Inhibition by puromycin of transneuronal transport in the mouse accessory olfactory bulb. Application of [3H]proline to the vomeronasal organ (VNO) in mice results in the transport of labelled material along the vomeronasal axons to their terminals in the glomerular layer of the accessory olfactory bulb (AOB). In addition labelled material leaves the vomeronasal nerve terminals and is found over the external plexiform layer (EPL), where a previous electron microscopic autoradiographic study showed that it is preferentially accumulated in mitral cells. Grain densities over the glomerular layer and the EPL were counted in light micrographs. After subtracting background, the overall density of grains in the EPL is about 10% of that over the glomerular layer at 6 h after administration of [3H]proline to the VNO (5 mice). In a further 7 mice, puromycin (or saline) was applied directly to the AOB at hourly intervals during the 6 h after [3H]proline administration. Under these circumstances the labelling in the EPL is only 2--4% of that in the glomerular layer (9% for the 2 saline controls). These observations are evidence that a major part of the transsynaptic transfer mechanism is dependent on protein synthesis, and also favour the view that free amino acids are an important component of the material transferred.


Thy-L antigen expression on rat brain cell lines. Cell lines derived from the central nervous system of rats were screened serologically for the presence of Thy-1 (theta), a cell surface differentiation antigen shared by brain and thymus of rats and mice. Both cytotoxicity absorption and indirect immunofluorescence assays were performed using a rabbit anti-rat thymocyte serum (ATS) with Thy-1 specificity. The complement-dependent cytotoxicity of ATS detected a mouse-rat cross-reacting determinant of the molecule bearing the Thy-1 antigen. Of 20 lines tested, 1 of 6 neuronal and 7 of 14 non-neuronal lines expressed Thy-1, as judged by their capacity to absorb ATS cytotoxicity for a Thy-1 positive thymoma line. Similar results were obtained in quantitative absorption assays of these lines employing a mouse anti-Thy 1.1 alloantiserum, but the xenoantiserum (ATS) was more sensitive for detecting the rat molecule bearing Thy-1. Indirect immunofluorescence, which was performed on several of the lines, yielded results in complete agreement with the cytotoxicity absorption assays, and revealed a generalized distribution of antigen in a speckled or patchy pattern over the membrane of cell bodies and processes.


Inhibitory effect of 1-beta-D-arabinofuranosylthymine 5'-triphosphate and 1-beta-D-arabinofuranosylcytosine 5'-triphosphate on DNA polymerases from murine cells and oncornavirus. This report compares the effect of the newly synthesized 1-beta-D-arabinofuranosylthymine 5'-triphosphate with 1-beta-D-arabinofuranosylcytosine 5'-triphosphate on the activity of DNA polymerases from mouse cells and oncornavirus. 1-beta-D-Arabinofuranosylthymine 5'-triphosphate inhibited all the activities of DNA polymerase alpha, beta, and gamma and viral DNA polymerase. The mode of inhibition of 1-beta-D-arabinofuranosylthymine 5'-triphosphate as well as 1-beta-D-arabinofuranosylcytosine 5'-triphosphate was competitive to the deoxynucleoside triphosphate with the same base. The inhibition constant (Ki) and the mode of inhibition of nucleotide incorporation varied with changes in the combination of the inhibitor, substrate(s), and enzyme species.


Distribution of alpha-fetoprotein- and albumin-containing cells in the livers of Fischer rats fed four cycles of N-2-fluorenylacetamide. Immunofluorescent localization of alpha-fetoprotein (AFP)- and albumin-containing cells was determined in the livers of Fischer rats fed 0.05% N-2-acetylfluorenamide for four 2-weeks-on, 1-week-off cycles. After this exposure multiple changes in the liver include over 1000 neoplastic nodules/liver, as well as extensive production of so-called oval cells and focal zones of atypical hepatocellular hyperplasia. Approximately 1% of the oval cells contain AFP, and about half of the zones of atypical hyperplasia include cells that contain AFP, but none of the neoplastic nodules or normal hepatocytes have any AFP-containing cells. Since up to 60% of the hepatocellular carcinomas developing from this regimen will predictably produce AFP, it is tentatively concluded that hepatocellular carcinoma may arise not only from "premalignant" neoplastic nodules but also from oval cells or the atpyical differentiation of hepatocytes.


Quinacrine fluorescence of Merkel cells in Xenopus laevis. It has been shown by electron microscopy that, in Xenopus laevis, Merkel cells are usually situated near the ducts of the skin glands. Cells which fluorescence in ultra-violet light after treatment of the skin with quinacrine can be identified with these Merkel cells by their position, shape and size. The method indicates the presence of purine nucleotides, probably ATP. This result is consistent with the view that "large opaque vesicles" are sites of ATP storage.


Analytical particle electrophoresis applied to immunologic research. The technique of analytical particle electrophoresis has been applied to the study of membrane phenomena associated with lectin stimulation of lymphocytes. It has been observed that such treatment results in a significant increase in the electrophoretic mobility of murine T-cells. Moreover, it has been demonstrated that the mobility changes in most cells are attributable to the release of a factor from relatively few cells. Preliminary evidence suggests that the murine factor has a comparatively low relative molecular mass, is heat-labile, and will alter the mobility of cultured human T-cells.


Quantitative ultramicro-scale immunoenzymic method for measuring Ig antigenic determinants in single cells. The Fluoro Ultra Micro Enzyme Linked Immuno Assay (FUMELIA) allows one routinely and quantitatively to measure a few thousand antigenic determinants on single cells. Highly purified Escherichia coli beta-galactosidase has been coupled to specific antibodies. By use of the Parafilm microcuvette techniuqe, the activity of the antibody-conjugated beta-galactosidase is assayed with a conventional spectrophotofluorometer. Attempts were undertaken to sensitize FUMELIA even further, so as to be able to detect a very few antigenic sites. It seems that even in its present state of development FUMELIA is more sensitive for the quantitation of cell-associated antigens than are techniques in which radiolabeled reagents are used. The potential of FUMELIA is illustrated by the quantitative measurement of membrane-bound immunoglobulins on single lymphocytes. It could be shown that T-cells as well as C-cells can synthesize Ig antigenic determinants. Thus it seems likely that T-cell receptors will, after all, be found to be immunoglobulins.


Quantitative cytochemistry: the basis of sensitive bioassays, for comparison of bio-and immuno-reactive hormone values. Cytochemistry now extends biochemistry down to the single-cell level. Special procedures have been developed for cutting sections of uniform thickness with no measurable or detectable artifact. The cytochemical bioassays use chromogenic reactions with the immediate precipitation of the resulting chromophore so that biochemical activity can be related to the individual cells constituting a tissue. This activity, in individual cells, is measured by scanning and integrating microdensitometry which also increases the sensitivity of cytochemical measurement over that of conventional biochemistry. A hormone, or indeed any biologically active substance, acting on its target cell, causes a change in the chemical activity of that cell that mediates the physiological effect of the hormone. By cytochemical methods one can assess such changes in the target cells even if these constitute only a small part of the target-organ; thus such methods are ideally suited to measuring chemical changes of this sort induced by the hormone. Such cytochemical bioassay of polypeptide hormones, done as "within-animal" assays, are about 1000-f0ld more sensitive than the equivalent radioimmunoassays and are as precise. Thus they have two advantages: (a) bioreactive hormone is measured rather than a composite of antigenic determinants characteristic of part of the hormone molecule and (b) their increased sensitivity allows discrimination between low normal and subnormal concentrations of the circulating hormone.


Plasma vitamin D-binding globulin in vitamin D deficiency, pregnancy and chronic liver disease. Plasma concentrations of vitamin D-binding globulin were measured by radial immunodiffusion in healthy subjects, pregnancy, and during oestrogen therapy. Subjects with disorders of vitamin D metabolism (dietary deficiency, malabsorption, anticonvulsant therapy, chronic liver disease) were also studied. Neither sex nor age influenced the plasma vitamin D-binding globulin concentration in healthy subjects, but there was a significant increase in concentration during pregnancy and oestrogen therapy. Elevated levels were found in vitamin D deficient elderly but not younger subjects, while levels in subjects with chronic liver disease were significantly reduced. Normal levels of vitamin D-binding globulin were present in hypervitaminosis D and no vitamin D-binding globulin was detected in human milk. No correlation was observed between plasma 25-hydroxycholecalciferol levels and plasma vitamin D-binding globulin concentrations.


The rapid quantitation of serum alpha-fetoprotein by two-site micro enzyme immunoassay. A simple, inexpensive and rapid micro two-site enzyme immunoassay procedure for serum alpha-fetoprotein is described. The assay is eminently suitable for the screening of neural tube defects and for the quantitation of AFP as a tumour marker.


Purification of placental alpha-1-foetoprotein. Quantitation of placental alpha-1-foetoprotein was done by the radioimmunoassay technique and gave a mean value of 6060 +/- 22.2 ng/g fresh tissue. The purification process included three methods. (1) Protein precipitation was performed using ammonium sulphate at 50 and 70% saturation. Elution on a Concanavalin A-sepharose column was used to diminish the interference of albumin with alpha-foetoprotein. (2) A coupling immunoadsorption technique using CNBr-activated Sepharose 4-B, antialbumin and antitransferrin, was found to be more reproducible. (3) Counter-immunoelectrophoresis and discontinuous gel electrophoresis gave a 60% yield with a 400-fold purification.


Further observations on isoelectric focussing of serum proteins using modified cellulose acetate gel membranes, and direct isoenzyme staining. 1. Treatment of cellulose acetate gel strips by boron trifluoride in methanol has been shown previously to produce a material suitable for isoelectric focusing. Further observations are reported here on the changes in the strips induced by the reagent. It has shown by conventional protein electrophoresis, for example, that not only is the degree of electroendosmosis reduced, but also that other properties are altered. 2. Minor modifications are introduced for the general isoelectric focussing method including improved electrode solutions. 3. Isoenzyme detection following isoelectric focussing of serum proteins is discussed in the light of experience gained using alkaline phosphatase and lactate dehydrogenase as examples.


A simple method for production of slides of CT images from multiformat radiographs. Sixteen on 1 multiformat images of CT scans can be mounted directly into special "super slide" 2 by 2 in. mounts. Use of special photographic equipment is thus avoided.


Stimulus and response related components of the late positive complex in visual discrimination tasks. Visual event-related potentials (ERP) were recorded from 6 subjects aged 10--15 in response to target stimuli in 2 vigilance tasks which differed in their processing requirements. Stimulus- and response-synchronized averages were computed for each subject for each quartile (Q) of that subject's reaction time (RT) distribution. Late positive complexes (LPC) of identical morphology were seen in the brain potentials from each task. In Q1, the LPC was an undifferentiated, large amplitude, positive-going potential with a parietal-maximum topography. As RT increased, the LPC became increasingly differentiated so that by Q3 3 components in the latency range of P300 were visible: two constant-latency peaks, P286 (fronto-central), P341 (parietal); and one variable-latency deflection, P539 (parietal), whose peak latency lengthened as RT increased. An additional constant-latency peak, P198, was also recorded. Principal component analyses of the averaged wave forms confirmed the visual identification of these components. The fixed-latency peaks identified in the brain potentials of these subjects were similar in timing and topography to previously reported late positive components. This study demonstrated. however, that the LPC can be segregated into stimulus-related and response-related components.


The frequency of bone abnormality in patients on anticonvulsant therapy. Biochemical and bone morphometric measurements were evaluated in 12 patients who were on long-term anticonvulsant therapy with barbiturates. Half of the patients had no symptomatic bone disease, and half presented with bone disease and pain. Serum biochemical values were normal except for a few patients who had an elevated serum level of parathyroid hormone; the concentration of serum 25-hydroxy vitamin D was decreased in the majority of patients in whom it was measured. Bone absorptiometric values were normal but proved to be misleading: the Singh Index and videodensitometric measurements indicated that bone mass was below normal in all patients. Bone morphometric data indicated that bone resorption was 3 times greater than normal, and there was no evidence of osteomalacia. Vitamin D and possibly calcium have been suggested as potentially useful agents in the treatment of the bone disease associated with chronic anticonvulsant therapy.


Sleep patterns in the intensive care unit and on the ward after acute myocardial infarction. Twelve patients aged 33--70 years (mean 49.5) underwent nightly recordings in the ICU and subsequently on the ward following acute myocardial infarction. Sleep patterns were analyzed according to night after infarct and ICU versus ward environment. Significant differences in nocturnal sleep patterns from matched controls initially after infarction included greater wakefulness, low REM sleep per cent, long REM latency, fewer REM periods, more awakenings, more stage shifts and decreased sleep efficiency. The usual circadian variation in HR was absent, and there was an estimated 8--10 h of unrecorded daytime sleep, which together suggested a quite generalized disruption of biological rhythms. With time, there was loss of daytime sleep, lowered nocturnal wakefulness and increased REM sleep. Slow-wave sleep (sometimes with very long duration delta waves) increased above normal over post-infarction nights 3--9, and sleep was otherwise renormalized by post-infarction night 9. No sudden sleep changes occurred with transfer from ICU to ward. The altered sleep patterns appeared mainly attributable to infarction itself. Twelve nocturnal anginal attacks occurred. Ten began in NREM sleep and two in REM periods without particularly intense phasic activity. Post-infarction nocturnal angina therefore appears to differ in pathogenesis from angina outside this period, which usually occurs in REM sleep. ECG changes could occur during sleep before awakening with pain, and overall decrease in ECG amplitude sometimes accompanied angina. Most attacks (10 of 12) occurred on post-infarction nights 4 and 5, indicating that undetermined that undetermined factors produce a secondary period of heightened risk at that time.


Conduction time in central somatosensory pathways in man. Simultaneous recording of the somatosensory evoked potential (SEP) from the neck and from the scalp allows investigation of conduction of somatosensory impulses within the central nervous system alone. The early components of the SEP produced by stimulation of the median nerve at the wrist were recorded from standardized electrode locations on the scalp and neck in 21 normal subjects. The peak latency of both the initial negative potential from the scalp, N20 (19.4 +/- 1.1 msec), and the major negative negative potential from the neck, N14 (13.8 +/- 0.9 MSEC), CORRElated positively with arm length and with height. The difference between the peak latencies of N20 and N14 (5.6 +/- 0.5 msec) was independent of both arm length and height. As the latency and distribution of N14 indicate that this potential probably arises from the dorsal column nuclei, the N20--N14 latency difference provides a measure of conduction time within central pathways which is independent of conduction time in the limbs and spinal cord. Recording of the SEP from the neck, simultaneously with that from the scalp, also facilitates clinical investigation of the somatosensory system.


The effect of experimental 'scotomata' on the ipsilateral and contralateral responses to pattern-reversal in one half-field. The averaged cortical responses to a reversing checkerboard pattern presented monocularly in either left or right visual half-fields have been recorded from the occipital scalp using a transverse chain of widely spaced electrodes referred to a common mid-frontal electrode. The half-field responses showed a consistent asymmetry, the dominant feature of which was a positive wave (P100) that was widespread on the ipsilateral scalp and maximally recorded from the midline and ipsilateral electrodes. This formed part of the triphasic negative-positive-negative complex, the other two components being an N75 and an N145. On the contralateral scalp it was generally possible to record a triphasic complex of opposite polarity, but this was usually of smaller amplitude and its components (P75, N105, P135) showed greater variation in latency and morphology than the ipsilateral components. With progressive occlusion of the pattern stimulus from the central regions of the visual half-field, the ipsilateral positive wave (P100) was increasingly attenuated, while components of the contralateral complex were relatively unaffected, or, in some cases, enhanced. By contrast, reducing the radius of the stimulated area had relatively little effect on the ipsilateral P100, while the contralaterally recorded response was attenuated. These differential effects on the half-field response components are discussed in relation to the anatomy of the central and paracentral cortical representation of the visual field. The implications for the interpretation of evoked potential recordings in patients with field defects are considered.


Asymmetry in EEG alpha coherence and power: effects of task and sex. Male and female subjects performed two spatial and two verbal tasks while EEG was recorded from right and left temporal and parietal sites. Measures of alpha band power asymmetry, coherence and phase angle were derived. Spatial tasks produced increased coherence within the right hemisphere. Interhemispheric coherence was higher for females, and at parietal than at temporal sites. Power asymmetry effects confirmed previous findings, reflected upon sex differences in cerebral lateralisation and upon strategies associated with functional cerebral organisation for task performance. The relation to performance studies and the value of the method in investigating dynamic functional organisation are discussed.


Suprachiasmatic nuclei lesions in the rat: alterations in sleep circadian rhythms. After continuous control recording (24 h/day) during 5 days, 14 rats received sham, complete or incomplete lesions of the suprachiasmatic nuclei (SCN). Sleep records were analysed from the 22nd to the 27th day after the lesion. In rats with complete lesions of the SCN, there was a strong decrease in the amplitude of the circadian variation in slow wave (SWS) and paradoxical (PS) sleep. The acrophase of SWS remained unaltered whereas that of PS shifted towards the end of the illumination (7.00--19.00 h) period. The importance of these changes in sleep organization seems to be correlated with the size of the SCN lesion.


A correlation between hippocampal responses to interhemispheric stimulation, hippocampal slow rhythmic activity and behaviour. The spontaneous electrical activity of the hippocampus as well as its responses to commissural stimulation were recorded in waking rats. Changes in magnitude of the early and late components of the response to commissural stimulation were associated with the behavioural state of the rat. The magnitude of the initial potential was higher in non-theta 'states' than in theta 'states', whereas the opposite was true for the late potential.


A long term time-lapse video system to document the patients spontaneous clinical seizure synchronized with the EEG. A long term audio/video clinical seizure monitoring system is described which is complementary to a previously described EEG seizure monitoring system. The video unit is mobile and based on time-lapsed video recording techniques to extend the unattended continuous video recording of the patient to many days if necessary. The simultaneous coverage of the EEG and behavior of the patient particularly during his seizure has been very useful in the workup of intractable epileptic patients being considered for neurosurgical treatment.


EEG field mapping.

EEG field mapping. A method for plotting isopotential maps of EEG fields is described. Forty-eight EEG channels are recorded, computer-processed, and used to plot a chart of the potential distribution every msec. This technique shows the usual characteristics of the EEG, and also some parameters that are normally hidden, such as the shape, surface and slope of the waves. The components of the visual evoked potential, for example, appear to be 'saltatory', as they do not move from one place of the scalp to another.


Sequence of 129 nucleotides at the 3'-terminus of encephalomyocarditis virus RNA. The sequence of 129 nucleotides next to the poly(A) tail of encephalomyocarditis virus RNA has been determined by rapid gel sequencing of cDNA synthesized with DNA polymerase I or reverse transcriptase and a phasing primer, [5'-32P]p(dT)8dC. The sequence is in accord with (a) the pyrimidine tracts which were mapped in blocks along the cDNA, (B) the sequences of seven characteristic T1 RNase oligonucleotides in the RNA transcribed from the cDNA with RNA polymerase, and (c) a limited amount of sequence deduced by partial spleen phosphodiesterase digestion and depurination of endonuclease IV oligonucleotides. The 3' end shows little secondary structure on its own. Ten nonsense codons block all three reading frames such that at least 26 nucleotides do not code for protein. The possible function of a homology A-A-U-A-A-A with other polyadenylated RNAs is discussed.


alpha1-Antitrypsin and alpha2-macroglobulin in newborn infants. I. The influence of perinatal complications. The levels of alpha1-antitrypsin and alpha2-macroglobulin in the plasma of 129 newborns were determined. The infants were divided into 3 groups according to their perinatal history. In healthy newborns with an uneventful perinatal history the normal values for alpha1-antitrypsin were 1.97 +/- 0.44 g/l, and for alpha2-macroglobulin 3.11 +/- 0.69 g/l. No changes in these levels were found during the first week of life. The levels of alpha1-antitrypsin and alpha2-macroglobulin showed significant correlation to each other. In healthy newborns with different complications in the obstetric history the levels of alpha1-antitrypsin were not influenced, whereas alpha2-macroglobulin decreased slightly during the first week of life. The levels of alpha1-antitrypsin and of alpha2-macroglobulin showed no further correlation to each other. In sick term and preterm newborns (n = 18) alpha1-antitrypsin was increased in 5 of 7 babies suffering from bacterial infections and lowered in 4 of 9 cases with respiratory disturbances. Alpha2-macroglobulin was lowered in 15 babies. These results indicate different kinetics of the two antiproteases in vivo.


Cellular immunity to encephalitogenic protein in multiple sclerosis. Correlations with other laboratory characteristics at different disease courses. A very suggestive, although non-significant, correlation was found between cellular immunity to encephalitogenic protein in multiple sclerosis and the genetic marker HLA-B7 in patients with no disease activity during at least the past 5 years and in those with a chronic progressive disease course during recent years, but not in patients studied longitudinally after a relapse. Only the latter group showed a significant correlation with parameters indicating intrathecal synthesis of immunoglobulin. No correlation was demonstrable with a parameter indicating intrathecal synthesis of measles antibodies.


Electrofocusing and electrophoresis of cerebrospinal fluid proteins in CNS disorders of known or probable infectious etiology. The CSF and sera of 17 patients with meningoencephalitis or encephalitis, 10 with meningitis or meningomyelitis, 5 subjects with postencephalitic sequelae and 2 with arachnoiditis were studied. Diseases involving the intracranial CNS parenchyma were found to give CSF electrofocusing findings differing from those of the other diagnostic subgroups. Differences regarding subfraction patterns of the gamma-globulin region seemed to be influenced of whether supra- or infratentorial structures were affected. The present study seems to indicate that the abnormal CSF-protein patterns found on electrofocusing were not only influenced by temporal but also by spatial factors.


Modulation of immune responses by suppressor T cells. The activity of suppressor T cells has been demonstrated in almost every phase of the immune response. These regulatory cells modulate both humoral and cell-mediated immunity utilizing antigen-specific and nonspecific mechanisms. For comparative purposes two murine models are described, the nonspecific suppressor T cell stimulated by the mitogen concanavalin A and the antigen-specific suppressor T cell stimulated by injection of the synthetic terpolymer acid 60-L-alanine30-L-tyrosine10 (GAT) in nonresponder mice. These two T cells are similar to expression of Ly alloantigens, ability to inhibit antibody responses, and the mediation of suppression, at least in part, by soluble products. However, differences in radio-resistance and antigenic specificity of the suppressor T cells, as well as differences in molecular characteristics of the soluble factors and their targets suggest that these T cells regulate the immune response by different mechanisms. The relationship of these two suppressor T cells to other nonspecific and antigen-specific suppressor T cells is discussed.


Lymphocyte transformation induced by autologous cells. Human peripheral blood T lymphocytes are stimulated to proliferate when cultured with autologous B-lymphoblastoid cell lines, autologous mitogen-induced lymphoblasts, or autologous non-T blood lymphocytes. This reaction, the autologous mixed lymphocyte reaction, has attributes of an immune response possessing both memory and specificity. The capacity to stimulate autologous T lymphocyte proliferation depends on the lineage of the lymphoid cell and not on its establishment in continuous culture or carriage of the EB viral genome. The determinant on non-T lymphocytes which stimulates the autologous mixed lymphocyte reaction appears to be an Ia determinant. Thus, allogeneic graft rejection and the allogenic mixed lymphocyte reaction are very likely extensions of an immune response expressed within the host.


Treatment of patients bitten by rabid or suspected rabid wolves with inactivated tissue culture rabies vaccine and rabies gammaglobulin. During 1972-1976 46 persons in 9 foci were bitten by wolves. 39 of them were immunized with antirabies gammaglobulin and tissue culture rabies vaccine; 7 received culture vaccine only. Rabies in wolves was confirmed clinically or in the laboratory in 8 foci. Bites of dangerous localization: face, head or fingers of the hands, predominantly multiple, were noted in 25 humans; 5 of them were young, 7 to 16 years old. Antirabies gammaglobulin was given to 9 people, predominantly in the dose of 0,5 ml per kg of weight, once on the 1st day after exposure (381-538 IU per kg of weight) to 14 people, once on the 2nd day (706-773 IU) to 3 people, twice on the 2nd and 3rd to 3 people, once on the 3rd and 5th day to 10 people, twice on the 2nd and 3rd day or on the 5th day after exposure. Vaccination course was started 24 hours after administration of gammaglobulin and predominantly in the dose of 5 ml; it lasted for 25 days and was followed by 3 booster injections on the 10th, 20th and 30th day. Titres of virus neutralizing antibody were tested in dynamics in 39 people immunized with gammaglobulin and tissue culture vaccine. Antirabies gammaglobulin induced some inhibitory effect, but 2-3 booster injections of the tissue culture rabies vaccine completely compensated this effect. During the observation period of 10 months to 5 years all the exposed people remained healthy.


alpha-Fetoprotein screening in patients with idiopathic hemochromatosis and liver cirrhosis. The serum alpha-fetoprotein level was measured by radioimmunoassay in 200 patients when admitted to hospital, 63 with idiopathic hemochromatosis and 137 with liver cirrhosis. In addition, repeated controls were performed in 19 subjects of each group for a mean period of 11 months (range 3--18 months). Elevated alpha-fetoprotein levels were observed initially or during the study period in 15 patients, a malignant liver tumor being demonstrated in 12 of them. In 4 of these patients, the abnormal alpha-fetoprotein concentration was the clue to the diagnosis of an unsuspected malignant hepatoma, but in none of these cases could the tumor be resected. The present results indicate that screening the serum alpha-fetoprotein level may contribute to the detection of malignant hepatoma in high-risk clinical groups, but the practical interest of such screenings may keep limited until more efficient therapeutic methods are developed.


alpha-Fetoprotein levels in normal individuals. Relative stability over time. Samples of serum were collected at 6- to 12-month intervals in two groups of healthy Africans and the amount of AFP measured by the radioimmunological method. The amounts present were found to remain fairly constant over time. The significance of this stability is discussed with particular reference of the highest levels of AFP observed in normal individuals in relation to primary liver carcinoma and the potential use of the method as a tool for early detection purposes.


Specific cell-induced transformation of Paramecium surface antigens. Paramecia may induce in other paramecia specific changes in the expression of genes determining surface protein synthesis, probably via cell-to-cell contact. Transformation from one antigenic type to a newly induced one is stimulated by the pretreatment of a cell with 5-fluorouracil. These observations are considered in terms of the regulation of the activity of genes in stable cells and in unstable cells which tend to transform spontaneously.


Repair of potentially lethal damage after single injection or continuous infusion of bleomycin. The repair of potentially lethal damage after administration of bleomycin by a single injection or by continuous infusion was studied in vivo. Experimental tumors were the fifth generation isotransplants of a squamous cell carcinoma which arose spontaneously in a C3Hf/He female mouse. Bleomycin was administered intraperitoneally by a single injection or by 24-hr continuous infusion. Cell survival was assayed by TD50 method. Dose-survival curve after a single injection exhibited a biphasic or upward concave curve. Surviving fraction increased rapidly if tumors were left in situ after treatment and reached a plateau at 5 hr, indicating that tumor cells were able to repair the potentially lethal damage induced by bleomycin. Dose-survival curve after continuous infusion was also biphasic, but had a small shoulder. The repair of potentially lethal damage was slight, if any, when tumors remained in situ for 6 hr after the end of 24-hr continuous infusion. This indicates that potentially lethal damage was being repaired during drug infusion.


Tube introducer and modified Celestin tube for use in palliative intubation of oesophagogastric neoplasms at fibreoptic endoscopy. A new method for palliative intubation of inoperable neoplasms at or near the cardia is described. A guidewire is passed through the stricture, which is dilated using Eder Puestow metal olive dilators. The tube to be inserted is mounted on an introducer, which grips its distal end from inside, and is slid into position along the wire under radiological control. Twenty-five patients have been intubated with one death directly resulting from the procedure. The method provides a simple and relatively safe means of relieving dysphagia and improving nutrition.


Intramitochondrial rodlet-like inclusions in human leukemic lymphocytes. Some mitochondria in leukemic cells in two of nine investigated patients with chronic lymphocytic leukemia and in one patient with morphologically less usual form of this disease contained unusual rodlet-like inclusion bodies. These structures were always present in the matrix of frequently dilatated intercristal spaces. The width of rodlet-like inclusions ranged between 10 and 20 nm, their maximal length was 114 nm. All observed rodlet-like inclusion bodies appeared to be composed of fine filaments 2...3 nm in width and resembled the DNA containing structures in mitochondria of few other cell types. In mitochondria of leukemic cells, however, these inclusions represent a further structural abnormality.


[Phagocytic granulocyte function in hemoblastoses]. Judging from the spontaneous NBT reduction, the indices of phagocytosis and NBT, there is a moderate, but statistically significant diminution of these parameters in leukaemia and malignant lymphoma including plasmocytoma. Moreover, further diminutions could be identified during the acute stage of the disease (first diagnosis or recidive) in acute leukaemia and lymphogranulomatosis, but not for chronic myeloic leukaemia.


[Complement activity in leukemic patients]. The in vitro test of heterologous anti-leukaemic sera against human leukaemic cells resulted in the serum of leukaemic patients being able to produce a lysis of leukaemic cells in the leukaemic phase as well as in remission when specific heterologous antibodies were present.


[Varying course of pancytopenia after busulfan treatment of chronic myelocytic leukemia (CML)]. In 84 patients with chronic myeloid leukaemia receiving a cytostatic monotherapy with busulfan, an aplastic syndrome developed which was confirmed by a biopsy of the pelvis crest and examination of the sternal marrow. The time interval until pancytopenia was detected varied considerably in each case, ranging between 6 and 126 months. There are no correlations to the initial doses of busulfan. 3 patients died of the immediate effects of the bone-marrow damage caused by busulfan. In 4 from 6 of the following pancytopenic patients the leukocyte values lay between 12,800/microliter and 80,400 microliter when busulfan adminstration was interrupted. Thus, it is scarcely possible to give any reliable informations about a leukocyte limit value as a standard for an interruption of therapy in order to prevent bone-marrow aplasia. Taking this into account, the conclusion may be drawn that relatively short control intervals have to be made in this monochemotherapy of CML which often can be used successfully for many years.


[Transplantation of bone marrow cells into lethally irradiated mice. Morphology of megakaryocytes]. We described morphological changes of megakaryocytes in the bone marrow and spleen of lethally irradiated mice, dependent on the time lapse following bone marrow transplantation. The functional active megakaryocytes of various morphological types were found to predominate in the tissues on about day 20 to 25.


Deficiency of beta-glucuronidase in neutrophils from patients with precancerous states of the larynx. Activity of beta-glucuronidase (GR) and acid phosphatase (AP) has been determined in peripheral blood neutrophils from 24 men with precancerous states of the larynx that is leukoplakia papillomas and pachydermia by means cytochemical methods described by Hayashi et al., and Barka and Anderson, respectively. The results obtained were expressed in terms of absolute counts of enzyme-positive and enzyme-negative cells with regard to enzyme activity variation within the enzyme-positive neutrophil population; the enzyme activity index score has been calculated. The control group consisted of 20 healthy subjects of the same sex. No significant alterations were found so far as AP activity is concerned between the group studied. In contrast, activity of GR in patients with precancerous states exhibited significant lowering. The most striking feature was in almost complete absence from the blood of GR-positive neutrophils with high activity of the enzyme. Majority of these cells showed only traces of the GR activity. According to authors opinion the deficiency of GR in neutrophils of patients with precancerous lesions pertains to problem of neutrophil-mediated cytotoxic effect against mammalian tumour cells.


A further contribution on nucleoli of human lymphocytes. Cultured human lymphocytes were investigated by means of light as well as electron microscopic procedures to provide more information on the structural organization of their nucleoli. The transformation of ring shaped nucleoli to nucleoli with less or more distinct nucleolonemata in PHA stimulated cells was characterized by a marked increase of granular RNP components in number indicating the activation of their production. This phenomenon seems to be related not only to the activation of the ribosomal RNA synthesis but also to its processing. The appearance of fibrillar RNP components in the central area of the ring shaped nucleoli apparently represents the first sign of the nucleolar RNA synthesis in these cells. The proportion of fibrillar and granular nucleolar RNP comonents in PHA inresponsive lymphocytes was similar to that in lymphocytes from patients with the usual type of lymphocytic leukemia. The intranucleolar chromatin areas appeared to be larger in PHA stimulated lymphocytes but the proportion of these areas to the nucleolar body did not show substantial difference as compared to the resting cells.


Haematological changes in experimental hydralazine-induced collagen-like syndrome in guinea pigs. Haematological studies were carried out in hydralazine-induced collagen-like syndrome in guinea pigs. 37.5 per cent of animals were found to be LE-positive. It was found that long-term administration of hydralazine caused a decrease of erythrocyte count, a decrease of haemoglobin concentration and a decrease of haemoglobin content in individual red blood cell as well as a decrease of a single erythrocyte volume. A significant leukopenia was shown in LE-positive subgroup of hydralazine-treated guinea pigs. The obtained results confirmed the similarity of hydralazine syndrome to systemic lupus erythematosus.


Unsaturated vitamin B12 binding capacity and serum lysozyme activity during the marrow granulocytes reserve tests. Unsaturated vitamin B12 binding capacity (UBBC) and serum lysozyme activity (LZM) were estimated durinng the endotoxin, prednisone and hydrocortisone marrow granulocyte reserve (MGR) pool tests. Our results showed, that no additional mechanism except the shift of MGR from marrow caused granulocytosis after typhoid vaccine administration. While the prednisone, when given orally diminished additionally the number of the physiologically destroyed neutrophils. The hydrocortisone, however, showed the results very similar to those obtained after typhoid vaccine adminstration. Thus the hydrocortisone test seems to be most useful. It gives as good information as typhoid vaccine test but does not show its side-effects.


Activity of lysosomal beta-glucuronidase in leukocytes of rats exposed to benzene and sodium selenate. Chronic exposure to benzene results in rats in the decrease of the lymphocyte count in the peripheral blood, the decrease of the beta-glucuronidase (BG) activity both in lymphocytes and neutrophilic granulocytes as well as in the damage to lysosomal apparatus of lymphocytes expressed in diffusion of the enzyme within the cell cytoplasm. Administration of selenium (sodium selenate) in dosis of 1.0 microgram/Kg during consecutive 10 days prior the exposure to benzene resulted in prevention of benzene-induced decrease of the BG activity in granulocytes and of a damage to lymphocyte lysosomes. Application of selenium in dosis of 5.0 microgram/Kg during the same time prior the exposure to benzene prevented the benzene-induced lymphocytopenia, induced the reactive increase of the granulocyte number, and caused, moreover, the prevention of the BG activity decrease in granulocytes. Simultaneously the increase of the BG-positive lymphocyte percentage was noted which was related to the increase of cells exhibiting the cytoplasmatic and extralysosomal localization of the enzyme. The results suggest that only smaller doses of sodium selenate prevented the damage to lysosomal membrane of lymphocytes induced by toxic effect of benzene.


[Blood morphology of the European eel (Anguilla anguilla). IV. Thrombocytes and their developmental stages]. By the aid of histological special stainings, cytochemical proofs, phase contrast and electron microscopical methods the thrombocytes (spindle cells) of the European ell (Anguilla anguilla) and their stages of development were described as an independent series of cells. After an artificial blood loss thrombocytoblasts, prothrombocytes and mature thrombocytes could distinctly be demonstrates. Besides the kidney the spleen was found to be the main place of the thrombopoiesis. Phase contrast investigations provided evidence for the relatively strong locomotion of these cells. By means of cytochemical proofs a further differentiation of the thrombocytes from other white blood cells can be made. Electron microscopical investigations of the thrombocytes yielded a great similarity to the thrombocytes of the amphibians. The endoplasmatic ring and particular inclusions could be shown to be characteritstic features of the fine structure.


Serum copper investigations in children with acute lymphoblastic leukemia. Serum copper level (SCL) was studied in 57 children with acute lymphoblastic leukemia (ALL) by atomic absorption spectrophotometry. It was found that changes in serum copper are to a greater extent related to the clinical course of ALL than to the age of the children examined. The highest mean SCL were obtained in untreated patients (261.2 microgram/100ml). Normalization of myelograms during treatment is accompanied by decrease of SCL. The significant increase of SCL in comparison with SCL in remission (129.8 microgram/100ml) occurred in extramarrow localizations (163.8 microgram/100 ml) and in the group of cases where after 1/2...2 months of maintenance therapy the recurrence of ALL was noted (168.8 microgram/100 ml). These observations suggest that SCL estimation may be useful as a auxiliary test in the clinical evaluation of the disease, for efficiacy of therapy and in predicting recurrences of ALL.


Primary sideroblastic anemia masked by bleeding. A patient with characteristic features of iron deficiency was unexpectedly found to have circulating siderocytes. Bone marrow iron stain at this time showed absence of both hemosiderin and ringed sideroblasts; electron microscopy revealed absence of mitochondrial iron loading but presence of cytoplasmic ferritin in normoblasts. Replenishment of iron stores led to development of typical sideroblastic anemia. These observations suggest that increased percentage of siderocytes in otherwise typical iron deficiency anemia may signify the presence of a sideroblastic process masked by iron deficiency due to bleeding.


[Demonstration of the anti-thrombocyte effect of anti-human lymphocyte serum]. A report is given how the antithrombocytic effect of 13 charges of human anti-lymphocyte globulin from the horse can be identified. The investigations were performed in mice and with the thrombocyte agglutination test. The test findings in mice correspond with those of the human compatibility control. Therefore, the test in mice is recommended to be used for the laboratory examination of anti-human lymphocyte globulins.


[ATP-content and shape of erythrocytes during preservation with adenine and nucleosides]. ACD blood with additions of adenine (A, 0.5 mM in blood), ademine + guanosine ((AG, 0.5 mM each) and adenine + guanosine + inosine (IAG, 0.5: 0.5: 18 mM) was stored for 6 weeks at 4 degrees C and the morphological changes in connection with the ATP content were observed. After a storage of 6 weeks 2--3% of the cells were present as diskocytes, 60% as echinocytes, and 40% as spherocytes. The delayed morphological alterations in the ACD-AG blood in comparison with ACD-A blood were also reflected by a higher ATP content of the ACD-AG blood during its storage. The alterations in the form of erythrocytes recorded in the morphological index Im (a subdivision was made according to 6 different stages of form) correlated with the ATP content. The coefficient of correlation amounted to r = 0.85. Thus, Im is a reliable criterium for evaluating possible storage damages of stored erythrocytes.


[Studies of 57-co-bleomycin in patients with carcinoma colli uteri (author's transl)]. The kinetics of 57Co-Bleomycin was studied in twelve patients with histologically proven carcinoma colli uteri and in ten patients with benign gynaecological conditions. Measurements were carried out on the whole body, on parts of the body and on urine and blood using a whole body counter. Excretion of the tumour-seeking radio-pharmaceutical is predominantly through the kidneys and can be described by a double compartment model. After 48 hours, 2-4% of the applied activity is still present in the body. Radiation exposure to the body, the kidneys and bladder was calculated from the measurements and MIRD tables as 0.02 rad/mci (total body), 0.4 rad/mCi (kidneys) and 0.2 rad/mCi (bladder). In view of the rapid elimination of 57Co-Bleomycin, these figures are lower than data data quoted in the literature. There was no evidence of significantly higher uptake of 57Co-Bleomycin in the carcinomatous epithelium of the collum uteri compared with benign diseases, using the total body counter in the scan node. As possible explanations for this, the authors suggest inadequate geometric resolution of the system and a low tumour/non-tumour distribution ratio.


Metabolic response to short periods of starvation in hypo- and hyper-thyroid male rats. 1) Thyroidectomized rats were fed with a low iodine diet, injected daily with 0, 0.1, 1.8 or 25 microgram of L-thyroxine/100 g body wt., and compared with intact controls. 2) Plasma protein-bound iodine was decreased in the rats given the 0 and 0.1 microgram doses, unchanged in those given the 1.8 microgram doses, unchanged in those given the 1.8 microgram dose increased in those given the 25 microgram one. 3) The liver content of DNA-P, phospholipid-P, proteins and fatty acids was decreased in the rats that did not receive thyroxine, practically recuperated in those receiving 0.1 microgram and normal in those given 1.8 or 25 microgram of thyroxine. 4) 3 h of starvation produced a reduction in the liver content of total fatty acids that disappeared after 24 h. 5) When fed, liver glycogen concentration was low in the rats given 25 microgram of thyroxine. 6) With starvation, the fall in liver glycogen and blood glucose, and the rise in liver acetyl-CoA and citrate and blood glycerol concentrations were faster in the thyroidectomized rats that did not receive thyroxine than in the other groups. 7) The rise in plasma free fatty acid and blood ketone bodies concentrations were similar in all the groups, the greater level of the first parameter being observed after 6 h of starvation in the rats given 25 microgram of thyroxine and in the second one after 24 h in the rats given either 0.1, 1.8 or 25 microgram of thyroxine. 8) The rapid decrease in the availability of carbohydrate stores with starvation in the thyroidectomized rats could be responsible for their fast call for lipid utilization. The slower response to fasting in the hyperthyroid animals is probably a consequence of their reduced amount of endogenous substrates to be mobilized.


Structural properties of the human MN blood group antigen receptor sites. It is shown that the MN blood group antigen determinant of the major human erythrocyte membrane (MN) sialoglycoprotein is located on its N-terminal octaglycopeptide. The only analytically detectable difference between peptides from MM and NN cells are Ser/Leu and Gly/Glu polymorphisms at the first and fifth positions, respectively. Destruction of the antigens by removal of the N-terminal residues suggests that these amino acids represent a part of the receptor areas for various anti-M or -N reagents. Evidence is presented that the N-terminal structure of the Ss glycoprotein is identical with that of MN glycoprotein from NN red cells up to the fifth residue. This provides an explanation for the 'N' antigen on this molecule and direct support for the earlier proposal that the MNSs locus is represented by homologous genes.


A trichrome stain for the intrahepatic localization of the hepatitis B surface antigen (HBsAg). A modified trichrome stain is described for the intrahepatic localization of the hepatitis B surface antigen; HBsAg containing cells exhibit specific green metachromasia contrasting with the granular brown colour of non infected hepatocytes and with the deep eosinophilic colour of ground glass cells of HBsAg-negative alcoholic or drug hepatitis. The technique is simple and reliable for routine screening of HBsAg positive material; its sensitivity is greater than H & E, similar orcein and inferior to immunohistochemistry as performed on frozen sections. Histological diagnosis can be made on the same slide, since several other morphological details are provided in the trichrome stained preparations. With this technique 387 biopsies from HBsAg seronegative individuals were negative; full cytoplasms metachromasia was mostly seen in asymptomatic HBsAg carriers, focal or partial staining in patients with histological evidence of liver cell necrosis. The presence and the staining pattern of HBsAg were of no help in predicting transition to chronicity or a transition from chronic persistent to chronic active hepatitis.


Methyl-methacrylate as an embedding medium in histopathology. Methyl-methacrylate embedding makes it possible to obtain semi-thin sections rich in detail and without tissue shrinkage. The procedure requires considerable time and labour, and great care must be taken to prevent heat damage to the tissue during the exothermic polymerization process. The original method given by Burkhardt (1966) and later modifications are described and discussed, with special attention to the practical problems encountered and their solutions.


Immunofluorescence on resin-embedded material. Some antigenic determinants can be preserved in tissues after plastic embedding. In the present study liver tissue was fixed with glutaraldehyde or paraformaldehyde, dehydrated with ethanol and toluene and embedded in araldite which was then polymerized at 37 degrees C. Immunofluorescence was performed on semi-thin sections etched with hydrogen peroxide. This procedure allows correlation with light microscopy (on the same stained semi-thin section) and with electron microscopy (on adjacent ultra-thin section). Good results were obtained with anti-nuclear, anti-mitochondrial, anti-microsomal and anti-alpha1-antitrypsin sera.


Sequence of action of genes at the secretor, H, ABO and Lewis loci. It is argued that Lewis genes are responsible for adding specificity to glycoprotein molecules after the activities of the secretor, H and ABO genes, respectively, have been expressed. This conclusion is based on the results of studies on the expression of ABO and Lewis antigens in salivas from Australian aborigines, and on biochemical results. A simple figure illustrating the antigens determined on red cells and in body secretions as a result of the action of these genes, in their correct order--secretor, H, ABO, Lewis--is presented.


Gypsy moth cell lines divergent in viral susceptibility. I. Culture and identification. A series of cell lines unique in insect virus susceptibility pattern have been isolated from the ovaries of the gypsy moth (Lymantria dispar: Lepidoptera: Lymantriidae) on a synthetic medium with mammalian and avian serum supplementation. Growth curves showed the poorest growth occurring on peptone-based media with somewhat better growth on amino-acid-based media. The best growth was obtained with combined media. Serological study distinguished the present cell lines from one another and from cell lines derived from other insect species grown routinely in the same laboratory. Baculovirus susceptibility among the new lines varied from no response to a specific complete replication response upon challenge by the homologous (gypsy moth) nuclear polyhedrosis virus.


Normal immunosuppressive protein: inhibitory effect on immune response against tumour cells. Normal immunosuppressive protein isolated from human plasma was found to inhibit the generation of primary cytotoxic effector lymphocytes against allogeneic tumour cells in vitro. Total inhibition was observed when NIP was present during the early stages of the sensitization process. In contrast, the generation of secondary cytotoxic lymphocytes in vitro was only slightly inhibited even though large amounts of NIP were used. The inhibition of target cell lysis by sensitized lymphocytes required long preincubation of a relatively small number of effector cells with large amounts of NIP and was most significant when tested at low effector: target cell ratios. Under the same conditions NIP showed no inhibitory effect on the cytotoxic activity of immune macrophages. The present in vitro experiments suggest that NIP exerts its effect through inhibition of DNA synthesis and cellular proliferation and to a limited extent only, by inducing specific suppressor cells.


Generation of immunological memory in tolerant mice. Adult mice pretreated with dinitrophenylated isologous gamma globulins respond very poorly to the hapten presented in immunogenic form. However, the responses of such tolerized and primed animals, or the spleen cells from such mice on adoptive transfer, to small doses of soluble antigen revealed normal secondary responsiveness in terms of size of response, relative affinities of the antibodies produced and antigenic sensitivity. In contrast, mice tolerized in early life and subsequently primed showed diminished responsiveness to secondary challenge, both in the original animals and on adoptive transfer. In this instance cells producing high affinity antibodies seemed to be preferentially deleted.


Independent mobility of cholera toxin binding sites and Thy-1 alloantigen on mouse thymocytes. The hypothesis that Thy-1.2 carries a carbohydrate antigenic determinant with the same specificity as the monosialoganglioside GM1 was tested by attempting to co-cap Thy-1.2 and GM1 in CBA thymocytes using anti-Thy-1.2 alloantiserum and cholera toxin. Co-capping of these determinants was not observed suggesting that they are on separate molecules. We do not therefore confirm the previous reported association between Thy-1 and GM1 (Thiele, Arndt & Stark, 1977).


Induction and specificity of delayed hypersensitivity to Staphylococcus aureus in mice. The induction and specificity of delayed hypersensitivity (DH) to Staphylococcus aureus in mice was evaluated in vivo by the footpad (FP) assay and in vitro by spleen cell stimulation. Repeated infections result in a biphasic DH response. The first DH response, observed following three subcutaneous injections, was route and antigen specific, required viable organisms, and could not be enhanced by the incorporation of bacteria in adjuvants. Footpad reactivity was transferred to non-injected recipients by spleen cells but not serum and was inhibited by anti-thymocyte serum but not by cyclophosphamide. Spleen cell stimulation was maximal with homologous antigen, but, some cross reactivity was observed when cells were stimulated with hererologous gram-positive antigens. No cross reactivity was observed when antigens from gram-negative bacteria were used to stimulate spleen cells. The FP reactivity to homologous antigen following 7 injections, the second DH response, is of longer duration than that following 3 injections. Mice given seven injections exhibit a greater degree of cross reactivity to heterologous gram-positive but not gram-negative bacterial antigens. Similar results were observed when spleen cells from mice receiving 7 injections were simulated with gram-positive antigens. Furthermore, the degree of spleen cell stimulation following three of seven injections could be increased by elicitation prior to the vitro experiments.


Reaginic antibody to contact sensitizing agents. Occurrence of cells which supress IgG and not reagin responses. Mice painted with the contact sensitizing agent picryl chloride produce IgG and reaginic or IgE-type antibody. They concomitanly produce suppressor cells which can inhibit IgG responses of normal mice to picryl chloride. These cells were produced between 3 and 7 days after primary sensitization (skin painting) and were T cells, as evidenced by their sensitivity to anti-theta serum and by their passage through nylon wool columns. Although they inhibited IgG responses they did not significantly change IgE titres. This correlates with a previous observation that repeated painting with picryl chloride increased IgE and not IgG responses.


On the mechanism of action of adjuvants. Evidence is presented that one a way which adjuvants exert their effect is by initiating an enduring increase in the localization of labelled cells in draining lymphoid tissues. Agents not generally considered to be adjuvants (carbon, latex, sheep erythrocytes) but capable of altering lymphocyte recirculation are shown to prevent the induction of tolerance by soluble bovine gamma globulin (BGG). These properties fulfil the criteria for adjuvanticity as defined by Dresser (1968) and link the expression of adjuvanticity to alterations in lymphocyte circulation.


Serological and chromatographic analyses of antigenic structures detected in human brain, thymus and lymph nodes. In the present paper it is shown that the non-species specific determinant of the antigenic thymus-brain system previously detected in murine and human brain also exists in human thymus. However, in contrast to the situation in mice and rats this antigenic structure is not expressed on suspended thymic lymphocytes, but is associated with thymic tissue largely depleted of thymic lymphocytes. Moreover, this determinant is also detectable in human lymph nodes. Besides the non-species-specific antigenic determinant human thymus and brain also share a species-specific determinant. In contrast to the non species specific determinant this structure is also displayed by suspended thymic lymphocytes, certain peripheral blood lymphocytes and bone marrow cells. The non-species-specific determinant detected in human thymus is borne by a structure, which physico-chemically resembles the thymus-brain antigen isolated from murine brain and thymus as well as from human brain. Although the structure bearing the species specific antigenic determinant has a similar apparent molecular weight both antigens could be separated by ion exchange chromotography indicating their molecular diversity.


Quantitation of antibodies to IgM, IgD and beta2-microglobulin in antisera to chronic lymphatic leukemia lymphocytes. Rabbit antisera were prepared to chronic lymphatic leukemia (CLL) lymphocytes and were tested for their reaction with radioiodinated, solubilized, cell surface proteins of normal and CLL lymphocytes. A pooled, hyperimmune antiserum precipitated at least 16 polypeptides from both normal and CLL lymphocytes as shown by sodium-dodecyl sulfate polyacrylamide gel electrophoresis. These polypeptides varied in molecular weight from 11,000 to 180,000. None was prominent or unique to the CLL lymphocytes, although four peptide bands in the CLL cells usually showed more radioactivity than their counterparts in normal cells. Precipitation with antisera of known specificity showed that the cell surface proteins from CLL and normal lymphocytes included HLA antigens and beta2-microglobulin. IgM and IgD were found in preparations of normal cells and in cells of 3 of 5 CLL patients. Of cells from the other 2 patients, one showed only IgD and the other no Ig. An antigen-binding capacity test was employed to quantitate the antibody content of a pooled anti-CLL lymphocyte serum. The antiserum reacted with all Ig classes; however, after absorption with light chains and F(ab')2 fragments, the serum retained activity only for IgM and IgD. The absorbed antiserum bound 45 microgram IgM, 1.5 microgram IgD and 4.5 microgram beta2-microglobulin per milliliter. These data indicate that rabbit anti-CLL lymphocyte sera fail to detect a qualitatively unique tumor-specific polypeptide on the surfaces of CLL cells. However, such antisera contain antibodies to many cell surface proteins including IgM, IgD, HLA antigens and beta2-microglobulin.


IgE-mediated cardiac hypersensitivity reactions. An experimental model. To investigate the involvement of the heart in acute allergic reactions in a system immunologically analogous to that of humans, a model of cardiac anaphylaxis mediated by IgE antibodies was developed in the guinea pig. Hearts obtained from guinea pigs, passively sensitized with homologous antidinitrophenyl IgE antibodies, were perfused and challenged in vitro with antigen. Challenge resulted in sinus and ventricular tachycardia, atrioventricular conduction block and substantial histamine release. The results demonstrate that IgE antibodies can sensitize the heart and that the severity of cardiac dysfunction, which follows challenge with specific antigen, directly correlates with the magnitude of histamine released. Since myocardial ischemia and similar arrhythmias occur during immediate hypersensitivity reactions in humans, this experimental model will be helpful in the investigation of cardiac involvement in acute allergic reactions.


Tumor-specific and tumor-associated membrane antigens of Rous sarcoma virus transformed hamster fibroblasts. Hamster fibroblasts transformed by an env- strain of Rous sarcoma virus (RSV) express at their surface tumor-associated antigens of unknown origin and a tumor-specific antigen (VCSA) which is not expressed by hamster fibroblasts transformed by unrelated DNA or RNA oncogenic viruses. This antigen was detectable by rabbit antibodies and a complement-dependent 51Cr-release cytotoxicity assay and is common to RSV-transformed cells of different animal species. By comparing the anti-VCSA serum which antisera directed against purified gp85, gs-proteins, reverse transcriptase or detergentlysed virus particles, it was shown that VCSA is not a known virion structural protein. Moreover, VCSA expression does not correlate with viral replication since it is not detectable in chick embryo fibroblasts productively infected with the transformation-defective virus RAV-1 which shares virus structural genes with RSV. Finally, in hamster cells transformed by an RSV mutant, temperature-sensitive for the ability to transform the host cell, VCSA expression at the cell surface correlates with the expression of the transforming gene.


Angiotensin induced changes in blood glucose level. Angiotensin given by intravenous route in rabbits caused a marked rise in blood glucose within 45 min. Guanethidine, bretylium, alpha methyl DOPA, adrenalectomy and reserpinization significantly influenced the changes in blood glucose induced by angiotensin II. It is suggested that the rise in blood glucose level induced by angiotensin II appears to be mediated by facilitation of catecholamine release or due to inhibition of reuptake.


Relationship of tumor-specific transplantation antigens to the histocompatibility complex: dissociation of in vitro alloantigen expression and in vivo alloimmunity from tumor-specific transplantation antigen strength. The studies reported here explore the relationship between tumor-specific transplantation antigens (TSTA) and (1) the expression of serologically defined specificities, and (2) the immunogenicity of H-2K- or H-2D-determined alloantigens on methylcholanthrene (MCA)-induced murine fibrosarcomas. Expression of H-2K and H-2D serologically-detected private specificities on groups of tumors raised in B10.A, B10.BR or B10.D2 strains varied greatly among tumors. No regular reciprocal or direct relationship to tumor TSTA strength was found. Each tumor, however, expressed H-2K or H-2D alloantigens stably as determined by direct cytotoxicity or absorption techniques. Even those tumors expressing little or no detectable alloantigen by serologic analysis were immunogenic in H-2K or H-2D incompatible congeneic hosts. This was assayed in terms of capacity to evoke primary or secondary tumor resistance, and to induce allo-antibody toward private H-2K or H-2D end specificities. An exception was that B10.BR tumors failed to induce anti-H-2Dk antibody despite strong surface alloantigen expression. While TSTA strength did not correlate with serologically detected alloantigen expression, TSTA strength did tend to correlate inversely with capacity to resist primary growth in the H-2K different host, and directly with resistance in the H-2D different host.


Two methods of demonstrating leprosy bacilli in smears. Two methods, the carbol fuchsin with acetic acid differentiation and the periodic acid-carbol pararosaniline, were used for demonstrating leprosy bacilli in skin smears. Bacillary smears from 200 long-treated patients with tuberculoid, borderline and lepromatous leprosy were stained with periodic acid-carbol pararosaniline. There were significantly greater BI and MI determinations than with classic carbol fucsin staining. With the former stain bacilli were found in 69 of 96 skin smears in which no bacilli could be seen by the latter stain. It is suggested that under the action of antileprosy drugs some leprosy bacilli may lose their acid-fastness and become chromophobic; chromophobic bacilli can be restored to their staining with periodic acid pretreatment. Leprosy bacilli in their chromophobic form can survive in healing and apparently healed lesions even after prolonged chemotherapy and can be a possible source of relapse.


Properties of delipidated hepatitis B surface antigen (HBsAg) and preparation of its proteolytic cleavage fragments carrying HBsAg-specific antigenic determinants. Treatment of hepatitis B surface antigen (HBsAg) with either chloroform-methanol (2:1, v/v) or 50% 1,1',3,3'-tetramethylurea did not affect the morphological integrity of the particles (about 20 nm in diameter), although the major portion of lipids was released as indicated by their increased buoyant density in CsCl (1.27 g/cm3 as compared with 1.20 g/cm3 for intact HBsAg). The antigenicity and polypeptide composition of HBsAg was not altered by delipidation. The carbohydrate chains of HBsAg contain penultimate beta-D-galactosyl residues. HBsAg was cleaved by chymotrypsin into fragments which were smaller than intact HBsAg by two orders of magnitude and which contained both the a and d determinants.


Gas-liquid chromatographic determination of kepone residues in finfish, shellfish, and crustaceans. Finfish, shellfish, and crustacean samples are extracted with isopropanol and benzene; the extract is filtered and then concentrated. The extract, dissolved in hexane, is treated with oleum and extracted with aqueous alkali. The aqueous phase is acidified and extracted with petroleum ether-ethyl ether (1 + 1). The Kepone residue is determined by electron capture gas-liquid chromatography (GLC). Recoveries obtained by 8 laboratories from 15 species of finfish fortified at 0.02-0.23 ppm ranged from 37 to 107% with a mean +/- relative standard deviation of 79.4 +/- 14.5%. For oysters fortified at 0.01-0.10 ppm, recoveries range from 63 to 129% with a mean of 78.8 +/- 20.8%. For crustaceans fortified at 0.05-0.26 ppm, recoveries ranged from 52 to 110% with a mean of 78 +/- 16.4%. The approximate limits of quantitation for finfish and for shellfish and crustaceans are 0.02 and 0.05 ppm, respectively, under the GLC conditions used in this study.


The preferential adsorption of hemoglobin to polyethylene. Hemoglobin adsorption to foreign surfaces has not previously been considered in studies of blood-material interactions, despite the fact that hemoglobin is the most abundant protein present in blood. A hemoglobin-like protein was detected on a number of surfaces exposed to blood plasma, serum, and red cell suspensions. Hemoglobin adsorption to polyethylene from plasma was found to approximately equal the amount of adsorption of albumin and fibrinogen. The high relative affinity of hemoglobin for polyethylene was further confirmed by adsorption isotherm and direct competition experiments. The data from all four experimental methods support the following ranking of plasma protein affinity for polyethylene: Hemoglobin greater than fibrinogen greater than albumin congruent to gamma-globulin.


The heterogeneity of human Gc-globulin.

The heterogeneity of human Gc-globulin. Gc-globulin or group-specific component, also known as the vitamin D-binding protein, was investigated by the combined use of electrofocusing and immunofixation. Serum of the Gc 2-2 type was found to contain a single protein band whereas serum of the Gc 1-1 type shows two bands with a lower isoelectric point. The Gc 1-2 type contains all three bands known as Gc-2 (pI 5.10), Gc-1Slow (pI 5.03), and Gc-1Fast (pI 4.95). Each apoprotein shows an anodal shift of about 0.07 pH unit after incubation with an excess of 25-hydroxycholecalciferol. After treatment with sialidase Gc-1Fast focuses in the position of Gc-1Slow, whereas the position of Gc-2 remains unchanged.


Calcium-binding protein of the chick chorioallantoic membrane. II. Vitamin K-dependent expression. A simple method was devised for the maintenance of the chorioallantoic membrane (CAM) of chick embryos in organ culture. Explants of CAM survived for up to 5 days in this system and retained the characteristic three-layered morphology (ectoderm, mesoderm, and endoderm). Induction of the CAM calcium-binding protein (CaBP) by effectors of calcium metabolism was studied in these organ cultures. Vitamin K was found to elicit a seven- to eightfold increase in CaBP, whereas no increase in CaBP activity occurred on supplementation with vitamin A, parathyroid hormone, an analogue of vitamin D, vitamin D and its hydroxylated metabolites, or with elevated calcium levels. The vitamin K-mediated induction of CaBP was dose-dependent, inhibited by the vitamin K antagonists warfarin and dicoumarol, selective for vitamin K5, and maximal at the developmental stage (13-15 days of incubation) corresponding to the onset of calcium transport by the CAM in vivo. CaBP levels increased after 60-70 h in cultures of 13-15 day CAM supplemented with vitamin K and reached maximal levels around 80-90 h of culture. The CAM ectoderm underwent extensive proliferation and often assumed a villuslike morphology in the vitamin K cultures.


Gustatory pathways in the bullhead catfish. II. Facial lobe connections. The second order projections of the gustatory system in catfish were examined using both retrograde (HRP) and anterograde (degeneration and autoradiographic) hodological methods. Golgi-stained material was used to demonstrate the different cell types in the primary gustatory sensory area, the facial lobe. Efferents from the facial lobe gather into ascending and descending secondary gustatory tracts. The descending tract terminates largely in the medial funicular nucleus and the commissural nucleus of Cajal in the region of the obex. A small portion of the descending tract continues caudally to terminate in the dorsal horn of the spinal cord. The ascending secondary gustatory tract terminates mostly in the ipsilateral superior secondary gustatory nucleus in the isthmic region. A small portion of the ascending tract continues rostrally to terminate in the posterior thalamic nucleus and in the region of the nucleus lobo-bulbaris. Sparser contralateral projections are also seen in the posterior thalamus and isthmic gustatory regions. Three cell types can be discerned in the facial lobe: small, medium and large. The small cells are intrinsic neurons, the medium cells project to the isthmic gustatory nucleus, and the large cells send fibers to  the other terminal areas described above, as summarized in figure 19.


An autoradiographic study of the projections of visual cortical area 1 to the thalamus, pretectum and superior colliculus of the rabbit. The projections of visual cortical area 1 (vl) to the thalamus, pretectum and superior colliculus of the rabbit have been studied by Giolli and Guthrie ('67, '71) using the  Nauta and Fink-Heimer methods to determine the course and distribution of degenerating nerve fibers. The present study represents a reinvestigation of these same projections utilizing the tracing method of autoradiography. An injection of 3H leucine was produced within a small region of vl in each of 18 adult albino rabbits, and the brains were subsequently processed for autoradiography by the method of Cowan et al. ('72). The results have confirmed the observations of Giolli and Guthrie ('67, '71) (1) by showing that vl of the rabbit projects to the thalamic reticular nucleus, the ventral lateral geniculate nucleus, the dorsal lateral geniculate nucleus; the pulvinar, the anterior and posterior pretectal nuclei and the superior colliculus and (2) by showing that a particular retinotopic organization is present in each of these projections. However, unlike Giolli and Guthrie ('67, '71), the present autoradiographic study has further revealed (1) that both the ventrolateral and the posterior thalamic nuclei receive inputs from vl and (2) that the nucleus of the optic tract is not innervated by axons originating from vl.


Cerebellar afferents in teleost catfish (Ictaluridae). The cerebellar afferents in the bullhead catfish (Teleostei) were labeled by relying on the retrograde transport of horseradish peroxidase (HRP). Retrogradely labeled neurons were seen in: spinal cord, lateral cuneate nucleus, inferior olive, reticular nuclei, vestibular nuclei, nucleus subeminentialis, n. lateralis valvulae, locus coeruleus, n. mesencephalicus dorsalis, the basal optic nuclei, and a nucleus at the isthmic level which may be equivalent to the pontine nuclei of birds and mammals. Inputs to the molecular layer arise from the inferior olive, locus coeruleus and n. lateralis valvulae in addition to subjacent granule cells. No projections to cerebellum were seen to arise from either the optic tectum or from the inferior lobe. The so-called "lobo-cerebellar" tract in teleosts was reported by Goldstein, amongst many others, who mistook the superior secondary gustatory nucleus for a deep cerebellar nucleus.


A simple method for reproducing Polaroid images as transparencies. A method to produce transparencies on radiographic subtraction film from Polaroid images using equipment available in most radiology departments is presented.


Chronic lymphatic leukemia presenting as atypical rosacea with follicular mucinosis. A case is described in which the initial clinical and histological features were suggestive of rosacea. Subsequent progress revealed the diagnosis to be chronic lymphatic leukemia. The initial biopsy specimens were reviewed and small but definite quantities of follicular mucinosis were noted after appropriate staining. If this had been noted earlier the diagnosis would have been made more rapidly. This case demonstrates the importance of detecting follicular mucinosis. We are not aware of follicular mucinosis having been described in association with chronic lymphatic leukemia previously. It is also rare for leukemias other than the monocytic type to present with cutaneous lesions, though skin lesions may occur in established cases.


Differential patterns of elastic fibers in scarring and non-scarring alopecias. Elastic fiber stain (acid alcoholic orcein) reveals diagnostically significant differences between several types of alopecia of the scalp. A short outline of elastic fiber distribution on the normal hair follicle emphasizes the elastic coat of the follicular isthmus, the sparsity of elastic fibers on the cyclic lower portion of the hair root, and the presence in the neck of the dermal papilla of an elastin-like body which is formed anew with each hair cycle. This body provides a marker of the gradual shortening of successive anagen hairs in male pattern alopecia. Patterns of elastic fibers in the perifollicular and interfollicular dermis are helpful in differentiating idiopathic pseudopelade of Brocq from pseudopeladic states secondary to lupus erythematosus and other disease processes. Within the idiopathic group, the development of elastic fibres on the lower cyclic portion of the hair root identifies a sub-group that may have a different, non-inflammatory pathogenesis and is provisionally designated as fibrosing alopecia.


Cytochemical and autoradiographic study of nuclei in zinc-deficient rat buccal epithelium. Ammoniacal silver stain showed that there was a decrease in nucleohistones in zinc-deficient rat buccal hyperplastic epithelium as compared to control. Autoradiographic study showed that there was decreased incorporation of 3H-arginine and 3H-lysine in the hyperplastic epithelium.


A contribution to the histological study of hypercementosis using metal staining. After studying hypercementosis, in detail using metallic staining procedures, the authors reached the following conclusions: (1) The histology of hypercementosis, observed by metallic staining reveals remarkable and characteristic microscopical alterations. (2) The Rio Hortega12 procedures, the mitochondria technique of Polak,10 and the method of Achucarro-Hortega13 are of great value for the study of the morphology of cementum. (3) Among the technical procedures used, we consider the 1916 method of Achucarro-Hortega as affording the best view of cementocytes. (4) The Sharpey fibers, revealing great irregularity in course, shape and size, and frequent interruptions could be clearly observed; heavily stained lacunae were frequently observed along the cementum-dentin border; a direct deposit of cellular cementum on the dentin in the middle third of the root was frequently observed. Curiously, an acellular cementum deposit next to a cellular cementum layer could be observed; we also noted numerous cementocytes of different shapes and sizes, at times exhibiting alterations of a degenerative type, particularly fragmentation of protoplasmic prolongations. (5) The techniques used also showed the frequency of blood vessels in the texture of hyperplastic cementum and the presence of capillaries near cementum-dentin junction.


In vitro cytotoxicity of periodontal dressings. The cytotoxicity of three periodontal dressings was evaluated by two in vitro cell culture techniques. Fresh and stored materials were found to be cytotoxic by both techniques and it appeared that the mechanism of cytotoxicity was different for the three dressing materials tested.


Enhancement of antigen-induced leukocyte histamine release by a mononuclear cell--derived factor. Supernatant fluids of phytohemagglutinin-stimulated mononuclear cells from ragweed-sensitive patients significantly enhanced the release of histamine from antigen-triggered leukocytes of ragweed-sensitive as well as control individuals. Supernatants of mononuclear cells from control individuals did not reveal this enhancing effect, nor was it found with the use of supernatants of unstimulated mononuclear cells of ragweed-sensitive patients or culture media with PHA alone. Supernatant fluids of phytohemagglutinin-stimulated mononuclear cells of patients sensitive to trees and grass also revealed this enhancing effect. The factor(s) responsible for the enhancement of antigen-induced histamine is heat labile and has a molecular weight of less than 10,000 daltons. The mechanism and site of action of the enhancing factor could involve initiating and/or modulating steps of the leukocyte histamine release reaction. This factor, presumably a lymphokine or a monokine, may constitute a regulating link between cell-mediated immunity and histamine-mediated hypersensitivity reactions in allergic patients.


Anaphylactoid reactions to iodinated contrast material. A review of the literature involving anaphylactoid reactions to iodinated contrast material (ICM) suggests that the reactions are nonantibody-mediated but that a complex activation of inflammatory mediators occurs. Histamine release and/or complement activation has been demonstrated in both in vitro and in vivo experimental systems. It appears that pretreatment of selected cases (those patients previously exhibiting an anaphylactoid reaction) is effective in reducing the frequency and severity of subsequent reactions when readministration is necessary.


Afferent fiber connections from the lower brain stem to the rat cerebellum by the horseradish peroxidase method combined with MAO staining, with special reference to noradrenergic neurons. The origin of cerebellar noradrenaline (NA) was investigated by means of the horseradish peroxidase (HRP) method, combined with the monoamine oxidase (MAO) staining method to identify the NA neurons. The present study gives clear cut evidence that the main source for cerebellar NA in the rat is the locus coeruleus (Lc), both the dorsal and ventral parts which innervate the entire cerebral cortex and hypothalamus respectively. The present study clearly demonstrates the pontine NA neurons such as the nucleus subcoeruleus (sc) NA neurons also send their axons to the cerebellum. The fact that some neurons in the nucleus dorsalis nervi vagi and the nucleus commissuralis were labeled by HRP indicates strong possibility that A2 NA neurons also innervate the cerebellum, though direct proof was lacking in this study. Furthermore, some unknown afferent fibers to the cerebellum were found in this study. These are as follows: (1) non-noradrenergic neurons in the Sc area, (2) nucleus reticularis gigantocellularis and nucleus reticularis parvocellularis, (3) group 1 (of Meessen and Olszewski), (4) a cell group extending to the ventrolateral region of the reticular formation of the pons, (5) nucleus ambiguug, (6) nucleus nervi facialis.


Allogeneic restriction of the delayed inflammatory reaction in the rat. Delayed-type hypersensitivity (DTH) to Listeria monocytogenes was measured in rats that were recipients of syngeneic, semisyngeneic, and allogeneic immune thoracic duct lymphocytes (TDL). DTH could be transferred only to recipients that shared at least one haplotype with the TDL donors. The restriction was expressed in an inability of sensitized lymphoblasts to localize efficiently at antigen injection sites in the pinna of the ear and peritoneal cavity. Failure of allogeneic lymphoblasts to extravasate in more than trace numbers into Listeria-antigen-induced exudates was reflected in an absence of other lymphocyte-mediated expressions of DTH. Thus, lymphocyte-dependent MCA was not detected in Listeria-antigen-induced peritoneal exudates borne by recipients of allogeneic immune TDL and blood monocytes were not recruited in increased numbers into such exudates as they were in exudates borne by syngeneic rats. But allogeneic restriction of the delayed inflammatory response to Listeria antigen was overcome, at least in part, when antigen-presenting macrophages of the same MHC type as the immune TDL donors were implanted in the peritoneal cavity. The results encourage the belief that the observed failure of immune TDL to transfer DTH to allogeneic recipients is related to the inability of sensitized donor T cells to recognize antigen displayed by allogeneic macrophages.


Subclass restriction of murine anti-carbohydrate antibodies. Examination of the subclass distribution of murine antibodies directed against groups A and C streptococcal carbohydrate, alpha-(1 leads to 3) dextran and phosphocholine yields the surprising observation that these carbohydrate antigens stimulate IgG responses largely restricted to the rare IgG3 subclass. This subclass restriction is particularly impressive in light of the low circulating levels of IgG3 in nonimmune mouse serum and the failure of a variety of other antigens including proteins and aromatic haptens to stimulate IgG3 antibody production. Attempts to alter the subclass restriction of antibodies with carbohydrate specificity by immunization with carbohydrate-coupled protein have been unsuccessful and indicate that immunoregulation of subclass expression probably occurs at the level of the antibody forming (B) cell. It is therefore conceivable that VH regions of murine immunoglobulins may be restricted to particular IgG subclasses. A similar type of subclass restriction has been reported in human and rat anti-carbohydrate antibodies. This recruitment of a minor immunoglobulin isotype by carbohydrate antigens in several species further supports the concept of immunoregulation at the level of subclass, and suggests that these and other mammals may share a structurally similar isotype with perhaps a common evolutionary origin.


Human monocyte cytotoxicity to tumor cells. I. Antibody-dependent cytotoxicity. Recent investigations examining mononuclear cell antibody-dependent cell-mediated cytotoxicity against tumor cell lines suggest that K lymphocytes and not monocytes are active in this cytotoxic reaction. We have found, however, that in an allogeneic assay system, human monocyte monolayers as well as lymphocytes mediate substantial lysis of 51Cr-labeled antibody-coated CEM lymphoblast tumor cells. This cytotoxicity is temperature-dependent and rapid, with most 51Cr release occurring in the first 4 hr of co-incubation. Interaction between target cell-bound antibody and the monocyte Fc receptor is necessary as demonstrated by the marked fall in antibody-dependent cell-mediated cytotoxicity (ADCC) produced by staphylococcal protein A, high concentrations of nonspecific immunoglobulin, and dilution of the target cell antiserum. Morphologic and functional characteristics of the monocyte-monolayer preparations establish their relative purity (greater than 95%) and indicate that monocytes and not contaminating lymphocytes are responsible for tumor cell lysis. Furthermore, preincubation of monocyte and lymphocyte preparations with latex particles or low concentrations of immunoglobulin distinguished monocyte from lymphocyte ADCC. Thus, normal human monocytes have the capacity to carry out antibody-dependent cytotoxicity against nucleated malignant target cells.


Antigen-specific T cell-mediated suppression. II. In vitro induction by I-J-coded L-glutamic acid50-L-tyrosine50 (GT)-specific T cell suppressor factor (GT-T8F) of suppressor T cells (T82) bearing distinct I-J determinants. The induction of new suppressor T cells (Ts2) by suppressive extracts (TsF) from L-glutamic acid50L-tyrosine50 (GT) nonresponder mice was examined. Incubation of normal spleen cells with allogeneic GT-TsF for 2 days in vitro led to the generation of Ts2 cells able to suppress subsequent responses to the immunogen GT-methylated bovine serum albumin (GT-MBSA) in vivo. This induction occurred efficiently when TsF donor and target cells differed at all of H-2, including the I-J subregion. B10.BR (H-2k) GT-TsF, adsorbed on, then acid eluted from GT-Sepharose and anti-I-Jk [B10.A (3R) anti-B10.A (5R)]-Sepharose in a sequential fashion could induce BALB/c (H-2d) spleen cells to become Ts2 only if nanogram quantities of GT were added to the purified GT-TsF. This indicates a requirement for a molecule or molecular complex possessing both I-J determinants and antigen (GT)-binding specificity, together with GT itself, for Ts2 induction. The induced Ts2 are I-J+, since their function can be eliminated by treatment with anti-I-Jk plus C. These I-J determinants are coded for by the precursor of the Ts2 and do not represent passively adsorbed, I-J coded TsF, since anti-Ijk antiserum [(3R X DBA/2)F1 anti-5R] which cannot recognize the BALB/c (I-Jd) TsF used for induction still eliminates the activity of induced A/J (I-Jk) Ts2. These data provide further evidence for and information about the minimum of two T cells involved in antigen-specific suppressor T cell systems.


Immunity to Plasmodium berghei yoelii in mice. II. Specific and nonspecific cellular and humoral responses during the course of infection. The kinetics of various specific and nonspecific immunologic responses were examined in BALB/c mice infected with 17X nonlethal Plasmodium berghei yoelii (a self-limiting infection). The sequence of events after infection was characterized by rapid sensitization of splenic T cells to malaria antigen and polyclonal B cell activation, followed by a period of depressed splenic proliferative responses in vitro to mitogens (PHA and LPS) and malaria (specific) antigen. At the same time, suppressed primary in vitro splenic PFC responses to trinitrophenyl-aminoethylcarbamylmethyl-Ficoll (TNP-F) were seen. This suppression was an active process requiring adherent cells. During this period, levels of antimalarial antibody also increased exponentially. As the infection was cleared, splenic malaria antigen-specific proliferative responses were again observed and splenic PFC and in vitro mitogen responses returned to preinfection levels after variable periods of time. Both splenic proliferative responses to malaria antigen and antimalarial antibody responses remained persistently elevated. In addition, some responses were examined in mice infected with 17X lethal P.b. yoelii (a fatal infection); in comparison to the early responses of mice infected with the nonlethal substrain, there was a decrease and delay in the development of a splenic T cell response to malaria antigen and a blunted antimalarial antibody response.


Sharing of Ia antigens between species. II. Molecular localization of shared Ia determinants implies the existence of more than one I sublocus of the rat MHC. A mouse alloantiserum B10.D2 anti-B10.BR (H-2d anti-H-2k) cross-reacted with rat lymphocyte surface glycoproteins with characteristics of Ia antigens. Sequential precipitation analysis on solubilized radiolabeled LEW rat lymphocyte antigens with this cross-reactive mouse alloantiserum and the rat alloantiserum BN anti-LEW (Ag-B3 anti-Ag-B1) revealed that the Ia-like antigen detected by the mouse alloantiserum also reacted with the rat anti-Ia antibodies. It was further shown that the rat alloantiserum also detected another set of Ia-like antigens that did not cross-react with the mouse alloantibody. Precipitation analysis with congenic rat strains confirmed that all Ia-like antigens precipitated by the rat alloantibody were encoded by Ag-B linked genes. Thus the shared Ia-like antigen must also be the product of Ag-B-linked gene(s) or be physically associated with such products. In addition, molecules bearing shared antigenic determinants were separable from at least some of the Ia-like antigens detected by the rat alloantiserum, possibly suggesting the existence of more than one sublocus coding for Ia antigens within the rat MHC.


Macrophage-lymphocyte interaction in migration inhibition factor (MIF) production against soluble or cellular tumor-associated antigens. I. Characteristics and genetic control of two different mechanisms of stimulating MIF production. For MIF production in response to 3 M KCl extracts of tumor, viable and metabolically active macrophages have been shown to be required to interact with the soluble tumor antigens and then come in contact with immune lymphocytes. The Mphi-lymphocyte interaction for MIF production was found to be under the control of genes mapping in the IA subregion of the H-2 complex. However, when intact tumor cells were used as antigen, Mphi were not required for immune lymphocytes to produce MIF. In addition, the interaction of immune lymphocytes with tumor cells for MIF production did not require H-2 compatibility. These and other observations strongly suggest that there are two different mechanisms for MIF production and that these may be mediated by two separate subpopulations of immune lymphocytes.


"Silent" alleles at the HLA-C locus.

"Silent" alleles at the HLA-C locus. Defined specificities of the HLA-C locus possess a significantly lower immunogenicity as compared with the HLA-A and HLA-B specificities. HLA-C "blanks" are not immunogenic at all. Among the possible explanations, the most likely one appears to be a low phenotypic expression of these alleles. C locus antigens as a whole are thus likely to be of minor relevance in donor/recipient matching for transplantation.


Characterization of suppressive immunoglobulin-binding factor (IBF). III. Biochemical and immunochemical characteristics of IBF produced by activated T cells. The biochemical characteristics of immunoglobulin binding factor produced by activated cells (ATC) were investigated. For this purpose, supernatants of ATC were purified by affinity chromatography on insolubilized IgG and the eluted material was iodinated (125I), treated with mercaptoethanol; and run on SDS polyacrylamide gels. The radioactivity was found in two peaks corresponding to m.w. of 38,000 d and 18,000 d. This result extends and confirms our previous findings that IBF produced by ATC is identical to IBF produced by L-5178-Y internally labeled thymoma cells. The effect of various pH, temperatures, and proteolytic and glycolytic enzymes on the binding properties of 3H-leucine-or H-fucose-labeled IBF to IgG and on the polyacrylamide gel profiles was also studied. By all these criteria, IBF appeared to be a glycoprotein in which the presence of the 38,000 to 40,000 d chain is necessary for the binding to IgG. In the attempt to study the relationships between IBF and I-region products, purified IBF produced by ATC was incubated with anti-Ia immunoadsorbent, and the eluted material was iodinated and run on gels. The 38,000 d and 18,000 d chains characteristic of IBF were found to be specifically retained on the relevant immunoadsorbent. These data favor the hypothesis that IBF bears or is associated with Ia determinants.


Direct solid-phase radioimmunoassay for measuring antigenic differences between the hemagglutinins of influenza viruses. A direct solid-phase radioimmunoassay, which involves binding of purified radio-labeled IgG to influenza virus hemagglutinins immobilized on Sepharose 4B gel, is described.


Staining of eosinophil granulocytes in chemotactic studies. A fast and reproducible method for the staining of eosinophils on millipore filters for the study of chemotaxis is described. This procedure allows for the easy differentiation of eosinophils from other granulocytes both at the intra-filter level and on the filter surface.


The role of basophils in inflammatory reactions. This review demonstrates that basophils reflect skin and lung mast cell reactivity and show characteristic changes in mediator release associated with clinical disease. Although the numbers of IgE molecules and IgE receptors on basophils have been enumerated, these have, in most instances, little influence on the release of histamine after challenge. There is, rather, a parameter of "releasability" that may be a major variable in allergic disease states. Basophils contain and release histamine, the eosinophil chemotactic factor of anaphylaxis (ECFA), a slow reacting substance of anaphylaxis (SRS-A), and a kallikrein. The release process is controlled by hormone-basophil receptor interactions that determine the cyclic AMP level; plasma and tissue adenosine levels appear prominent in this control. Histamine feeds back to negatively modulate basophil and mast cell release through a specific histamine 2-receptor; it also inhibits lymphocyte and neutrophil function. Like neutrophils, basophils contain beta-glucuronidase while neutrophils contain SRS-A and a low-molecular-weight ECF. The stimuli for primary basophil and neutrophil release are, however, quite different, although phagocytic stimuli, which fail to cause basophil mediator release, potentiate the IgE response. It is concluded that basophols play a significant in vivo role in inflammation by acting as an interface between foreign antigens, the serum cascade systems, and other inflammatory cells.


Epidermal cytoplasmic antigens: II. Concurrent presence of antigens of different specificities in normal human skin. Cytoplasmic antigens of 3 different specificities are expressed concurrently in the epidermis of normal human skin. They differ by their location in distinct layers of the epidermis. One type is present in all epidermal cells, another only in cells of the superficial layers, and the third only in basal cells. All 3 antigens were present in 10 specimens of normal human skin. The superficial and basal cell antigens were absent in 4 basal cell epitheliomas and 3 squamous cell carcinomas. This finding, together with the stratification of the antigens in different layers of the skin, suggests epidermal cytoplasmic antigens are differentiation antigens.