-lymphocyte proliferative response. In every case but one this classification corresponded to the responder status given the strain based on its ability to mount an in vivo antibody response to the same antigen. For two of the antigens, poly-(Tyr,Glu)-poly-D,L-Ala--poly-Lys and TEPC-15, the immune response genes controlling the T-lymphocyte proliferative response were mapped to the K region or I-A subregion of the major histocompatibility complex, as had previously been shown for the control of the antibody responses to these antigens. This tight linkage of the two phenotypic responses very strongly suggests that the same immune response gene controls the expression of both the proliferative and antibody responses. Since there is essentially no contribution from B lymphocytes in the T-lymphocyte proliferation assay, it seems reasonable to conclude that none of the seven immune response genes studied are expressed solely in B lymphocytes.


Participation of the H-2 antigens of tumor cells in their lysis by syngeneic T cells. Cytotoxic T lymphocytes were generated in vitro against H-2 compatible or syngeneic tumor cells. In vitro cytotoxic activity was inhibited by specific anti-H2 sera, suggesting that H-2 antigens are involved in cell lysis. Two observations directly demonstrated the participation of the H-2 antigens on the tumor cells in their lysis by H-2-compatible T cells. First, coating of the H-2 antigens on the target tumor cell reduced the number of cells lysed on subsequent exposure to cytotoxic T cells. Second, when cytotoxic T cells were activated against an H-2 compatible tumor and assayed against an H-2-incompatible tumor, anti-H-2 serum that could bind to the target cell, but not to the cytotoxic lymphocyte, inhibited lysis. H-2 antigens were also shown to be present on the cytotoxic lymphocytes. Specific antisera reacting with these H-2 antigens, but not those of the target cell, failed to inhibit lysis when small numbers of effector cells were assayed against H-2-incompatible target cells or when effector cells of F1-hybrid origin and bearing two H-2 haplotypes were assayed against a tumor cell of one of the parental strains. These findings suggest that it is the H-2 antigens on the tumor cell and not those on the cytotoxic lymphocytes that are important in cell-mediated lysis of H-2-compatible tumor cells.


Factors regulating macrophage production and growth: identity of colony-stimulating factor and macrophage growth factor. The activities of a colony-stimulating factor (CSF), which stimulates granulocyte-macrophage colony formation by mouse hemopoietic cells, and macrophage growth factor (MGF), which stimulates proliferation of activated peritoneal macrophages, have been demonstrated by various criteria to reside in the same molecular species. These criteria include occurrence in various sources and copurification of the activities in mouse L-cell-conditioned medium as well as the biological, physicochemical, and antigenic properties of the activities of L-cell-conditioned medium. CSF and MGF activities of L-cell-conditioned medium are ascribable to a glycoprotein of mol wt approximately 60,000 which migrates electrophoretically with alpha-globulin. Human urinary CSF, which also possesses MGF activity, has similar properties and can be neutralized by antiserum to highly purified L-cell medium CSF. A procedure is described for the partial purification of material from L-cell medium that has activity at 1 ng/ml in both MGF and CSF assays.


The capacity of mitogen-responsive T cells to "home" to their tissue of origin, analyzed by means of chromosome markers. Lance and Taub (1) showed that when radioactively labeled lymphocytes were injected into a syngeneic mouse and the lymph node cells of this animal transferred to a second syngeneic recipient, the proportion of radioactivity found in the lymph node relative to the amount present in the spleen of the secondary recipient had increased markedly. The interpretation of this result was that some lymphocytes have the capacity to "home" to their organ of origin. The purpose of the experiments described here was to test the "homing" copacity of T cells by a method that did not involve radioactive labeling. It has been shown elsewhere that some or all mouse T cells are stimulated to divide in culture by the mitogens phytohemagglutinin (PHA) and concanavalin A (Con A) (2). We therefore elected to inject karyotypically distinct lymphocytes into syngeneic recipients and to follow their subsequent distribution by culture of lymph node and spleen cells of the recipient with PHA or Con A. In this manner the homing capacities of spleen and lymph node T cells could be determined, and furthermore, the effects of labeling with chromium-51 ((51)Cr) could be assayed with respect to the persistence of mitogen responsiveness in the injected cells.


Evidence for direct linkage between antigen recognition and lytic expression in effector T cells. The relationship between antigen recognition and lytic expression by effector T cells was examined by coculturing two cytolytically active lymphoid cell populations. When antigen recognition between the populations could occur only in one direction, then cytotoxicity was expressed only in that direction and the population whose antigens were recognized lost its lytic activity. In contrast, the cocultured effector cell population fully maintained its lytic potential. This lack of reciprocal inactivation was taken as evidence that T-cell receptor accomodation by surface antigen is linked to the expression of cytolytic activity by effector T lymphocytes.


Anatomical observations of the subarachnoid cisterns of the brain during surgery. Subarachnoid cisterns and their contents are described briefly in the light of observations made during microsurgical explorations. The concept of intracranial surgery in terms of moving from one cistern to another is presented here with particular emphasis on the cisterns in surgical approaches to intracranial vessels and nerves for the treatment of aneurysms, arteriovenous malformations, and for surgery of basal tumors.


Graded activation of myofibrils and the effect of diameter on tension development during contractures in isolated skeletal muscle fibres. If the space constant of the T-system (lambdaT) its not large in comparison with the radius (a) of a muscle fibre, different levels of depolarization should activate different proportions of the cross-section. This possibility was tested in isolated muscle fibres with isotonic and isometric K  contractures. 2. During isonic contractures with more than 40 mM-K, wavy myofibrils appeared in the centre of the fibre. The sarcomere spacings (s) of the wavy myofibrils, measured parallel to the long axis of the myofibrils, were 1-9-1-95 mum. However, the superficial myofibrils could shorten to or below s=1-5 mum without becoming wavy. 3. In the same muscle fibre where myofibrils became wavy during K contractures, no waviness appeared during repetitive electric stimulation in normal Ringer (50 shocks/sec, 12 degrees C), although s decreased below 1-5 mum. Wavy myofibrils were interpreted as not activated. 4. With isometric contractures it was found that the amount of depolarization needed to obtain maximal tension was smaller for fibres of shorter radius. The degree of depolarization for producing maximal tension is related to a by 6 mV/10mum. 5. These results strongly suggest that in K contractures lambdaT is not large in comparison with a.


Electromechanical studies on the inotropic effects of acetylstrophanthidin in ventricular muscle. Three phases in the inotropic response of acetyl strophanthidin (AcS) on the electromechnical activity of the frog ventricular myocardium were identified and studied using a single sucrose voltage-clamp technique and other conventional electrophysiological methods. 2. the positive inotropic response of the drug was accompanied by a shift in tension-voltage relation, so that more tension developed with every depolarization step above the mechanical threshold (-50mV). Only at higher drug concentrations or with long exposure times did the mechanical threshold shift to more negative membrane potentials (-60 to-70 mV). 3. In tetrodotoxin-treated muscles AcS produced marked potentiation of twitch tension and an appropriate shift in the tension-voltage relation. 4. the positive inotropic response of the drug was not related to the magnitude of the direction of the fast or slow Na current. 5. in tetrodotoxin-treated ventricular strips the direction or the magnitude of the secondary inward current (ICa or INa) were not related to the inotropic effect of AcS. 6. AcS shortens the action potential markedly during the later stages of its positive inotropic response. When Ca2+ is omitted from the bathing solution AcS not only fails to shorten the action potential, but often prolongs it. 7. The shortening of the action potential in the presence of AcS is accompanied by an increase in the "instantaneous" membrane conductance both at rest and during the time course of the plateau. 8. The decline in the positive inotropic response of the drug was accompanied by the shortening of the action potential. Electrical or chemical prolongation of the action potential restored the full positive inotropic response if the membrane had not depolarized. 9. Membrane depolarization and the development of diastolic tension always occurred at later stages of drug action. Elevation of [Mg+2]degrees to 5 or 10 mM prevented or suppressed the membrane depolarization and the diastolic tension. 10. KCl-induced contractures were potentiated throughout the duration of drug exposure. The tonic component of the contracture tension was markedly elevated especially at later stages of drug action. 11. The experimental evidence suggests that no unitary mechanism could account for multiple actions of acetyl strophanthidin. However, the contributions of the Na pump, the Ca+2 sequestering system, and the K-efflux system to the various stages of drug action are discussed.


Electrical responses to frog taste cells to chemical stimuli. 1. Cells inside a fungiform papilla of the frog tongue were impaled with a glass capillary micro-electrode filled with 3 M-KCl. Cells considered to be taste cells showed a resting potential of about -35 mV and an input resistance of 17 Momega on the average. 2. Application of chemical stimuli such as salts, acids and quinine produced a sustained depolarization in a taste cell, the magnitude of depolarization being dependent on the stimulus concentration. Water and weak NaCl solution yielded a hyperpolarization. The thresholds for depolarization as well as the concentration-response relationships for various chemical stimuli in taste cells are in approximate agreement with those determined from the glossopharyngeal nerve responses. 3. The magnitude of depolarization produced by 0-1 M-NaCl and 0-03 M-CaCl2 was dependent on the membrane potential level and reduced linearly with a rise in the latter. However, depolarizations generated by 0-001 M-HDl and 0-02 M quinine changed little in magnitude by a membrane potential change over a wide range. 4. During depolarizations induced by NaCl and KCl a marked reduction in the input resistance of a cell was observed, the amount of the reduction depending on the stimulus concentration. The reduction was also produced by CaCl2 and HCl, but it is small compared with those by NaCl and KCl. Quinine produced an increase in the resistance associated with a depolarization. Water and weak NaCl solution produced an increase in the resistance associated with hyperpolarization. 5. The receptive mechanisms for various kinds of chemical stimuli are discussed in relation to changes in the membrane potential and the membrane conductance of taste cells.