Experience in palliative treatment of univentricular heart including tricuspid atresia. Over a period of ten years 110 patients with univentricular heart, including cases with tricuspid atresia, received palliation. The overall hospital mortality was 14.5%. Late mortality during a mean follow-up period of two years, was 6%. A group of patients with univentricular heart ("complex" group: 21 cases) had numerous and significant associated anomalies which affected the surgical results (16 survivors). Eight patients were discovered to have univentricular heart only at open heart surgery (5 survivors). In the remaining patients hospital mortality was 7.4% (6/81). As compared with the natural history these results indicate that palliative surgery is still an effective type of treatment for these two severe cardiac malformations.


Neutralization kinetic studies with genital cytomegalovirus isolates, an antigenically variable group. Antisera were prepared in rabbits against four low-passage genital isolates, and against two established strains of human cytomegalovirus. With these sera, and seven strains of virus, 42 virus-antisera combinations were examined by kinetic neutralization procedures, and NK-values derived. No evidence accrued indicating that the five genital isolates constituted an antigenically distinct group. The findings support the view that the human cytomegaloviruses are antigenically heterogeneous, with different strains reflecting an antigenic mosaic, the elements of which are present in varying amounts.


Observations of antigenic relatedness between viruses of the herpes simplex "neutroseron". The antigenic relatedness of three viruses of the herpes simplex type 1 neutroseron - herpes simplex virus types 1 (HSV-1) and 2 (HSV-2) and bovine mammillitis virus (BMV) - has been examined by immune precipitation and virus neutralization tests. Many virus-specific infected-cell polypeptides were shown to possess antigenic sites shared by both HSV-1 and HSV-2. Cross-neutralization between the viruses is mediated through antibodies to at least two antigenic sites, one shared by HSV-1, HSV-2 and BMV and one shared by HSV-1 and HSV-2 but not BMV.


A novel surface antigen on lymphoid cells transformed by Epstein-Barr virus. A surface antigen (SA) was detected on EBV-carrying lymphoid cell lines by an indirect membrane immunofluorescence test with serum from rabbits immunized with Raji cells; the antiserum had been extensively absorbed with normal human blood and tonsil cells. The SA was not detected on normal human umbilical-cord and adult peripheral blood lymphocytes or EBV-negative cell lines. The incidences of the SA and EBV-determined membrane antigen (MA) on certain EBV-carrying cell lines were not compatible. Antibody against SA or MA was differentially abolished by absorption with the SA-positive but MA-negative cell line or the MA-positive but SA-negative cell line, respectively. The results of cross-absorption tests of antisera against either Raji cells or P3HR-1 cells suggested that SA is not a single but a complex antigen.


Apparent absence of virus-specific transplantation rejection antigen in herpes simplex virus-transformed hamster cells. HSV-1 sensitized hosts were not resistant to challenge with small numbers of HSV-1 transformed cells. Virus immunization did not inhibit or enhance the incidence of tumour metastases.


Anti-allergic activities of a new benzopyranopyridine derivative Y-12,141 in rats. Passive cutaneous anaphylaxis (PCA) mediated in rats by IgE-like antibodies against egg albumin or the benzylpenicilloyl determinant was inhibited in a dose-dependent manner by intravenous treatment with Y-12,141; the ED50 was 0.09--0.2 mg/kg. The inhibitory effect of Y-12,141 ON PCA was about 5 times as potent as that of disodium cromoglycate (DSCG). Oral treatment with Y-12,141 resulted in the inhibition of PCA, showing an ED50 of 2.5 mg/kg. This action of Y-12,141 on PCA was considered to be due to the inhibition of the release of allergic mediatros from mast cells in a manner similar to DSCG. The results suggest that Y-12,141 may have an anti-allergic activity.


Similarity of casein- and endotoxin-induced, myeloma- associated and aged SJL/J amyloid in various strains of mice. Amyloidosis was induced in a number of strains of mice by repeated injections of casein and endotoxin. Spontaneous amyloid was obtained from Balb/C mice bearing a myeloma tumor (IgG2a producing MOPC 173 tumor) and from aged SJL/J mice. Both the induced and spontaneous forms were similar in their size, immunological reactivity, peptide maps and in the susceptibility of histological sections to oxidizing agents with or without trypsin digestion. Since case-induced murine amyloid resembles the nonimmunoglobulin from of human amyloid, it is concluded that an immunoglobulin form in mice has yet to be characterized.


Antigen-induced release of histamine from passively sensitized guinea pig lung slices. I. Optimum conditions for in vitro passive sensitization and challenge with antigen. The effect of several parameters (pH, Ca++ concentration, time, temperature, lung slice size) on in vitro antigen-induced histamine release from passively sensitized guinea pig lung slices was investigated. With respect to pH, Ca++ concentration and time, the optimal conditions for the passive sensitization step (pH 7.8, Ca++ 1.5 mM, 2 h) were distinctly different from those for the antigen challenge step (pH 8.2, Ca++ 10mM, 15 min). Maximum antigen-induced release was obtained with 3 X 0.25 X 0.25 mm lung slices at 37 degrees C using air as gas phase.


Lateral and transmembrane redistribution of tissue-specific antigens in single cells and monolayers. Tissue-specific antigens in the membranes of corneal endothelial cells react with anti-tissue antibodies only in metabolically active monolayers and dispersed cells. After metabolic inhibition by exposure of these preparations to cold, the antigen-antibody complexes, like free antigens, undergo transmembrane redistribution leading to their internalization by the cells. This transmembrane redistribution is reversible and can be followed by using fluorescein-labeled antibodies. Reexpression of the complexes on the cell surfaces occurs after return from metabolic inhibition to metabolic activity. Dispersed corneal endothelial cells are also capable of lateral redistribution (capping) of the complexes although cells in monolayers do not share this capability. Capping in the dispersed cells occurs only at ambient temperatures and, because it results in shedding of the complexes, is irreversible. The data indicate that macromolecules in the membranes of cells organized in tissues are restricted in their movement as compared to the macromolecules of cells functioning in a dispersed state.


Preferential synthesis of IgE reaginic antibodies in rats immunized with alum-adsorbed antigens. The reaginic antibody response to alum-precipitated ovalbumin (OA) and the dialyzed water-soluble extracts of ragweed (DWSR) and Alternaria tenuis (DWST) in several strains of rats appeared to be wholly an IgE response. There was no evidence of a heat-stable (IgGa) antibody to OA, DWSR and DWST in the sera of the rats immunized with these antigens suspended in alum. Wistar-Furth and Lew inbred and hooded outbred rats produced comparable amounts of reaginic antibody after immunization with DWST, but BN inbred rats failed to generate a reaginic response to this antigen. The amount of antigen-induced histamine release from rat peritoneal mast cells did not always correlate with the level of circulating IgE-specific antibody.


The effect of hapten-specific suppression of IgE on antigen-induced histamine release from mouse peritoneal mast cells. Subcutaneous injections of a mixture of dinitrophenylated ovalbumin (DNP3-OA) and dextran sulfate into Swiss-Webster mice elicited short-lived primary and long-lasting secondary IgE antibody responses to both DNP and OA. Histamine was released on in vitro challenge with antigen (OA or DNP22-BSA) of washed peritoneal mast cells (PMC) obtained from mice during a primary or a secondary IgE response. Administration of an intravenous injection of a tolerogenic conjugate of DNP8-mouse gamma-globulin, either prior to immunizationor during an ongoing IgE response, resulted in almost complete disappearance of circulating anti-DNP IgE antibody and in a very marked decrease in histamine release from PMC on challenge with DNP22-BSA. However, the IgE response to OA of these mice and the histamine release from their PMC on challenge with OA were not affected. Moreover, the PMC of mice, which had been tolerized to DNP, could be passively sensitized with serum containing DNP-specific IgE antibody for the release of histamine on DNP22-BSA challenge. The most significant finding of this study is the observation that the time course for the loss of reactivity of PMC to DNP22-BSA, after administration of the tolerogen during an ongoing secondary response, paralleled the decrease in circulating anti-GNP IgE antibody.


Inhibition of rat mast cell degranulation and histamine release by histamine-rat gammaglobulin conjugate. Histamin-rat-gamma-globulin conjugate inhibited degranulation and histamine release of rat peritoneal mast cells to a greater extent than the rat globulin or histamine alone. Since mast cells contain histamine receptors, it may be assumed that the histamine bound to the gamma-globulin combines with the rat mast cell histamine receptor and inhibits the degranulation and histamine release by a feedback mechanism.


Impairment of acute protein reactivity in chronic renal failure. The acute phase protein response was studied after elective surgery in 13 normal subjects and 9 patients with severe  chronic renal failure. Total haemolytic complement reactivity (CH50) and serum concentrations of C1q, C1s, C4, C3, factor B, properdin, C5, C9, C-reactive protein (CRP), caeruloplasmin, alpha1-acid glycoprotein and haptoglobin were measured preoperatively and on days 2, 4 and 6 after operation. Abnormalities were seen in the group with chronic renal failure. Firstly, there was no significant acute phase response of C1s, C3, C5, C9 and CH50 and a significant reduction in the response of factor B. Secondly, CRP showed prolonged elevation in the post-operative period in contrast to the transient rise seen in the control group. With the possible exception of alpha1-acid glycoprotein, the behaviour of the non-complement proteins (caeruloplasmin and haptoglobin) was comparable for the two groups. These defects could impair the physiological response to infection in patients with severe chronic renal failure.


Acute inflammation following intraperitoneal injection of antigen into actively sensitised rats. Rats actively sensitised to bovine serum albumin (BSA) in Freund's complete adjuvant were challenged by intraperitoneal injection of BSA. By washing out the peritoneal cavities at different times it was possible to divide the changes that occurred into two phases. The first was characterised by the release of histamine and slow reacting substance of anaphylaxis (SRS-A) with an associated extravasation of plasma proteins. The second phase was characterised by an increase in neutrophilic polymorphonuclear cells in the peritoneal cavity. Extracellular enzyme activites were also investigated. The kinetics of these events were studied and are discussed in terms of the possible mechanisms involved.


Alkaline phosphatase, phosphoethanolamine, and ethanolamine content in benign prostatic hypertrophy. The amino acid patterns of benign prostatic hyperplasia specimens show that the contents of phosphoethanolamine and ethanolamine are very variable from one tissue sample to another. An increased phosphoethanolamine content is accompanied by a low ethanolamine content, and vice versa. The concentrations of the two substances correlate well. There exists a possible involvement of alkaline phosphatase in this inverse relationship between phosphoethanolamine and ethanolamine. In specimens with increased phosphoethanolamine and decreased ethanolamine levels alkaline phosphatase activity was low; however in specimens with decreased phosphoethanolamine and increased ethanolamine levels alkaline phosphatase activity was high.


Immunochemical identification of prostatic epithelial cells in culture. An indirect immunofluorescence technique for identifying prostatic acid phosphatase was used to evaluate primary prostatic cell cultures and established cell lines of prostatic origin. With the use of this technique, we positively identified acid phosphatase immunochemically of prostatic origin in cell lines EB 33 and MA 160. Other cell lines showing positive immunofluorescence reactions include H 494, H 575, and DU 145.


Qualitative analysis of proteinuria associated with bladder cancer. In the present investigation molecular components associated with the urines from bladder cancer patients and normal individuals are identified. Polyacrylamide gels of urines from bladder cancer, bladder papilloma, and normal individual exhibit clear differences in banding patterns. Urine from bladder cancer patients shows gels with increased quantities of low (less than 100,000) and increased and additional high (less than 100,000) molecular weight proteins when compared to gels with urine from papilloma and normal individuals. In order to localize and identify proteins in the urine from bladder cancer, papilloma, and normal individuals, proteins were separately fractionated on Sephadex G-200 columns and each elution fraction was reacted on Ouchterlony gel diffusion against various specific antisera. The qualitative analysis of proteins in urine from bladder cancer patients is discussed in relation to their molecular weight distribution in the Sephadex G-200 profiles and their possible role in tumor host relationships.


Side-specific effects of sodium on (Na,K)-ATPase. Studies with inside-out red cell membrane vesicles. Using inside-out vesicles of human red cell membranes, the side-specific effects of Na+ on phosphorylation of (Na,K)-ATPase have been studied using low concentrations of [gamma-32P]ATP (less than or equal to 0.1 microM). Phosphorylation is stimulated by Na+ at the cytoplasmic membrane surface (extravesicular Na+) alone and not by Na+ at the external surface (intravesicular Na+). At 37 degrees C, external Na+ (less than or equal to 10 mM) does, however, increase the steady state level (approximately 2 1/2-fold) of phosphoenzyme above that observed with cytoplasmic Na+ alone; hydrolysis is increased to only a small extent. Little stimulation by external Na+ is observed at 0 degrees C. As Na+ at the cytoplasmic side is decreased to very low levels (less than or equal to 0.2 mM) several kinetic changes are observed: (i) the apparent turnover of phosphoenzyme (ratio Na+-ATP-ase/phosphoenzyme level) is markedly increased (approximately 3-fold, (ii) Rbext sensitivity (inhibition of (Na)-ATPase at low ATP levels) is reduced, and (iii) the ratio of Na+ ions transported per molecule of ATP hydrolyzed is decreased. These results are compatible with a reaction pathway involving a transition from one form of phosphoenzyme, E1-P, to another, E2-P of which the hydrolysis is decreased by moderate levels of external Na+. It is suggested also that an alternate reaction pathway for Na+-ATPase occurs at very low cytoplasmic Na+, one via hydrolysis of E1-P and not associated with Na+ translocation.


The sialoglycoproteins of murine erythrocyte ghosts. A modified periodic acid-Schiff stain procedure staining nonsubstituted and O-acetylated sialyl residues on glycopeptides. Murine erythrocyte ghosts (from DBA/2, CD-1, and B6D2 strains) contain significant amounts of O-acetylated sialyl residues, which are slowly oxidized by periodate. Sialic acids have been purified from murine erythrocyte ghosts and the existence of O-acetylated sialic acids has been confirmed: 1) by assaying with the Warren procedure before and after de-O-acetylation with 0.1 N NaOH for 45 min at 4 degrees C; 2) by thin layer chromatography on cellulose; and 3) by gas-liquid chromatography. Because these sialyl residues are unevenly distributed on the sialoglycoproteins of murine erythrocyte ghosts, the periodic acid-Schiff staining detects only one major sialoglycoprotein. A modification of the periodic acid-Schiff stain method removes these O-acetyl groups after electrophoresis and reveals two additional sialoglycoproteins which bear the majority of the O-acetylated sialyl residues. Rat erythrocyte ghosts have similar residues on one of their two sialoglycoproteins. Ghosts of human, rabbit, and guinea pig erythrocytes do not contain detectable amounts of O-acetylated sialyl residues.


Studies with anti-growth hormone receptor antibodies. Antisera against a partially purified growth hormone receptor derived from rabbit liver were generated in guinea pigs. The antisera specifically inhibited the binding of 125I-ovine growth hormone (oGH) to liver membranes but had no effect on the binding of 125I-ovine prolactin to rabbit mammary gland receptors. These antisera did not bind or destroy 125I-oGH. Moreover, the binding of labeled growth hormone to membrane particles derived from liver of several species was also inhibited by the antisera, thus suggesting that immunological determinants of the growth hormone receptor of several species are similar. gamma-Globulin fractions derived from the antisera were responsible for the inhibition. In addition 125I-gamma-globulin derived from one antiserum bound to membrane pellets with a corresponding decline in 125I-oGH binding. Kinetic analysis of inhibition of 125I-oGH binding suggested a hyperbolic competitive inhibition, a point of view which is favored by the demonstration of a hormone receptor . antibody complex. The availability of the antireceptor sera confirmed previous data that differential affinity chromatography separated growth hormone and prolactin receptors in solubilized rabbit liver membrane preparations. The antireceptor sera will be useful probes in further characterization of the growth hormone receptor.


Response to hormones of cells cultured from human giant cell tumors of bone. Giant cell tumors of bone obtained from 7 patients were dispersed with clostridial collagenase and trypsin and adherent cells were maintained in culture. Early cultures contained both mononucleated and multinucleated cells presumably derived from the stromal and giant cells of the original tumor. The original multinucleated cells did not survive for greater than 7-10 days whereas the mononucleated cells persisted and could be passaged by trypsinization. In 5 of 7 early cultures exposed to parathyroid hormone (PTH) there was a rise in cAMP within 5-10 min in both cells and medium which averaged approximately 12-fold. None of the cells responded to calcitonin and a variable rise in cAMP was seen after incubation with prostaglandin E2. In cells cultured from 3 tumors the PTH response disappeared with passage of the cells, but in the remaining 2, PTH response persisted through multiple passages. The presence as well as the magnitude of the PTH-induced cAMP response in these cells is consistent with a skeletal origin.


Evaluation of T and B lymphocytes in liver infiltrates of patients with chronic active hepatitis. The proportions of T and B lymphocytes in the liver infiltrates of 23 patients with chronic active hepatitis have been determined. The results were compared with the values obtained from peripheral blood and with the presence of HB virus markers and alpha-fetoprotein in liver tissue. A group of patients with chronic liver disease other than chronic active hepatitis were studied as controls. In chronic active hepatitis the percentage of hepatic T cells was 49 +/- 8 SD (control patients 61 +/- 8) (P less than 0.01), whereas the percentage of B cells was 40 +/- 10 (control patients 18 +/- 8) (P less than 0.01). No correlation was observed between hepatic T and B cells and the presence of HB virus. The numbers of T cells in liver tissue was significantly higher, the numbers of B cells lower, in patients whose biopsies were positive for alpha-fetoprotein than in those whose biopsies were negative. In peripheral blood, only the patients with chronic active hepatitis and established cirrhosis presented lower absolute values of T cells, whereas surface immunoglobulin-positive lymphocytes were within the normal range.


Correction of Fallot's tetralogy after palliative operations. During the period 1960 to 1978, 98 patients underwent intracardiac repair of Fallot's tetralogy after palliative operations. Preoperative symptoms were cyanosis, dyspnea, increased fatigue with squatting and hypoxic spells. The hemoglobin concentration varied from 19 to 22 g/100 ml. At correction only 65 of 95 shunts were patent and needed surgical closure. Seventeen early deaths occurred (19%), the main causes being cardiac failure and arrhythmia. One patient died 3 years after correction from pneumonia. The subjective clinical result was excellent or good in all surviving patients. At repeat heart catheterization in 26 patients a high percentage of residual ventricular septal defects and pulmonary stenosis/insufficiency was found. However, the majority of defects were of minimal haemodynamic significance, and so far did not seem to do harm to the patients' subjective function.


Detection of bacterial phosphatase activity by means of an original and simple test. A new test for the detection of bacterial phosphatase activity has been devised. The test is performed using agar media containing both methyl green (MG) and phenolphthalein diphosphate (PDP); in these media phosphatase-producing strains grow deep-green-stained colonies whereas non-producing strains do not. A total of 739 different strains were tested, including 593 staphylococci, 95 micrococci, 11 streptococci, 10 corynebacteria, 14 enterobacteria, and 16 candidae. All strains found phosphatase-positive according to the conventional phosphatase test displayed deep-green-stained colonies on MG-PDP media, whereas all phosphatase-negative strains showed unstained colonies on the same media. The main advantages of the present phosphatase test as compared with other conventional ones are that it is more simple to perform, it can reveal the phosphatase activity of colonies grown in deep agar, and can be incorporated into commercial multitest kits.


Antibodies to germinating and yeast cells of Candida albicans in human and rabbit sera. Two major antigenic components, I and II, were detected by double immunodiffusion in sonic extracts of the germinating (G) or yeast (Y) cells of the dimorphis organism, Candida albicans group A. Component I may be a heterogeneous mixture of antigens which are stable to heating and phenol. Component II is more homogeneous but is labile to heat and phenol. Rabbit antisera, showing only precipitin to component II or certain human sera at high dilution, were found to react with G cells to give an immunofluorescence which was confined to the germ tubes. This suggested that component II is localised on the germ tubes, whereas no immunofluorescent reaction against the yeast cells could be detected under the same conditions although component II was as readily extracted from these cells as from G cells. This suggested that component II might exist in a cryptic state in the Y cells. In support of the latter contention it was shown that live Y cells did not absorb precipitin to component II nor were they capable of providing these antibodies in rabbits. Using both human and rabbit sera, it was shown that the antigenic specificity of the immunofluorescence assay where Y cells were used was related to component I and that where G cells were used it was related to both components I and II.


Intracellular marking with lucifer yellow CH and horseradish peroxidase of cells electrophysiologically characterized as glia in the cerebral cortex of the cat. Intracellular microelectrodes filled with either Lucifer Yellow CH, a highly florescent dye, or horseradish peroxidase (HRP) were used to electrophysiologically characterize and mark cells in the cerebral cortex of cat. Fifty-eight cells, characterized electrophysiologically as glia, were marked with Lucifer Yellow CH. All were identified as protoplasmic astrocytes, and included cells in the glia limitans of the molecular layer. An additional 54 cells, similarly characterized as glia, were labeled with HRP. The results were the same; only protoplasmic astrocytes were labeled. The "staining quality" of the glia labeled with HRP was superior to that of cells injected with Lucifer Yellow; greater lengths of individual processes were revealed, and they could often be followed to blood vessels where they ended on the walls of vessels with expanded perivascular end-feet. The observations indicate that the many previously reported studies on presumed glial cells in the cat cerebral cortex have characterized the behavior of protoplasmic astrocytes. Neurons were also marked during these experiments. The "staining" quality of the Lucifer Yellow filled neurons was excellent; dendritic spines, axons, and axon collaterals were clearly visible. These fine neuronal details were not as well revealed after HRP labeling. High resting membrane potentials (RMP's) were not a prerequisite for obtaining well-marked neurons (mean RMP of Lucifer Yellow filled neurons was -33.6 mV; mean RMP of HRP filled neurons was 42.3 mV). In contrast, the mean RMPs of Lucifer Yellow and HRP marked glia was -68 Mv and -75 mV respectively, and the quality of "staining" appeared to be more closely related to the RMP.


Trajectory of group Ia afferent fibers stained with horseradish peroxidase in the lumbosacral spinal cord of the cat: three dimensional reconstructions from serial sections. A reconstruction was made of the intramedullary trajectory of 23 physiologically identified Ia afferents from cat hind limb muscles (medial gastrocnemius, soleus, plantaris, flexor digitorum-hallucis longus, and hamstring). The afferents were stained by intra-axonally injected HRP. The axons of these afferents were traced over distances of 5.8 mm to 15.7 mm rostrocaudally. In the dorsal funiculus fibers from all the muscles showed a similar course and similarly bifurcated into an ascending and a descending branch. The mean diameters of stem axons, ascending branches, and descending branches were 6.6 micrometer, 5.8 micrometer, and 3.0 micrometer, respectively. Within the analyzed lengths of the spinal cord five to eleven collaterals were given off from the two branches. The distances between adjacent collaterals of the ascending and descending branches averaged 1200 micrometer and 790 micrometer, respectively. The collaterals as a rule passed through the medial half of the dorsal horn before they entered the deeper parts of the gray matter. The terminal distribution areas common to all Ia collaterals were: (1) the medial half of the base of the dorsal horn, mainly lamina VI: (2) lamina VII; and (3) lamina IX. The numbers of terminals were largest in lamina IX and smallest in lamina VII. The density of terminals in lamina IX was highest in the homonymous motor cell column. The terminal distribution areas of adjacent collaterals showed no overlap in the sagittal plane. Terminal branches carried one bouton terminal and up to six boutons en passage with an average of 1.8 terminals per terminal branch. Apparent axosomatic and axodendritic contacts were seen on small-sized and medium-sized neurons in laminae V-VI, medium-sized neurons in lamina VII, and large neurons in lamina IX. One motoneurons was contacted by an average of 3.3 terminals. In addition to the common features, Ia collaterals of various muscles of origin showed some differences in their trajectories in the ventral horn, and in their terminations in the gray matter.


AVLINE: a search resource for audiovisual instructional materials. This manuscript describes the historical development and present scope of AVLINE (AudioVisuals-on-LINE), the National Library of Medicine's computer data base of information on nonprint instructional materials in dentistry, medicine, nursing, and allied health. The manuscript outlines the early and present review processes and involvement of dentistry peer reviewers; and provides data describing AVLINE titles by health science discipline and media characteristics for the total data base and the subset of dentistry. The manuscript also includes information on how to access AVLINE and a sample AVLINE citation.


Early effects of corticosteroids on basophils, leukocyte histamine, and tissue histamine. The comparative effect in 11 atopic subjects of a single intravenous injection of methylprednisolone on sequential studies of blood eosinophils, basophils, leukocyte sensitivity to antigen for histamine release, leukocyte histamine content, and skin histamine was examined. No significant changes occurred in any parameter after placebo treatment. In contrast, 4 hr after intravenous treatment with steroid there were significant decreases in mean eosinophil counts (-95%), basophil counts (-72%), and histamine content of 1 X 10(7) leukocyte samples (-62%). Temporal changes in the latter paralleled alterations in circulating basophil levels. No significant changes occured in the antigen histamine release sensitivity, or the total skin histamine. Studies over a longer period after steroids in 4 subjects showed eosinophil and basophil levels at a nadir at 8 hr, remaining suppressed for 24 hr, and returned to pretreatment levels by 72 hr. Results suggest that corticosteroids induce a prominent decrease in leukocyte histamine due to a depletion of basophils without a decrease in histamine content per basophil, and that skin tissue histamine stores remain unchanged by such treatment.


Cytotoxicity of ionophore A23187 for basophils and other human blood cells. The calcium ionophore A23187(A23) at concentrations exceeding 1 microgram/ml has been shown to be progressively cytotoxic for human blood basophils, neutrophils, lymphocytes, and erythrocytes. Toxicity to basophils was considered to be manifested by the increasing inability of 2-deoxyglucose (2DG) to inhibit histamine release (HR) at increasing concentrations of A23. The toxicity to neutrophils and lymphocytes was demonstrated by decreased lactate production (LP) after incubation with A23 of Ficoll-Hypaque fractions greatly enriched in each respective cell type. Red cells present in dextran-sedimented leukocytes were increasingly susceptible to lysis during washing subsequent to exposure to increasing concentrations of A23. A concentration of 4 microgram A23/ml, which is cytotoxic at 37 degrees C, produced optimal and noncytotoxic HR at 22 degrees C. It was possible to reduce A23 concentrations required for optimal HR by increasing Ca++ from 0.6 to 3 mM.


Plasmodium-infected blood cells analyzed and sorted by flow fluorimetry with the deoxyribonucleic acid binding dye 33258 Hoechst. Red cells from Plasmodium berghei infected mouse blood can be sorted on the basis of their DNA content with the bisbenzimidazole dye 33258 Hoechst. The optimal conditions for dye uptake have been established and with these conditions uninfected cells are nonfluorescent and can be completely separated from infected cells which exhibit fluorescence in almost direct proportion to the number of parasite nuclei (i.e. DNA) they contain. The number of fluorescent cells detected and their fluorescence intensity is shown to be dependent on the dye concentration and the incubation medium being used. At least a proportion of the infected cells sorted from each fluorescence peak in the cell distribution retain their infectivity in vivo with some, but not all, conditions of labeling. This technique is being used to separate minor cell populations from infected blood for biochemical and immunochemical analyses and to screen human samples for malaria infected cells.


Specificities of killing by T lymphocytes generated against syngeneic SV40 transformants: studies employing recombinants within the H-2 complex. T lymphocyte effectors to syngeneic SV40-transformed cells, generated by secondary in vitro sensitization of immune spleen cells, lyse SV40 transformed targets that are syngeneic at the H-2 locus. In this study we have employed recombinants within the H-2 region to examine in detail this H-2 specificity. H-2b effectors were found to lyse SV40-transformed targets from recombinants bearing either H-2Kb or H-2Db.H-2k effectors recognized only SV40-transformed H-2Kk, and not H-2Dk target cells. By using the same protocol for sensitization, no effector cells could be detected in H-2d mice. Effectors generated in H-2 recombinant mice showed that the response capacity resides with K and D. For example, HTG, which is H-2d except at the D locus (H-2Db), produced effector cells specific for SV40-transformed H-2Db targets. Thus, the secondary in vitro response to SV40 transformants was found to depend only on the K and D alleles and not to be modified by the I region to any measurable extent.


Idiotypic analysis of anti-GAT antibodies. III. Determinant specificity and immunoglobulin class distribution of CGAT idiotype. The humoral response to poly-(L-glutamic acid60, L-alanine 30, L-tyrosine10), GAT, in mice is further characterized by both idiotype and fine specificity analyses. The common idiotype on murine anti-GAT antibodies (CGAT) was identified in anti-GAT antisera from seven additional strains of mice. These data confirm that the CGAT idiotype can be induced in all inbred strains of mice. Using a partially inbred strain of mice selected for their ability to respond to poly-(L-glutamic acid50, L-tyrosine50), GT, we demonstrated that the GT copolymer is capable of inducing antibodies that express the CGAT idiotype. In contrast, antisera directed against poly-(L-glutamic acid60, L-alanine40), GA, bind GAT but lack CGAT idiotype. These results indicate that GAT molecules contain determinants either similar or identical to those on GT molecules, which are responsible for the induction of CGAT idiotypic antibodies. We also demonstrated that both GAT responder and nonresponder strains of mice can produce anti-GAT antibodies with similar fine specificity patterns and CGAT idiotype. In addition, we demonstrate that antibodies of the IgM, IgG1, and IgG2 immunoglobulin classes express the CGAT idiotype.


Solid phase radioimmunoassay for quantitation of antigen-specific IgG in human sera with 125I-protein A from Staphylococcus aureus. Radiolabeled protein A from Staphylococcus aureus (Staph A) has been used to develop a solid phase, noncompetitive radioimmunoassay for quantitation of specific IgG antibody. The assay involves two incubations: First, agarose-insolubilized antigen is mixed with serum samples for 1 to 4 hr during which specific antibody is bound; second, after a washing procedure, the solid phase immune complexes are incubated for 4 to 18 hr with 125I-Staph A, during which the radiolabeled detection protein binds to the insolubilized specific IgG antibody. In a comparative study of the IgG antiphospholipase A antibody content of 23 human sera drawn from honeybee venom-sensitive patients, resulted of the Staph A assay correlated highly (r = 0.981, p less than 0.001, N = 23) with those obtained from a liquid phase, competitive radioimmunoprecipitation (double antibody) assay. The two assays demonstrated comparable precision, sensitivity, and reproducibility. In contrast, the use of 125I-Staph A in the solid phase radioimmunoassay was superior to 125I rabbit anti-human IgG because of lower negative serum (blank) values, shorter time required to reach equilibrium binding, and greater precision and reproducibility. In principle, the 125I Staph A assay may be applied ot IgG quantitation for crude allergen extracts as well as purified antigens. Furthermore, the sera of a number of mammalian species may be studied without further modification.


Synergistic cytotoxicity. I. Characterization of a heat labile plasma fraction that induces nonspecific cytotoxicity by human mononuclear cells. The mechanism by which non-immune mononuclear cells recognize invading foreign material and are activated for cytotoxic attack was studied in a model system employing human mononuclear cells, fresh plasma, and 51Cr-labeled xenogeneic target erythrocytes. In these experiments, fresh antibody-depleted plasma or mononuclear leukocytes alone were poorly cytotoxic to xenogenic erythrocytes. However, these target cells were rapidly lysed when both fresh antibody-depleted plasma and mononuclear cells were present in the assay. The plasma factor could not be removed by extensive absorption with the target cells, was present in plasma from hypogammaglobulinemic patients, was heat labile, and was sensitive to incubation with zymosan and cobra venom factor. The "antigen" specificity of this reaction was directed by the serum factor inasmuch as target cells autologous to the effector cells could be killed in the presence of antibody-depleted xenogeneic plasma, but not autologous plasma. These data suggest that an important mechanism for the recognition of "foreigness" by non-immune mononuclear cells is via interaction with a plasma component, possibly a factor related to serum complement.


Cell types required for H-2-restricted cytotoxic responses generated by trinitrobenzene sulfonate-modified syngeneic cells or trinitrophenyl-conjugated proteins. Murine spleen cells were fractionated over nylon wool or Sephadex G-10 columns, and the cell types involved in the generation of trinitrophenyl (TNP)-specific, H-2 restricted (TNP-self) cytotoxic effector cells were studied from cultures stimulated with trinitrobenzene sulfonate (TNBS)-modified syngeneic cells, TNP-conjugated soluble proteins such as bovine gamma-globulin (TNP-BGG), or bovine serum albumin (TNP-BSA). Unfractionated or nylon nonadherent responding cells generated such effectors, irrespective of whether the cultures were stimulated with TNBS-modified cells or TNP-conjugated proteins. TNP-modified T lymphocytes, B lymphocytes, and phagocyte-enriched spleen cells were all capable of stimulating TNP-self effectors. TNP-self effectors. TNP-self as well as allogeneic cytotoxic responses were dependent on the presence of a radioresistant non-T cell that was removed by Sephadex G-10 fractionation and was replaced by irradiated, Thy 1.2-negative, glass adherent spleen cells, enriched in phagocytic cells. Results obtained by using glass adherent cells that were allogeneic or semi-syngeneic to the responding cells indicated that H-2 homology was not required for efficient glass adherent cell function, and that the H-2 restriction of TNP-self effectors is not determined by these glass adherent cells.


A quantitative assay for experimental allergic encephalomyelitis in the rat based on permeability of spinal cords to 125I-human gamma-globulin. We have developed a quantitative assay for experimental allergic encephalomyelitis (EAE) in the rat based on permeability of the spinal cord to 125I-human gamma-globulin (HGG). This assay is highly reproducible and eliminates many of the drawbacks of assaying for EAE on the basis of clinical and/or histologic criteria. Using the assay, we have shown a direct correlation between onset of histologic changes in the spinal cord and onset of permeability changes in the spinal cord. No rat without histologic lesions manifest permeability alterations, and all rats with histologic lesions did manifest increased permeability to 125I-HGG. Furthermore, strains of rats susceptible to EAE demonstrated permeability changes, whereas resistant rats did not. In addition, we demonstrated by permeability and histologic criteria that guinea pig myelin basic protein emulsified with incomplete Freund's adjuvant is encephalitogenic in the Lewis rat. We also demonstrated that recipients of passive transfer of sensitized cells develop permeability changes along with histologic lesions. We conclude that measuring permeability to 125I-HGG in the spinal cords of rats is a valid assay for EAE, and its improves upon current indices of EAE in that it is readily quantifiable.


Detection of HLA-DRw (Ia-like) antigens on human T lymphocytes grown in tissue culture. Human T lymphocytes that proliferate in the presence of conditioned medium from PHA-stimulated allogeneic peripheral blood cells were shown to express IPA antigens after the 8th culture day. Ia antigens as detected by xenogeneic antisera were present on 80 to 90% of the cultured cells which were also strongly reactive with xenogeneic antisera defining a human T cell antigen and formed E rosettes. Control cultures with PHA or no conditioned medium expressed T cell but not Ia antigens. These cultured T cells also express the same HLA-DRw determinants as the B cells of the donor they were derived from. Absorption of xenogeneic Ia, and HLA-DRw alloantisera with cultured T cells completely removed the reactivity of these sera for enriched peripheral blood B lymphocytes from normal donors.


IgE-selective and antigen-specific unresponsiveness in mice. I. Induction of the unresponsiveness by administration of ovalbumin-pullulan conjugate. An experimental model was demonstrated in mice for the induction of IgE-selective unresponsiveness to ovalbumin, a protein antigen. An administration of ovalbumin, conjugated with pullulan, a linear polymer of glucose, (OA-pullulan) into mice resulted in the induction of a long lasting, IgE-selective unresponsiveness to the subsequent immunization with native OA in the form optimal to elicit IgE antibody response. The IgE-selective unresponsiveness is antigen specific and is infectious to normal mice by transferring the spleen cells from mice receiving OA-pullulan conjugate at least 2 weeks before. In contrast to other modified antigens, OA-pullulan was found to elicit good IgM and IgG antibody responses, but not an IgE response, without the aid of an adjuvant.


Cross-reactions between tryptic polypeptides of staphylococcal enterotoxins B and C. The strong cross-reactions demonstrated for staphylococcal enterotoxins B (SEB) and C1 (SEC1) by measurement of antigen-binding capacity were reflected in well defined polypeptides obtained by limited tryptic digestion from SEB and SEC1. Two antigenic determinants on each enterotoxin were capable of reacting with heterologous antibody, one on the first 57 amino acids and one on the last 150 residues of the polypeptide backbone. The larger, carboxyl terminal polypeptides bound efficiently to homologous antiserum but about two orders of magnitude less efficiently to heterologous antibody. The amino terminal peptides showed only weak homologous binding but nearly comparable heterologous binding. It is proposed that the determinant on the amino terminal polypeptides is largely responsible for the strong reciprocal binding of the intact enterotoxins and that their low antigen-binding capacity is due to a random or a structurally distorted conformation in solution.


Plaque-forming cell response in BALB/c mice to two preparations of LPS extracted from Salmonella enteritidis. The pattern of development of antibody-forming cells in BALB/c mice after immunization with PW-LPS or TCA-LPS was shown to be different. On days 10 and 20, the primary response to PW-LPS was characterized by a low level of IgM synthesis. The plaque-forming cell (PFC) response to TCA-LPS, however, increased from day 10 to day 20. Initially, IgM was the only detectable antibody synthesized but by day 20 a significant number of IgG-producing spleen cells had developed. After a secondary immunization with the appropriate lipopolysaccharide (LPS) preparation, IgG-producing spleen cells were detected in mice immunized with either PW-or TCA-LPS. Partial removal of the LAP or TCA-LPS with phenol or trypsin and pronase significantly reduced the PFC response, suggesting that the protein moiety played an influential role in the immunogenicity of TCA-LPS. The TCA-LPS contained the same antigenic dterminants as PW-LPS, so any difference observed between PFC response was not due to any associated immunogenic moiety.


The expression of Ia antigenic determinants on macrophages required for the in vitro antibody response. A subpopulation of antigen-presenting macrophages required for an in vitro antibody response to burro erythrocytes was deleted by pretreating the splenic macrophages with anti-Ia serum and complement (C). The in vitro response of the macrophage depleted T-B cell population could not be restored by the addition of macrophages resistant to anti-Ia antibodies and C (Ia-). The response of Ia- macrophages and the macrophage-depleted T-B cells was only reconstituted by the addition of Ia+ macrophages. Macrophages pretreated with anti-Ia antibodies restricted to react with determinants of one I subregion could not support the in vitro antibody response when added to cultures whose macrophages were pretreated with anti-Ia serum and C specific for the I-J subregion. These results confirmed that Ia determinants of the I-A, the I-E, and the I-C subregions were all expressed on the I-J+ macrophage required for an in vitro antibody response.


The activation of guinea pig T lymphocytes by anti-beta 2-microglobulin serum. In order to study further the role of beta 2-m in the regulation of the immune response, we have examined the effects of a goat anti-guinea pig beta 2-m serum on a number of T lymphocyte functions in vitro. Anti-beta 2-m serum produced a marked inhibition of the response of peritoneal exudate T cells to antigen and mitogen stimulation. Surprisingly, a marked activation of lymph node T lymphocyte proliferation was observed in the absence of antigen or mitogen stimulation. This stimulatory effect of anti-beta 2-m serum was shown to be specific for beta 2-m and required the presence of macrophages. The T cell proliferative response induced by anti-beta 2-m could not be blocked by antisera to the antigens of the guinea pig MHC. These studies suggest that beta2-m may play some critical role in the immune response at the level of T cell activation.


In vitro analysis of allogeneic lymphocyte interaction. III. Generation of a helper allogeneic effect factor (AEF) across an I-J subregion disparity. Allogeneic effect factors (AEF) were produced across an I-J subregion incompatibility. The helper activity of these AEFs is H-2 restricted since they help B cells only of the stimulator haplotype and of other haplotypes that carry the same I-J subregion gene(s) as the stimulator haplotype. Immunoadsorption studies demonstrate that they consist of I-J determinants derived initially from the GVHR host and MLR stimulator cells and not the GVHR donor and MLR responder cells used to generate AEF. It is postulated that the genetic restriction of AEF helper activity is mediated in part by the ability of the GVHR activated donor T cells to acquire, in vivo, recipient T cell and/or macrophage derived I-J determinants. Cellular adsorption studies indicate that AEF helper activity may be adsorbed by B cells, but neither T cells nor macrophages, of the stimulator haplotype. The results suggest that an I-J-positive AEF interacts with an I-J subregion controlled complementary recognition structure on a target B cell and, after antigenic stimulation, activates that B cell to IgG antibody synthesis.


Specific binding of T lymphocytes to macrophages. V. The role of Ia antigens on Mphi in the binding. The effect of anti-Ia alloantiserum on the capacity of selected peritoneal exudate lymphocytes (selected PEL) to bind to antigen-pulsed F1 (responder x nonresponder) macrophages was investigated. With the use of selected PEL for antigens under Ir gene control, it was shown that anti-Ia serum to the responder haplotype blocked adherence of selected PEL to antigen-pulsed macrophages whereas anti-Ia serum to the nonresponder haplotype did not. The target cell of the anti-Ia alloantiserum appeared to be the macrophage because anti-13 Ia in contrast to anti-2 Ia did not inhibit binding of F1 (2 x 13) DNP-GL selected PEL to DNP-GL pulsed strain-2 Mphi (responder strain). Taken together with previous experiments that indicate that an antibody to the native protein antigen employed is unable to block specific binding, the present results suggest that T cells may recognize fragments of exogenous antigen in association with Ia molecules.


Interferon-induced enhancement of IgE-mediated histamine release from human basophils requires RNA synthesis. Incubation of human leukocytes with certain viruses results in the enhancement of IgE-mediated release of histamine. This enhancement is produced by interferon. The present experiments show that an induction period of 6 to 9 hr and new RNA synthesis are required for interferon to enhance histamine release. This points to the possibility that interferon may exert its antiviral and histamine-release enhancing activities by acting through a common pathway.


Differences in HLA antigen recognition by human influenza virus-immune cytotoxic T cells. The specificity of in vitro induced human influenza-immune cytotoxic effector cells was analyzed with respect to recognition of HLA-A and -B-linked gene products. The influenza-immune cytotoxic activity observed on panels of virus-infected targets demonstrated that virus-immune effectors preferentially lyse targets with which they share HLA-A or -B specificities. Virus-immune effectors from certain donors recognized virus in conjunction with some, but not all, of their self HLA-A and -B antigens. Among donors who share a given HLA antigen (such as A2 or B7), there are differences in the ability of their virus-immune T cells to recognize the shared antigen. Virus-infected target cells from HLA-A2 or -B7 "nonresponder" donors could be lysed by virus-immune T cells obtained from other donors who shared only the HLA-A2 or -B7 antigen with these target cells. These observations suggest that the absence of cytotoxic T cell responses by some donors to influenza virus in conjunction with HLA-A2 or -B7 is not due to control by the structural genes that code for these HLA antigens, but rather may result from control by regulatory genes that act at the level of the responder and/or stimulator cell. The results are discussed in the context of Ir gene regulation of human T cell responses.


Mechanisms of mouse mast cell activation and inactivation for IgE-mediated histamine release. The IgE-mediated histamine release from mouse mast cells requires Ca++, is optimal at 37 degrees C, and is enhanced by phosphatidylserine. The rate of release is relatively slow. The mast cells can be activated to release histamine by either anti-IgE or anti-Fab antibodies and, in the case of cells from sensitized mice, by the immunizing antigen. The incubation of mast cells with antigen in the absence of Ca++ or phosphatidylserine fails to release histamine. Such cells are desensitized to the further addition under optimal conditions of the same antigen. Desensitization is antigen specific, requires optimal levels of antigen, and occurs at both 30 degrees and 37 degrees C. In contrast, anti-IgE desensitizes all IgE-mediated histamine release reactions.


Human immune responses to hapten-conjugated cells. II. The roles of autologous and allogeneic histocompatibility determinants in proliferative responses in vitro. Proliferative responses of human lymphocytes primed in vitro to autologous TNP-cells were found to be associated with autologous D-region determinants irrespective of HLA-B locus antigens. Family studies of secondary TNP-conjugate proliferative responses demonstrated a gene dosage effect in this phenomenon. Moreover, co-culture with allogeneic cells did not affect the net TNP-conjugate proliferative responses of primed responder cells, suggesting that HLA-D region preference was due to a requirement for representation of TNP-molecules in association or combination with autologous MHC structures. Alloantigens were found to influence the sensitization of lymphocytes to autologous hapten-conjugated cells. Co-culture of allogeneic and TNP-modified autologous stimulator cells in primary cultures enhanced the secondary TNP proliferative response. Sensitization of human lymphocytes to allogeneic cells alone did not prime responses to autologous modified cells. However, priming lymphocytes to modified autologous cells potentiated responses to allogeneic cells. The data suggest a complex relationship between responses to alloantigens and modified autologous cells.


The proliferative response of human lymphocytes to antigen is suppressed preferentially by lymphocytes precultured with the same antigen. Human blood lymphocytes activated in vitro with antigen to which the donor is reactive are capable of suppressing the secondary proliferative response of autochthonous fresh cells to antigen. Both antigen-specific and antigen-nonspecific suppression can be detected in each experiment. These suppressor cells act by decreasing the number of lymphocytes entering the proliferative response rather than by slowing or otherwise inhibiting ongoing proliferation. The suppressor cells must be added soon after fresh cells are stimulated with antigen to be effective, but the suppressor cells themselves need not proliferate to exert their effect. Suppressor cells are optimally effective when added in numbers equal to those of the responding population, but still exert a significant effect at one-eighth that number.


Immune suppression in vivo with antigen-modified syngeneic cells. II. T cell-mediated nonresponsiveness to fowl gamma-globulin. Intravenous administration of syngeneic spleen cells coupled with the palmitoyl derivative of fowl gammma-globulin (p-F gamma G) results in a profound state of F gamma G-specific tolerance in C57BL/6 mice. Administration of p-F gamma G coupled syngeneic cells specifically reduces both the primary and secondary hapten and carrier-specific PFC responses to TNP-F gamma G. Since the haptenic response is affected, the tolerance functions at the level of the F gamma G-specific helper T cell. As few as 10(3) p-F gamma G spleen cells carrying only 1 ng of p-F gamma G can induce tolerance. At least a 2-day-induction period is required. This nonresponsiveness is long lived, lasting over 120 days. Spleen cells from tolerized mice can transfer suppression to normal syngeneic recipients. Treatment of tolerant spleens with anti-Thy 1.2 antiserum + C eliminates the suppressor cell activity. In addition, thymocytes and purified splenic T cells from tolerized mice can transfer suppression to normal recipients. Thus, at least a component of this nonresponsiveness is mediated by suppressor T cells. The requirement of antigen association with cell membrane components and the general applicability of this method of inducing T cell nonresponsiveness are discussed.


Evidence for shared idiotypy expressed by the IgM, IgG, and IgA serum proteins of a patient with a complex multiple paraprotein disorder. The results of a comparative idiotypic analysis of multiple Ig paraproteins isolated from the serum of an individual patient, Ca, with Sjögren's syndrome and Waldenström's macroglobulinemia are reported. At initial presentation, Ca serum was found to contain two major paraproteins, an IgMkappa and an IgGkappa, together with a small elevation in the level of IgA protein. The patient's clinical course was characterized by dramatic and opposing changes in the respective serum levels of the IgMkappa and IgGkappa paraproteins over an extended time period that coincided in part with received chemotherapy. Idiotypic antigenic analysis of the IgMkappa and IgGkappa paraproteins revealed that the two monotypic proteins shared identical idiotypic determinants. The Ca IgA serum fraction, specifically isolated by an immunoabsorbent and free of any IgG and IgM, was shown to possess idiotypic determinants identical to the IgG and IgM proteins. In extensive tests of specificity, the idiotypic determinants shared by Ca IgM, IgG, and IgA proteins were not present in large excesses of heterologous IgM and IgG, nor on Ig molecules contained in a large number of normal and myeloma sera.


The activation of the alternative complement pathway by fetal calf serum and mouse thymocytes. Mouse thymocytes activated the alternative complement pathway of mouse serum in the presence of heated fetal calf serum. The activation required C3 from the fetal calf serum but was independent of antibody either in the murine or bovine serum. No other murine cells tested, including erythrocytes, peripheral blood lymphocytes, lymph node cells, spleen cells, and various cultured cell lines, activated the alternative complement pathway as effectively as thymocytes. In addition, sera from species other than cows could not substitute for fetal calf serum. The C3 deposited on thymocytes was in the form of both C3b (immune adherence positive) and C3bi (conglutinable). We propose that the basis of activation in this system is the specific protection of bovine C3b on mouse thymocyte surface.


Slow reacting substance (SRS) from ionophore A23187-stimulated peritoneal mast cells of the normal rat. I. Conditions of generation and initial characterization. When rat peritoneal mast cells were exposed to the ionophore A23187, a principle was released that possessed the biologic properties of slow reacting substance (SRS) from various sources. The response was dose, time, and temperature dependent with no activity being demonstrated in unstimulated cells. Supporting evidence that the mast cell product was similar or identical to SRS obtained from other sources include: 1) appropriate differential bioassay profile, 2) resistance to lipolysis and proteolysis, 3) acid lability and base stability, 4) inactivation by limpet arylsulfatase, and 5) inhibition by low concentrations FPL 55712. These data demonstrate that the isolated rat peritoneal mast cell contains the biosynthetic capacity to produce a bioreactive substance with the properties of SRS.


Regulation of IgE antibody production by serum molecules. V. Evidence that coincidental sensitization and imbalance in the normal damping mechanism results in "allergic breakthrough". Experiments presented in this paper were designed to test a new concept concerning the possible pathogenesis of the allergic phenotype. This concept, termed "allergic breakthrough" considers that one of the avenues toward the allergic phenotype involves coincidental sensitization combined with an imbalance in the normal damping mechanism that serves to limit IgE antibody production. The three predictions of this concept that can be tested experimentally are: 1) manipulations that are effective in heightening or re-establishing the damping mechanism should manifest persistence insofar as IgE antibody synthesis to the relevant allergen is concerned; 2) once allergic breakthrough has occurred, the height of production of IgE antibodies specific for the sensitizing agent should remain elevated at levels characteristic of the allergic phenotype, even after the threshold of damping activity has returned to a normal level; and 3) allergic breakthrough should display specificity in that breakthrough would occur in response to subsequent exposure to the specific antigen to which coincidental sensitization initially occurred, but not for other unrelated antigens. The studies presented herein confirm each one of these predictions, thereby providing substantial support for the validity of this concept as one possible distinguishing feature between individuals manifesting the nonallergic and allergic phenotypes, respectively.


Specific inhibition of human lymphocyte responses by primed autologous lymphocytes. I. Evaluation of MLR inhibition as a model for suppression. Human lymphocytes from person A, primed for 10 to 14 days in MLC against lymphocytes from person B, inhibit specifically the proliferative response to B by fresh (i.e., unprimed) lymphocytes of A. Gamma-irradiated (2000 R) primed lymphocytes likewise inhibit specifically, although less strongly. Cells of A, primed with cells of B and then irradiated, usually can inhibit the response of A to cells of any individual sharing HLA-D antigens with B, and the effect tends to be independent of the number of stimulating cells. We also often see inhibition of responses to cells sharing HLA-A and -B antigens with person B, but this effect tends to be lost when the number of stimulating cells is increased. Similarly, at low doses, cells primed for HLA-D antigen a appear not to inhibit the response to an irrelevant HLA-D antigen b on the same stimulating cell. At higher doses of primed cells, even the response to the irrelevant antigen is inhibited. These data suggest to us that at least two mechnaisms may be involved: one directed at the stimulating cell (most likely cell-mediated cytolysis), and predominant at high ratios of primed cells to stimulating cells; the other directed at specific clones of responding cells, and predominant at low ratios.


Suppressor cells for the afferent phase of contact sensitivity to picryl chloride: inhibition of DNA synthesis induced by T cells from mice injected with picryl sulfonic acid. Previous reports have shown that picryl sulfonic acid (PSA) induces suppressor T cells that inhibit the effector phase of contact sensitivity, whereeas its DNP counterpart, dinitrobenzenesulfonate (DNBS) induces cells that inhibit the afferent phase of sensitization. Accordingly, cells from mice injected with DNBS, but not PSA, could be shown to inhibit the DNA synthesis in the lymph nodes that occurs during sensitization. It is now shown that PSA does induce T cells that suppress DNA synthesis but this can only be detected with enriched T cells or by using a regimen of PSA injection different frm previously used to induce suppressor cells for the effector phase. The T cells did not affect responses to oxazolone or dinitrofluorobenzene (DNFB) and were distinguishable from suppressors of the efferent phase in that they could be produced in adult thymectomized but not cyclophosphamide-treated mice. T cells from mice injected with DNBS that inhibited DNA synthesis to DNFB had the same properties.


Binding studies with antibodies having phosphorylcholine specificity and fragments derived from their homologous Streptococcus pneumoniae type 27 capsular polysaccharide. The capsular polysaccharide from Streptococcus pneumoniae type 27 (S-27) has been fragmented into oligosaccharides by controlled degradations. Binding of these derived small ligands with heterogeneous rabbit anti-S-27 antibody of selected phosphorylcholine affinity indicates that a major immunodeterminant of S-27 is the phosphorylcholine moiety, but that these anti-PC. antibodies are also capable of binding to the polysaccharide backbone from which the P.C. has been removed.


Neoantigen of the complement membrane attack complex of cytotoxic human peripheral blood lymphocytes. Neoantigenic determinants (neoAg) specific for the assembling membrane attack complex (MAC) of complement were detected by immunofluorescence microscopy on the surface of cytotoxic lymphocytes during the antibody-dependent cellular cytotoxicity (ADCC) reaction. This study employed antibody-sensitized chicken erythrocytes as target cells, human peripheral blood lymphocytes as effector cells, and RITC-conjugated rabbit F(ab')2-anti-neoAg. NeoAg was present on 60% of ADCC plaque-forming lymphocytes (PFL). Eight out of 182 neoAg-positive PFL were observed in direct contact with their target cells. In these cases MAC-specific neoAg was visualized at the zone of contact between the cells. Anti-neoAg Ig was found to inhibit ADCC plaque assays up to 62%; and 51Cr-release assays up to 79%. Stimulation of lymphocytes by PHA or mixed lymphocyte culture increased the expression of neoAg. In the case of PHA, increased neoAg expression was correlated with an increased incorporation of 14C-leucine into C5, C6, C7, and C8 antigens, which was detected by immunodiffusion and autoradiography.


Properties and applications of monoclonal antibodies directed against determinants of the Thy-1 locus. Fusion of cells of the mouse myeloma line, P3/X63-Ag8 with spleen cells from AKR/J mice immunized against C3H thymocytes or from (BALB/c x BALB.K)F1 mice immunized against AKR/J thymocytes gave rise to hybrid cell lines that continuously secrete antibodies specific for the Thy-1.2 and Thy-1.1 antigens, respectively. Monoclonal antibodies from four such cell lines were analyzed in detail. All were 19S IgM, and, in the presence of complement (C), had high lytic titers on T cells of the appropriate antigenicity. Their specificity was shown by lysis of thymocytes from Thy-1 congenic mouse strains, A/J(Thy-1.2) and A. Thy 1.1. Furthermore, they lyse only 60 to 70% of lymph node cells, suggesting cytotoxicity for mature T cells and not B cells. Treatment of peripheral lymphocyte populations with monoclonal antibody plus C eliminated effector cytotoxic T lymphocytes, their precursors, and the mitogenic response to Con A, but did not affect the response to LPS. Purified, fluorescein-labeled monoclonal anti-Thy-1 antibody could be used to distinguish T and B cells. Purified antibody coupled to Sepharose 6MB was used to separate viable T and B cells. Two independently isolated anti-Thy-1.2 hybridomas are indistinguishable and bind the same determinant whereas a third is unique and may bind a separate site.


In search of alpha 1-microglobulin on the lymphocyte surface. alpha 1-Microglobulin was found by immunofluorescence not to be associated with human lymphoid and nonlymphoid cell lines. No accumulation of alpha 1-microglobulin was detected in culture media of these cell lines. A weak membrane fluorescence with anti-alpha 1-microglobulin on peripheral lymphocytes could not be blocked by the purified protein. No release of alpha 1-microglobulin into the growth medium was seen by normal cultured leukocytes. Treatment of normal lymphocytes, erythrocytes, and various cell lines with solubilization techniques did not yield any alpha 1-microglobulin. alpha 1-Microglobulin and protein HC display immunologic and biochemical identity. However, anti-protein HC stained almost all of the tested cell lines and normal lymphocytes. Blocking experiments with the purified protein were not successful. Antibodies reacting with a minor impurity (50,000 d) in the alpha 1-microglobulin or protein HC preparations could be absorbed from anti-alpha 1-microglobulin with normal leukocytes and a lymphoid cell line.


Genetic control of the IgG2a response to sheep erythrocytes in mice: isotype- and antigen-specific T cell-mediated suppression in low responders. The IgG2a response to sheep erythrocytes is examined in different congenic strains of mice. B10, B6, and C57BL/Ks animals produce a low level of IgG2a antibodies to SRBC during the primary response in vivo. They remain low responders after secondary challenge in vitro. Total spleen cells or nylon-purified T cells from these low responders inhibit the IgG2a response of H-2 compatible-responding mice in a mixed culture system. This suppression is mediated by Thy-1+, Ly-1-, Ly-2+, and I-J+T cells only present in the spleen of low responding animals. These suppressor T cells appear to be IgG2a- and SRBC-specific. Function of non-H-2-linked genes as regulators of suppressor T cells differentiation is discussed.


Maturation of the lymphoid system. I. Induction of tolerance in neonates with a T-dependent antigen that is an obligate immunogen in adults. A/J mice displayed a striking ontogenetic difference in the capacity to respond to DNP-Ficoll, a T-independent antigen, and to aggregated human gamma-globulin (AHGG), a T-dependent antigen. Thus, whereas responses to DNP-Ficoll of 4-day-old mice were similar in magnitude to those of adult animals, responses to AHGG did not become pronounced until mice were some 30 to 40 days of age. The inability of young animals to respond to AHGG was reflective of a negative consequence of lymphocyte/antigen interaction, since such mice became specifically unresponsive to subsequent challenges with AHGG. Unresponsiveness induced by neonatal injection of AHGG lasted 50 to 60 days, in contrast to that induced by deaggregated HGG, which persisted some 100 days longer. The unresponsive state induced by injection of neonates with AHGG maintained itself upon adoptive transfer and did not appear to be linked to suppressive factors associated with either serum or lymphoid cells for its maintenance. Finally, AHGG was also shown to be capable of inducing unresponsiveness in neonatal, athymic mice. These results demonstrate that AHGG, the normally immunogenic form of HGG in adult mice, can serve as an effective tolerogen when administered into a neonatal environment.


Maturation of the lymphoid system. II. Characterization of the cellular levels of unresponsiveness induced in neonates by a T-dependent antigen that is an obligate immunogen in adults. It has previously shown that AHGG, a form of HGG that is highly immunogenic in euthymic adult mice, is capable of inducing specific unresponsiveness when injected into neonatal animals. This report extends this finding and indicates that such a neonatal treatment results in the induction of tolerance in T as well as B cells. Furthermore, a similar conclusion was reached regarding specific T lymphocyte function in animals treated as neonates with OVA. The ability of LPS to modulate responses of neonatal animals to AHGG or DHGG was also examined. It appeared that such mice were not susceptible to the adjuvant effects of LPS until the 4th week of life. Furthermore, LPS was incapable of inhibiting the unresponsiveness induced in mice by either DHGG or AHGG until the 3rd or 4th week of life.


In vitro analysis of allogeneic lymphocyte interaction. IV. Dual recognition of B cell-associated Mls locus and I-region determinants by a helper allogeneic effect factor (AEF) generated across a minor H locus disparity. A helper allogeneic effect factor (AEF) was produced across an incompatibility at the minor histocompatibility loci. This AEF is genetically restricted in its activity since it helps B cells only of the stimulator haplotype and of haplotypes that share both an Mls and I-region identity with the stimulator haplotype. The I-region genes involved here map to the I-A and/or I-C subregions. An anti-LyM immunoadsorbent column but neither an anti-H-2 nor an anti-Ia column absorbed AEF helper activity. It is suggested that the activation of T helper cells by a positive allogeneic effect across a minor H locus difference and their genetically restricted interaction with allogeneic B cells may in part be due to the acquisition by alloactivated T cells of stimulator cell-derived LyM and/or Mls determinants. The data presented indicate that helper T cells recognize Mls locus alloantigens in the milieu of self Ia antigens.


The action of strontium on basophil leukocytes and its use to probe the relationship between immunologic stimulus and secretory response. Strontium will substitute for calcium in the activation of histamine secretion from human basophil leukocytes stimulated by an immunologic reaction or by the ionophore A23187. Strontium is required in 10-fold higher concentration (1 to 10 mM) to activate histamine release compared with calcium (0.1 to 1.0 mM). In terms of maximum release obtainable for a particular immunologic stimulus, strontium is more effective than calcium. Results are presented to show that calcium and strontium act at the same site but strontium is a more sensitive probe for that site. Strontium can be used to demonstrate that immunologic stimuli activate calcium-binding sites in basophils even when no secretion is observed in the presence of calcium. It is suggested that the degree of secretion observed from basophils depends on the number of occupied Fc receptors for IgE and the coupling of these Fc receptors to calcium transport sites.


Thymic control of secretory antibody responses in the rat. The effect of neonatal thymectomy on salivary and serum antibody responses was studied in rats. Local immunization of thymectomized rats with a T-dependent antigen (DNPBGG) elicited negligible amounts of IgA anti-DNP antibody in saliva. In contrast, both normal and sham-thymectomized animals demonstrated substantial levels of salivary IgA antibody. All thymectomized rats locally injected with a T-independent antigen (DNP-Lys-Ficoll) exhibited salivary IgA antibody production. Salivary IgG antibodies were somewhat decreased in thymectomized rats injected with either antigen; however, the final effect of T cell deprivation on IgG synthesis was not as pronounced as on IgA synthesis. Serum IgA antibody was induced in control rats injected with DNPBGG, whereas this Ig class of antibody was absent in thymectomized rats. The results suggest that thymus-derived cells exert a regulatory influence on both serum and secretory IgA responses to antigens.


The genetics of PLT response. II. HLA-DRw is a major PLT-stimulating determinant. PLT response is restricted by the HLA-D region. The present study was undertaken to help define the role of HLA-DRw in PLT restimulation. Haplotype-primed intrafamily PLT cells were made against specificities HLA-DRw1, HLA-DRw3, and HLA-DRw7; each PLT was then restimulated with cells from a 35-member unrelated panel. Restimulation values for each PLT were subjected to bimodal clustering analysis. In addition, blocking experiments were performed with other intrafamily and homozygous typing cell PLT after preincubation with B cell alloantisera. The results show a high correlation (0.881 less than or equal to r less than or equal to 1.00) between the HLA-DRw specificity of the priming haplotype and the HLA-DRw specificity of unrelated panel cells that restimulate in PLT. When stimulating cells were absorbed with the corresponding DRw alloantisera or p29,34 heteroantiserum (against B cell specific antigens), PLT restimulation was significantly blocked. However, the PLT cells treated with antisera showed no effect. The results strongly suggest that HLA-DRw is the principal PLT-stimulating determinant.


A serologic comparison of Moloney lymphoma cell surface and Moloney oncornavirus antigens. Moloney lymphomas and Moloney sarcomas share strong tumor antigens. In this report we analyze the cell-surface antigens on a Balb/c Moloney lymphoma, LSTRA, using hyperimmune sarcoma regressor sera (alphaMo) as a primary reagent. We also use heterologous anti-viral p30 and gp70 sera for a direct analysis of virion protein antigens on the LSTRA surface. Using radiolabeled alphaMo-binding assays, we demonstrate that LSTRA tumor antigens detected by these sera are all Moloney viral antigens; approximately 1/3 of these antigenic determinants are expressed on the intact virus, and the other determinants are revealed by detergent lysis of the virus. The major viral antigens expressed on the LSTRA cell surface are viral env gene products, whereas gag gene products are only sparsely represented. We conclude that alphaMo sera detect almost exclusively viral antigens on LSTRA cells, and these antigens are almost exclusively virion env gene products.


Specific inhibition of the antibody-mediated activation of a defective beta-D-galactosidase by circulating activating epitope-binding molecules. Activation of a defective Escherichia coli beta-D-galactosidase by specific activating antibody is inhibited competitively by a molecule with immunoglobulin properties but devoid of activating capacity. This molecule is found in the serum of nonimmunized rabbits and is no longer detectable after beta-D-galactosidase administration, but can be demonstrated in rabbits injected with antigens other than the enzyme. The data show that the inhibitory molecule recognizes and interacts specifically with the activating epitope of the activatable enzyme and that, although unable to activate the latter, it competes with the activating antibody and inhibits activation.


Immune response gene control of determinant selection. II. Genetic control of the murine T lymphocyte proliferative response to insulin. The genetic control of the murine T cell proliferative response to insulin was examined. It was found for two responder strains of mice that each recognizes a different determinant on the insulin molecule. H-2b mice recognize a determinant in the A chain loop of insulin whereas H-2d mice recognize a determinant that resides in the B chain, possibly in the last eight amino acids. Using H-2 recombinant strains of mice, the location of Ir gene control of the response to both determinants was mapped to the K region and/or I-A subregion of H-2. The possibility of non-MHC regulation of MHC-controlled immune responses is suggested by studies of recombinant inbred strains of mice.


Effects of ethanol on lipid metabolism.

Effects of ethanol on lipid metabolism. Alcohol promotes accumulation of fat in the liver mainly by substitution of ethanol for fatty acids as the major hepatic fuel. The degree of lipid accumulation depends on the supply of dietary fat. Progressive alteration of the mitochondria, which occurs during chronic alcohol consumption, decreases fatty acid oxidation by interfering with citric acid cycle activity. This block is partially compensated for by increased ketone body production, which results in ketonemia. Thus, mitochondrial damage perpetuates fatty acid accumulation even in the absence of ethanol oxidation. Alcohol facilitates esterification of the accumulated fatty acids to triglycerides, phospholipids, and cholesterol esters, all of which accumulate in the liver. The accumulated lipids are disposed of in part as serum lipoprotein, resulting in moderate hyperlipemia. In some individuals with pre-existing alterations  of lipid metabolism, small ethanol dose may provoke marked hyperlipemia which responds to alcohol withdrawal. Inhibition of the catabolism of cholesterol to bile salt may contribute to the hepatic accumulation and hypercholesterolemia. The capacity of lipoprotein production and hyperlipemia development increases during chronic alcohol consumption, probably as a result of the concomitant hypertrophy of the endoplasmic reticulum and Golgi apparatus. However, this compensation is relatively inefficient in ridding the liver of fat. This inefficiency may be linked to alterations of hepatic microtubules induced by ethanol or its metabolites, which interfere with the export of protein from liver to serum, promoting hepatic accumulation of proteins as well as fat. As liver injury aggravates, hyperlipemia wanes and liver steatosis is exaggerated. Derangements of serum lipids similar to those found in other types of liver disease also become apparent. The changes in serum lipids may be a sensitive indicator of the progression of liver damage in the alcoholic.


Mechanisms of regulation of cell-mediated immunity. IV. Azobenzenearsonate-specific suppressor factor(s) bear cross-reactive idiotypic determinants the expression of which is linked to the heavy-chain allotype linkage group of genes. T-cell derived suppressor factor(s) (SF) specific for azobenzenearsonate (ABA) were prepared by the mechanical disruption of suppressor cells. Such suppressor factors were adsorbed to and recovered from immunoadsorbents prepared from the F(ab')2 fragments of rabbit immunoglobulin directed against the cross-reactive idiotype of A/J anti-ABA antibodies. These ABA-suppressor factors were not retained on Sepharose 4B immunoadsorbent columns which had been coupled with F(ab')2 fragments or normal rabbit immunoglobulins prepared from prebleeds of rabbits used to make anti-idiotypic antiserum. The specificity of the F(ab')2 rabbit anti-idiotypic serum was established by direct idiotypic-binding assays and by affinity purification over an immunoadsorbent consisting of CRI+ anti-ABA immunoglobulin from A/J mice. ABA-suppressor factors were shown to be specifically absorbed and eluted from F(ab')2 anti-idiotypic columns. Futhermore, the eluted suppressor factor can be specifically reabsorbed and recovered from a second anti-idiotypic immunoadsorbent. The concordance between antigen-binding specificity and the presence of idiotypic determinants was demonstrated by adsorbing ABA SF to antigen columns and then fractionating the ABA-specific factor on anti-idiotypic immunoadsorbents. ABA-suppressor factors were shown to be specifically retained on immunoadsorbents directed against major histocompatibility complex (MHC) determinants. Factor eluted from anti-MHC columns could then be specifically adsorbed to anti-idiotypic immunoadsorbents. This suggests that the same molecular complex that is recognized by the H-2 alloantiserum is specifically adsorbed to an anti-idiotypic immunoadsorbent. Genetic analysis of the expression of CRI+ suppressor factor was performed using the C.AL-20 mouse strain which has the AL/N allotype and produces CRI+ anti-ABA immunoglobulins. The implication of these findings to the nature of T-cell-derived regulatory molecules is discussed.


Tolerance induction during ontogeny. I. Presence of active suppression in mice rendered tolerant to human gamma-globulin in utero correlates with the breakdown of the tolerant state. A specific state of T- and B-cell tolerance to human gamma-globulin (HGG) was induced in utero by intravenous administration of the deaggregated antigen to pregnant BALB/cCr mice. Tolerance persisted in the offspring until the 12th-wk of age and then began to gradually disappear. Suppressor cells could only be found when responsiveness to HGG ultimately appeared in the in utero-treated animals but not when they were completely unresponsives. In contrast, HGG-specific suppressors found in animals made unresponsive to HGG as adults appear to be associated with either the establishment and/or maintenance of the unresponsive state. To the extent that these experiments are consistent with natural self-tolerance to a serum protein, we conclude that active suppression is not a prerequisite from maintenance of unresponsiveness to self.


Induction and mode of action of suppressor cells generated against human gamma globulin. II. Effects of colchicine. The ability of colchicine (Col) to interfere with suppressor cells specific for the soluble protein antigen human gamma globulin (HGG) has been examined. This interference may be the mechanism of the adjuvanticity promoted by Col. When injected into A/J mice at the appropriate time and concentration, both Col and cyclophosphamide promoted an adjuvant increase in the plaque-forming cell response to 100 micrograms of immunogenic, aggregated HGG. Col abrogated both the induction of suppressor cells when injected with 3 h of tolerization with deaggregated (DHGG) and the expression of previously induced suppressor cells when injected with the antigenic challenge. Interference with the generation and expression of antigen-specific suppressor cells had no detectable effects on the immunologic unresponsive state to HGG. Col did not interfere with the induction of tolerance at a dose (1 mg/kg) that abolished the generation of suppressor cells. Furthermore, the absence of colchicine-sensitive-suppressor cells during the establishment of tolerance had no observable effect on the duration of unresponsivness in either helper T- or B-lymphocyte populations. Finally, Col was not able to terminate the unresponsive state established by DHGG even when responsive splenic B cells could be demonstrated in tolerant animals. These data indicate that suppressor cells are not required for the establishment and maintenance of the unresponsive state to this antigen.


Active suppression of 1-fluoro-2,4-dinitrobenzene-immune T cells. Requirement of an auxiliary T cell induced by antigen. We investigated T-T cell interactions in the suppression of contact sensitivity. Suppressor cells that block the efferent limb of sensitivity (Ts-eff) can inhibit the passive transfer of contact sensitivity mediated by 1-fluoro-2,4-dinitrobenzene immune cells (T DH). But, Ts-eff cannot block the passive transfer of TDH which comes from cyclophosphamide (Cy) pretreated sensitized mice. We interpret these results to indicate that lymph node cells from sensitized mice contain not only TDH but also another intermediate cell which is required for the suppression of TDH by Ts-eff. This intermediate cell is sensitive to cyclophosphamide and requires antigen activation for its development. It is sensitive to adult thymectomy and anti-brain associated theta serum and is therefore designated as an auxiliary T-suppressor cell (Ts-aux). It is not sensitive to splenectomy and it carries I-J determinants. Ts-aux are required for the activity of suppressors of the efferent limb (Ts-eff) but not of suppressors of the afferent limb (Ts-aff). Thus, in the feedback loops in contact sensitivity, the generation of Tdh is coordinated with the development of auxiliary Ts which are essential for the suppression of those TDH.


Mechanisms of idiotype suppression. I. In vitro generation of idiotype-specific suppressor T cells by anti-idiotype antibodies and specific antigen. Normal BALB/c spleen cells are unresponsive in vitro to the phosphorylcholine (PC) determinant in the presence of anti-idiotype antibodies specific for the TEPC-15 myeloma protein (T15) which carries an idiotypic determinant indistinguishable from that of most anti-PC antibodies in BALB/c mice. The possibility that idiotype-specific suppressor cells may be generated during the culture period was examined by coculturing the cells with untreated syngeneic spleen cells. Cells that had been preincubated with anti-T15 idiotype (anti-T15id) antibodies and a PC-containing antigen, R36a for 3 d, were capable of specifically suppressing the anti-PC response of fresh normal spleen cells, indicating that idiotype-specific suppressor cells were generated during the culture period. The presence of specific antigen also appeared to be necessary because anti-T15id antibodies and a control antigen, DNP-Lys-Ficoll, were not capable of generating such suppressor cells. Suppressor cells were induced only in the population of spleen cells nonadherent to nylon wool and the suppressive activity was abrogated by treatment with anti-Thy 1.2 serum and complement. These results indicate that anti-idiotype antibodies and specific antigen can generate idiotype-specific suppressor T cells in vitro. These in vitro results may reflect in vivo mechanisms of idiotype suppression.


Different D end-dependent antigenic determinants are recognized by H-2-restricted cytotoxic T cells specific for influenza and Bebaru viruses. Two different BALB/c anti-CBA (H-2k)monoclonal antibodies that bind to Kk and Dk antigens blocked Tc cell-mediated lysis of L929 (Kk, Dk) target cells, but with quite different specificity. One antibody (30R3) powerfully blocked Kk-specific lysis mediated by alloreactive or Kk-restricted Tc cells immune to ectromelia, Sendai, or influenza viruses. The other antibody (27R9) blocked these anti-Kk Tc cells much less than 30R3, but in contrast, 27R9 blocked anti-Dk lysis much more than 30R3. Most importantly, 27R9 strongly blocked Dk-restricted anti-influenza Tc cells, but did not significantly block Dk-restricted anti-Bebaru (BEB) lysis. This result indicated that different H-2 determinants coded in the D end of H-2k were recognized by influenza-and BEB-immune Tc cells. These determinants may be carried on two different molecules coded by the H-2D and H-2L loci, but other possibilities are not yet excluded.


Two Plasmodium knowlesi-specific antigens on the surface of schizont-infected Rhesus monkey erythrocytes induce antibody production in immune hosts. Purified schizonts (6--10 nuclei) and membranes of schizont-infected erythrocytes from the Malaysian and Philippine strain of Plasmodium knowlesi are analyzed immunochemically using immunoglobulin of rhesus monkey hyperimmune sera against schizonts and of sera from naturally immune monkeys. The anti-schizont Ig identifies less than 20 immune components in Triton X-100-solubilized schizonts and membranes of infected cells. Of these antigens, 9 (component 1, 3, 4, 5, 6, 10, 11, 18, and 20) are common to parasites and membranes of infected erythrocytes, and 12 (2A,B, 6, 8, 9, 12, 13p, 14, 16A,B, 19 A,Bp, 21, 22p, and 23) are predominantly found in the parasite; 4 components (13i, 19A,Bi, 22A, B, and 24) are unique to the membrane of infected erythrocytes. Only three parasite-specific components (1, 13, and 19) are exposed on the surface of parasitized erythrocytes as revealed by both lactoperoxidase-catalyzed radioiodination and extensive absorption of anti-schizont Ig using intact infected erythrocytes. Two plasmodium-specific antigens (1 and 13) on the surface of infected erythrocytes are recognized by sera of rhesus monkeys rendered naturally immune against P. knowlesi infections and, therefore, represent antigens in vivo. Analyses of schizonts and membranes of parasitized erythrocytes of the two different strains of P. knowlesi yields only some minor quantitative, but no qualitative differences when analyzed with both types of antisera. Importantly, components 1 and 13 appear identical in both strains.


Isoelectric focusing of CSF proteins in known or probable infectious neurological diseases and the Guillain-Barré syndrome. The CSF and serum proteins of 120 patients with known or probable infectious neurological diseases or the Guillain-Barré syndrome were examined with thin-layer IEF. All but two of these patients exhibited one or combinations of different CSF-protein aberrations in the acidic and alkaline range. Aberrant non-Ig fractions (including transferrin, the tau-fraction and gamma-trace protein) were found in frequencies varying between 4 and 48%. CSF Ig components of restricted heterogeneity, i.e. oligoclonal bands and/or regional increases of gamma-globulins, were more frequent in patients with (meningo-)encephalitic or (meningo-)-myelitis/radiculitic disorders (respectively 69 and 48%) than in subjects with meningitis or Guillain-Barré syndrome (17%). The occurrence of such Ig abnormalities was higher in subacute or chronic than in acute disease and in subjects examined greater than 4 weeks after the onset rather than earlier. Ig-band spectra with marked anodal extension were found predominantly in (meningo-)encephalitic disorders with infratentorial symptoms. Age and sex were not found to influence the occurrence of abnormal Ig fractions. Such components could be detected in spite of pronounced blood-CSF barrier defects.


Measles antibodies and kappa-lambda light chain distribution in immunoglobulins of patients affected with multiple sclerosis. The presence of measles antibodies in serum immunoglobulin G fractions from seven patients affected with multiple sclerosis was investigated with HI technic. The kappa-lambda light chain ratios of all samples under investigation were evaluated. Three multiple sclerosis patients, who displayed either fractionation or a tendency towards fractionation in their serum, had slightly elevated measles antibody titers associated to increased kappa/lambda ratios.


Pattern and flash visual evoked responses in multiple sclerosis. In multiple sclerosis (MS), increase of delay may be important, but the earlier change is the behavior of the wave shape; amplitude is always lower even in MS patients without visual subjective symptomatology, provided not only flash but also pattern stimulation is performed. Normal amplitude with pattern stimulation does not necessarily mean normal with flash. Combining both stimulation methods, we obtained lowering of amplitude and change of wave shape in 100% of cases while latency was increased in only 77% of cases. Sectorial ischemic neuropathy, and open angle glaucoma patients show the same findings, which emphasizes that the method is not specific for demyelinating diseases, nor selective for papillomacular bundle involvement, but that VER, considering only its first components, reflects axonal damage somewhere in the visual pathway. Flash and pattern VER might be even more pathognomonic in MS than oligoclonal gammaglobulin fractionation on agar, positive in 75% of cases. It is suggested that subclinical MS cases should be submitted to full electro-ophthalmographic investigation.


Rapid diagnosis of viral neuroinfections by immunofluorescent and immunoperoxidase technics. The results of immunofluorescent (IF) and immunoperoxidase (IP) technics applied for the detection of antigen in cerebrospinal fluid (CSF) cells in patients with mumps, herpes zoster and herpes simplex meningitis and meningoencephalitis are presented. Thirty patients were under study. The detection of mumps and herpes zoster viral antigen in CSF cells was possible in 100% of cases investigated. Herpes simplex virus antigen was detected in four of seven cases with symptoms of severe meningoencephalitis. Complement fixation (CF) antibodies to herpes simplex virus (type I) and positive seroconversion were detected in the four latter patients. The diagnostic value of the methods used for the detection of mumps, herpes simplex and herpes zoster viral antigens in CSF cells of patients is discussed.


A histological and histochemical study of changes of fiber types in experimental myotonia. In both fast and slow muscles of rats treated with 20--25 diazacholesterol there were qualitative alterations, such as changes of fiber outlines, numerous moth-eaten fibers and rare ring fibers. In addition there were generally larger groups of Type I and intermediate fibers than in normal controls ("type-grouping" tendency) in the preparations for oxidative enzymes in the extensor digitorum longus (EDL) of myotonic animals. Quantitative evaluations of EDL and soleus of myotonic rats revealed moderate hypothrophy of Type I and Type II fibers with an increase in the numbers of Type I and of Type III fibers in the EDL and a significant decrease of the nondominant fibers in the soleus muscle. The data are discussed in the light of a neurally mediated and/or direct action of the drug on the muscle fiber.


Serum cortisol levels of multiple sclerosis patients during ACTH treatment. The adrenocortical function in 52 multiple sclerosis patients was studied and compared to that of ten healthy control subjects. The basic secretion rate of cortisol of the MS patients was within the range of normal. Reactivity of the adrenal cortex to stimulation by synthetic ACTH was markedly reduced independent of the duration of the disease with the exception of four patients who were in an early stage of the disease. No difference could be demonstrated between patients undergoing a bout and patients in a stage of remission. The pathogenetic and prognostic significance of the results of the present study are discussed.


On the mechanism of sudden death with subarachnoid hemorrhage. The mechanism of death in some patients with subarachnoid hemorrhage is cardiac arrhythmia. Prevention of cardiac arrhythmias by suitable drugs might save the life of patients whose brain is still good.


Neutral lipid and sphingolipid composition of the brain of a patient with membranous lipodystrophy. Lipids were extracted from brains of a patient with membranous lipodystrophy (ML) and three normal patients and the neutral lipid and sphingolipid constituents were investigated. The storage of a large amount of free fatty acid was observed in the ML brain, but no cholesterol ester was found. Total lipid, cholesterol and cerebroside contents were slightly decreased in the white matter of the ML brain. The composition of free and sphingolipid fatty acids did not differ between ML and normal brains.


Somatosensory evoked potentials following stimulation of the trigeminal nerve in man. Somatosensory evoked potentials following trigeminal nerve stimulation can regularly be recorded from the contralateral scalp on C5/C6 (10--20 system), a region which overlies the primary face region of the somatosensory cortex. From the first three peaks analyzed (N 13, P 19 and N 26), the first positive peak (P 19) is most prominent and reliable and therefore is recommended for the routine measurements of neurophysiological examination.


Adherence of multiple sclerosis lymphocytes to measles infected cells. Adherence of lymphocytes from multiple sclerosis (MS) patients and control subjects to cells persistently infected by measles and control cells was studied. No statistically significant difference between MS patients and controls could be found in this test. Lymphocytes adhered more readily to cells infected by measles than to control cells. The cell to cell contacts are made by the tips of the microvillii of the Lu cells.


The response of the acute-phase plasma protein alpha2-macroglobulin to vitamin A deficiency in the rat. Alpha2-macroglobulin (alpha2-M) was determined in the sera of vitamin A-deficient and pair-fed control rats. Alpha2-macroglobulin levels were assayed by radial immunodiffusion using a purified standard sample from sera from turpentine-stressed rats. There was no significant increase in the serum level of alpha2-M in vitamin A-deficient rats relative to pair-fed controls, indicating that there was no acute response to the stress of the deficiency. When turpentine was injected to elicit an acute response, the serum level of alpha2-M rose equally (by almost 100-fold) in both deficient and normal rats. It was concluded that the response to vitamin A deficiency of serum glycoprotein synthesis shows some specificity in that the synthesis of alpha2-M in response to stress was not affected by vitamin A, whereas we previously found that levels of alpha1-M, which is very similar to alpha2-M, decline with vitamin A deficiency.


The effect of full-thickness skin grafts on the actomyosin content of contracting wounds. In evaluating the myofibroblast theory of wound contraction, comparisons were made between grafted and nongrafted 15-day wounds. Pregnant rabbit uteri and normal rabbit skin were used as controls. Small amounts of actin and myosin were present in contracting open wounds but not in noncontracting grafted wounds, which suggests some participation for contractile proteins in wound contraction. The current study refutes the widely accepted contention that wound granulation tissue is as rich as smooth muscle in its contractile protein content.