bluuebunny/crossref_metadata_2025 huggingface dataset:
Ontogeny of B Cells in the Chicken
Authors: Grossi Carlo E. , Lydyard Peter M. , Cooper Max D.
Published: December 2022

B lymphocytes simultaneously expressing surface and cytoplasmic IgM were first detected in the bursa of Fabricius from 12-day embryos lacking in detectable Ig-positive cells elsewhere. Bursal lymphocytes showed characteristic patterns of cIgM4 distribution which were related both to age and anatomic localization within the bursal follicles. Cells with diffuse cytoplasmic IgM (type 1) were the first to appear and later became the predominant cell type in the cortex. Cells containing cIgM localized in perinuclear and in Golgi regions (type 2) were the most abundant cell type in the bursa between the 16th day of incubation and the 3rd week after hatching. Cells with dispersed IgM-containing vesicles (type 3) appeared soon after hatching, and were prevalent in the medulla of mature follicles; release of IgM from type 3 cells observed at physiologic temperature was prevented by cold and by cytochalasin B. A rough correlation between the immunofluorescence patterns of cIgM distribution and the susceptibility of sIgM to be modulated by anti-µ antibodies was demonstrated. Type 1 cells required lower concentrations of anti-µ antibody for modulation of their sIgM than did type 2 and 3 cells. Except in very young birds, cells having the characteristics of bursal lymphocytes were not found in extrabursal tissues, including the bone marrow.

These results suggest that during normal development of the chicken, B cells are generated exclusively within the bursa and at least a subpopulation of B cells undergoes an unusual IgM secretory stage in differentiation. The data also suggest that tolerance or clonal abortion of B cells should be most easily accomplished in ovo.
Suppressor Cells in Mice Infected with Trypanosoma Brucei
Authors: Eardley Diane D. , Jayawardena Anil N.
Published: December 2022

Within 2 to 3 days of infection with Trypanosoma brucei strain S42, the ability of spleen cells from infected CBA mice to mount a primary in vitro antibody response to sheep red blood cells (SRBC) is profoundly reduced, and suppressor cells are generated as detected by cell mixture experiments. Suppressor cell activity lies in the T and adherent cell compartments of spleens from infected mice, but not in the B cell compartment, although antibody responses to a thymus-independent antigen, DNP-Ficoll, are significantly reduced. Suppression of antibody responses of normal spleen cells depends on viable cells from infected mice. The trypanosome, itself, plays no direct role in suppression, and we have ruled out the possibility of antigenic competition as a mechanism of suppression.

Our data is consistent with the model of suppressor T cells induced by concanavalin A mitogenesis. We hypothesize that trypanosome antigens may directly stimulate T cells with the concomitant release of factors with affinity for macrophage surfaces thus becoming suppressive for T and B cell responses.
Immunosuppression by Spleen Cells from Moloney Leukemia
Authors: Cerny Jan , Grinwich Kazimiera D. , Stiller Ronald A.
Published: December 2022

Spleens of mice bearing MuLV (Moloney)-induced leukemia contain cells that inhibit the antibody response of normal syngeneic lymphocytes to sheep RBC in Marbrook cultures. In order to determine whether these immunosuppressive cells are virus-infected tumor cells or normal cells we pretreated leukemic spleen cell suspensions with syngeneic mouse antiserum to Moloney leukemia antigen(s) (plus complement) and with rat anti-Moloney serum (plus complement). The cytotoxic treatment killed approximately 20% to 30% and 60% to 70% of the cells, respectively. The remaining viable cell population was tested for MuLV production (in an infectious center assay on S+L- fibroblasts), for lethal effect on newborn mice, and for immunosuppressive activity. After the treatment with anti-Moloney sera the number of MuLV-releasing cells decreased 10-fold and the leukemogenic potential in vivo decreased 100-fold as compared to leukemic spleen cells pretreated with nonimmune mouse and rat sera (plus complement). In contrast, the ability of the antisera-treated cells to inhibit anti-SRBC reponse remained undiminished. This indicates that, in part, the immunosuppressive cells in the leukemic spleen are normal, noninfected cells, involved, perhaps, in immune regulation.
Negative Allogeneic Effects in Vitro
Authors: Swain Susan L. , Dutton Richard W.
Published: December 2022

When splenic T cells from normal mice recognize alloantigens on primed spleen cells there is a dramatic suppression of the secondary antihapten PFC response of the latter cells. From studies using intra-MHC recombinants it appears that differences at K or D alone can elicit allosuppression. There is sometimes suppression to differences in the I region. Even small differences in H-2K, as seen in the H-2b mutants, are sufficient to lead to such negative allogeneic effects. Thus, as far as has been determined, the specificity of allosuppressive T cells is indistinguishable from that of cytotoxic effector cells.

Negative allogeneic effects and positive allogeneic effects differed in the degree to which they influenced the primary and secondary responses. In our experiments no evidence for positive allogeneic effects was seen in the secondary response to TNP-carrier even with cells differing at the I region only. In contrast the primary response to SRBC was much enhanced by allogeneic cells and little suppression of that response was seen.

It is suggested that the allosuppression is distinct from cytotoxicity and may be a profitable model for the study of suppressor T cells.
Reduced Severity of Lesions in Mink Infected Transplacentally with Aleutian Disease Virus
Authors: Porter David D. , Larsen Austin E. , Porter Helen G.
Published: December 2022

Inoculation of mink late in the second trimester of pregnancy with Aleutian disease virus (ADV) produces a persistent infection in the offspring. When these mink were analyzed at 83 days of age and compared with adolescent mink infected for a similar length of time, the transplacentally infected mink show: 1) a marked reduction in plasmacytosis, immunoglobulin level and specific ADV antibody; 2) increased amounts of infectious ADV and numbers of cells containing viral antigen; 3) a marked reduction in immune complex glomerulonephritis and absence of immune complex arteritis; 4) free ADV antigen in the glomeruli; and 5) a striking accumulation of eosinophils in the tissues. The findings suggest that the degree of ADV expression is partially immunologically controlled.
Serum Lipoproteins and C5̄6̄7̄-INH Activity
Authors: Lint Thomas F. , Behrends Carole L. , Gewurz Henry
Published: December 2022

Serum contains an activity termed C-INH which inhibits the attachment of the C complex of complement to bystander cells. The serum constituents which express this activity have not yet been defined. The present studies were initiated to examine the possibility that serum lipoproteins have C-INH activity. Lipoproteins were prepared by sequential flotations and were free of other proteins detectable by immunochemical procedures; they comprised 3% of the serum proteins but accounted for approximately 25% of the serum C-INH activity. The lipoproteins selectively inhibited the attachment of C to bystander cells by interacting with C in solution; they did not inhibit either the formation of C from C and C7 or the lysis of preformed EC by limiting amounts of C8 and C9. Furthermore, the lipoproteins did not inhibit the complement-mediated hemolysis of EA or EAC. Inhibitory activity was found in each of the three major lipoprotein density classes, with the high density lipoproteins having the greatest specific activity. Poly-L-lysine, previously shown to counteract serum -INH also partially blocked the inhibitory activity of the lipoproteins. These experiments point to lipoproteins as a major constituent of serum -INH, and emphasize that lipoproteins exercise a potentially important modulating effect upon the complement membrane attack mechanism.
The Role of T Cells in Anti-Herpes Simplex Virus Immunity
Authors: Pfizenmaier Klaus , Jung Hartmut , Starzinski-Powitz Anna , Röllinghoff Martin , Wagner Hermann
Published: December 2022

Mice infected with herpes simplex virus develop little or no cytotoxic T lymphocyte (CTL) response. However, in lymph nodes (LN's) draining a local site infected with HSV, antigen-specific CTL precursors are sensitized, which upon transfer to in vitro culture conditions develop within 72 hr into effective CTL. The in vivo blockade of CTL differentiation can be overcome by cyclophosphamide, suggesting that a cyclophosphamide-sensitive mechanism blocks the in vivo generation of HSV-immune CTL.

The cytolytic activity of HSV-immune CTL is H-2 restricted and antigen specific. Thus CTL sensitized toward HSV type 1 discriminate between syngeneic targets infected with either the immunologic HSV variant type 1 or type 2 (and vice versa). H-2-matched target cells exposed for 30 min to infectious HSV are lysed within 60 min of contact with CTL. Since HSV replication is believed to require more than 4 to 5 hr, the data suggest that either the expression of HSV-dependent “early proteins” takes place within 30 to 90 min or cell membrane-integrated HSV virion represents the target antigen of CTL.
Paul-Bunnell Antigen in Lymphoma and Leukemia Spleens
Authors: Kano Kyoichi , Fjelde Audrey , Milgrom Felix
Published: December 2022

Lymphoid cells obtained from spleens of patients with lymphomas or leukemias were studied for the presence of heterophile (Paul-Bunnell (P-B)) antigen. A mixed agglutination (MA) test was established utilizing monolayers of cells attached to poly-L-lysine-coated wells of plastic U plates. After incubation of the monolayers with infectious mononeucleosis (IM) sera, indicator cells, sheep, or trypsinized bovine erythrocytes were added. The results were assessed according to sedimentation patterns of the indicator cells on the monolayers. Positive MA reactions were shown to be due to specific binding of P-B antibodies to the corresponding antigens on the spleen cells. Positive results were obtained with 15 of 37 spleens from patients with Hodgkin's disease, 5 of 8 lymphoma spleens, 4 of 15 chronic myelocytic leukemia spleens and 2 of 4 chronic lymphocytic leukemia spleens. Only 2 of 25 spleens from patients with various other diseases and 1 of 26 apparently normal thymus specimens gave positive results. This study confirmed demonstration of P-B antigen in lymphoma and leukemia by means of absorption experiments, which was reported previously.
Antibody-Dependent Killing of Erythrocyte and Tumor Targets by Macrophage-Related Cell Lines: Enhancement by PPD and LPS
Authors: Ralph Peter , Nakoinz Ilona
Published: December 2022

Five murine macrophage or monocyte-related tumor cell lines in culture were compared for antibody-dependent lysis (ADL) of a tumor target (T lymphoma EL4) and lysis and phagocytosis of sheep erythrocytes (RBC). Some lines were effective only with RBC targets, while others were capable of lysing these and tumor targets. Both murine alloantisera to H-2 and Thy 1.2 antigens on the target cells and rabbit anti-mouse spleen and anti-mouse thymus sera directed target lysis, while normal mouse or rabbit sera were inactive. Preincubation of macrophage line RAW264 with lipopolysaccharide (LPS) or purified protein derivative from Mycobacteria (PPD) for 2 days resulted in an average of 76% or 86% increase, respectively, in ADL of RBC, although there was no stimulation in RBC phagocytosis or in nonspecific or antibody-dependent lysis of tumor targets.

These results indicate that macrophage cell types are capable of antibody-dependent tumor lysis, that macrophages are probably heterogeneous in effector cell activities, and that they can be stimulated to increased ADL in the complete absence of other cell types.
Membrane Receptors for IgG and Complement (C4, C3b and C3d) on Human Eosinophils and Neutrophils and Their Relation to Eosinophilia
Authors: Anwar A. R. E. , Kay A. B.
Published: December 2022

Receptors have been shown, on human eosinophils and neutrophils, for rabbit IgG, human IgG, and human C4, C3b and C3d. Both types of granulocyte formed rosettes with EArabG. This was directly related to the amount of sensitizing antibody. No rosettes were formed with E alone or EArabM Receptors for human IgG were also demonstrated by using heat-aggregated IgG and fluorescent-labeled anti-human IgG. In addition, aggregated human IgG inhibited both eosinophil and neutrophil rosettes with EArabM.

Red cell intermediates were prepared with EArabM (EA) and functionally pure human complement components. Eosinophil and neutrophil rosette formation was demonstrable with EAC14 and EAC1423b, prepared with limited amounts of C4 (EAC3b), and was related directly to the input of either C4 or C3. No granulocyte rosettes were formed with EAC1 or EAC142. By decay experiments it was shown that C2, in addition to forming the classical pathway C3 convertase with C1 and C4, also inhibited eosinophil and neutrophil EAC14 rosettes. EAC3d intermediates (prepared by treating EAC3b cells with the C3b inactivator and so rendering them immune adherence negative, unsusceptible to lysis with C5–C9 and more agglutinable with anti-C3d) formed rosettes with eosinophils and neutrophils in comparable numbers to EAC3b cells. In general, the percentage of neutrophils bearing receptors for rabbit IgG, human IgG, or human C4, C3b, and C3d was approximately two and one-half times greater than eosinophils.

There was a significantly reduced percentage of eosinophils forming rosettes with EAC14 or EAC3b when patients with eosinophilia of various etiology were compared to controls; however, no differences were found with EArabG. When neutrophils from patients with eosinophilia were compared to controls in terms of percentage of EArabG, EAC14, or EAC3b rosettes, there were no significant differences. These results, taken together with previously published findings on the monocyte, suggest that all circulating phagocytic cells have similar membrane receptors for promoting adherence of opsonized particles and that with eosinophils the numbers of complement receptors may be altered in disease.
Characterization of Suppressive Immunoglobulin-binding Factor (IBF)
Authors: Fridman Wolf H. , Guimezanes Annick , Gisler Roland H.
Published: December 2022

L-5178-Y, a θ-positive, Fc receptor-bearing mouse thymoma cell line spontaneously releases immunoglobulin-binding factor (IBF) upon short-term incubation in vitro. IBF produced by L-5178-Y cells is identical in its biologic activity with IBF produced by Fc receptor positive alloantigen-activated T cells. It suppresses the in vitro plaque response of mouse spleen cells to sheep erythrocytes by interfering mainly with the late phase of the generation of antibody-forming cells. Therefore, L-5178-Y thymoma affords a homogeneous source of IBF in sufficient quantities for the study of its biochemical nature and the mechanism by which it interferes with cells participating in antibody synthesis.
Spontaneous Cell-Mediated Cytotoxicity by Human Peripheral Blood Lymphocytes in Vitro
Authors: Nelson David L. , Bundy Bonita M. , Strober Warren
Published: December 2022

With isolated subpopulations of human peripheral blood mononuclear cells, spontaneous cell-mediated cytotoxicity (SCMC) and antibody-dependent cellular cytotoxicity (ADCC) were demonstrated to be functions of a population of surface immunoglobulin (sIg)-negative, Fc receptor-positive lymphocytes. Organ distribution studies revealed that SCMC and ADCC effector cell activity occurred in parallel, being present in spleen and absent in tonsils. Treatment of effector cells with trypsin significantly inhibited SCMC but not ADCC and this inhibition was not reversed by short-term incubation in autologous plasma. These results demonstrate that SCMC and ADCC involve different cytotoxicity mechanisms and are consistent with two subsets of effector cells with separate functional capabilities or a single effector cell mediating both forms of cytotoxicity through separate mechanisms.
A Mycoplasma Mutant Resistant to Lysis by C: Variations in Membrane Composition and Altered Response to the Terminal C Complex
Authors: Dahl Jean S. , Hellewell Susan B. , Levine R. P.
Published: December 2022

Mutant strains of Acholeplasma laidlawii resistant to immune lysis by the classical C pathway have been obtained after the exposure of wild-type cells to nitrosoguanidine. The lysis of one mutant strain, comr-328 is less than 20% of that of the wild-type strain when incubated with antiserum prepared against wild-type cells and guinea pig serum as a source of C. The osmotic stability of comr-328 is similar to that of the wild-type strain. Absorption of anti-wild-type antiserum with mutant or wild-type cells results in the complete loss of C fixing antibody as shown by the inability of the absorbed antiserum to sensitize wild-type cells for immune lysis. Both strains convert 125I-labeled C3 to C3b to the same extent and bind equivalent amounts of 125I-C3b. Whereas the wild-type strain is sensitive to lysis by reactive lysis, comr-328 is resistant. Although both cell types are lysed equally well by amphotericin B, comr-328 is less sensitive than the wild-type strain to lysis by melittin.

The membrane lipids of both strains are qualitatively identical, as judged by thin-layer chromatography of chloroform-methanol extracts, but the amount of each lipid varies according to the strain. The most significant variation is found in the glycolipids; the monoglucosyldiglyceride to diglucosyldiglyceride ratio of comr-328 is 2-fold higher than that of the wild-type strain. The difference in the phospholipids is less marked. While more than 35 Coomassie Blue-stained polypeptides are detected by SDS-polyacrylamide gel electrophoresis of solubilized membranes, major quantitative differences occur in only three of these bands. Band 11 is more prominent in comr-328 than in the wild-type strain whereas bands 14 and 26 appear to be less abundant in the mutant.
Characterization of Antigen-Binding Receptors in Vitro
Authors: Benfari Mary Jane F. , Cooperband Sidney R. , Moolten Frederick L.
Published: December 2022

This study reports a novel method for studying the binding of soluble antigen (bovine serum albumin, BSA) to surface receptors on lymphoid cell populations under equilibrium conditions in the presence of 10% normal rabbit serum or 20 mg/ml of ovalbumin. Virtually no nonspecific uptake was demonstrated to nonlymphoid tissues. Detectable quantities of BSA could be found on both nonimmune and immune lymphoid populations. The binding of BSA was demonstrated to be antigen specific and to be proportional to the numbers of binding cells. The quantity of antigen bound was proportional to the free antigen exposed to the population and saturation could be achieved with 1 to 2 µg of antigen/1.2 ml of culture, per 10 × 107 cells. The kinetics of antigen binding was very rapid and occurred within 10 min at 4, 27, and 37°C. The binding was independent of the viability of cells. Binding was antigen specific and could be partially blocked by cross-reacting serum albumins, but not by non-cross-reacting albumins. Binding was independent of cytophylic antibody concentrations in the serum.
Abnormal Polyclonal B Cell Activation in NZB/NZW F1 Mice
Authors: Cohen Philip L. , Ziff Morris
Published: December 2022

Spleen cells from autoimmune (10-month-old) NZB/NZW (B/W) mice failed to generate appreciable numbers of antibody-forming cells (AFC) in vitro to TNP-substituted sheep erythrocytes in response to the polyclonal B cell activators (PBA), LPS and PPD, despite normal DNA synthetic responses to these agents and normal AFC responses to TNP-Ficoll. The failure to respond to PBA in old B/W mice was not due to suppressor T cells since anti-brain-associated-θ-treated spleen cells still failed to generate AFC in response to PBA. The defect was age-related since cells from young B/W mice generated vigorous AFC responses to PBA. It is suggested that the failure of the spleen cells of old B/W mice to generate AFC is a result of in vivo polyclonal B cell activation in the course of autoantibody formation.
In Vitro Reactivity in Allograft Tolerance: Persistence of Mixed Leukocyte Culture Reactivity in Highly Tolerant Rats
Authors: Wright Peter W. , Loop Stephen M. , Bernstein Irwin D.
Published: December 2022

Mixed leukocyte culture reactivity was studied in adult W/Fu rats judged to be highly tolerant after the inoculation of (W/Fu × BN)F1 hybrid spleen and bone marrow cells at birth. Reactivity was observed in a majority of tolerant donors tested and documented by quantitative dose-response and kinetic studies. Allograft tolerance cannot be explained by a complete lack of specific immune reactivity to tolerated alloantigens.
Two Cytotoxic Cells in Peritoneal Cavity of Virus-Infected Mice: Antibody-Dependent Macrophages and Nonspecific Killer Cells
Authors: Macfarlan Roderick I. , Burns William H. , White David O.
Published: December 2022

Two new types of cell-mediated immune cytolysis of togavirus-infected cells are compared. The peritoneal cavity of mice 2 days after infection contains a nonadherent, non-phagocytic, non-θ-bearing, trypsin-resistant, EDTA-sensitive cell displaying broadly specific cytotoxicity for uninfected or virus-infected syngeneic or xenogeneic cell lines. Peritoneal macrophages from normal mice are cytotoxic to infected target cells sensitized with minute amounts of homologous antiviral IgG.
An in Vitro Model of Lymphocyte Homing
Authors: Woodruff Judith J. , Katz Ira M. , Lucas Linda E. , Stamper Hugh B.
Published: December 2022

Rat thoracic duct lymphocytes (TDL) are capable of selective adherence to the endothelium of high-endothelial venules (HEV) when overlaid onto glutaraldehyde-fixed sections of lymph nodes. The data presented indicate that lymphocyte adherence is an energy-dependent, calcium-requiring event that involves membrane determinants on TDL which are sensitive to trypsin. Surface sialic acids on lymphocytes are not essential and treatment of the cells with neuraminidase does not interfere with their attachment to HEV. There was no evidence that microtubule-associated functions play a role in binding. Adherence, however, is abolished by cytochalasin B, indicating that the cytoplasmic contractile microfilament system exerts an important effect. The results imply that lymphocyte surface membrane modulation is involved in the development of strong adhesive forces that bind the cells to the endothelium. In addition, lymphocyte-HEV adherence is reduced by ionophore A-23187, an agent known to inhibit surface membrane receptor movement. It is suggested that specific binding of recirculating lymphocytes to HEV is not a passive event, but that activation of cytoplasmic contractile forces in the lymphocyte is required for the formation of stable lymphocyte-HEV binding.
Effects of Ionic and Nonionic Detergents on Antigen-Antibody Reactions
Authors: Qualtiere Louis F. , Anderson Amil G. , Meyers Paul
Published: December 2022

Using highly sensitive and quantitative radioimmunoassay procedures we have measured the effects of different concentrations of three commonly used detergents, SDS, DOC, and Triton X-100, on antibody-antigen reactions. Triton X-100 had a relatively mild effect on primary antigen-antibody bindings, the precipitin reaction, and a double antibody RIA as evidenced by only an 8 to 10% inhibition of binding or precipitation. These results were not detergent concentration dependent, as Triton concentrations ranging from 5 to 0.1% had virtually no differential effects. Sodium deoxycholate (DOC) had a more profound effect on both primary antigen-antibody binding and the precipitin reaction than did Triton X-100, and its effects, unlike those of Triton X-100, were concentration dependent. There was a direct relationship between concentration of DOC and degree of inhibition of both primary binding and immune precipitation especially in antigen excess. Sodium dodecylsulfate (SDS), at concentrations 10- to 100-fold less than either Triton X-100 or DOC, had profound inhibitory effects on primary antigen-antibody binding, the precipitin reaction, and a double antibody radioimmunoassay. Generally, at concentrations greater than 0.01% SDS, almost all immunochemical reactivity is destroyed.
Increase in Titer of the Naturally Occurring Human Antibody to Neuraminidase-Treated Lymphocytes After Influenza
Authors: Rogentine G. N. , Doherty C. M. , Pincus S. H.
Published: December 2022

Nine of 25 (36%) humans suffering from naturally acquired influenza A infection developed significant increases in the titer of a “naturally” occurring antibody to neuraminidase-treated human lymphocytes. Only two of 43 normal and noninfluenza respiratory infection controls showed titer changes of this antibody, p < 0.001. The antibody was not directed at influenza virus C fixation, hemagglutination inhibition, or neuraminidase antigens. Three of 10 normals given a highly immunogenic, formalin-killed influenza A vaccine developed significant titer rises. These results suggest that influenza virus, live or dead, can provoke an increase in antibody to a cross-reacting antigen present on neuraminidase-treated human lymphocytes.
Kinetics of the in Vitro Lymphocyte Response to Myelin Basic Protein in the Lewis and Brown Norway Strains of Rat
Authors: Sheffield Warren D. , Gonatas Nicholas K. , Kim Seung U.
Published: December 2022

Several parameters of the in vitro lymphocyte proliferative response to myelin basic protein (BP) in Lewis (Le) and Brown Norway (BN) rats were examined. The results demonstrate that BN rats, a strain normally resistant to BP-induced experimental allergic encephalomyelitis, and Le rats, a strain readily susceptible to the disease, have similar patterns of the proliferative response to BP. An important difference, however, is that BN lymphocytes, although responding significantly to BP, are unable to proliferate to the same level as Le lymphocytes. In experiments measuring the lymphocyte response as a function of antigenic stimulus, days of culture, or type of adjuvant used, the BN rat peak response was in general 70% or less of the Le rat peak response. Furthermore, the BN lymphocyte response was reduced when B cells were removed whereas there was no effect in the Le rat. A negative feedback mechanism, possibly suppressor cells, has been suggested to explain these differences.
Suppressor T Cells: Presence in Mice Rendered Tolerant by Neonatal Treatment with Anti-Receptor Antibody or Antigen
Authors: Du Clos Terry W. , Kim Byung S.
Published: December 2022

Specific tolerance to phosphorylcholine (PC) was induced in BALB/c mice by two methods. Neonatal mice received a single injection of either: 1) PnC, the C-polysaccharide from S. pneumoniae, R36a vaccine which has PC as a major antigenic determinant or 2) ARA, an homologous antibody directed against the receptor for PC. Spleen cells from animals treated as neonates with either PnC or ARA were specifically suppressed for the response to PC antigens in vitro. In addition, cells from either group of unresponsive animals co-cultured with spleen cells of normal BALB/c mice markedly suppressed the response of the normal cells to PC. Greater than 90% inhibition of the plaque-forming cell response was obtained when unresponsive cells were mixed with normal cells in ratios of 1:1 or greater. Equal numbers of cells from animals made unresponsive by PnC or ARA produced an equivalent degree of suppression. Neither supernatants of cultures nor sera of animals of either unresponsive group suppressed the response of normal spleen cells to PC. Suppression by cells from both groups of tolerant mice was eliminated by treatment with anti-Thy 1.2 serum and C. Presumably, a common cell is responsible for suppression caused by cells from mice made unresponsive by either procedure.
Suppression of Stimulation in Mixed Leukocyte Culture by Newborn Splenic Lymphocytes in the Mouse
Authors: Bassett Mary , Coons Teresa A. , Wallis Wayne , Goldberg Ellen H. , Williams Ralph C.
Published: December 2022

Recent evidence (1, 2) has suggested that newborn human lymphocytes exert a suppressive effect on proliferation of adult lymphocytes in mixed luekocyte culture (MLC). Using mitotic indices as a measure of cell proliferation Olding et al. (1) have shown that 80% of the dividing cells in MLC containing male cord blood lymphocytes and adult female peripheral blood lymphocytes are of cord blood origin. An attractive explanation for this phenomenon is that newborn lymphocytes are capable of suppressing proliferation of the adult lymphocyte population. More recently Olding and Oldstone (2) showed that newborn cells which were responsible for suppressing proliferation of maternal lymphocytes resided in the T lymphocyte population. The objective of the present study was to determine in a murine system the effect of newborn versus adult stimulatory cells on the proliferation of responder cells in MLC.
Immunogenic Properties of Liposomal Model Membranes in Mice
Authors: Yasuda Tatsuji , Dancey George F. , Kinsky Stephen C.
Published: December 2022

Liposomal model membranes, which were prepared with sphingomyelin, cholesterol, and dicetylphosphate, and sensitized by incorporation of either dinitrophenyl-ε-aminocaproylphosphatidylethanolamine (DNP-Cap-PE) or fluoresceinthiocarbamylphosphatidylethanolamine (Fl-PE), can induce a hapten-specific humoral response when administered to AKR mice in saline. The magnitude of the response (as measured by the appearance of direct plaque forming cells in the spleen and/or serum antibody titer) is dependent on liposomal dose, the epitope density of the PE derivative in the liposomes, and the time after immunization.

In the case of DNP-Cap-PE sensitized liposomes, only a minor IgG response (as measured by the production of indirect plaques or mercaptoethanol-resistant antibody) could be detected after either primary or secondary immunization. Together with the finding that mice deficient in, or depleted of, thymus-derived (T) lymphocytes respond to liposomes containing DNP-Cap-PE, the available data indicate that these liposomes are primarily T cell-independent immunogens. The liposomes are also immunogenic in other inbred mice strains (differing in H-2 haplotype), although the magnitude of the response varies sufficiently to suggest the existence of high, intermediate, and low responders. On the basis of this preliminary survey, further examination of the possibility that the immunogenicity of liposomes may be under genetic control seems warranted.
Enhancement of Liposomal Model Membrane Immunogenicity by Incorporation of Lipid A
Authors: Dancey George F. , Yasuda Tatsuji , Kinsky Stephen C.
Published: December 2022

Incorporation of lipid A into sphingomyelin-cholesterol-dicetylphosphate liposomes stimulates the humoral response of AKR mice (and other inbred strains) to model membranes that are also sensitized with the synthetic amphipathic antigen, dinitrophenyl-ε-aminocaproylphosphatidylethanolamine (DNP-Cap-PE). This effect is observed with liposomes containing varying amounts of DNP-Cap-PE and the magnitude depends on the quantity of lipid A that is incorporated. No significant increase in the frequency of direct anti-DNP plaque-forming cells (PFC) is apparent in the spleens of mice immunized with a mixture of DNP-Cap-PE sensitized liposomes (lacking lipid A) and liposomes prepared with lipid A (lacking DNP-Cap-PE). Thus, enhancement of liposomal immunogenicity by lipid A is contingent on its presence in the same bilayers that contain DNP-Cap-PE. These results further indicate that covalent attachment of lipid A to an antigenic determinant is not required to obtain an increased hapten-specific response.

Three lines of evidence support the conclusion that stimulation can be attributed to the known mitogenic effect of lipid A on bone marrow-derived (B) lymphocytes. 1) Incorporation of alkali-treated lipid A, which is nonmitogenic, does not enhance the immunogenicity of DNP-Cap-PE sensitized liposomes in AKR mice. 2) Conversely, lipid A incorporation had no effect on anti-DNP PFC production in C3H/HeJ mice, a strain possessing B cells that are not responsive to this mitogen. 3) In contrast, lipid A did increase the response to DNP-Cap-PE sensitized liposomes in either nude or thymectomized mice that are deficient in thymus-derived (T) lymphocytes.

The available data suggest that modulation of liposomal immunogenicity by lipid A may be an effective method for enhancing antibody formation against lipid antigens.
Immunologic Properties of Purified Sendai Virus Glycoproteins
Authors: Örvell Claes , Norrby Erling
Published: December 2022

Egg-grown Sendai virus was used for purification of the glycoproteins of the virus. The two glycoproteins, HN glycoprotein (VP 2) and F glycoprotein (VP 4), were separated by filtration on DEAE-Bio-Gel A columns and by sucrose density electrofocusing and were used for preparation of rabbit hyperimmune sera. Rabbit hyperimmune sera directed against the HN and F glycoproteins were immunologically distinct. Antiserum directed against the HN glycoprotein inhibited hemagglutination (HA) and neuraminidase (NA) activity and had high neutralizing capacity. Antiserum against the F glycoprotein inhibited hemolysis but did not inhibit HA, NA activity, or virus infectivity. Rabbit hyperimmune sera against untreated, Tween 80-ether, and formalin-treated purified virions were also studied. These sera contained antibodies against both glycoprotein structures, but antibodies against the F glycoprotein blocking hemolysis could not be demonstrated. The two specific glycoprotein antisera were used to estimate the relative amount of hemagglutinin and hemolysin exposed on the envelope of Sendai virions from Vero cells by means of competition studies with 125I-labeled purified rabbit γ-globulin directed against egg-grown whole virions. Anti-HN and anti-F sera gave 60 to 70% and 30 to 40% inhibition, respectively, under conditions of antibody excess. Passive immunization with antisera against the two glycoproteins protected mice against natural Sendai virus infection.
Histocompatibility-2 System in Wild Mice
Authors: Klein Jan , Zaleska-Rutczynska Zofia
Published: December 2022

Sixteen B10.W congenic lines carrying wild derived H-2 haplotypes on C57BL/10Sn or B10 background were typed by the allogeneic cell-mediated lymphocytotoxicity (CML) assay; in addition, selected lines were also typed by the TNP-CML assay and by skin grafting. The analysis revealed similarity or identity of two strain pairs: SNA57 (H-2w21) seems to carry a similar haplotype as B10.SM (H-2v), and STA10 and STA12 seem to share the same H-2K and H-2D alleles. All other B10.W strains were different from each other and from B10 congenic lines carrying inbred-derived H-2 haplotypes. These results agree with the results of the serologic typing with two exceptions: the KPA42, KPA132, and SNA57 lines, which were serologically indistinguishable from each other and from B10.SM, were distinguished by histogenetic typing. The presence among wild mice of a haplotype (H-2w21) that appears to be very similar to a haplotype (H-2v) carried by an inbred strain (B10.SM) has some interesting implications for considerations of H-2 gene mutability. The finding that haplotypes derived from different localities are different provides further evidence that the H-2 polymorphism is extensive, indeed.
Functional Differentiation of B Lymphocytes in Congenital Agammaglobulinemia
Authors: Percy Maire E. , Dosch Hans-Michael , Gelfand Erwin W.
Published: December 2022

Cultures of peripheral blood lymphocytes (PBL) in which specific hemolytic plaque-forming cells (HcPFC) had been induced were labeled with 14C-amino acids. Antigen-specific products in the culture supernatants were characterized by using indirect immune precipitation in conjunction with specific immunoabsorbents and/or gel filtration followed by SDS-polyacrylamide gel electrophoresis. After 5 days of culture with antigen (sheep red blood cells or ovalbumin) newly synthesized IgM and specific IgM antibody were demonstrated in culture supernatants from normal donors and from four out of five patients with congenital agammaglobulinemia (cAγ). Secreted products bound specifically to antigen and pretreatment of labeled supernatants with anti-µ and anti-L chain antisera, but not with anti-γ anti-serum, prevented binding. Typical µ- and L chains constituted only a proportion of the antigen-binding peptides recognized by the anti-µ reagents. Induction of IgM antibody synthesis was dependent on the presence of antigen and was correlated with the generation of HcPFC. No major differences between the antigen-induced products of cAγ and normal PBL were observed. These findings suggest that in the absence of terminal B cell differentiation in vivo, certain patients with cAγ possess precursor cells that can respond to antigen in vitro with the synthesis of specific humoral products, including IgM antibody.
Different Sensitivities to Hydrocortisone of Natural Killer Cell Activity and Hybrid Resistance to Parental Marrow Grafts
Authors: Hochman Paula S. , Cudkowicz Gustavo
Published: December 2022

Natural killer cells and cells mediating F1 anti-parent responses in vivo and in vitro differ in their sensitivity to hydrocortisone acetate. NK cell activity is sharply decreased after in vivo drug administration whereas induction of specific F1 anti-parent or anti-allogeneic cytotoxicity and hybrid resistance to parental marrow grafts are not impaired. Because of several other properties shared by the NK and anti-Hh host reactivities, it is still suggested that the effector cells are generated from a single differentiation pathway, but that they differ with respect to specificity and sensitivity to hydrocortisone.
The SC5b-7 Complex: Formation, Isolation, Properties, and Subunit Composition
Authors: Podack Eckhard R. , Kolb William P. , Müller-Eberhard Hans J.
Published: December 2022

Activation of C in C8-depleted serum results in the formation of a soluble complex containing C5, C6, and C7. The complex has an electrophoretic mobility of an α-globulin, an s-rate of 18.5S, and a m.w. of 668,000 daltons. This complex was isolated and upon SDS polyacrylamide gel electrophoresis it was found to contain, in addition to C5b, C6 and C7, an 88,000 dalton glycoprotein. The protein was identified as the band V protein of the soluble C5b-9 complex. It is referred to as SIIIs-protein, or S-protein. Since the S-protein does not bind to C5b-6, it is concluded that it is incorporated during the fusion of C5b-6 with C7. The SC5b-7 complex exhibits the same neoantigen as the SC5b-9 complex, but compared to the C5b-6 complex it appears to contain an additionally qualitatively distinct neoantigen.
Surface Immunoglobulin and Fc Receptors on Murine B Lymphocytes: Loss of Receptor Interaction after Cell Activation
Authors: Scribner Duncan J. , Weiner Howard L. , Moorhead John W.
Published: December 2022

Although surface immunoglobulin (sIg) and Fc receptors (FcR) are independent receptors in the membrane of B lymphocytes and are assumed not to interact under normal conditions, it has been shown that anti-Ig-induced redistribution, i.e., patching and capping, of sIg results in the co-capping of FcR, suggesting some type of receptor interaction. We have investigated this phenomenon by comparing the interaction of these receptors on normal vs activated B cells under a variety of conditions. It was found that, in contrast to unstimulated control cells, sIg and Fc receptors on blast cells induced by either Fab′2 anti-Ig or endotoxin protein do not interact, i.e., little if any co-capping was seen. This loss of receptor interaction was not due to increased cell size since cells which were stripped of their sIg and FcR and allowed to re-express both receptors for 24 hr, but which had not increased in size during this period, also failed to show co-capping behavior. Additional experiments showed that the Fc receptors on activated B cells could be capped directly by using complexes of FITC-KLH-anti-KLH, indicating that the loss of sIg-FcR co-capping on activated cells is not due to any restriction in the mobility of the Fc receptor in the membrane. These results strongly suggest that B lymphocyte activation is accompanied by membrane alterations that result in the loss of sIg and FcR interaction and that these changes do not require receptor redistribution and re-expression.
In Vivo Responses to Inhaled Proteins
Authors: Willoughby Judith Barbaras , Willoughby William F.
Published: December 2022

Quantitative studies describing the in vivo behavior of an inhaled protein antigen are presented. Aerosol mists were generated from bovine serum albumin (BSA) solutions without detectable alteration in the physical or antigenic properties of the protein molecule. The fate of inhaled 125I-BSA in vivo was studied in rabbits by using a paired-isotope technique to quantitate antigen sequestration within organs, as distinct from circulating antigen label present within the plasma compartment of such organs. Total body uptake could be controlled and averaged 3.20 ± 0.26% of the protein introduced into the aerosol chamber. Accessibility of such aerosolized antigen to the alveolar sacs was confirmed by ultraviolet microscopy with fluorescein-tagged BSA. Immediately after a 90-min aerosol exposure, over 90% of the in vivo label was localized within organs (stomach, 72%; lungs, 4%; intestine, 7%). Maximal concentrations of antigen in the intestines occurred 30 min later. The subsequent decline in gastrointestinal antigen concentrations was accompanied by the appearance of degraded antigen peptides in the serum, peaking at 24 hr. By 48 hr, over 99% of the inhaled antigen had been degraded, and the radiolabel excreted into the urine (93%) or incorporated into the thyroid (6%); less than 0.1% remained in the lung. Similarly, with repeated daily administrations the antigen was rapidly degraded, and no accumulations of antigen in the lung were observed. The presence of humoral immunity induced by systemic immunization did not affect the fate of inhaled antigen. Biweekly exposures of previously unimmunized animals to BSA aerosols resulted in vigorous humoral antibody responses representing IgG and IgM immunoglobulin classes. Despite the presence of good Arthus skin reactivity in animals immunized by systemic depot injections or by inhalation routes, subsequent repeated challenges with antigen aerosol failed to induce Arthus-like injury in the lung. This study suggests that antigen inhalation may function primarily as an important physiologic means by which natural immunity to environmental antigens is established. Humoral antibody, although capable of readily mediating cutaneous Arthus reactions, appears to require an initiating event or mechanism, as yet undefined, before it can interact with inhaled antigen to form pathogenic immune complexes.
The Influence of Anaphylactic Shock on Fluid in the Peritoneal Cavity
Authors: Fleisher Moyer S. , Mayer Leo L. , Wilhelmj C. M.
Published: December 2022

Conclusion

It appears that the difference in osmotic pressure of the recovered peritoneal fluid of shocked and normal animals is due to a higher chlorine ion concentration in the former.

Protein constituents in the two sorts of fluids are apparently not different and cannot therefore be factors in the difference in osmotic pressure.

Phosphates are also probably not responsible for the difference although they may play some part.

We cannot at the present time offer an explanation as regards this difference in the chlorine ion concentration in shocked animals as compared to normal animals.
The Hypersensitiveness Following the Administration of Placental Extract
Authors: Smith George H. , Musselman L. K.
Published: December 2022

In a previous paper (Jour. Immunol., 1926, 12: 7) it has been shown that employing either the actively or passively sensitized guinea pig, the administration of an extract of human placental tissue confers upon the animal a hypersensitive state toward female human serum while hypersensitivity toward male human serum is markedly less. The following note deals with a demonstration of the same reaction in vitro, using the uterine horn.

The guinea pigs used in this experiment were sensitized by the intraperitoneal injection of the serum of a rabbit which had received repeated injections of an extract of human placental tissue. Within a period of from twenty-four to ninety-six hours after such a passively sensitizing injection the guinea pigs were sacrificed, the uterine horns were suspended in oxygenated Ringer-Locke solution and adjusted to record muscular activity upon a kymograph. The results of one such experiment are presented in the following graphs.
Studies in Complement Splitting with Special Reference to the Activation of Yeast Absorbed and Complement Deficient Guinea Pig Serum
Authors: Parsons Elizabeth I.
Published: December 2022

Guinea pig complement was split into two pieces by means of CO2. The hemolysis resulting from a mixture of the two fractions showed a loss of about two-thirds of the complement activity. The results of earlier workers as to the quantitative relationships of the two pieces and the modification of the midpiece when standing in sodium chloride were confirmed. Both midpiece and endpiece were found to be labile at 55°C. for thirty minutes. Guinea pig complement was split into three pieces by means of (NH4)2SO4 as described by Browning and Mackie. Our results, although less satisfactory quantitatively, confirmed those of the above workers. In light of the difficulties encountered this difference is probably due to the water supplies used for dialysis. Attempts were made to split complement into four pieces by means of CO2 and (NH4)2SO4 as described by Browning and Mackie, but with no success. It is highly probable that the water supply affected these results. Guinea pig complement absorbed with yeast was activated by both the midpiece and endpiece from the CO2 fractionation, but more consistently and to a greater extent by the former. Heating of the pieces at 55°C. for thirty minutes reduced their ability to reactivate. The reactivation of the yeast absorbed serum by means of the three pieces from the (NH4)2SO4 splitting shows this function to lie entirely in the pseudoglobulin fraction. It is partially destroyed by heating at 56°C. for thirty minutes, due to its isolated condition. Serum from the complement deficient guinea pig was always activated by the endpiece from the CO2 splitting in high dilutions, and by the midpiece frequently, but in lower dilutions. Heating at 55°C. for thirty minutes destroyed the activating ability entirely. The data available indicate that both the pseudoglobulin and the albumin from the (NH4)2SO4 splitting will activate the deficient serum to a small extent, but a mixture of the two is essential for the most satisfactory activation. The component removed from complement by the absorption with yeast is not identical with that removed by nature in the complement deficient guinea pig serum, since it is impossible to activate them in an identical manner with the fractions obtained by splitting guinea pig serum with CO2 and (NH4)2SO4.