bluuebunny/crossref_metadata_2025 huggingface dataset:
Complement-Mediated Release of Histamine from Human Leukocytes
Authors: Grant J. Andrew , Dupree Elton , Goldman Armond S. , Schultz Duane R. , Jackson Anne L.
Published: December 2022

Activation of either the alternative or classical pathway of complement generated a factor which induced release of histamine from both non-allergic and allergic human basophils. This factor probably is derived from the complement system since 1) its formation was associated with loss of C3 activity in human serum, 2) chemotactic factor, probably also a complement product, was generated simultaneously, 3) heat inactivation blocked its formation, 4) anti-C3 and anti-C5 blocked formation of the factor, and finally 5) anti-C5 inhibited the activity of the factor once it had been formed. It appears that both complement-mediated and allergen-mediated release of histamine from basophils are secretory, non-cytolytic pathways since both were maximal at 37°C, required the presence of divalent cations, and were inhibited by theophylline. One consistent difference between these two mechanisms was noted: complement-initiated release of histamine occurred more quickly.
Antigenic Analysis of Three Strains of Mycoplasma Arginini by Two-Dimensional Immunoelectrophoresis
Authors: Thirkill Charles E. , Kenny George E.
Published: December 2022

The occurrence of species-specific and strain-specific antigens in three strains of Mycoplasma arginini (G-230, leonis and 23243) was studied by two-dimensional immunoelectrophoresis. Approximately 20 antigenic components could be detected in each strain. It was possible to analyze 6 to 7 major and distinct components from each strain by two techniques: “enhancement” where antigen to an additional strain is added to the first phase of the electrophoresis which increases the size of common peaks and “suppression” where antiserum to an additional strain is incorporated in the second phase whereby peak size of components to which both sera have antibody are decreased. A total of 10 distinct antigens were recognized. Electrophoretic mobilities relative to bovine albumin ranged from 0.2 to 1.08. Three components were common to all strains; two of these represented major amounts of material. Four components represented strain-specific components. Unique fast components were found both in strains 23243 and G-230. Three antigens were distributed into only two of the three strains. The electrophoretic mobilities of some common antigens were quite different between strains.
Antigen-Specific and Nonspecific Mediators of T Cell/B Cell Cooperation
Authors: Marrack (Hunter) Philippa C. , Kappler John W.
Published: December 2022

Evidence is presented for two types of helper T cells in the mouse specific for a protein antigen, keyhole limpet hemocyanin (KLH). The first cell is able to help B cells respond to the hapten, trinitrophenyl (TNP), when coupled to KLH presumably through the participation of KLH-specific T cell factors. The second helper cell when stimulated with KLH is able to help B cells respond to red blood cell antigens, but not TNP-KLH, through the production of a non-antigen-specific factor.

Both types of helper cells were shown to be present in the spleens of KLH-primed mice, although they occurred with different frequencies. They were shown to be different T cells by their segregation from each other in vitro at limiting dilution.
Listeria Cell Wall Fraction: A B Cell Mitogen
Authors: Cohen J. John , Rodriguez Gilberto E. , Kind Phyllis D. , Campbell Priscilla A.
Published: December 2022

A crude cell wall-rich fraction of Listeria monocytogenes contains a substance or substances which cause blast transformation and thymidine incorporation by mouse spleen cells. The substance, Listeria cell wall fraction (LCWF), is a B cell mitogen in the mouse. It is not endotoxin, in that it stimulates B cells in C3H/HeJ mice. It is weakly mitogenic for rat spleen cells and not at all mitogenic for human peripheral blood or tonsil lymphocytes. Thus, it is dissimilar to both endotoxin and purified protein derivative (PPD) in its spectrum of activity. A cell wall-associated mitogen may play an early role in Listeria resistance.
Adult Murine Lymph Node Cells Respond Blastogenically to a New Differentiation Antigen on Isologous and Autologous B Lymphocytes
Authors: Ponzio Nicholas M. , Finke James H. , Battisto Jack R.
Published: December 2022

Cells derived from lymph nodes (LN) of adult CBA mice respond blastogenically to mitomycin-treated autologous, as well as isologous spleen cells. This isogeneic LN-to-spleen (mixed lymphocyte culture) is best obtained when both responder and stimulator cells are derived from donors greater than 10 weeks of age. Responsive cells appear restricted to LN since they could not be detected in adult spleen, marrow, or thymus. LN cells do not require the presence of spleen in order to differentiate into responder cells since those derived from neonatally splenectomized mice are fully active.

Stimulator cells appear in the spleen, bear Ig on their surfaces, and can be detected in spleens of irradiated, bone marrow-reconstituted mice. Experiments comparing the responsiveness of adult LN cells and that of neonatal T cells toward mitomycin C-treated lymphoid cells from a variety of sources suggest the presence of two iso-antigens on B lymphocytes. Since both antigens apparently are absent on precursor bone marrow cells and develop with time, they have been classified as murine differentiation antigens 1 and 2 (MDA-1, MDA-2). Whereas both appear in the spleen, only one, MDA-1, is also detectable by this methodology in LN. Both MDA-1 and MDA-2 activate neonatal T cells, but MDA-2 triggers only adult LN. Whereas MDA-2 developed in an x-irradiated, bone marrow-reconstituted spleen, MDA-1 did not over a 9-week interval.
Macrophage Heterogeneity in Receptor Activity: The Activation of Macrophage Fc Receptor Function in Vivo and in Vitro
Authors: Rhodes John
Published: December 2022

The Fc receptor activity of single macrophages and of distinct macrophage populations was examined in terms of cellular avidity for IgG antibody by using a rosette assay which permits characterization of receptor activity at the cellular level and quantitation of any changes in such receptor activity. Marked heterogeneity was shown to exist within normal populations of macrophages. Both normal peritoneal and alveolar macrophage populations exhibit a normal or logistic distribution of cellular avidities for IgG antibody. The mean avidity of the peritoneal population, however, was found to be approximately 3 times greater than that of the alveolar population. Moreover, alveolar macrophages possess a range of cellular avidities 3 times greater than that of peritoneal macrophages.

Comparison of normal unstimulated peritoneal macrophages and induced inflammatory exudate macrophages revealed a 6-fold increase in the proportion of high avidity cells in the latter population. Normal peritoneal and normal alveolar macrophages were shown to undergo a striking increase in Fc receptor activity in vitro. Under certain conditions of culture these cells acquire the antibody-binding capacity characteristic of high avidity cells in the inflammatory exudate population. Fresh guinea pig serum was shown to prevent this activation of Fc receptor function in vitro. Aged guinea pig serum was somewhat less effective. The serum factor(s) responsible appears to be consumed or exhausted in some way by macrophages. It is heat stable and has a molecular weight of less than 100,000 daltons. Serum IgG was found not to be involved in the effects of fresh serum on receptor activation in vitro.

The activation of Fc receptor function described here may facilitate both the presentation of antigen to specific T lymphocytes and the antibody dependent killing of tumor cells by macrophages. Changes in receptor activity of the kind described may thus play an important regulatory role in the induction of an immune response and in the effector mechanisms of immunologic surveillance.
Immunologic Functions of Isolated Human Lymphocyte Subpopulations
Authors: Sondel Paul M. , Chess Leonard , MacDermott Richard P. , Schlossman Stuart F.
Published: December 2022

The studies presented herein have evaluated both the specificity and cellular basis of cell-mediated lympholysis (CML) in man. An efficient and quantitative 51Cr release assay was utilized to study the role of highly purified human T and B cells in CML. After in vitro sensitization human T cells develop the capacity to kill specifically allogeneic cells to which they were sensitized. In contrast, B cells were neither triggered to proliferate nor activated to kill allogeneic targets. B cells were not activated to kill even when sensitized in the presence of potentially “helper” T cells, nor did they block T cells from killing during the effector phase. Cell-free supernatants taken from active in vitro sensitization cultures were not lympholytic and did not modulate T cell killing. Hence, these studies show that both the afferent and efferent phases of human CML are T cell functions.
Studies on the Cytophilic Properties of Human β2 Microglobulin
Authors: Anderson Clark L. , Kubo Ralph T. , Grey Howard M.
Published: December 2022

Human β2microglobulin is cytophilic for mouse, rat, and to a lesser extent guinea pig lymphocytes and polymorphonuclear leukocytes but not for human, rabbit, chicken, frog, and turtle cells, nor for any mature erythrocytes tested. Mouse splenic lymphocytes bind 5- 7-fold more β2microglobulin than thymocytes although hydrocortisoneresistant thymocytes resemble spleen cells in this regard. The β2microglobulin binding by B and T cells in mouse spleen is similar. The structure responsible for β2 microglobulin binding to the surface of cells is distinct from the Fc receptor specific for aggregated IgG.
Detection of Membrane-Associated Antigens on Lymphoid Cells by Antibody Coupled to Staphylococcal Protein A
Authors: Kearney Raymond , Chia Eunice , Basten Antony
Published: December 2022

A simple technique is described for the detection of membrane-associated antigens on lymphoid cells. It is based on the observation that the protein A component of staphylococci binds to the Fc pieces of IgG molecules. Lymphocytes from various sources (mouse, rat, and human tissues) were incubated with hyperimmune antisera directed against surface determinants. Subsequent treatment with a suspension of staphylococci containing protein A permitted visualization of both the presence and distribution of determinants on the cell membrane. The method had comparable sensitivity to the fluorescent sandwich technique and could be used to detect a variety of membrane antigens on both T cells and B cells.
Tolerance and Contact Sensitivity to Dnfb in Mice
Authors: Phanuphak Praphan , Moorhead John W. , Claman Henry N.
Published: December 2022

Contact sensitivity to 2,4-dinitro-1-fluorobenzene (DNFB) in mice could be rapidly and specifically suppressed by a single i.v. injection of DNBSO3 just before the elicitation skin test. The effects of desensitization were also demonstrated in vitro by the diminished response of cultured desensitized lymph node cells to specific antigenic stimulation (DNBSO3). The effect was dependent on the dose of DNBSO3 injected, and it occurred immediately and disappeared within 2 days. We believe that the mechanism is a functional defect of antigen-reactive cells which have been triggered to proliferate by the desensitizing antigen. These rapidly cycling cells may not migrate properly or respond adequately to the skin test challenge and are also refractory to additional antigen stimulation in vitro. The mechanism so postulated is derived from the finding that there was an inverse relation between the proliferative rates of the cells and their degree of specific reactivity to antigen. When the desensitizing antigen raised the proliferative rate of lymph node cells, the specific reactivity of both the whole animal and of the isolated lymphoid cells was low and vice versa.
Effect of Cortisone on the Disappearance Kinetics and Tissue Localization of Soluble Immune Complexes
Authors: Haakenstad Alan O. , Case Jean B. , Mannik Mart
Published: December 2022

The effect of cortisone treatment on the disappearance kinetics, the hepatic uptake, and the glomerular deposition of i.v. administered, soluble immune complexes (HSA-anti HSA) was examined in mice. An initial rapid disappearance of complexes from the circulation occurred after the injection of complexes into control mice, which was caused by increased vascular permeability. This phase was absent in the cortisone-treated group. The half life of complexes composed of more than two antigen and two antibody molecules (τ; Ag2Ab2) was prolonged from 1.93 hr in control mice to 4.71 hr in cortisone-treated mice while the half life of Ag2Ab2 complexes was unchanged (11.40 hr vs 12.04 hr). Although the clearance velocity of τ; Ag2Ab2 complexes was suppressed in cortisone-treated mice, the quantity of complexes specifically located in the liver at 1, 2, and 4 hr was not significantly different in the two groups. Persistence of circulating τ; Ag2Ab2 complexes was associated with enhanced and prolonged glomerular deposition of complexes in the cortisone-treated mice.
Complement-Induced Histamine Release from Human Basophils
Authors: Hook William A. , Siraganian Reuben P. , Wahl Sharon M.
Published: December 2022

The incubation of zymosan, endotoxin, or immune aggregates with normal human serum activates a factor which induces release of histamine from autologous basophils. The reaction can be divided into two steps: in the first, complement must be activated and in the second, the histamine-releasing factor interacts with basophils. The generation of histamine-releasing activity in serum occurs at 17 to 37°C but not at 0°C, is inhibited by heating the serum at 56°C for 30 min, or by the addition of EDTA to the serum. Once generated, the histamine-liberating activity is stable to heating at 56°C for 30 min. Gel filtration of the activated serum demonstrated that this factor eluted in the same region as a factor with chemotactic activity. Both factors have a molecular weight of about 16,000 daltons and their activities were inhibited by antibody to human C5. This is therefore a pathway for histamine release by C5a where the activation of the basophil is unrelated to the membrane bound IgE.
Non-Random Distribution of Surface Immunoglobulin on Murine B Lymphocytes
Authors: Abbas Abul K. , Ault Kenneth A. , Karnovsky Morris J. , Unanue Emil R.
Published: December 2022

An indirect labeling technique was used to map the surface immunoglobulin (Ig) on murine splenic B lymphocytes by freeze-etching. Cells were labeled first with fluorescein conjugates of monovalent (papain-digested) anti-Ig antibody followed by monovalent anti-fluorescein antibody coupled to ferritin. The technique avoids crosslinking and aggregation of surface Ig. Freeze-etched replicas of cells labeled at 4°C, as well as of cells prefixed with paraformaldehyde showed that surface Ig was distributed in small clusters with interconnecting networks. The observed pattern was analyzed statistically by comparing it with the expected random (Poisson) distribution and shown to be non-random to a high degree of statistical significance. The deviation from randomness could be explained by the presence of clusters and relative excesses of bare membrane. Such an observed distribution of surface Ig suggests that this membrane macromolecule may be organized in a specific manner. The distribution may also play a role in the function of surface Ig as the antigen receptor on B lymphocytes.
Serologic and Biochemical Studies of the Ia System of the Mouse H-2 Gene Complex. Further Evidence for an I-C Subregion
Authors: David Chella S. , Cullen Susan E. , Murphy Donal B.
Published: December 2022

Three antisera were made against B10.A(5R) tissues to show the presence of an I-Cd subregion in that strain: 1) B10 anti-B10.A(5R) [anti-Dd, I-Cd]; 2) (B10 × C3H.OL) F1 anti-B10.A(5R) [anti-Dd]; 3) (B10 × HTI)F1 anti-B10.A(5R) [anti-I-Cd]. Serologic analyses of the antisera clearly showed the presence of an Id segment in B10.A(5R) and provided further evidence for the existence of a new I-C subregion. These anti-I-Cd sera define the Ia.7 specificity and analyses of several recombinants map Ia.7 within the I-C subregion. Ia.7 is present in I-Ck, I-Cd and Ip strains. Biochemical analyses showed that the Ia.7 molecule has a molecular weight of 31,000 and is distinct from the 46,000 m.w. H-2K or D molecules. The possible roles of the I-C region in immune response, mixed lymphocyte reaction, and graft-vs-host reaction are discussed.
Capping of the Lymphocyte C3 Receptor and Temperature-Dependent Loss of C3 Rosettes
Authors: Gormus Bobby J. , Shands Joseph W.
Published: December 2022

The percentage of mouse spleen cells rosetted by bacteria-antibody-complement complexes (BAC) was found to be temperature dependent. Maximal rosetting was observed when BAC and spleen cells were incubated in ice (0.5°C), whereas less rosetting resulted from 37°C incubations. When rosetted cells were incubated at 37°C, cap formation was found to occur, and indirect evidence suggested that the capped bacteria were subsequently shed from the lymphocyte surface. Neither capping nor shedding occurred at 0.5°C, suggesting that these phenomena were responsible for the decreased percentage of rosetted cells found after incubation at 37°C. An incidental observation indicated that a second factor potentially contributing to the loss of rosetted cells at 37°C was the removal of BAC from the surface of cells by phagocytes.
Malaria (Plasmodium Knowlesi) Merozoites: Immunity and the Surface Coat
Authors: Miller Louis H. , Aikawa Masamichi , Dvorak James A.
Published: December 2022

Immune serum agglutinated merozoites of Plasmodium knowlesi in culture. Agglutinated merozoites attached to erythrocytes but were usually unable to invade. Marked aggregation of merozoites was associated with reduced invasion of erythrocytes in these cultures. The agglutination and reduced invasion were immunologically specific, for the effect of serum was greatest against homologous strains of P. knowlesi. Merozoite agglutination was caused by the binding of surface coats on adjacent parasites. This coat appeared on the plasma membrane of merozoites after it was exposed to culture medium, both with and without immune serum. The coat consisted of protein or glycoprotein, since it was susceptible to trypsin treatment. It appears that antibodies directed against this surface coat are crucial for reduced invasion of erythrocytes, at least in vitro.
Pathogenesis of Experimental Allergic Uveitis Induced by Retinal Rod Outer Segments and Pigment Epithelium
Authors: Meyers Roberta L. , Pettit Thomas H.
Published: December 2022

Experimental allergic uveitis (EAU) was produced in strain 13 guinea pigs after immunization with purified guinea pig retinal rod outer segments and pigment epithelium (PE) in mycobacterial adjuvant emulsion, The lesions of EAU appear as inflammatory infiltrates of the iris, ciliary body, and choroid, often with photoreceptor degeneration. Specific antibodies are frequently detected in the serum of some but not of all animals with clinical uveitis. Immunohistochemical staining of normal and inflamed eyes with serum from the immunized animals with clinical disease demonstrated specific antigens localized in the retinal photoreceptor layers, whether or not circulating precipitating antibodies were also present in the serum. All immunized animals demonstrated delayed-type hypersensitivity (DTH) characterized by skin reactions of mononuclear cells and by the in vitro inhibition of migration of cells from the sensitized animals in the presence of specific antigen whether or not clinical uveitis occurred. However, stronger DTH reactions were observed during clinical uveitis. Cellular immunity appears to correlate with clinical EAU, whereas, no correlation was found with serum antibody. These findings suggest that the retinal photoreceptor cell and PE are the source of the significant antigens in autoimmune uveitis and retinitis.
Antibody Production and Interaction with Lymphoid Cells in Relation to Tumor Immunity in the Moloney Sarcoma Virus System
Authors: Harada Minoru , Pearson Gary , Redmon Lena , Winters Elaine , Kasuga Sidney
Published: December 2022

The temporal development of antibodies to Moloney sarcoma virus (MSV)-induced antigens in relation to tumor progression was followed with the membrane immunofluorescence (MF) and antibody-dependent normal lymphocyte cytotoxicity (ADNLC) assays. Antibody was detected at 14 days following virus infection by MF. In mice that developed primary tumors which regressed, MF titers developed to high levels following a period between 2 and 4 weeks post-inoculation during which the titers remained at low and constant levels. The increase in MF titers corresponded closely on a temporal basis to the initiation of the regression process. In contrast, antibody remained at low levels in those mice that developed progressively growing tumors. ADNLC was detected in the sera of regressor mice approximately 6 weeks post-inoculation but was not detectable at any of the time intervals in sera from mice with progressively growing tumors. Attempts to enhance the cytotoxic activity of normal spleen cells directly with immune serum were unsuccessful. The synergistic effect of serum on the cytotoxic activity of immune lymphocytes was less pronounced than with normal lymphocytes in this system. Temporal studies indicated that there was an inverse relationship between the development of cellular immunity and the capability of these cells to be activated in the ADNLC assay.
Presence of Thymic Antigen on Rabbit Basophils
Authors: Day Richard P. , Singal Dharam P. , Bienenstock John
Published: December 2022

A goat heteroantiserum specific for a rabbit thymus lymphocyte antigen (RTLA) reacted with rabbit basophils resulting in the release of histamine. This activity was equally absorbed out by thymocytes or basophils. Absorption with either of these cell types also resulted in equal loss of thymocytotoxicity. No effect in either system occurred after absorption with either granulocytes or rabbit fibroblasts. These results could not be explained by the presence of immune complexes or aggregated globulin present in the RTLA antiserum. Further, the RTLA antiserum had no anti-IgE activity, as demonstrated by its lack of reactivity with mast cells that were otherwise capable of releasing histamine normally after challenge with antigen. We have thus shown a basic difference between mast cells and basophils. We conclude that the rabbit basophil may bear a thymic marker.
Determination of Antibody-Hapten Association Kinetics: A Simplified Experimental Approach
Authors: Skubitz Keith M. , Smith Thomas W.
Published: December 2022

A simplified method has been developed for the determination of antibody-hapten association kinetics that permits the study of high affinity interactions with second order forward rate constants of the order of 107 to 108 M-1 sec-1. Use of tritiated haptens of high specific activity and antibodies of high affinity allows reactions to be run at initial hapten and antibody concentrations of the order of 10-9 to 10-10 M, well below the level at which mixing becomes the rate-limiting step. Separation of antibody-bound from free hapten by the use of dextran-coated charcoal can be carried out with sufficient rapidity (2 sec) that the systems under investigation are not appreciably disturbed.

With this technique, the association of 3H-ouabain with rabbit ouabain-specific antibody was found to occur with a rate constant of 0.8 × 107 M-1 sec-1, similar to association rates of dye haptens with antibodies of substantially lower affinity. The ratio of this association rate constant to the independently determined dissociation rate constant was 5.4 × 109 M-1, in satisfactory agreement with a k0 value of 3.5 × 109 M-1 determined by Sips analysis of data obtained under equilibrium conditions. This approach should be applicable to the direct kinetic assessment of numerous high affinity antibody-hapten systems of current interest.
Release of Cyclic AMP from Macrophages by Stimulation with Prostaglandins
Authors: Gemsa Diethard , Steggemann Ludwig , Menzel Jörg , Till Gerd
Published: December 2022

The effect of various prostaglandins (PG) on the generation of cyclic AMP in rat peritoneal macrophages has been studied in vitro. PGE1 produced a rapid intracellular accumulation of cyclic AMP which was followed by its release into the extracellular space. More cyclic AMP was released with prostaglandins of the E-type than with A- and F-types. It is suggested that release of cyclic AMP from macrophages may participate in the modulation of leukocyte function.
Reactions of Murine Myeloma Cells with Infectious Mononucleosis Sera
Authors: Yoshida Hiroshi , Kano Kyoichi , Milgrom Felix
Published: December 2022

Reactions of murine myeloma cells with infectious mononucleosis sera were studied by means of cytolysis in agarose gel. Of 75 sera tested, 30 lysed IgM myeloma cells, MOPC-104E. The antibodies responsible for the lysis of the myeloma cells were shown to be different from Paul-Bunnell antibodies and other antibodies found in infectious mononucleosis sera. Three types of antibodies acting upon the myeloma cells were identified serologically on the basis of absorption experiments with bovine erythrocytes, θ-positive murine lymphoma cells and guinea pig kidney cells. Antibodies of the first group could be absorbed with none of these antigens, antibodies of the second group could be absorbed only with lymphoma cells, and antibodies of the third group could be absorbed with any of these three antigens.

Evidence was presented that the antibodies under study combine with antigenic cell membrane components of a subpopulation of IgM-producing murine B cells.
Inhibition of Lymphocyte Stimulation by Measles Virus
Authors: Sullivan John L. , Barry David W. , Albrecht Paul , Lucas Susan J.
Published: December 2022

The effect of measles virus on phytohemagglutinin-(PHA) induced stimulation of human peripheral blood mononuclear cells was investigated to delineate possible mechanisms for viral suppression of cell-mediated immunity (CMI). It was noted that medium which had several days contact with uninfected monolayers as well as unpurified measles virus preparations produced significant inhibition of 3H-thymidine incorporation by PHA-stimulated lymphocytes. When partially purified measles virus preparations were used, however, marked inhibition was observed and the inhibitory effect of cell-derived factors could be separated easily from the virus-induced inhibition.

Experiments to determine the mechanisms of this virus-induced inhibition of 3H-thymidine incorporation showed the following: 1) live measles virus and not UV-irradiated or heat-inactivated virus produced inhibition; 2) the inhibitory effect observed was not the result of a viral-induced inhibitor being released from measles-infected lymphocytes; and 3) monocyte depletion had no effect on the ability of measles virus to inhibit 3H-thymidine incorporation by PHA-stimulated lymphocytes.

Since it was found that measles virus-infected lymphocytes display an impaired response to in vitro PHA stimulation, perhaps this dysfunction may be extended to mediator release and other functions associated with delayed cutaneous hypersensitivity (DCH) in vivo.
Genetic Control of Responses to Bacterial Lipopolysaccharides in Mice
Authors: Watson James , Riblet Roy
Published: December 2022

C3H/HeJ mice contain a defect in a single autosomal locus which is not linked to the H-2 histocompatibility or the heavy chain allotype loci that restricts immune, mitogenic, and polyclonal responses to bacterial lipopolysaccharides (LPS). Adult thymectomized C3H/HeJ mice that have been irradiated and reconstituted with C3HeB/FeJ bone marrow cells respond well to LPS. Cell-mixing experiments using C3H/HeJ-C3HeB/FeJ spleen cultures show that the failure of C3H/HeJ spleen cells to support responses to LPS is not due to nonspecific or LPS-induced suppressive events, or the lack of accessory cell types. C3H/HeJ and C3HeB/FeJ spleen cells bind LPS and respond to other B cell mitogens equally well. We suggest that the B lymphocytes of C3H/HeJ mice have a defect in a membrane component that is activated via interaction with LPS, and initiates the intracellular events that lead to cell proliferation.
Immune Mechanisms in Leukemia: Evaluation of Immunocompetent Cell Populations
Authors: Lukasewycz Omelan A. , Martinez Douglas , Murphy William H.
Published: December 2022

Dose-response curves of the cellular immune response of C58/wm mice to syngeneic line Ib malignant lymphoid cells (Ib cells) were computer analyzed by the PROBT subroutine in the IBM Scientific Subroutine Package. An analysis of the relative immunogenicity of various admixtures of x-irradiated (XIb) and viable Ib cells (VIb) after i.p. injection showed that the ratio had to be approximately 100:1 to be immunogenic. Viable Ib cells contained in immunogenic mixtures multiplied in vivo at a logarithmic rate up to 5 or 6 days but were eliminated immunologically by 8 or 9 days.

Adoptive cell transfer techniques were used to quantify the protective effect of immune spleen cells (ISC). Essentially a constant dose of ISC (106.4) protected mice against a challenge dose of 102 to 105 VIb cells; more than 105 VIb cells were lethal. Two techniques were used to quantify immunity even though mice ultimately died of transplanted leukemia, viz., mean survival time (MST) with a fixed challenge dose of VIb cells, or MST with a fixed time for death. The sensitivity and statistical limitations of these assays are presented.

To amplify the sensitivity of assays for adoptive cellular immunity a technique of antigenic stimulation was used, viz., 1 day after x-irradiated mice (600 R) received an i.p. injection of normal or immune spleen, bone marrow or thymic cells they received an i.p. injection of XIb cells containing an admixture of VIb cells. The technique amplified the sensitivity of ISC transfer tests approximately 100-fold and made it possible to detect protective effects of bone marrow and thymic cell populations.
Regulation of the Release of Colony-Stimulating Activity from Mitogen-Stimulated Lymphocytes
Authors: Ruscetti Francis W. , Chervenick Paul A.
Published: December 2022

Lymphocytes stimulated by Concanavalin A (Con A) as well as other mitogens release colony stimulating activity (CSA) which stimulated granulocyte and mononuclear cell formation from hematopoietic progenitor cells in vitro. Removal of Con A from lymphocyte cultures by the competitive inhibitor, α methyl mannoside, resulted in the cessation of CSA release regardless of the time the inhibitor was added to the cultures. However, removal of Con A after 6 hr of incubation had no effect on cellular DNA synthesis and lymphocyte transformation. In addition, vinblastine and cytosine arabinoside completely inhibited DNA synthesis but had no effect on CSA release. CSA release is dependent on an active energy process since it was completely and reversibly inhibited by the inhibitors of protein synthesis puromycin and cycloheximide. These results suggest that CSA release from stimulated lymphocytes requires continuous binding of mitogen and active protein synthesis but is not dependent on DNA synthesis and cell proliferation.
Interaction of Plant Lectins with Purified Human Lymphocyte Populations: Binding Characteristics and Kinetics of Proliferation
Authors: Boldt David H. , MacDermott Richard P. , Jorolan Esther P.
Published: December 2022

With six purified plant lectins as probes of cell surface structure, human T, B, and Null lymphocytes have been examined for cell surface differences. The results of binding experiments with radioiodinated lectins indicate that these three lymphocyte populations, although functionally different, bind equivalent amounts of E- and L-PHA, lentil PHA, Con A, RCA-I, and WGA. Since these lectins are capable of binding to at least four different cell surface oligosaccharides, these data indicate that human T, B, and Null lymphocytes do not differ with respect to surface content of these carbohydrate structures. In addition, all six lectins were investigated for their ability to stimulate DNA synthesis in these human lymphocyte populations. Four of these lectins (E- and L-PHA, lentil PHA, and Con A) were mitogenic for human lymphocytes, whereas RCA-I and WGA failed to stimulate significant DNA synthesis in any lymphocyte population. When incubated with the mitogenic lectins, both B and Null cells displayed a delayed peak of DNA synthesis in comparison to T cells. Thus, despite virtually identical binding characteristics, the lectin-induced mitogenic response exhibited by B and Null cells was markedly different kinetically from that of T cells.
Structural Studies on Induced Antibodies with Defined Idiotypic Specificities
Authors: Capra J. Donald , Tung Amar S. , Nisonoff Alfred
Published: December 2022

Amino acid sequence analysis has been performed on three groups of heavy (H) chains of A/J mice. H chains derived from unimmunized animals were compared to anti-p-azophenylarsonate (anti-Ar) antibodies which were further subdivided into those possessing and those depleted of a cross-reacting idiotype (CRI).

It was found that anti-Ar antibodies bearing the CRI are homogeneous through the first hypervariable region of the H chain. The same sequence was obtained for pooled antibody isolated from the ascites fluid of 18 A/J mice or from a single mouse. The H chains appear to belong to a minor VH subgroup. In the first 30 positions Anti-Ar antibodies depleted of the CRI had the same sequence as those containing the CRI (with small amounts of heterogeneity at some positions), but contained a mixture of sequences in the first hypervariable region of the H chain.

These studies indicate that antibodies with similar specificity and with identical framework sequences, but which differ in their hypervariable regions, contain different idiotypic determinants, and support the concept that the idiotypic determinants reside primarily within hypervariable regions.
Suppressor Cells in Tumor-Bearing Mice Capable of Nonspecific Blocking of in Vitro Immunization Against Transplant Antigens
Authors: Eggers Arnold E. , Wunderlich John R.
Published: December 2022

Spleen cells from mice with progressively-growing methyl-cholanthrene-induced tumors, when immunized in vitro against transplant alloantigens, developed less cytotoxic activity against these antigens as measured by a short-term chromium-release assay than did spleen cells from normal mice. The hyporesponsiveness of spleen cells from the tumor-bearing mice seemed to be due to the presence of suppressor cells which could be removed by nylon-column passage but not by anti-θ treatment and which, in mixture experiments, could inhibit the response of normal spleen cells. The suppression appeared to occur at the sensitization stage and not at the effector stage of the in vitro tests. No evidence was found for mediation of the suppression by soluble factors. These observations emphasize the growing importance of suppressive mechanisms in tumor immune systems.
Induction of Histamine Release and Desensitization in Human Leukocytes
Authors: Petersson Bengt-Åke , Nilsson Ann , Stålenheim Gunnemar
Published: December 2022

Hog anaphylatoxin (AT) in concentrations from 0.5 to 5 µg/ml gives a dose-dependent histamine release from human leukocytes. Concentrations of 100 µg/ml AT give the same high histamine release as 5 µg/ml. This is in contrast to the histamine release obtained with anti-IgE or allergen, which give low histamine release with high doses. The histamine release obtained with AT is completed in 20 sec and the reaction is temperature- and calcium-dependent. Treatment of cells with AT in the presence or absence of calcium makes them insensitive to another challenge with AT. Such treated cells are fully responsive, however, to challenge with anti-IgE if the pretreatment has been performed in the absence of calcium. This, together with the calcium- and temperature-dependence indicates that the AT-induced histamine release is nontoxic. Treatment of cells with AT in the presence of calcium induces, besides histamine release, decrease in sensitivity to anti-IgE, indicating that both AT and anti-IgE release histamine from the same cells. We discuss to what extent AT and cell-bound Ig share intracellular mechanisms for induction of histamine release.
Regulation of the Immune Response: Production of a Soluble Suppressor by Immune Spleen Cells in Vitro
Authors: Thomas David W. , Roberts Walden K. , Talmage David W.
Published: December 2022

The addition of ovalbumin-immune spleen cells (Ova ISC) to sheep erythrocyte-immune spleen cells (SRBC ISC) in Mishell-Dutton-type cultures resulted in a dramatic reduction of PFC to SRBC and was dependent upon the addition of soluble Ova at low concentrations of Ova ISC. The suppressing cells in the Ova ISC were shown to be irradiation sensitive, depleted by anti-θ antiserum and complement treatment, and did not absorb to glass bead columns. Ova ISC-induced inhibition also occurred in culture chambers across a cell-impermeable membrane and a soluble inhibitor was recovered in chambers opposite the Ova ISC. This suppressor factor was sensitive to trypsin treatment and to heating at 80°C, but not to 70°C, for 30 min. The molecular weight, as determined by sucrose gradient analysis, was between 55,000 and 60,000 daltons. This suppressor factor appears to be distinct from a T-cell “helper” factor which was found to be sensitive to heating at 70°C for 30 min. We propose that this suppressor factor participates in the termination of most immunologic responses and is responsible for the antigenic competition phenomenon.
C3 Receptors on Lymphoid Cells: Isolation of Active Membrane Fragments and Solubilization of Receptor Complexes
Authors: Dierich Manfred P. , Reisfeld Ralph A.
Published: December 2022

Complement receptor activity for cell bound C3b and C3b was detected on plasma membrane fragments prepared by nitrogen cavitation from cultured human lymphoid cells. The activity of the membrane fragments reflected the activity of the whole cells in that cells which did not form rosettes (P3J and RPMI 4098) resulted in inactive membranes and cells with high rosette formation (NC37 and Raji) yielded highly active membrane fragments. Two test systems were devised to detect these receptor activities, namely a rosette inhibition and a hemagglutination assay. Solubilization of C3 receptors was accomplished by extraction of active plasma membrane fragments with 2 M KBr. dissociation and reassociation experiments suggest C3b and C3d receptors to be highly complex molecular structures. It appears that these complement receptors on plasma membranes rely on both protein and lipid moieties for the expression of their activity.
Migratory Patterns of B Lymphocytes
Authors: de Kruyff R. H. , Gilmour D. G. , Thorbecke G. J.
Published: December 2022

Homing of 3H-adenosine-labeled bursa cells to follicles in the chicken spleen was greatly reduced by prior incubation of the transferred cells with rabbit anti-chicken-immunoglobulin sera. This effect was transient, being present 6 hr after cell transfer but gone by 18 hr, when incubation was at 0°C without complement. The inhibition persisted until at least 18 hr after incubation at 37°C with complement.
Cell Interactions Between Histoincompatible T and B Lymphocytes
Authors: Dorf Martin E. , Katz David H. , Graves Mary , DiMuzio Henry , Benacerraf Baruj
Published: December 2022

The genetic restrictions for successful carrier-primed T cell and hapten-primed B cell cooperative interactions were established in a totally in vivo double adoptive transfer system. In complementation experiments, we have demonstrated that genes localized in the K-end of the H-2 complex control successful T-B cell cooperative interactions. Thus, DBA/2 KLH-primed T cells cooperated with (C3H × C3H.OH)F1 and (C3H × C3H.OL)F1 DNP-primed B cells to make a secondary response to DNP-KLH, but not with C3H or (C3H × C3H.A)F1 DNP-primed cells. The presence of foreign histocompatibility determinants on the F1 B cells did not prevent successful physiologic cell cooperation provided the T and B cells shared common cell interaction genes coded for in the K-end of the H-2 complex.
Conformation-Dependent Antigenic Determinants in the Toxic Lectin Ricin
Authors: Olsnes Sjur , Saltvedt Erling
Published: December 2022

The major part of the ricin-precipitable antibodies in sera produced by immunizing rabbits with formaldehyde-treated ricin is precipitated also by the isolated ricin A and B chains. In contrast, in antisera produced by immunizing with formaldehyde-treated ricinus agglutinin only a small part of the antibodies cross-reacting with ricin can be precipitated by the isolated A and B chains, or bound to immunoabsorbents containing the isolated ricin chains. In immunodiffusion studies with anti-ricinus agglutinin sera, a star-shaped precipitate was formed when isolated A and B chains recombined to form intact ricin.

Both anti-ricin and anti-ricinus agglutinin sera neutralized effectively the ability of ricin to inhibit protein synthesis in HeLa cells. Anti-ricin serum also neutralized the inhibitory effect of the isolated A chain on protein synthesis in a cell-free system and the ability of the isolated B chain to induce indirect hemagglutination. In contrast, anti-ricinus agglutinin serum did not neutralize the biologic activities of the isolated ricin A and B chains.

Anti-ricinus agglutinin serum formed a precipitate with the hybrid ricin A chain/abrin B chain, and protected against the toxic effect on HeLa cells of this hybrid, indicating conformational changes of ricin A chain upon binding to the B chain.

It is concluded that the anti-ricinus agglutinin serum contains antibodies directed against conformational determinants present on intact ricin, but not present or exposed in the isolated A and B chains. At least part of these conformational determinants appears to be carried by the A chain.
Immunoglobulin Biosynthesis in Myeloma-Associated and Casein- and Endotoxin-Induced Murine Amyloidosis
Authors: Baumal Reuben , Ackermann Alex , Wilson Bill
Published: December 2022

Amyloidosis was induced in C57BL mice by daily injections of casein and in BALB/c mice by daily injections of endotoxin. There was no obvious disorder of immunoglobulin biosynthesis by spleen lymphocytes in these mice either before, during the development of, or in the amyloidotic stage. The pattern of immunoglobulin synthesis, assembly, and secretion was unaltered, the relative amount of heavy and light chains produced was normal, and there was an absence of immunoglobulin polypeptide chain fragments. Small amounts of amyloid were present in only 1 of 19 BALB/c and C3H mice (the IgG2a producing MOPC 173 tumor) bearing immunoglobulin-producing myeloma tumors and variants of these tumors. There was no relationship between excess light chain production by tumor plasma cells or spleen lymphocytes and the development of amyloidosis and there were no light chain fragments demonstrable. Antiserum prepared against casein-induced amyloid cross-reacted by immunofluorescence with the amyloid present in the MOPC 173 tumor-bearing mice, indicating the presence of common antigenic determinants in these two forms of amyloid. Attempts to study the biosynthesis of amyloid with incorporation of radioactively labeled amino acids were unsuccessful.
The Immune Response of Mice Treated with Anti-µ Antibodies: The Effect on Antibody-Forming Cells, Their Precursors and Helper Cells Assayed in Vitro
Authors: Gordon Julius , Murgita Robert A. , Tomasi Thomas B.
Published: December 2022

Previous studies have shown that mice treated from birth with heterologous anti-µ antiserum are severely immuno-suppressed with respect to numbers of splenic plaque-forming cells (PFC) to sheep red blood cells (SRBC) in all immunoglobulin classes. In this study we have investigated, using in vitro techniques, the cellular site of the deficit created by anti-µ. The primary PFC response of spleen cells originating from anti-µ-treated mice was completely suppressed in vitro. The response was restored by the addition to the cultures of B cells but not T cells. T cells were derived from normal spleens which had been depleted of B lymphocytes and adherent cells by filtration through cotton wool columns, or by educated thymus cells obtained from the spleens of lethally irradiated mice injected with syngeneic thymocytes and SRBC.

Restoration of the PFC response of spleen cells from anti-µ-treated mice by normal B cells suggested that a cellular deficiency rather than an activity process by inhibitory cells was the cause of the imunosuppression. Also, co-culturing spleen cells from normal and suppressed mice did not reveal the presence of inhibitory cells. Spleen cells from x-irradiated mice, injected with bone marrow from anti-µ-suppressed mice, gave rise to PFC cells when cultured with SRBC and normal T cells, suggesting that stem cells giving rise to B cells were not affected by anti-µ treatment. Similarly, educated thymus cells derived from suppressed mice could provide helper function when reconstituted in vitro with normal B cells. Exposure of normal bone marrow and spleen cells to anti-µ serum prior to passage through syngeneic x-irradiated recipients demonstrated that spleen cells were much more sensitive than were bone marrow cells to suppression by anti-µ antibodies. It is concluded that the target of anti-µ antibody is a µ-chain bearing B cell precursor to the IgM-, IgG-, and IgA-producing cell.
The Isolation and Partial Characterization of Neutrophil Chemotactic Factors from Escherichia Coli
Authors: Schiffmann Elliott , Showell Henry V. , Corcoran Barbara A. , Ward Peter A. , Smith Eric , Becker Elmer L.
Published: December 2022

Heat-stable, chemotactically active peptides have been obtained from Escherichia coli culture filtrates. They range in size between 150 and 1500 daltons and are anionic at neutral pH. Free carboxyl groups but not free amino groups appear to be required for activity. The N-terminal group may be blocked. There do not appear to be internal aromatic or basic residues in the chemotactically active fractions. A highly purified, not completely characterized, fraction was found to contain aspartic acid, serine, glutamic acid, alanine, and glycine.
A BALB/c Mouse IgA Myeloma Protein That Binds Salmonella Flagellar Protein
Authors: Mason Smith A. , Potter Michael
Published: December 2022

A BALB/c IgA mouse myeloma protein in ascites (MOPC-467) had been found to bind an antigen in heat extracts from Salmonella, Pasteurella, and Herellea cultures. MOPC-467 ascites was shown to immobilize motile S. adelaide, agglutinate formalin fixed S. milwaukee, and purified IgA (M467) was capable of precipitating column-purified flagella and flagellin from S. milwaukee.