bluuebunny/crossref_metadata_2025 huggingface dataset:
Release of Slow Reacting Substance of Anaphylaxis from Human Leukocytes
Authors: Grant J. Andrew , Lichtenstein Lawrence M.
Published: January 2023

Mediators of immediate hypersensitivity were released from human leukocytes by means of allergen challenge. Leukocytes from 12 consecutive donors were challenged with allergens to which these donors were clinically sensitive. In all, histamine and a second substance (SRS-Aleuk) which caused prolonged contraction of guinea pig ileum, were released. No SRS-Aleuk was obtained without allergen-challenge, and none was observed from non-atopic leukocytes challenged with the same allergens. The time of release after addition of allergens and the allergen dose-response curves for release of both histamine and SRS-Aleuk were quite similar. SRS-Aleuk was synthesized de novo after allergen-challenge since this factor was not demonstrated in disrupted cells. SRS-Aleuk was soluble in 80% ethanol and was not destroyed by base hydrolysis. The smooth muscle contracting activity of SRS-Aleuk resembled that of slow reacting substance of anaphylaxis (SRS-A) obtained from human lung and rat peritoneal cells, but was different from histamine, bradykinin, serotonin, and prostaglandin E1 and F2α. Preliminary studies suggest that both histamine and SRS-Aleuk are synthesized by human basophils.

We conclude that SRS-A is generated by human leukocytes as well as human lung and nasal polyps, and suggest that this mediator may play an important role in the pathogenesis of human immediate hypersensitivity reactions.
Preparation and Properties of Rabbit Anti-Mouse Plasmacyte Serum Purified by in Vivo Absorption: Further Evidence for a New Antigenic Determinant on Plasmacytes
Authors: Yutoku Masamichi , Grossberg Allan L. , Pressman David
Published: January 2023

A new surface antigenic determinant on both normal and malignant plasma cells of mice has been detected with rabbit antisera, obtained by immunization with mouse myeloma cells and purified by in vivo absorption. One IgG- and one IgA-producing myeloma was used. Although the determinant was detected on plasma cells and one lymphoma and not on other lymphoid cells, it was also found on cells of liver, kidney, and brain. Thus the antigen is not a plasmacyte-specific antigen. In spite of an apparent similarity between the tissue representation of the determinant and that of the PC.1 alloantigenic determinant, the present determinant is not identical with the PC.1 determinant, since it is found on the respective tissues of PC.1-negative strains of mice.
Acid Lability of Rhinoviruses: Loss of C and D Antigenicity after Treatment at pH 3.0
Authors: Hughes John H. , Chema Samson , Lin Nancy , Conant Robert M. , Hamparian Vincent V.
Published: January 2023

Serologic evidence is presented for the existence of rhinovirus antigens with C and D type reactivity, and for their loss after acid treatment. No neutralizing antibody could be detected in animal antisera prepared with pH 3.0-treated rhinovirus antigens. In contrast, antisera prepared with untreated (native) rhinovirus antigens, contained high levels of neutralizing antibody. In immunodiffusion tests, no precipitin lines were detectable when native antigens were diffused against antisera prepared with pH 3.0-treated antigens. Acid treated antigens also were non-reactive by immunodiffusion when reacted against antisera prepared with treated or untreated antigens. The diffusion of native antigens against hyperimmune animal sera prepared with untreated antigens gave one precipitin line. With normal human sera, two precipitin lines were observed with untreated antigens. Rhinoviruses and enteroviruses were found to share a common group reactive antigen.
Studies on the Structural and Conformational Basis for the Relative Resistance of Serum and Secretory Immunoglobulin a to Proteolysis
Authors: Underdown Brian J. , Dorrington Keith J.
Published: January 2023

The structural features which make secretory IgA relatively resistant to proteolysis, compared to the major serum immunoglobulin IgG, were investigated.

Secretory IgA, serum IgA, and IgG were incubated with pre-activated papain (4 hr) and Pronase (2 hr) at slightly alkaline pH in the absence of reducing agents. Digest mixtures were analyzed by Sephadex G-200 gel filtration and immunodiffusion. Under conditions which resulted in essentially quantitative cleavage of IgG1 myeloma proteins to Fab and Fc fragments, approximately 85% of secretory IgA, normal human serum IgA, and IgA myeloma proteins of both A1 and A2 subclasses remained undigested. Thus, both secretory IgA and serum IgA were relatively resistant to proteolysis compared to the major serum immunoglobulin, IgG1. Secretory IgA dimer was more resistant than IgA myeloma dimers since incubation with Pronase for 3 hr caused the depolymerization of the latter to a monomer-like fragment whereas secretory IgA dimer remained intact.

Reduction with 10 mM dithiothreitol (DTT) and alkylation of serum IgA caused this immunoglobulin to become susceptible to proteolysis. Secretory IgA required a greater concentration of reducing agent (35 mM DTT) for conversion to the digestible form than did IgA myeloma dimers. Circular dichroism was used to detect conformational changes in both serum and secretory IgA which occurred after mild reduction and alkylation. Part of the conformation change in IgA proteins after mild reduction and alkylation was localized to the Fc fragment.

It was concluded that secretory IgA was more resistant than the major serum immunoglobulin IgG1 and that this relative resistance was due in part to the greater resistance of immunoglobulin A per se, and in part due to additional stability of the secretory IgA form.
In Vivo Suppression by Cholera Toxin of Cell-Mediated and Foreign Body Inflammatory Responses
Authors: Warren Kenneth S. , Mahmoud Adel A. F. , Boros Dov L. , Rall Theodore W. , Mandel Mark A. , Carpenter Charles C. J.
Published: January 2023

Cholera toxin (CT) injected i.v. into mice resulted in a rise in cyclic AMP levels in splenic white cells. This was associated with and followed by a marked and relatively prolonged decrease in both the spleen cells and circulating lymphocytes. The effect of CT on a series of cell-mediated immunologic reactions related to the pathogenesis of schistosomiasis was then tested. CT profoundly suppressed dermal footpad swelling to soluble schistosome egg antigens, granuloma formation around schistosome eggs injected into the pulmonary microvasculature of unsensitized and sensitized mice, production of macrophage migration inhibition factor by intact granulomas (obtained from the livers of infected mice) maintained in vitro in tissue culture, and it ameliorated portal hypertension and esophageal varices in hepatosplenic schistosomiasis. In addition CT significantly prolonged allograft survival in mice even though it was first administered 6 days after grafting. Finally, CT completely eliminated foreign body granuloma formation around plastic beads (used as controls for the egg granulomas) which is a nonimmunologic reaction due to activation of chemical mediators of inflammation.
Conformational changes in C1q after binding to immune complexes: detection of neoantigens with monoclonal antibodies.
Authors: Golan M D , Burger R , Loos M
Published: January 2023

The formation of neoantigens within the C1q molecule after the binding of C1r and C1s to C1q and the binding of C1q to immune complexes is described. The neoantigens were detected by different monoclonal anti-C1q antibodies. This immunochemical study supports the hypothesis drawn from functional studies that the activation of the classical C pathway results from conformational changes within the C1q molecule leading to the activation of C1r and subsequently C1s.
Differential effect of activated T amplifier cells on B cells responding to thymus-independent type 1 and type 2 antigens.
Authors: Braley-Mullen H
Published: January 2023

Concanavalin A (Con A), antilymphocyte serum (ALS), and the allogeneic effect have all been shown to enhance antibody responses to thymus-independent (TI) antigens by a T cell-dependent mechanism. Enhancement presumably involves an effect on the responding B cells by amplifier T cells (TA) that are activated by the particular agent used, i.e., Con A, ALS, or allogeneic cells. In the present study the ability of these agents to augment antibody responses to TI-1 and TI-2 antigens was compared. It was found that antibody responses to TI-2 antigens were markedly enhanced by allogeneic cells or ALS given with antigen. Antibody responses to TI-1 antigens were in all cases unaffected. Because TI-1 and TI-2 antigens primarily stimulate separate subpopulations of B cells, these results suggest the B cell subset that responds to TI-1 antigens, i.e., Lyb-5-negative B cells, may not be responsive to signals from nonspecifically activated TA. This conclusion was further supported by experiments demonstrating that responses to TI-2 antigens could not be enhanced in mice that have only Lyb-5-negative B cells, i.e., CBA/N x BALB/c F1 males, and that responses to a T-dependent antigen (HRBC) could be augmented by the allogeneic effect in normal mice but not in mice lacking Lyb-5-positive B cells.
Expression of Lyt-1 by a subset of B lymphocytes.
Authors: Manohar V , Brown E , Leiserson W M , Chused T M
Published: January 2023

Using two-color flow cytometry and multiparameter data analysis, we have shown that the IgM bright, large subset of mouse splenic B lymphocytes express Lyt-1. This is not due to B cell uptake of immune complexes of Lyt-1 and antibody from T cells. The IgM bright cells of autoimmune NZB mice express more Lyt-1 than normal controls. This is because IgM containing plasmablasts, which are greatly increased in NZB spleens, are Lyt-1+. NZB spleen also contains more cells that are Lyt-1+ (but perhaps Lyt-1.2-), Thy-1.2 dull, and smaller in size than cells in normal mice. Thus, Lyt-1 is common to the T and B cell precursor or is induced independently during the ontogeny of T and at least one subset of B cells. We suggest that it be called Lyt-1.
A phorbol ester tumor promoter induces changes in the expression of immunoglobulins and DR antigens in human lymphoblastoid cells.
Authors: Roth P , Halper J P , Weinstein I B , Pernis B
Published: January 2023

The present study indicates that 12-O-tetradecanoyl phorbol 13-acetate (TPA) can induce either increases or decreases in the relative expression of membrane-associated IgM and IgD as well as the expression of other markers of lymphocyte differentiation on human B lymphoblasts, depending on the clone studied. The changes in membrane Ig expression are apparent within 24 hr and are maximal at about 48 hr after the addition of the compound. In clone BL, TPA decreases the membrane IgM/IgD ratio as determined by the percentage of cells bearing these isotypes and by their densities on single cells. In clone MW-E, TPA has the opposite effect. Although these changes are followed by an increase in Ig secretion by MW-E cells, measured by immunofluorescence and the protein A plaque assay, there is no induction of Ig secretion in BL cells. In addition, TPA increases the expression in BL of an additional B lymphocyte marker, the D region-related (DR) antigens, while having the opposite effect in MW-E. In two sublines of Daudi, a membrane IgM-positive, IgD-negative, nonsecreting cell line, TPA treatment resulted in the expression of membrane IgD without the induction of Ig secretion. Thus, the TPA-induced changes in the IgM/IgD ratio and DR antigen expression are inversely related and are not due to differential proliferation of subpopulations. Taken together, it appears that, depending on the particular clone of B lymphoblasts studied, TPA can induce or inhibit a coherent program of B lymphocyte maturation.
Production of macrophage-activating factor by T lymphocyte clones and correlation with other lymphokine activities.
Authors: Kelso A , Glasebrook A L , Kanagawa O , Brunner K T
Published: January 2023

The production of macrophage-activating factor (MAF) by antigen-stimulated murine T lymphocyte clones has been compared with their cytolytic function and release of other lymphokines. MAF activity was measured by the capacity of peptone-induced peritoneal exudate cells or bone marrow-derived macrophages to lyse 51Cr-labeled tumor cells after incubation with supernatant from the stimulated T cells and a nonactivating, amplifying dose of lipopolysaccharide. Of 72 clones generated against H-2, MIs, H-Y, or Moloney leukemia virus-associated antigens, 68 were found to produce detectable quantities of MAF. Release of MAF by clones 1) occurred within 1 to 12 hr of exposure to antigen, 2) required stimulation with cells of the relevant antigenic specificity, and 3) could also be induced by concanavalin A, indicating that the cloned cells were the source of the activity. The capacity of a clone to produce MAF was independent of its antigenic specificity, cytolytic activity, or ability to produce interleukin 2 or granulocyte-macrophage colony-stimulating activity. In contrast, production of interferon and MAF was not dissociated for any of the clones tested.
Estimate of the light chain repertoire size of fetal and adult BALB/cJ and CBA/J mice.
Authors: LeJeune J M , Briles D E , Lawton A R , Kearney J F
Published: January 2023

The potential light chain repertoires expressed by IgM antibodies of fetal liver and adult spleen cells were compared. B cells from these sources were stimulated by the polyclonal mitogen bacterial lipopolysaccharide and after 3 days were fused to construct hybridomas. The light chains synthesized by these hybridomas were isolated by electrophoresis and their heterogeneity was examined by isoelectric focusing. A total of 109 BALB/c and 77 CBA/J hybridoma-derived light chains obtained from fetal liver and adult spleen cells were analyzed in this way. Fifty-three and 45 unique light chains were obtained from BALB/c and C57BL/6 hybridomas, respectively. Some of these were repeated more frequently than others, but no significant differences were seen in the heterogeneity of the light chain spectrotypes of fetal and adult CBA/J and BALB/cJ mice. Furthermore, the kappa:lambda ratio of light chains were the same in fetal liver as adult-derived hybridomas. These results show that the heterogeneity of light chains as determined by IEF and the kappa:lambda ratio of IgM immunoglobulins is established very early in ontogeny.
Viral antigens act as helper determinants for antibody responses to cell surface antigens.
Authors: Bromberg J S , Lake P , Brunswick M
Published: January 2023

Thy-1 alloantigens on murine thymus cells are weak immunogens in vivo for PFC responses in the absence of other antigenic disparities between the donor and recipient. Our previous work showed that non-H-2 alloantigens acted as helper determinants to augment anti-Thy-1 PFC responses. In this report we demonstrate that strong helper antigens are also produced by infection of donor thymus cells with viruses such as HSV-1, NDV, or vaccinia. This helper effect (as much as 30-fold) for a cellular antigen, requires linked recognition (expression of Thy-1 and virus in the same cell membrane), is T-dependent, antigen- (virus) specific, and is Thy-1-specific. The recognition of the viral helper sites is not restricted by the MHC genotype of the thymus cell donor, indicating that host reprocessing of antigen occurs. These are the first results that show that adventitious antigens may function as helper determinants for antibody responses to native membrane antigens and may be the mechanism that initiates several forms of acute post-viral autoimmune disease.
Induction of transplantation tolerance in guinea pigs by spleen allografts. II. Responses in the mixed leukocyte reaction correlate with the tolerant state.
Authors: Shevach E M , Chan C , Bitter-Suermann H
Published: January 2023

We have developed a model for the induction of transplantation tolerance in the guinea pig by vascularized spleen allografts. Spleen allografts from strain 13 to strain 2 hosts frequently survived in healthy recipients without clinical GVHD or induced clinical GVHD. (2 x 13)F1 to strain 2 spleen allografts survived indefinitely without inducing GVHD. In contrast, strain 2 spleen allografts were rejected by strain 13 hosts. An excellent correlation was observed between the clinical course and the degree of reactivity to donor strain stimulator cells in the MLR. Animals that had rejected their grafts had normal or enhanced proliferative responses in the MLR. Strain 2 hosts with long-term surviving strain 13 or (2 x 13)F1 grafts had markedly suppressed anti-13 responses. Animals with GVHD had a suppressed MLR toward donor strain stimulator cells with simultaneous reactivity to host strain stimulator cells. Cells capable of suppressing the response of normal host strain cells to donor strain stimulators were present in some long-term surviving animals and may be responsible in part for the maintenance of the tolerant state.
Autoimmune effector cells. III. Role of adjuvant and accessory cells in the in vitro induction of autoimmune encephalomyelitis.
Authors: Killen J A , Swanborg R H
Published: January 2023

The present investigation shows that autoreactive effector cells that transfer experimental allergic encephalomyelitis (EAE) can be activated from spleens and lymph nodes of Lewis rats given a single injection of 25 micrograms myelin basic protein (BP) in incomplete Freund's adjuvant (IFA), despite the fact that the cell donors do not develop EAE. Rather, these donor rats are unresponsive to EAE when given an encephalitogenic emulsion of BP in complete Freund's adjuvant (CFA). Lymphoid cells from rats given a single injection of BP-IFA were almost as effective as cells from BP-CFA-treated rats with respect to transferring EAE after in vitro activation with BP or concanavalin A (Con A). Irrespective of whether donors received BP in IFA or CFA, BP-cultured spleen and lymph node cells (SpC and LNC, respectively) transferred EAE, whereas Con A-cultured SpC but not LNC exhibited effector cell activity. Con A-cultured LNC were able to transfer EAE if the cultures were reconstituted with irradiated adherent phagocytic cells (which could be obtained from normal Lewis rat spleens) or with conditioned medium from these adherent SpC. These findings indicate that accessory cells are required for in vitro induction of this T cell-mediated autoimmune response.
Rabbit anti-human CSF IgG. I. Characterization of anti-idiotype antibodies produced against MS CSF and detection of cross-reactive idiotypes in several MS CSF.
Authors: Tachovsky T G , Sandberg-Wollheim M , Baird L G
Published: January 2023

Serum from rabbits injected with IgG prepared from the CSF of patients with multiple sclerosis were rendered anti-idiotypic by adsorption on sequential columns of pooled human gamma-globulin, free light chains and free heavy chains. Adsorbed antisera precipitated between 15% and 45% of the autologous ligand and were inhibited by autologous CSF and serum and F(ab')2 fragments prepared from serum IgG. Idiotypic cross-reactivities were detected among the CSF IgG of several MS patients using three of these antisera. Two antisera demonstrated low levels of cross-reactivity with CSF IgG obtained from two of six heterologous MS patients. The third antiserum detected highly cross-reactive idiotypic determinants in the CSF IgG of two family members both having MS.
The cellular basis for resistance to induction of tolerance in BXSB SLE male mice.
Authors: Hang L , Izui S , Slack J H , Dixon F J
Published: January 2023

The autoimmune BXSB male mouse early in life displays a resistance to tolerance induction that is common with the other autoimmune strains (MRL/1, NZB x W, and NZB) whereas its female counterpart, which in early life is immunologically normal, is easily tolerized. The cellular basis for the BXSB male's resistance to tolerance induction with DHGG was investigated by using adoptive male-female cell transfer systems. We found that irradiated female recipients of male T-depleted bone marrow cells, plus either male or female thymus cells, resisted tolerance induction, had hyperactive splenic Ig-secreting cells, and developed early glomerulonephritis. Recipients of male thymic cells plus T-depleted female bone marrow cells could be tolerized, did not have enhanced Ig secretion, and did not develop glomerulonephritis. Furthermore, donor male T-depleted bone marrow cells--but not male thymic, female thymic, or female bone marrow cells--resisted induction of tolerance before their transfer to produce lymphoid chimeras.
Biosynthesis of platelet-activating factor (PAF)acether). III. Formation of PAF-acether from synthetic substrates by stimulated murine macrophages.
Authors: Mencia-Huerta J M , Roubin R , Morgat J L , Benveniste J
Published: January 2023

Rat adherent macrophages (M phi) isolated from 2.5 x 10(6) peritoneal cells release 5.7 +/- 1.3 ng/ml platelet-activating factor (PAF-acether: 1-O-alkyl-2-acetyl-sn-glyceryl-3-phosphorylcholine) when stimulated by zymosan (Z). The yield of PAF-acether was increased to 13.8 +/- 1.8 ng/ml when stimulation occurred in the presence of 0.1 mM acetyl-CoA. That M phi used the acetyl moiety of acetyl-CoA to increase PAF-acether biosynthesis was demonstrated by the following criteria 1) as opposed to sodium acetate, malonyl-CoA, butyryl-CoA, palmitoyl-CoA, and CoA were inefficient in increasing the yield of PAF-acether; 2) radiolabeled PAF-acether was obtained after exposure of Z-stimulated M phi to 3H acetyl-CoA; actually, both radioactivity and biologic activity were eluted from HPLC with the same retention time as synthetic or natural PAF-acether; 3) after treatment with phospholipase A2, both radioactivity and biologic activity disappeared from these fractions. The yield of PAF-acether was further increased to 23.6 +/- 2.9 ng/ml by adding 0.5 microM totally synthetic 2-lyso PAF-acether (1-O-octadecyl-sn-glyceryl-3-phosphorylcholine) to Z-stimulated M phi cultured in the presence of acetyl-CoA. Again, using 3H 2-lyso PAF-acether, the formation of radiolabeled PAF-acether could be demonstrated. These results indicate that an acetyl-transferase is capable of synthesizing PAF-acether from 2-lyso PAF-acether and acetyl-CoA in intact M phi.
Identification of Treponema pallidum antigens: comparison with a nonpathogenic treponeme.
Authors: Lukehart S A , Baker-Zander S A , Gubish E R
Published: January 2023

The major antigens of Treponema pallidum, Nichols strain were examined by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting techniques. Organisms were extracted from rabbit testes, purified by differential centrifugation, washed, and disrupted by sonication before solubilization in SDS buffer, SDS-PAGE analysis on 12.5% gels revealed at least 35 distinguishable polypeptides with molecular weights between 14,000 and 100,000. Proteins were electrophoretically transferred from gels to nitrocellulose paper and were stained with pooled rabbit anti-T. pallidum serum (IRS) and 125I-protein A. At least eight distinct antigenic molecules were revealed, with the major component residing at 48,000 daltons. Extensive washing of T. pallidum before solubilization did not alter the antigenic profile. Five molecules with common treponemal antigenic determinants were identified both by staining blots of T. phagedenis, biotype Reiter with IRS and by staining blots of T. pallidum with rabbit anti-Reiter antiserum. Three putative T. pallidum-specific antigens were identified by staining T. pallidum with IRS that had been extensively adsorbed with Reiter treponemes.
HLA restriction of secondary mumps-specific cytotoxic T lymphocytes.
Authors: Kress H G , Kreth H W
Published: January 2023

Mumps virus-specific cytotoxic T lymphocytes (CTL) were induced by incubating peripheral lymphocytes from healthy blood donors with autologous mumps virus-infected stimulator cells. Maximum cytotoxicity was generated after 5 to 7 days in culture. No cytotoxic activity was induced in the absence of viral antigens. The cytotoxicity induced in vitro was a secondary immune response, because no specific lytic activity could be generated in lymphocyte cultures from mumps seronegative donors. The effector cells were restricted by the HLA complex in a "hierarchical" pattern. The major restriction antigens for secondary CTL were certain HLA B determinants such as B18, B27, Bw25, Bw62, and Bw63. Mumps responses in spite of high [3H]-thymidine uptake in sensitized cultures. The present data suggest that mumps virus-specific CTL may be directly regulated by certain genes of the HLA B locus.
Induction of high affinity IL 2 receptors on B cells responding to anti-Ig and T cell-derived helper factors.
Authors: Prakash S , Robb R J , Stout R D , Parker D C
Published: January 2023

We have reported that IL 2 is one of the essential helper factors in culture supernatants from concanavalin A-activated spleen cells or T cell hybridomas that support proliferation and immunoglobulin secretion in B cell cultures responding to anti-immunoglobulin. Here we show that cells in such cultures consume IL 2 and bear high affinity IL 2 receptors detected by binding of purified, radiolabeled IL 2. Induction of high affinity IL 2 receptors depends on addition of both anti-immunoglobulin and helper factors, and does not occur in cultures given only anti-Ig, only helper factors, concanavalin A plus helper factors, or LPS. The majority of IL 2 receptors are on cells that also bear endogenous membrane immunoglobulin, because they are found in the membrane immunoglobulin-positive fraction when cultured cells are separated by fluorescence sorting after overnight culture in the absence of anti-immunoglobulin to allow reexpression of membrane immunoglobulin.
Differential abilities of human peripheral blood monocytes quantitatively or qualitatively differing in HLA-DR and HLA-DS expression to support B cell activation in liquid and semisolid cultures.
Authors: Whisler R L , Newhouse Y G , Lachman L B
Published: January 2023

The present investigation was performed to determine whether the activation of human B cells by Staphylococcal protein A (SpA) in liquid and semi-solid cultures might be dependent on distinct subsets of peripheral blood mononuclear-phagocytes (M phi) defined by the expression of HLA-DR and HLA-DS determinants. Highly pure HLA-DR- M phi functioned as effectively as HLA-DR+ MO in supporting B cell liquid proliferative responses when SpA was continuously present in cultures. However, HLA-DR+ M phi were two to three times more effective than HLA-DR- M phi in promoting B cell proliferative responses when either M phi or B cells were pulsed with SpA and were then cultured without supplemental SpA. Similarly, B cell activation in semisolid cultures was crucially dependent on HLA-DR+ M phi because colony responses were reduced fivefold in the presence of M phi expressing low/intermediate HLA-DR levels compared to M phi-containing cells with high HLA-DR levels. HLA-DS- M phi isolated by two different techniques were more effective than HLA-DS+ M phi in supporting both liquid proliferative and colony responses of B cells. Flow microcytofluorometry analysis of the dual expression of HLA-DR and HLA-DS on highly pure HLA-DR- M phi and HLA-DR+ M phi revealed that both HLA-DR- and HLA-DR+ M phi expressed low levels of HLA-DS. Importantly, the expression of HLA-DS on HLA-DR- M phi was bimodal, with an HLA-DR-, DS+ subset and an HLA-DR-, DS-subset being present. Other experiments supported the conclusions that the differential abilities of the HLA-DR-, -DS-defined subsets of M phi to support B cell activation did not represent M phi suppressive effects or differences in IL 1 production. Collectively, these results indicate that B cell activation can be directly supported by M phi whose predominant phenotype is HLA-DR+, -DS-. Thus, the accessory cell pathway of B cell activation described here is distinct from the pathway known to be required for T cell responsiveness, and could serve to provide early alternative or ancillary signals for triggering B cells.
Differential effects of recombinant human interferon-gamma and interleukin 2 on natural killer cell activity of peripheral blood in early human development.
Authors: Ueno Y , Miyawaki T , Seki H , Matsuda A , Taga K , Sato H , Taniguchi N
Published: January 2023

Ontogenic development and the lymphokine responsiveness of human NK cell activity against K562 target cells in peripheral blood lymphocytes were evaluated in fetuses, premature infants, and term neonates by using a 4-hr 51Cr-release assay. Basal NK activity and NK boosting by lymphokines were comparatively assayed after an 18-hr incubation with medium alone, recombinant human IFN-gamma (1000 U/ml), and recombinant human IL 2 (25 U/ml), respectively. Lymphocytes from 20-wk-old fetuses lacked NK cell activity even after the pretreatment with IFN-gamma. Low, but significant levels of NK activity and NK boosting by IFN-gamma were observed in premature infants after 27 wk of gestation, with a progressive intrauterine maturation of these activities. Both basal NK activity and NK boosting by IFN-gamma in term neonates were still lower than those of adult controls. The grade of NK boosting by IFN-gamma appeared to depend on the development of basal NK activity. Contrary to IFN-gamma, IL 2 could induce marked NK activity even in 20-wk-old fetuses who lacked both basal and IFN-gamma inducible NK activities. NK boosting by IL 2 was much more efficient than that by IFN-gamma at any period of human life. The facts that IL 2-induced NK boosting could occur without any appreciable production of IFN-gamma in neonatal lymphocytes, and that ample neutralizing doses of anti-IFN-gamma antibody hardly suppressed IL 2-mediated NK boosting even in adult lymphocytes, indicated that the effect of IL 2 on NK boosting might be independent of IFN-gamma production. On the basis of the ontogenic differences in the development of the lymphokine responsiveness of NK cell activity and on the different NK boosting mechanisms of these lymphokines it was suggested that so-called human "pre-NK cells" might be divided into IFN-gamma sensitive and IL 2-sensitive cells. Whether these cell populations belong to different cell lineages or different maturation stages of the same cell line, however, remains unsettled.
Detection of cross reactive anti-DNA antibody idiotypes on tissue-bound immunoglobulins from skin biopsies of lupus patients.
Authors: Isenberg D , Dudeney C , Wojnaruska F , Bhogal B S , Rauch J , Schattner A , Naparstek Y , Duggan D
Published: January 2023

Cross-reactive anti-DNA antibody idiotypes have been identified on tissue-bound immunoglobulins from skin biopsies of patients with systemic lupus erythematosus (SLE) and discoid lupus erythematosus (DLE). Four polyclonal and two monoclonal anti-idiotypic reagents were used to screen biopsies from 24 patients with SLE, 23 patients with DLE, and 15 other patients with IgM-positive skin biopsies. Up to 46% of the SLE patients and 30% of the DLE patients were found to share idiotypes present on immunoglobulins deposited at the dermal-epidermal junction. Inhibition studies in four patients indicated that the idiotypes were on anti-DNA antibodies. In contrast, none of the anti-idiotypic antibodies bound to any of the control biopsies. These findings imply that some tissue-bound autoantibodies are derived from related families of high-frequency germ-line genes that are expressed in both SLE and DLE.
Low molecular weight C1q in systemic lupus erythematosus.
Authors: Hoekzema R , Hannema A J , Swaak T J , Paardekooper J , Hack C E
Published: January 2023

In sera of patients suffering from an exacerbation of systemic lupus erythematosus (SLE), increased amounts of abnormal C1q were detected, contrasting with decreased or even undetectable levels of normal C1q in these sera. When analyzed immunochemically by double immunodiffusion, this low m.w. C1q (LMW-C1q) appeared to be identical with the defective C1q in serum of individuals with an inherited, homozygous inability to produce functional plasma C1q. These persons show a tendency to develop SLE-like syndromes. Like the genetically defective C1q, the abnormal C1q molecule in SLE sera was hemolytically inactive, did not incorporate in C1, was found in the supernatant of euglobulin-precipitated serum, and appeared in the break-through fraction of a cation-exchange column. Sucrose gradients and gel filtration analyses supported the putative identity of the molecules. SDS-PAGE and immunoblots revealed the presence of subunits that reacted with antibodies against C1q and confirmed the C1q-like nature of LMW-C1q. Low levels of LMW-C1q were also detected in serum and plasma of normal individuals. A radial immunodiffusion technique was used to measure LMW-C1q in the serum of 54 patients. Although these patients were not selected for parameters of disease activity, their levels of LMW-C1q were significantly higher than those of normal individuals and children with decreased C3 levels due to acute glomerulonephritis.
The induction of tolerance to thyroglobulin significantly reduces the severity of thyroiditis in obese strain chickens.
Authors: Sanker A J , Clark C R , Sundick R S
Published: January 2023

Obese strain chickens develop severe spontaneous autoimmune thyroiditis several weeks after hatching, characterized by mononuclear cell infiltration and antibodies to thyroglobulin (Tg). The presence of antibodies to Tg suggests that Tg is an important antigen in this disease, but it does not provide definitive evidence. To clarify this point, Obese strain chicks were tolerized at hatching with Tg and then examined up to 6 wk later for antibodies to Tg, thyroid pathology, and function. Various tolerance regimens were tested. The optimal conditions were i.v. injection of Tg within 24 hr of hatching, and injection of at least 1 mg. Tg isolated from normal thyroid glands was satisfactory, and it did not have to be deaggregated. Tolerance induced by the above procedure significantly retarded all parameters of autoimmunity, although by 6 wk of age some of the tolerized chicks had severe thyroiditis. Multiple weekly injections of Tg were no more effective than a single injection at hatching. Interestingly, a single injection at hatching was very effective, yet it was cleared from the circulation within 24 hr. In summary, tolerance induced with Tg had a profound effect on the disease and thus provides good evidence for the role of Tg in this disease.
Interferon-like activity in an anti-interferon anti-idiotypic hybridoma antibody.
Authors: Osheroff P L , Chiang T R , Manousos D
Published: January 2023

We describe here an anti-idiotypic hybridoma antibody directed against affinity-purified rabbit idiotypic antibodies (Rb-Id) to a homogeneous protein, the recombinant human leukocyte A interferon (rIFN-alpha A). The supernatant of the hybridoma, designated 3-1B, was able to inhibit the neutralization of rIFN-alpha A activity by the idiotypic antibodies. An in vivo passage of uncloned 3-1B cells yielded hybridoma cells (presumably a subclone), designated 3B1, the supernatant of which exhibited interferon-like antiviral activity with both bovine kidney (MDBK) cells and human amnion (WISH) cells. This activity could be absorbed by polymer-bound goat anti-mouse immunoglobulin serum and by Rb-Id coupled to Affi-Gel 10, and could be partially eluted from the latter at pH 2.5. The anti-idiotypic hybridoma antibody was able to compete with 125I-rIFN-alpha A for binding to the Rb-Id and also to interferon receptor-bearing MDBK cells. The clinical significance of an interferon-like anti-idiotypic antibody is discussed.
Role of interleukin 1 in promoting human monocyte-mediated tumor cytotoxicity.
Authors: Onozaki K , Matsushima K , Kleinerman E S , Saito T , Oppenheim J J
Published: January 2023

Human peripheral blood monocytes from normal donors obtained by separation on a Percoll gradient showed considerable cytotoxicity against tumor cells when preincubated in vitro for 24 hr with human monocyte-derived interleukin 1 (IL 1). In contrast, monocytes after pretreatment in medium alone had low cytotoxic activity. All the IL 1 preparations, including IL 1 which was purified by high-performance liquid column chromatography (HPLC), as well as crude culture supernatant from human monocytes promoted monocyte-mediated cytotoxicity in the same dose-dependent manner as the thymocyte growth-promoting activity. There was no endotoxin or interferon (IFN) activity in the highly purified IL 1, suggesting that IL 1 itself was the active moiety. The effect of IL 1 on monocyte-mediated cytotoxicity was partially inhibited by indomethacin, whereas pretreatment of monocytes with prostaglandin (PG) E1 or E2 rather than IL 1 also resulted in substantial monocyte cytotoxicity. Thus, the effect of IL 1 on monocyte-mediated cytotoxicity is presumably mediated by PGE. Since fresh monocytes that were not preincubated exhibited levels of spontaneous cytotoxic activity similar to that of monocytes preincubated with IL 1, it seemed likely that the effect of IL 1 was to maintain the spontaneous level of activity rather than to induce cytotoxic activity. To elucidate this possibility, monocytes were first preincubated in medium alone for a longer period, and after losing their spontaneous activity they were further incubated with or without IL 1. Such "aged" monocytes did not develop cytotoxic activity in response to IL 1 but did in response to other agents known to induce macrophage cytotoxicity, such as endotoxin or lymphokine-containing supernatants. Therefore, the major effect of IL 1 actually seemed to prolong the cytotoxic state of monocytes. These results also suggest that IL 1 released by macrophages or monocytes may play a role in host defense against neoplastic cells by acting on monocytes as an autostimulating factor.
Generation of anti-peptide monoclonal antibodies which recognize mature CSF-2 alpha (IL 3) protein.
Authors: Conlon P J , Luk K H , Park L S , March C J , Hopp T P , Urdal D L
Published: January 2023

The colony-stimulating factor CSF-2 alpha (IL 3) has been purified to homogeneity, the protein sequenced, and the gene encoding this lymphokine cloned. Knowledge of the protein sequence permitted the synthesis of peptides corresponding to the amino terminus of the molecule. These peptides, after conjugation to palmitic acid, were used to immunize mice. Spleen cells from mice immunized with one of these peptides (CSF-2 alpha 1-14) were fused with the myeloma cell line NS-1. The fusion resulted in the isolation of two hybridoma cell lines, designated 6A5 and 4D4, that secreted antibodies that were specific for the immunizing peptide. The antibodies did not react with a closely related peptide CSF-2 alpha 7-16. The antibodies were capable, however, of recognized CSF-2 alpha protein as judged by the ability of the antibodies to remove CSF-2 alpha activity from culture medium of PHA-stimulated LBRM-33-5A4 cells, to immunoprecipitate radiolabeled CSF-2 alpha protein, and to detect CSF-2 alpha protein bound to nitrocellulose membranes.
Functional properties of two human B cell growth factor species separated by lectin affinity column.
Authors: Dugas B , Vazquez A , Geŕard J P , Richard Y , Auffredou M T , Delfraissy J F , Fradelizi D , Galanaud P
Published: January 2023

The supernatants from PHA-activated normal human T cells (conditioned medium) were fractionated by differential adsorption on Con A-Sepharose columns. The effluent contained both IL 2 activity (tested on the CTL-L2 cell line) and BCGF activity (tested on anti-mu-activated normal B-enriched cells). Absorption of such effluent on PHA-activated T cell blasts removed the IL 2 activity without affecting the BCGF activity. The eluate also displayed BCGF activity (tested on anti-mu-activated B-enriched cells) without detectable IL 2 activity. The two BCGF species isolated by this criteria synergize with semipurified IL 1 to support the anti-mu-induced proliferation of highly monocyte-depleted B cells. Both BCGF species required the presence of anti-mu Ab to support the proliferation of small B cells (at the G0 stage). However, in contrast to crude conditioned medium and to the BCGF present in the effluent, the BCGF present in the eluate was unable to induce the proliferation of unfractionated or large (at the G1 stage) B cells in the absence of anti-mu Ab. These results support the existence of two functionally different human BCGF species, differing at least in their relative sugar content and distinct from IL 2.
T3 monoclonal antibody activation of nonspecific cytolysis: a mechanism of CTL inhibition.
Authors: Mentzer S J , Barbosa J A , Burakoff S J
Published: January 2023

The T3 antigen is expressed on all cytotoxic T lymphocytes (CTL). Monoclonal antibodies (MAb) to the T3 antigen previously have been shown to inhibit CTL-mediated killing of cells expressing the relevant target antigens. The mechanism of T3 MAb inhibition, however, remains undefined. In this report, we describe a novel effect of the T3 MAb: the stimulation of allospecific CTL clones to kill target cells that do not express the relevant HLA antigens. The stimulation of nonspecific killing was seen only with MAb to the T3 antigen; MAb to other function-associated antigens (e.g., LFA-1, LFA-2, LFA-3, T4, T8, HLA-A,B,C, and DR) had no effect. T3 MAb stimulated nonspecific killing by CTL clones expressing both the T4+ and T8+ phenotype and by CTL clones specific for both class I and class II HLA alloantigens. Target cell susceptibility to T3 MAb stimulated killing was variable. CTL clones lysed some target cell lines very efficiently (e.g., K562, Daudi, and M124.1) but lysed other cell lines much less efficiently (e.g., 23.1, Mann, and L cells). In CTL-mediated cytotoxicity assays with target cells expressing the relevant HLA antigens, T3 MAb demonstrated the expected inhibition of cytolysis. Thus, the ability of T3 MAb to stimulate and inhibit CTL-mediated cytolysis suggests that both effects may be the result of a common mechanism of activation.
Modulation of macrophage-mediated tumoricidal activity by neuropeptides and neurohormones.
Authors: Koff W C , Dunegan M A
Published: January 2023

Recent evidence has suggested that stress may suppress the immune system and increase the frequency and severity of viral and neoplastic disease. The mechanisms for stress-induced modulation of immune function are unclear, but several neuropeptides are thought to be involved. Because macrophages play an important role in the host defense against infection and neoplasia, several stress-related neuropeptides were screened in efforts to determine whether these substances affect macrophage-mediated tumoricidal activity. Adrenocorticotropin and noradrenaline each completely blocked the capacity of mouse recombinant interferon-gamma (INF-gamma) to activate murine peritoneal macrophages to a tumoricidal state as measured by the lysis of 125I-UdR-labeled melanoma target cells. Vasoactive intestinal peptide significantly potentiated the suppressive effects of noradrenaline. In contrast, neurotensin markedly enhanced the cytolytic capability of peritoneal macrophages activated with INF-gamma. Several other neuropeptides, including substance P, alpha-endorphin, beta-endorphin, Leu-enkephalin, and Met-enkephalin, had no effect on macrophage activation. These findings demonstrate that selected stress-related neuropeptides and neurohormones significantly modulate the capacity of macrophages to attain a tumoricidal state and suggest that alteration of macrophage function by neuropeptides may be a prominent feature of stress-induced enhancement of neoplastic disease.
E receptor-related immunosuppressive factor in malignant pleural fluid and plasma: molecular mechanism of action on DNA-polymerase-alpha.
Authors: Oh S K , Lapenson D P
Published: January 2023

We previously purified a potent serum suppressor factor from malignant ascites fluid and showed that it had serologic cross-reactivity with E receptor of human T lymphocytes. We termed this factor "suppressive E receptor factor" (SER). Subsequent studies on SER showed that SER interfered with the production of interleukin 1 and 2 as well as interfering with their activities on target cells. However, SER was not directly cytotoxic to lymphocytes. In this study, we compared the inhibitor of DNA-polymerase (IDP) activity with the suppressive activity on phytohemagglutinin-induced DNA synthesis on intact cells. These two activities were closely correlated (with a linear correlation coefficient of 0.988) even with the whole plasmas derived from cancer patients. Fractionation and purification of IDP activity identified it with SER of a similar potency. Therefore, SER appeared to exhibit its potent immunosuppressive effect via its direct interference on DNA-polymerase activity. Furthermore, the DNA-polymerase inhibitory activity of SER appeared to be specific to DNA and it did not affect the RNA-polymerase activity. SER inhibition of DNA polymerase activity with respect to DNA primer as well as with the nucleotide substrate. Direct inhibition on DNA-polymerase-alpha activity may be one of the possible mechanisms of action of SER on lymphocyte proliferation.
A role for helper cell subpopulations in a genetically controlled cytolytic T lymphocyte response to the H-2Db antigen.
Authors: French D L , Plate J M
Published: January 2023

The primary cytolytic T lymphocyte (CTL) response to the H-2Db antigen in several strain combinations appears to be under genetic control. Our studies were undertaken to determine the mechanisms involved in responders that were absent from the nonresponders and that led to CTL generation. In these studies, H-2Dk-anti-H-2Db combinations served as CTL responders, and H-2Dd-anti-H-2Db combinations served as CTL nonresponders. The nonresponsiveness of the H-2Dd-anti-H-2Db strain combinations appeared to be due to the inability of the Db antigen to activate a helper cell subpopulation. The activation of this helper cell subpopulation, as demonstrated in the H-2Dk-anti-H-2Db response, resulted in the production of factors that led to the induction of CTL differentiation. Antigen-specific pre-CTL for Db were present in the nonresponders as well as in the responders. Interleukin 2 (IL 2)-producing cells were also present in both nonresponders and responders, because all strain combinations tested in this system produced detectable levels of IL 2. Additional analysis of these data suggested that different determinants on the H-2Db antigen were recognized by distinct populations of cells. The activation of pre-CTL and IL 2-producing cells by the recognition of determinants on the H-2Db antigen was not sufficient for the generation of effector CTL, as demonstrated in the H-2Dd-anti-H-2Db response. We suggest that determinants on the H-2Db antigen that are distinct from those recognized by pre-CTL and IL 2-producing cells are recognized by a helper cell subpopulation in the H-2Dk strains, and that activation of this subpopulation is required for the production of factors that mediate CTL differentiation. This is the first description of the role that CTL differentiation factors play in a genetically controlled response.