bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparative Effects in Mice of Purified Bacterial Cell-Wall Components on Granulocyte-Macrophage Progenitor Cells and Their Regulators
Authors: Staber Fritz G.
Published: December 2022

The effects were investigated in mice of the injection of chemically pure preparations of three structurally unrelated cell-wall components from Gram-negative bacteria (BCWC), lipid A, outer-membrane lipoprotein, and murein on humoral and cellular parameters of granulocyte-macrophage production. Analysis of granulocyte-macrophage progenitor cells (GM-CFC) from normal C57BL/6 mice by velocity sedimentation showed that these cells sedimented as a relatively uniform population with a peak sedimentation velocity between 4.2 and 4.5 mm/hr. Pretreatment of mice with murein caused no significant alteration of this profile. However, GM-CFC from mice injected with either lipid A or lipoprotein sedimented more rapidly than normal. The whole profile was broadened and more than one peak was observed. Sera from mice injected with any of the three components stimulated the growth of GM-CFC in vitro, and at high serum concentrations, the absolute number of colonies and the proportion of pure granulocytic, pure macrophage, and mixed colonies were very similar. However, serial dilution of the sera revealed a significant qualitative difference between post-lipid A serum (PLAS) and post-lipoprotein serum (PLPS) on the one hand and post-murein serum (PMS) on the other hand. Thus, with serial dilution the number of colonies induced decreased more rapidly in the case of PMS than in the case of PLAS or PLPS. At dilutions insufficient to induce any colony growth by themselves both PLAS and PLPS potentiated the growth of GM-CFC stimulated by semi-purified mouse lung-conditioned medium (MLCM), whereas PMS, like normal mouse serum (NMS), had no significant effect.
Enhanced Uptake of 3H-Glucosamine by Macrophages Stimulated by Macrophage-Activating Factor
Authors: Sober Arthur J. , Haynie Marilynn , David John R.
Published: December 2022

The enhanced uptake and incorporation of 3H-glucosamine by guinea pig peritoneal exudate macrophages stimulated by macrophage-activating factor (MAF) were studied. MAF was produced by concanavalin A- (Con A) stimulated lymphocytes. Two systems were developed to assay this phenomenon, one using microtiter plates, the other using suspension cultures. Enhanced uptake of 3H-glucosamine by macrophages was found after 48-hr incubation in microtiter plates and after 24 hr in suspension cultures. Increased uptake of 14C-glucose and 3H-galactose was also observed under these conditions. The enhanced uptake of 3H-glucosamine was markedly reduced or abolished by the addition of glucose to the cell cultures or by replacement of the medium with fresh medium before pulsing with labeled sugar. These observations indicate that the mechanism of enhanced uptake reflects in part increased glucose utilization by the stimulated macrophages.
Inhibition of Antibody Production in Plasmacytoma Cells
Authors: Abbas Abul K.
Published: December 2022

Previous studies have demonstrated that highly multivalent DNP conjugates of γ-globulins, but not of KLH or BSA, inhibit the production of anti-DNP IgA antibody by the mouse plasmacytoma line, MOPC 315, in vitro. The present experiments were designed to compare the fate of various DNP antigens, trace-labeled with 125I, in MOPC 315 cells. After continuous exposure of MOPC 315 cells to various antigens for up to 20 hr, suppressive DNP-globulin conjugates are extensively internalized by the cells whereas nonsuppressive DNP-KLH and DNP-BSA remain largely bound to the surface. These differences are not attributable to cooperative binding of DNP-globulins to Fc receptors, and, at least with DNP-KLH, are not due to major differences in the molecular charge of the antigens. Moreover, all internalized antigens persist within cells and are not extensively hydrolyzed for up to 18 hr of antigen-free culture. Electron microscopic studies show that internalized antigens are not enclosed within pinocytotic vesicles, and are often found close to the endoplasmic reticulum or Golgi complex. It is proposed that prolonged intracellular persistence of antigens may be a central mechanism in the inactivation of antibody-secreting cells as well as in B lymphocyte tolerance.
Genetic Control of the Cytotoxic T Cell Response to SV40 Tumor-Associated Specific Antigen
Authors: Knowles Barbara B. , Koncar Mirjana , Pfizenmaier Klaus , Solter Davor , Aden David P. , Trinchieri Giorgio
Published: December 2022

The H-2 gene complex controls the ability of mice to generate cytotoxic T cells to the SV40 tumor-associated specific antigen (TASA). Mice of six H-2 haplotypes (H-2b, d, f, k, q, and s) were challenged in vivo with syngeneic simian virus 40 (SV40)-transformed cells. By testing splenic lymphocytes in a 51Cr cytotoxic test, only H-2b mice were able to generate cytotoxic T cells specific for syngeneic SV40-transformed target cells. In contrast, immune lymphocytes from both H-2b and H-2k mice assayed after secondary in vitro restimulation lysed SV40-transformed target cells of appropriate H-2 genotype. Popliteal lymph node cells from H-2b, k, and d mice, immunized by footpad injection of SV40 and then incubated for 14 days in vitro before assay, are cytotoxic for syngeneic SV40-transformed cells. With this immunization protocol, H-2b mice respond to SV40 TASA in association with both the H-2 K and D locus-coded molecules, whereas H-2k mice respond to SV40 TASA in association with the H-2 K molecule only and H-2d mice respond to SV40 TASA in association with the H-2 D gene product. SV40 TASA-immune lymphocytes from various F1 (responder × nonresponder) mice were also unable to lyse syngeneic H-2Kd or H-2Dk SV40-transformed target cells. Inhibition of the response to SV40 TASA in association with H-2Dd is found when lymphocytes from SV40 TASA-immune F1 hybrid (H-2b × H-2d) or H-2 congenic recombinant (H-2KbDd) mice are analyzed. A dominant suppression or recessive helper mechanism is hypothesized to account for these results.
Enhancing Role of IgG in Lysis of Rabbit Erythrocytes by the Alternative Pathway of Human Complement
Authors: Nelson Ben , Ruddy Shaun
Published: December 2022

Adsorption of human serum with rabbit erythrocytes (E) or stromata at 0°C removes about 30% of its alternative pathway activity as measured by lysis of rabbit E in MgEGTA buffer. The activity is restored by adding back a low pH eluate from the stromata. Hypogammaglobulinemic sera have much lower levels of eluate activity than do normals. The activity of the eluate is totally abrogated by treating it with anti-IgG immunoabsorbant or Staphylococcal protein A. Reciprocal experiments comparing eluates from rabbit E stromata or zymosan revealed that eluate from zymosan had no restorative effect on the lysis of rabbit E by stromata-adsorbed serum and that the eluate from the stromata effected no increase in C3 consumption when zymosan-adsorbed serum was exposed to zymosan at 37°C. Conversely, each of the eluates enhanced alternative pathway activity when tested with their respective adsorbed sera. That this activity is not essential for alternative pathway activation was demonstrated by the ability of hypogammaglobulinemic sera to lyse rabbit E despite depletion of IgG to levels of less than 80 ng per ml as detected by radioimmunoassay. The results of these experiments suggest that specific IgG antibodies enhance, but are not essential for, lysis of rabbit E by the alternative pathway of human complement.
Humoral and Formed Elements of Blood Modulate the Response of Peripheral Blood Monocytes
Authors: Musson Robert A. , Henson Peter M.
Published: December 2022

Human and rabbit peripheral blood monocytes normally adhere to plastic tissue culture plates in vitro when they are suspended in Hanks' media. Increasing amounts of autologous serum or heat-inactivated plasma in the cell suspensions prevented the adherence of both monocytes and lymphocytes. The inhibitory effect of plasma was separated into three areas of activity by chromatography on Sephacryl S-200. The profile of inhibitory activity did not coincide with the protein elution profile, suggesting that inhibition was not a nonspecific protein effect. A layer of adherent platelets overcame the inhibitory effect of plasma on monocyte adherence. Platelets selectively increased monocyte as opposed to lymphocyte adherence and this was specific for platelets in that neither neutrophils nor fibroblasts could substitute for platelets. Both plasma and platelets acted directly on monocytes.
Genetic and Cellular Requirements for Primary Antibody Immune Response to Thy-1 Antigens in Mice
Authors: Zaleski Marek B. , Gorzynski Timothy
Published: December 2022

An effective anti-Thy-1 response requires at least three conditions to be simultaneously fulfilled—1) The responder has to share an H-2 haplotype, or at least K and D alleles, with the donor of the Thy-1 disparate thymocytes, 2) The responder has to differ from the donor by some non-H-2 cell-surface antigens, and 3) The responder has to possess functionally capable T cells. All these requirements are consistent with the hypothesis that Thy-1 antigens and the non-H-2 antigen(s), the latter serving as carrier, form a complex that is viewed as a variant of self and then recognized in the context of the H-2 molecules by the responder's T cells. An attractive feature of the proposed hypothesis is that it could be applicable to a variety of cell-surface and also exogenous antigens.
Production of Human Immunoglobulin E Antibody In Vitro
Authors: Tjio Anthony H. , Hull William M. , Gleich Gerald J.
Published: December 2022

The discovery of immunoglobulin E (IgE) (1) has contributed significantly to the understanding of hypersensitivity reactions mediated by reaginic antibody (2) and has led to the development of new diagnostic procedures in the field of allergology (3, 4). Recent studies have demonstrated the production of IgE protein by peripheral blood lymphocytes in vitro and have examined the kinetics and pharmacologic control of IgE biosynthesis (5, 6). The purpose of this communication is to describe our observation of in vitro production of IgE protein and antigen-specific IgE antibodies by peripheral blood mononuclear cells obtained from atopic and nonatopic individuals.

A modification of the lymphocyte culture method of Waldmann et al. (7) was used in our studies. Blood, 60 ml, was obtained from 11 patients, eight of whom are atopic (geometric mean serum IgE, 247 ng/ml; range 61 to 1079) and three of whom are nonatopic (geometric mean serum IgE, 96 ng/ml; range 73 to 143).
The Effect of Mitogens on the Expression of Epstein-Barr Virus Antigens in Human Lymphoid Cell Lines
Authors: Tovey Michael G. , Lenoir Gilbert , Begon-Lours Jacqueline , Tapiero Haim , Rochette-Egly Cécile
Published: December 2022

Treatment of human lymphoblastoid cells with either phytohemagglutinin (PHA), concanavalin A, Staphylococcus protein A, or polyinosinic acid-polycytidylic acid, in combination with 5-iodo-2′ deoxyuridine (IUdR) markedly increased the expression of Epstein-Barr virus (EBV) early antigen (EA) relative to IUdR alone. Such treatment did not, however, modify the production of virus capsid antigen in any of the lymphoid cell lines tested. The effect of PHA on EA induction in Raji cells was not accompanied by changes in the incorporation of labeled precursors into cellular DNA, or in the intracellular concentration of either adenosine 3′5′ cyclic monophosphate or guanosine 3′5′ cyclic monophosphate. However, those mitogens that stimulated EA expression in Raji cells also increased the fluorescence polarization of 1,6 diphenyl 1,3,5-hexatriene-labeled Raji cells. The possible role of cell surface changes in the mitogen activation of latent EBV in human lymphoblastoid cells is discussed.
Thymic Control of Secretory Antibody Responses in the Rat
Authors: Ebersole Jeffrey L. , Taubman Martin A. , Smith Daniel J.
Published: December 2022

The effect of neonatal thymectomy on salivary and serum antibody responses was studied in rats. Local immunization of thymectomized rats with a T-dependent antigen (DNPBGG) elicited negligible amounts of IgA anti-DNP antibody in saliva. In contrast, both normal and sham-thymectomized animals demonstrated substantial levels of salivary IgA antibody. All thymectomized rats locally injected with a T-independent antigen (DNP-Lys-Ficoll) exhibited salivary IgA antibody production. Salivary IgG antibodies were somewhat decreased in thymectomized rats injected with either antigen; however, the final effect of T cell deprivation on IgG synthesis was not as pronounced as on IgA synthesis. Serum IgA antibody was induced in control rats injected with DNPBGG, whereas this Ig class of antibody was absent in thymectomized rats. The results suggest that thymus-derived cells exert a regulatory influence on both serum and secretory IgA responses to antigens.
IgE Antibody Formation and Aging
Authors: Fujiwara Misa , Kishimoto Susumu
Published: December 2022

The IgE, IgG, and IgM antibody responses to DNP-As were assessed in young adult and aged C57BL/6J mice. The primary IgE anti-DNP antibody response was significantly reduced in aged mice; however, the secondary response of 54-week-old mice was comparable to that of young adult mice, although the decline of the antibody level was more rapid in aged mice compared to young adult mice. Multiple booster injections could not restore the depressed immune response of 66-week-old mice to the level of the immune response of young adult mice in terms of IgE antibody responses to DNP-As. The IgE anti-carrier response was considerably lower than the IgE anti-hapten response, and more transient and significantly lower in aged mice than in young adult mice. The avidity of IgE antibody for DNP-determinant was invariably higher in young adult mice than in aged mice. Booster injections increased the avidity of IgE antibody in both groups of mice. Moreover, the hemagglutinating antibody response was also impaired in aged mice, which mounted minute or no amounts of IgG antibody. Upon transfer of the thymocytes from young syngeneic donor mice, the aged mice could restore substantially the IgE and hemagglutinating antibody responses to DNP-As. It became clear in the study of individual mice that the variability of the IgE anti-DNP antibody response increased with advancing age, but the variability within aged mice decreased and deviation from young adult mice was less after the secondary immunization.
Lack of Correlation between Neutralizing Antibody Production and Suppression of Coxsackievirus B-3 Replication in Target Organs: Evidence for Involvement of Mononuclear Inflammatory Cells in Host Defense
Authors: Woodruff Jack F.
Published: December 2022

CD-1 mice inoculated with coxsackievirus B-3 i.p. developed a generalized infection involving the heart, pancreas, and liver. The disease was nonlethal and viral growth in the target organs was terminated in about a week. Administration of cortisone acetate 30 min to 2 hr before infection markedly enhanced the severity of disease. Abnormally high titers of virus were found in the target organs between days 3 and 7 with persistence of infectious virus in the heart for at least 2 weeks. In addition the extent of necrosis of myofibers, pancreatic acini, and hepatic parenchyma was increased and a high percentage of the animals died. There was no evidence that the anti-viral antibody response was impaired in steroid-treated mice since concentrations of neutralizing antibody in the circulation were normal. In contrast, immigration of mononuclear inflammatory cells into the hearts of these animals was depressed and when present, foci of inflammation contained some polymorphonuclear leukocytes. The data indicate that inhibition of coxsackieviral growth cannot be attributed to the sole effects of neutralizing antibody and suggest that mononuclear inflammatory cells infiltrating the heart play a role in primary host defense.
Analysis of the Diversity of Murine Antibodies to Dextran B1355: N-Terminal Amino Acid Sequences of Heavy Chains from Serum Antibody
Authors: Schilling J. , Hansburg D. , Davie J. M. , Hood L.
Published: December 2022

The N-terminal amino acid sequences of two γ and two µ chains from normally induced serum antibodies to dextran in BALB/c mice are presented. These heavy chains are derived from antibodies with three distinguishable idiotypes. These variable region (VH) sequences are all identical as far as they have been analyzed (27 to 53 residues). The light chains from these antibodies are all of the λ type and are identical by isoelectric focusing analysis. Accordingly, the diversity of dextran antibodies appears to reside primarily in the heavy chains. The implications of these observations for antibody diversity are discussed.
A Serologic Comparison of Moloney Lymphoma Cell Surface and Moloney Oncornavirus Antigens
Authors: Humphrey David , Tsukamoto-Adey Ann , Witte Owen M. , Fox Robert , Jerabek Libuse , Weissman Irving L.
Published: December 2022

Moloney lymphomas and Moloney sarcomas share strong tumor antigens. In this report we analyze the cell-surface antigens on a BALB/c Moloney lymphoma, LSTRA, using hyperimmune sarcoma regressor sera (αMo) as a primary reagent. We also use heterologous anti-viral p30 and gp70 sera for a direct analysis of virion protein antigens on the LSTRA surface. Using radiolabeled αMo-binding assays, we demonstrate that LSTRA tumor antigens detected by these sera are all Moloney viral antigens; approximately ⅓ of these antigenic determinants are expressed on the intact virus, and the other determinants are revealed by detergent lysis of the virus. The major viral antigens expressed on the LSTRA cell surface are viral env gene products, whereas gag gene products are only sparsely represented. We conclude that αMo sera detect almost exclusively viral antigens on LSTRA cells, and these antigens are almost exclusively virion env gene products.
Requirement of Histocompatible Macrophages for the Induction of a Secondary Cytotoxic Response to Syngeneic Tumor Cells in vitro
Authors: Taniyama Tadayoshi , Holden Howard T.
Published: December 2022

The role of macrophages in the induction of secondary cytotoxic responses to syngeneic tumors was evaluated. When spleen cells from MSV-immune mice were extensively depleted of macrophages, they failed to mount an in vitro secondary cytotoxic response to syngeneic tumors. The response could be reconstituted by the addition of nonimmune macrophages, with syngeneic macrophages being much more efficient than allogeneic macrophages. The relative inefficiency of the allogeneic macrophages to restore the response was shown not to be due to: 1) induction of suppression by allogeneic cells, since in mixing experiments they did not inhibit the response; 2) differences in the kinetics of the response when allogeneic macrophages were employed; 3) requirements for different concentrations of syngeneic or allogeneic macrophages for this accessory function. Neither heat-killed nor frozen and thawed macrophages were capable of restoring the response. However, macrophages could still act as accessory cells if DNA synthesis was prevented. Furthermore, identity between macrophages and T lymphocytes at the I-A subregion of the MHC appeared to be necessary but perhaps not sufficient for an optimal response, and at this time it is not clear whether compatibility at other non-H-2 loci may also be required. The results of the present study suggest that macrophages serve an accessory function in the generation of a secondary cytotoxic response to tumor-associated antigens even when intact tumor cells are used as the stimulators.
Specific Inhibition of the Antibody-Mediated Activation of a Defective β-D-Galactosidase by Circulating Activating Epitope-Binding Molecules
Authors: de Macario Everly Conway , Macario Alberto J. L.
Published: December 2022

Activation of a defective Escherichia coli β-D-galactosidase by specific activating antibody is inhibited competitively by a molecule with immunoglobulin properties but devoid of activating capacity. This molecule is found in the serum of nonimmunized rabbits and is no longer detectable after β-D-galactosidase administration, but can be demonstrated in rabbits injected with antigens other than the enzyme. The data show that the inhibitory molecule recognizes and interacts specifically with the activating epitope of the activatable enzyme and that, although unable to activate the latter, it competes with the activating antibody and inhibits activation.
IgE Responses to Synthetic Polypeptide Antigens
Authors: Dessein Alain , Germain Ronald N. , Dorf Martin E. , Benacerraf Baruj
Published: December 2022

The serum IgE response to the synthetic polypeptide antigen L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT) was investigated. The present study details the experimental conditions required to elicit optimal IgE anti-GAT antibody titers, the genetics controlling this response, and the ability of anti-suppressor cell treatments to affect the magnitude and duration of IgE anti-GAT levels in responder and nonresponder strains of mice. Significant IgE anti-GAT titers were obtained in BALB/c (IgG responder) mice after administration of 1 to 100 µg of GAT in adjuvant (Maalox or Maalox-pertussis), with optimal secondary IgE titers after challenge with 1 or 10 µg doses. All IgE responses were transient in comparison to IgG antibody levels. IgE anti-GAT immunity was found to be regulated by a gene(s) in the I-A/I-B subregion of the H-2 complex, in agreement with earlier studies on serum IgG or cell-mediated immunity. Mice bearing the H-2ja, p, q, s haplotypes were nonresponders, and treatment of SJL (H-2s) or DBA/1 (H-2q) mice with cyclophosphamide or low dose irradiation to remove suppressor cells failed to permit IgE anti-GAT responses, while markedly augmenting IgE anti-TNP responses in the same mice, and IgE anti-GAT responses in BALB/c mice. However, the presence of functional GAT-specific IgE B cells in nonresponder strains was established by demonstrating IgE anti-GAT responses to GAT complexed with methylated serum albumin in these mice. The implications of these data with respect to H-2 linked Ir gene function and class-specific regulation of antibody responses are discussed.
Cell-Mediated Lympholysis of Fluorescein Isothiocyanate-Conjugated Autologous Human Cells: Evidence for Hapten-Specific Altered-Self Reactive Human Cytotoxic T Lymphocytes
Authors: Friedman Steven M. , Irigoyen Oscar , Kuhns Janet , Chess Leonard
Published: December 2022

The combined stimulus of FITCI-conjugated autologous cells plus either allogeneic cells, soluble antigens, or T cell-derived helper factor(s) triggers the in vitro differentiation of FITCI altered-self-reactive human CTL. Fractionation of responder cells into highly purified lymphocyte subpopulations and complement-mediated lysis of FITCI altered-self-reactive killer cells with rabbit anti-human T cell antibodies demonstrated that both the precursor and effector cell responsible for FITCI altered-self-cytolysis are T lymphocytes. Killer cell specificity was investigated in three experimental protocols: 1) the CTL generated were found to efficiently lyse FITCI but not TNP-modified autologous targets, whereas in reciprocal experiments, TNP altered-self-reactive CTL lyse TNP but not FITCI-derivitized autologous cells; 2) T cell-mediated lysis of 51Cr-labeled FITCI-derivitized autologous targets was specifically inhibited by nonradiolabeled FITCI but not TNP-modified autologous cells; and 3) FITCI altered self-reactive CTL specifically lyse some but not other FITCI-modified allogeneic targets. Taken together, these results show that although the FITCI altered-self-reactive human CTL generated are hapten specific, they appear to recognize more than hapten alone on the target cell surface. Moreover, the observation that soluble antigens, allogeneic cells, and T cell helper factor(s) can facilitate hapten-specific CTL responses to both TNP and FITCI-conjugated autologous cells suggests that these helper stimuli act by preferentially triggering the differentiation of CTL precursors that have specifically interacted with chemically modified autologous stimulators.
Ontogeny of the Ferret Humoral Immune Response
Authors: Suffin Stephen C. , Prince Gregory A. , Muck Katie B. , Porter David D.
Published: December 2022

Infant ferrets are born with nearly undetectable immunoglobulin levels, but by 9 days of age the infant ferret serum contains 77, 29, and 13% of adult mean serum levels of IgG, IgA, and IgM. Transmucosal uptake of IgG by the infant ferret occurred for the first 30 days of life. The specific anti-respiratory syncytial virus neutralizing titer of whole milk was 5.5 times higher than maternal serum despite a lower concentration of immunoglobulins in the milk.
Aging Increases Expression of LPS-Induced Autoantibody-Secreting B Cells
Authors: Meredith Patricia J. , Kristie James A. , Walford Roy L.
Published: December 2022

Escherichia coli lipopolysaccharide (LPS), a polyclonal B cell activator, has been employed to achieve in vitro stimulation of autoantibody-secreting B cells in young adult and aged mice of long-lived strains as assayed in a hemolytic plaque technique to syngeneic mouse erythrocytes. Aged 21- to 24-month-old C57BL/6J and (C57BL/10Sn × C3H/HeDiSn)F1 mice were found to express 3 to 4 times as many LPS-induced plaque-forming cells (PFC) to autologous erythrocytes than did younger 6-month-old animals. With the use of cyclophosphamide (CY), a significant enhancement of auto-PFC production in young mice occurred, approaching levels found in non-CY-treated old mice. Thus, autoreactive clones of lymphocytes exist in the spleens of young adult mice, but under normal circumstances produce little autoantibody. The situation in aged members of these strains, therefore, does not seem to involve an actual increase in numbers of autoreactive B cells, but may possibly involve some form of deregulation, permitting increased age-related expression of autoreactive lymphocyte clones.
Characterization of Anti-Forssman (ANTI-Fs) Antibodies in Human Sera: Their Specificity and Possible Changes in Patients with Cancer
Authors: Young, William W. , Hakomori Sen-Itiroh , Levine Philip
Published: December 2022

The majority of normal human sera hemolyze sheep erythrocytes in the presence of complement (“hemolyzers”), whereas sera of the remaining population do not lyze sheep erythrocytes (“nonhemolyzers”). The antibodies responsible for this lytic activity are now identified as being specific to Forssman (Fs) glycolipid, predominantly of the IgM class, and only weakly cross-reactive with blood group A glycolipid. These conclusions are based on the following observations: 1) “Hemolyzer” sera produced complement-dependent lysis of liposomes containing Fs glycolipid, whereas “nonhemolyzer” sera reacted very weakly with the same Fs liposomes; the level of hemolytic activity generally paralleled the reactivity of the sera with Fs liposomes. 2) Human “anti-Fs” did not cross-react with liposomes containing either globoside, the precursor to Fs, or blood group A glycolipid. 3) The complement-dependent hemolysis of sheep erythrocytes by hemolyzer sera was specifically inhibited by Fs liposomes but not by either A- or globoside liposomes. 4) In five of six cases tested, hemolysin activity was present in the IgM fraction but not the IgG fraction, whereas in the remaining case the activity was present in the IgG fraction.

We have previously described the presence of Fs glycolipid in the normal gastrointestinal mucosa of a minor percentage of Taiwanese cancer patients, whereas in the majority of cases Fs was absent in the normal mucosa but present in the tumors derived therefrom. The sera of cancer patients displayed a decreased anti-Fs reactivity as compared with sera of a control group suggesting that Fs-positive tumor tissue may affect serum anti-Fs levels.
Limited Trypsin Cleavage Distinguishes MHC and Thymus-Leukemia Antigens
Authors: Pischel Ken D. , Little J. Russell
Published: December 2022

A simple technique is presented for the identification of particular cell membrane antigens. The method employs labeled membrane antigens that are isolated immunospecifically and subjected to limited trypsin digestion followed by polyacrylamide gel electrophoresis in detergent. A large “core” peptide is produced by proteolysis of murine thymus-leukemia antigens (TLA) and from antigens of the major histocompatibility complex (MHC). The tryptic cores from H-2K and H-2D are regularly distinguishable from the thymus-leukemia antigens (TLA) by gel electrophoresis in one dimension. This chemical distinction is particularly important in the analysis of antigen mixtures isolated with antisera specific for β2 microglobulin. These techniques have been used to identify thymus-restricted β2 microglobulin-associated antigens on cell membranes from mouse, man, guinea pig, and monkey. In appropriate inbred mouse strains, these are the TLA and it is proposed that in the three other species examined they may be analogues, although not necessarily homologues, of TLA. The broad species distribution of these thymus-restricted cell membrane antigens suggests that they are involved in the differentiation of thymus-dependent lymphocytes (T cells).
Lysosomal Enzymes in Inflammatory Synovial Effusions
Authors: Hadler Nortin M. , Spitznagel John K. , Quinet Robert J.
Published: December 2022

The concentrations of several polymorphonuclear neutrophilic lysosomal constitutents were quantitated by immunochemical and enzymatic assays in 28 inflammatory and 9 noninflammatory synovial fluids. The quantities of lactoferrin, myeloperoxidase, and enzymatically determined lysozyme were covariate with the neutrophil count. Enzymatic activities measured with synthetic substrates developed for the assay of chymotryptic-like cationic protein (cathepsin G) and elastase, along with immunochemically determined lysozyme, were independent of the neutrophil count. Although the latter assays were developed and standardized with human neutrophilic lysosomal constitutents, they measure different activities in inflammatory synovial effusions. No elastase was detected if elastin was used as the substrate.

Regardless of the source of the enzymes, there was a negative correlation between their concentration and the degree of radiographic destruction of the joint from which the fluid was obtained. Lysosomal enzymes in solution in synovial fluid are not likely to be primarily involved in cartilage destruction.
Histamine-Releasing Activity (HRA)
Authors: Thueson David O. , Speck Linda S. , Lett-Brown Michael A. , Grant J. Andrew
Published: December 2022

Supernatants from 1- to 2-day cultures of human mononuclear cells induced the release of histamine from basophils. Generation of this histamine-releasing activity (HRA) was stimulated by addition of concanavalin A to the cell cultures. Mononuclear cells were also cultured with SKSD and Candida albicans antigens. Stimulation of HRA production by these antigens was correlated with positive delayed skin reactions. Serial dilutions of supernatants assayed for HRA provided a semiquantitative determination of the level of HRA in mitogen- or antigen-stimulated samples. Antigen increased HRA production when added during the first or second day of culture. Generation of HRA probably requires active protein synthesis, since puromycin was inhibitory, and since preformed HRA could not be recovered from lysed cells. HRA was detected in supernatants after 4 hr, and the effects of antigen stimulation were apparent after 8 hr of culture. The rate of HRA synthesis diminished after 18 hr of culture. Replacement of supernatants with fresh culture medium allowed continued synthesis of substantial quantities of HRA during the second day of culture. A linear correlation was observed between the amount of HRA produced and the mononuclear cell concentration. Our findings provide evidence for the interaction of lymphocytes and basophils via a soluble mediator.
Histamine-Releasing Activity (HRA)
Authors: Thueson David O. , Speck Linda S. , Lett-Brown Michael A. , Grant J. Andrew
Published: December 2022

We have recently reported the production of a histamine-releasing activity (HRA) by 1- to 2-day cultures of human mononuclear cells. Synthesis of HRA was augmented by addition of the mitogen concanavalin A or by antigens that commonly elicit cell-mediated immune responses. Since we have previously shown that histamine was released from basophils by activated complement, some of the properties of HRA and C5a were compared. No correlation between the percentage of histamine released from normal basophils by HRA or C5a was found. Anti-C5 abolished C5a-induced release, but did not inhibit the release of histamine from basophils by HRA. HRA did not resemble C3a: HRA released a higher percentage of histamine than C3a and HRA was not inhibited by anti-C3a. Pretreatment of basophils with HRA in the absence of calcium, followed by challenge with C5a, caused an augmented release of histamine by C5a. Basophils desensitized to C5a and anti-IgE released normally to HRA challenge. Basophils pretreated with HRA released normally to rechallenge with HRA.

Release of histamine from basophils by HRA was calcium-dependent and magnesium-independent. The maximal percentage of intracellular histamine was released within a few minutes after exposure to HRA. Optimal release occurred at 37°C.

HRA had an apparent m.w. of 10,000 to 15,000 daltons as determined by gel exclusion chromatography. HRA was relatively resistant to the effects of elevated temperature but was destroyed by trypsin and α-chymotrypsin. The effect of trypsin was prevented by soybean trypsin inhibitor. Nucleases and neuraminidase had no effect on HRA.
Effects of C1̄ on the Size of Soluble Immune Aggregates and on Their Processing by Macrophages
Authors: Kijlstra Aize , van Es Leendert A. , Daha Mohamed R.
Published: December 2022

The effect of isolated C1̄, the first component of complement (C), on the processing of soluble IgG aggregates by mononuclear phagocytes was studied in vitro with a homologous guinea pig system and isolated stable IgG2 aggregates. The interaction of C1̄ with IgG2 aggregates was demonstrated by the inhibition of functional C1̄ hemolytic activity and was shown to be dependent upon the number of IgG2 molecules per aggregate.

Analysis of the interaction of C1̄ with the soluble aggregates by ultracentrifugation on sucrose gradients revealed dissociation of aggregates into smaller aggregates and monomeric immunoglobulin molecules. As a consequence of this effect the processing of C1̄-treated aggregates by peritoneal macrophages or Kupffer cells was significantly decreased compared to aggregates not treated with C1̄. The decreased processing was shown to be due to an inhibited binding of the dissociated aggregates to the cells. When the aggregates were incubated with macrophages in various concentrations of fresh serum containing isolated C1̄, the inhibitory effect of C1̄ was overcome and a stimulated degradation was seen, depending upon the ratio of C1̄ to the other serum proteins. On the other hand, when aggregates had been preincubated with C1̄, subsequent addition of fresh serum could not reverse the inhibitory effect. Although competition between C1̄ and Fc receptors for the Fc parts in the immune aggregates may explain the inhibitory effect of C1̄ on the degradation of soluble immune aggregates by macrophages, dissociation of these aggregates by C1̄ is the major mechanism involved.
The Chicken's Femoral-Lymph Nodules: T and B Cells and the Immune Response
Authors: McCorkle Fred M. , Stinson R. S. , Olah I. , Glick B.
Published: December 2022

Femoral lymph nodules (FLN), which are barely perceptible in normal birds, after a footpad injection of sheep red blood cells (SRBC) may either significantly enlarge (responder) or remain reduced in size (nonresponder). There were approximately 38% T cells and 53% B cells in the FLN of responder chickens. Significantly more plaque-forming cells (PFC) developed in the FLN than in the spleen after a footpad injection of SRBC. Total antibody, mercaptoethanol- (ME) resistant, and ME-sensitive fractions were significantly higher in birds given i.v. than in those given footpad injections. There were no differences in PFC and agglutinin titers between FLN-responders and nonresponders. The number of PFC in FLN exceeded the number of splenic PFC previously reported. The high PFC response of the FLN may reflect the large percentage of B cells in this lymphomyeloid tissue or the presence of antigen-experienced B cells in the FLN. Although FLN may influence a systemic immune response its major role appears to be restricted to a local response.
H-2-Linked Ir Gene Control of Antibody Responses to Insulin
Authors: Schroer Joyce A. , Inman John K. , Thomas James W. , Rosenthal Alan S.
Published: December 2022

The immune response to insulin in the mouse is under MHC-linked Ir gene control (only H-2d mice respond to pork insulin whereas H-2b, H-2d, and H-2v respond to beef insulin; H-2a, q, k, r, s respond to neither). In addition, species variants of insulin can be used to study the effects of specific amino acid exchanges on the immune response. To evaluate these effects at the cellular level, a new technique has been developed for covalently coupling insulin to SRBC. Two bifunctional reagents (methyl-4-mercaptobutyrimidate and m-maleimidobenzoyl N-hydroxysuccinimide ester) were used to prepare insulin-SRBC for use in a modified Jerne plaque assay. Nonresponder mice failed to make a significant anti-insulin IgM or IgG PFC response over the entire course of a primary response to beef (H-2k and H-2q) or pork (H-2b, H-2k and H-2q) insulin. Responder and F1 (responder × nonresponder) mice make mostly IgG anti-insulin PFC with about 50% being IgG1 and 50% IgG2. Extensive cross-reactivity between beef and pork insulin after immunization with one insulin was found at the antibody-secreting cell level. Control of the response of H-2b mice to DNP-beef insulin has been mapped previously in either the K or I-A region. The present study maps the control of the H-2b PFC response to native beef insulin in this same region(s).
Susceptibility of CBA/N Mice to Infection with Salmonella Typhimurium: Influence of the X-Linked Gene Controlling B Lymphocyte Function
Authors: O'Brien Alison D. , Scher Irwin , Campbell Gary H. , MacDermott Richard P. , Formal Samuel B.
Published: December 2022

CBA/N mice have an X-linked immune defect that is associated with the absence of a subpopulation of mature or late developing B lymphocytes. The effect of this B cell defect on susceptibility of mice to Salmonella typhimurium was investigated. The 50% lethal dose (LD50) of S. typhimurium for CBA/N mice was 1000-fold less than for the immunologically normal, histocompatible CBA/CaHN strain. Genetic analysis revealed that the susceptibility of the CBA/N strain to S. typhimurium, like the immune defect, was X-linked since immune-defective F1 male mice derived from crosses between CBA/N and immunologically normal strains were susceptible to S. typhimurium (LD50 ≤ 20), whereas immunologically normal F1 males and females derived from these matings were resistant (LD50 ≥ 5 × 103). By contrast, both immune-defective and normal F1 mice were resistant to another facultative intracellular parasite, Listeria monocytogenes. Deaths among the immune-defective mice occurred primarily late in the course of infection (>10 days) whereas deaths of immunologically normal, S. typhimurium-susceptible strains (e.g., C57BL/6 and BALB/c) occurred early in the infectious process. To determine if the increased susceptibility of B cell-defective mice to S. typhimurium and the expression of the X-linked immune-deficiency gene (xid) were actually linked, F2 and backcross mice were phenotyped for the immune defect by measuring serum IgM levels and then challenged with the organism. Although no formal linkage was established, there was a close correlation between low serum IgM levels and susceptibility, since 93% of such mice succumbed to infection. These studies suggest that B cells play an important role in those events that govern the development of resistance to S. typhimurium late in the course of infection.
Tumor-Specific Immunity Induced by Somatic Hybrids
Authors: Kim Byung S. , Liang Weitze , Cohen Edward P.
Published: December 2022

Hybrid clones were derived from fusion of TEPC-15 plasmacytoma cells of BALB/c mice with mouse L cells of C3H origin. The morphology, tumorigenicity, and immunogenicity of three representative clones were extensively studied. One clone (LTC-1) showed a morphology intermediated to that of either parental cell and possessed the highest tumorigenic and immunogenic properties. The other two clones displayed a “flat” morphology which differed significantly from that of either parent. One of these two, LTC-4, eventually induced tumors in some (BALB/c × C3H)F1 mice but failed to stimulate protective immunity against TEPC-15 tumor cells in BALB/c mice. The other hybrid clone, LTC-2, has a “very flat” morphology and did not induce tumors, although it was capable of stimulating a significant level of tumor immunity. Histologically, all the tumors induced by hybrid cells were fibrosarcomas rather than plasmacytomas. These results indicate that the morphology of hybrid cells may be correlated with the tumorigenicity as well as the histologic appearance of tumor. In addition, the degree of tumorigenicity of individual hybrid clones does not correspond to their immunogenicity in the host, suggesting that major antigens responsible for immunogenicity may not play an important role in induction of tumors.
Nature of the Stimulus Leading to Lymphocyte-Induced Angiogenesis
Authors: Auerbach Robert , Sidky Younan A.
Published: December 2022

Experiments are described that characterize the nature of the stimulus leading to lymphocyte-induced angiogenesis (LIA), a reaction previously shown to reflect a local in vivo graft-vs-host reaction. The studies demonstrate that circulating cells of the host animals provide the stimulation essential for activation of donor lymphocytes and that the major allogeneic stimulus in congenic lines of mice is correlated with I-region disparity, primarily associated with IA-controlled determinants. The results are readily compartible with the hypothesis that is proposed that LIA is in large part the consequence of the release of soluble mediators or lymphokines that may act either directly on endothelial cells or indirectly by activating macrophages, which in turn generate the vascular reaction.
Natural NK-Cell Targets in the Mouse Thymus: Characteristics of the Sensitive Cell Population
Authors: Hansson Mona , Kärre Klas , Kiessling Rolf , Roder John , Andersson Birger , Häyry Pekka
Published: December 2022

In a previous study thymocytes from young mice less than 3 weeks old, especially those of NK low reactive genotypes, were found sensitive for NK-mediated lysis. This phenomenon has been further explored in the present study in which the NK-sensitive thymocyte population of young strain A/Sn mice has been characterized.

Injection of suckling A/Sn mice with cortisone drastically decreased sensitivity of the remaining medullary thymus cells for NK lysis, whereas sensitivity for lysis by anti-H-2-educated CTL blasts was enhanced. The NK-sensitive population repopulated the thymus after cortisone treatment 2 to 4 days earlier than the majority of the thymocytes. In vitro treatment of thymocytes with fresh rabbit or guinea pig serum, known to kill cortical thymocytes, also abolished the NK sensitivity, while sensitivity for allospecific CTL was increased. Depletion of thymocytes bearing receptors for the peanut agglutinin (PNA) strongly reduced the sensitivity of the remaining thymocytes for NK lysis, as well as for lysis by guinea pig and rabbit serum. Fractionation of thymocytes on a 1 × G velocity sedimentation gradient revealed a distinct sensitive peak sedimenting approximately 5 mm/hr whereas the majority of the thymocytes sedimented at the 3 to 4 mm/hr region. Morphologic studies confirmed that the majority of the NK-sensitive cells were large and rapidly dividing, indicating also by the high spontaneous incorporation of 3H thymidine into these 1 × G fractions. Free-flow electrophoresis analysis revealed that the NK-sensitive thymocytes distributed between the majority of the thymocytes and the rapidly migrating, more negatively charged medullary thymocyte.

Taken together, these results show that the NK sensitive “natural” target is a minor population of the thymus, clearly distinguishable from the majority of the cortical as well as medullary thymocytes.
Preparation of an Antibody to Mouse Serum IgD
Authors: Finkelman Fred D. , Kessler Steven W. , Scher Irwin
Published: December 2022

Mice have recently been shown to have serum IgD. We have used affinity chromatography to partially purify mouse serum IgD, and have prepared a rabbit antiserum against this mouse Ig class. This antiserum, once adsorbed by IgD-depleted mouse serum bound to Sepharose, was isotype specific as determined by radioimmunoassay and fluorescence activated cell sorter analysis, and bound to the same splenic lymphocyte surface molecules as a hybridoma produced monoclonal anti-mouse δ antibody.
Ultrastructural Localization of J Chain in Human Intestinal Mucosa
Authors: Nagura Hiroshi , Brandtzaeg Per , Nakane Paul K. , Brown William R.
Published: December 2022

J chains were localized by immunoperoxidase cytochemistry in the intestinal mucosa of immunologically intact patients and of patients with immunoglobulin deficiency and associated intestinal nodular lymphoid hyperplasia. J chains were present in an aggregate of lymphocytes from one of two immunologically intact patients and in lymphoid nodules from two of five immunodeficient patients. Ultrastructurally, the J chains were localized to the perinuclear spaces and endoplasmic reticulum of the lymphocytes, but not to the plasma membranes. A few of the J chain-containing cells resembled activated lymphocytes, but most corresponded to less mature cells with IgM expressed on their surface membranes. J chains were present also in numerous plasma cells in the lamina propria of the immunologically intact patients and in a few plasma cells in the immunodeficient patients. The J chains in plasma cells were localized to the perinuclear spaces and endoplasmic reticulum, and infrequently to elements of the Golgi apparatus. Acid-urea treatment of the plasma cells intensified the reactions for J chains in the endoplasmic reticulum, but did not reveal additional evidence of J chains in Golgi organelles. J chains were not seen in columnar epithelial cells before acid-urea treatment, but afterward, J chains as well as IgA, IgM, and secretory component were found on basolateral plasma membranes and in intracellular vesicles.

The findings indicate that: 1) J chains in B lymphocytes may appear before distinct morphologic evidence of blast transformation; 2) J chains may combine with immunoglobulin polymers in the endoplasmic reticulum of lymphoid cells; 3) J chain-containing immunoglobulin polymers probably are continually in complex with secretory component, their membrane-bound receptor, during transit across intestinal epithelial cells.