bluuebunny/crossref_metadata_2025 huggingface dataset:
Stimulation of endothelial cell binding of lymphocytes by tumor necrosis factor.
Authors: Cavender D , Saegusa Y , Ziff M
Published: December 2022

Lymphokines and monokines have been reported to affect endothelial cell (EC) morphology and function. In experiments here described, we have demonstrated that recombinant tumor necrosis factor (TNF) stimulates the adhesion of T lymphocytes to confluent monolayers of human umbilical vein EC. The increase in adhesion induced by TNF was EC-specific inasmuch as preincubation of the lymphocytes with TNF did not alter binding, and preincubation of human dermal fibroblasts with TNF did not increase their inherently low adhesiveness for lymphocytes. Stimulation of T-EC binding occurred after treatment of the EC with as little as 0.01 U/ml (1 pg/ml) of TNF. In kinetic experiments, preincubation of EC with TNF for 4 hr resulted in optimal adhesion. TNF-treated EC retained their increased adhesiveness after fixation with paraformaldehyde, suggesting that TNF stimulated binding by increasing the expression or accessibility of EC surface receptors for lymphocytes. Although antibodies to the lymphocyte function-associated antigen 1 alpha- or beta-chains on the T cell markedly inhibited unstimulated T-EC binding, such antibodies had no effect on the increase in EC adhesiveness induced by TNF, indicating that the increased binding resulted from the generation of an alternate binding receptor on the EC membrane. These findings provide additional evidence that cytokines participate in the mobilization of mononuclear cells in the chronic inflammatory reaction by stimulation of the adhesiveness of endothelium for circulating lymphocytes.
The in vivo effects of interleukin 1. I. Bone marrow cells are induced to cycle after administration of interleukin 1.
Authors: Neta R , Sztein M B , Oppenheim J J , Gillis S , Douches S D
Published: December 2022

We have previously reported that interleukin 1 (IL-1) administration 20 hr before irradiation protects mice from lethal effects of radiation. The recovery of total nucleated bone marrow cells and of hematopoietic progenitor cells was enhanced in IL-1 treated, as compared to untreated, irradiated mice. This suggested that IL-1 administration may affect the cells in the bone marrow of normal mice. Intraperitoneal administration of recombinant IL-1 resulted in bone marrow cell enlargement and increased cycling of these enlarged cells. In addition, the capacity of bone marrow cells from IL-1 treated mice to proliferate in response to granulocyte macrophage-colony-stimulating factor (GM-CSF) in cell suspension cultures was enhanced. The above effects were not genetically restricted as C57BL/6, B6D2F1, C3H/HeN, and C3H/HeJ mice showed similar responses. A comparative study showed that 100 ng of IL-1 was much more effective in stimulating bone marrow cells by the above criteria than 5 micrograms GM-CSF. Since IL-1, unlike CSF, can not be demonstrated to have a direct in vitro stimulatory effect on bone marrow cells, the aforementioned in vivo effects of IL-1 are presumably mediated by other hematopoietic growth factors. We have previously shown that IL-1 induces the appearance of high titers of CSF in the serum. Consequently hematopoietic growth factors that are generated at local sites following IL-1 administration may mediate the observed cell cycling effect.
Induction of interleukin 2 receptor (TAC) by tumor necrosis factor in YT cells.
Authors: Lee J C , Truneh A , Smith M F , Tsang K Y
Published: December 2022

The effect of human recombinant tumor necrosis factor alpha (TNF-alpha) and interleukin 1 beta (IL-1 beta) on interleukin 2 receptor (IL-2R) expression on YT cells was examined. IL-2R expression was assessed by flow cytometric analysis with a monoclonal antibody to IL-2R (anti-TAC). TNF-alpha, like IL-1 beta, induced increased levels of IL-2R expression on YT cells with similar kinetics of induction. Maximum induction occurred at 20 to 30 hr. On a molar basis. TNF was less active than IL-1 beta. RNA isolated from TNF-alpha- or IL-1 beta-treated YT cells contained increased levels of IL-2R-specific mRNA as indicated by slot blot analysis by using an IL-2R-specific mRNA probe. Kinetic and IL-1 beta mRNA expression studies indicated that the TNF effect was a direct one. Because IL-2R expression is known to be associated with lymphocyte activation, the present results suggest that TNF-alpha may play a role in the regulation of immune responses.
Independent expression of a regulatory idiotype on heavy and light chains. A further immunochemical analysis with anti-heavy and anti-light chain antibodies.
Authors: Zanetti M , Rogers J
Published: December 2022

The heavy (H) and light (L) chains of murine monoclonal autoantibody 62 reacting with thyroglobulin independently express idiotypic (Id) determinants that are very similar if not identical with the Id62 expressed on the intact protein. In this report, we describe the production and characterization of rabbit antibodies to isolated H62 and L62 chains to further prove that individual chains express Id62 in an immunogenic form. The results demonstrate that both chains are capable of eliciting antibodies that, after appropriate adsorption, behave like conventional anti-Id62 antibodies prepared against the intact antibody molecule. By direct radioimmunoassay binding, competition of Id binding and Western blot anti-H62 and anti-L62 antibodies identify as Id-positive the same group of IgG1, bind in a reciprocal fashion to H- and L-chains of parental monoclonal antibody 62, and detect Id62-positive polyclonal serum autoantibodies to thyroglobulin. We conclude that monoclonal antibody 62 expresses independently a similar Id on both polypeptide chains and the intact antibody molecule, or its isolated chains, induce qualitatively similar anti-Id responses. These results are discussed in light of the possible structure/function implication such autoantibodies may have within the Id network.
Isotype concentrations of human antibodies to Haemophilus influenzae type b polysaccharide (Hib) in young adults immunized with the polysaccharide as such or conjugated to a protein (diphtheria toxoid).
Authors: Mäkelä O , Mattila P , Rautonen N , Seppälä I , Eskola J , Käyhty H
Published: December 2022

Antibody responses of young adults to Haemophilus influenzae type b polysaccharide (Hib) or its protein conjugate were studied with special attention to the isotype composition of the antibodies. Three conclusions of interest can be made: 1) Immunoglobulin G (IgG) antibodies in polysaccharide-immunized volunteers displayed the subclass pattern previously found in antibodies to meningococcal type A polysaccharide. IgG1 was the predominant subclass in IgG antibodies of some individuals, IgG2 in others. Still others had the two subclasses in varying but more even proportions. 2) The conjugate vaccine induced a geometric mean response 2 to 3 times higher and an IgG response 4 times higher to Hib than the polysaccharide vaccine. 3) Anti-Hib antibodies induced by the conjugate vaccine still had essentially the same IgG subclass composition as anti-Hib antibodies induced by the polysaccharide. This composition was strikingly different from the composition of the anti-diphtheria toxoid response induced by the same conjugate vaccine.
Intrahepatic, nucleocapsid antigen-specific T cells in chronic active hepatitis B.
Authors: Ferrari C , Penna A , Giuberti T , Tong M J , Ribera E , Fiaccadori F , Chisari F V
Published: December 2022

Hepatitis B core antigen (HBcAg)-specific T cell lines were established from hepatic lymphomononuclear cells derived from five patients with chronic active hepatitis B. No hepatitis B virus envelope antigen-specific cell lines were established. Proliferation in response to recombinant and native HBcAg, but not to native hepatitis B surface antigen containing the pre-S(2) region, confirmed the specificity of the five T cell lines. All cell lines represented mixed populations of CD4+ and CD8+ T cells. The CD4+ subset provided antigen-specific help to autologous B cells with respect to anti-HBc production and to CD8+ cells with regard to HBcAg-induced proliferation and suppressor activity. The CD8+ subset contained suppressor cells that selectively inhibited the proliferative response of autologous HBcAg-specific CD4+ cells without inhibiting CD4+ cells of unrelated specificity (tetanus toxoid). Moreover, the CD8+ cells were also capable of suppressing HBcAg-stimulated antibody to HBcAg production without showing inhibition of total immunoglobulin production stimulated by pokeweed mitogen. The cytotoxic potential of the T cell lines was established in a lectin-dependent cytotoxicity system; natural killer cytotoxicity was completely absent. Our data suggest that the lesional T cells present at the site of hepatocellular injury in chronic active hepatitis B are primarily HBcAg-specific lymphocytes of the helper and suppressor/cytotoxic phenotypes and that both are functionally competent.
Immunosuppressive activity of T cell clones generated from human T cells stimulated with autologous TPHA cells.
Authors: Gulwani B , Maio M , Imberti L , Melamede R J , Ferrone S
Published: December 2022

T cell clones were generated from human T cells stimulated with autologous phytohemagglutinin (PHA)-activated T (TPHA) cells. Characterization of three T cell clones originated from donor SF and one from donor JM showed that they proliferated when stimulated with autologous TPHA cells, non-T cells, and peripheral blood mononuclear cells, but did not proliferate when stimulated with allogeneic TPHA cells, non-T cells, and mononuclear cells, with autologous and allogeneic resting T cells, and with PHA. These results in conjunction with the blocking of the proliferation by anti-histocompatibility leukocyte antigen class II monoclonal antibodies indicate that these class II antigens are involved in the proliferation of T cell clones stimulated with autologous lymphoid cells. The four T cell clones are cytotoxic neither to autologous lymphoid cells nor to a panel of cultured human cell lines. The four T cell clones display immunosuppressive activity, since they inhibit the proliferation of autologous and allogeneic cells stimulated with antigens and mitogens and the secretion of immunoglobulin by B cells stimulated with pokeweed mitogen in presence of T cells. Furthermore, the four T cell clones display differential inhibitory activity on the proliferation of cultured human cell lines. The immunosuppressive activity is species-specific, since the T cell clones do not inhibit the proliferation of murine cells. The suppression is mediated by a factor(s) with an apparent m.w. of 13,000 to 16,000. The suppressor activity is labile at alkaline pH and is lost following incubation with pronase (100 U/ml) for 30 min at 37 degrees C.
Differential expression of nuclear proto-oncogenes in T cells triggered with mitogenic and nonmitogenic T3 and T11 activation signals.
Authors: Shipp M A , Reinherz E L
Published: December 2022

The relationship between induction of nuclear proto-oncogenes and cellular proliferation is not fully understood. To better define this relationship, we have studied c-fos, c-myc, and c-myb mRNA induction in T lymphocytes where early and late activation events have been clearly delineated. In T cells, initial activation from G0 to G1 results from stimulation of either the antigen/major histocompatibility complex receptor (T3-Ti) or the T11 structure; further cycle progression and proliferation follow interaction of interleukin 2 (IL-2) with the IL-2 receptor. These events can be dissected with monoclonal antibodies to T3 or T11 which cause early activation but differ in their ability to initiate IL-2-dependent cycle progression and proliferation. In T lymphocytes triggered through either T3-Ti or T11, c-fos is induced with a nonmitogenic activation signal whereas c-myb is only induced with a mitogenic signal capable of triggering IL-2 and IL-2 receptor expression. Furthermore, c-myc induction is biphasic and associated with both early and late activation events. Early c-myc, like c-fos, is induced with a nonmitogenic signal. In contrast, induction of late c-myc, like that of c-myb, requires a mitogenic signal. Thus, appearance of c-fos and initial c-myc mRNA seem to be early responses to membrane signaling whereas late c-myc and c-myb are more directly associated with actual cellular proliferation. That nonmitogenic stimulation of T cells via T3-Ti not only abrogates T11-mediated proliferation but also eliminates late c-myc and c-myb transcription further supports this notion.
Murine interstitial nephritis. VI. Characterization of the B cell response in anti-tubular basement membrane disease.
Authors: Clayman M D , Michaud L , Neilson E G
Published: December 2022

In the present study we have examined the murine B cell response in anti-tubular basement membrane (alpha TBM) disease. Whereas only certain strains of mice are susceptible to the development of interstitial lesions after immunization with heterologous renal tubular antigen, all strains make anti-tubular basement membrane antibodies (alpha TBM-Ab), and all express the 3M-1 kidney antigen which is the target of disease. The magnitude of the alpha TBM-Ab response in serum and renal eluates, measured by radioimmunoassay against crude tubular antigen or affinity-purified 3M-1, also mapped independently of susceptibility. The fine specificity of epitope binding was further analyzed using a rat monoclonal alpha 3M-1 antibody to competitively inhibit the binding of renal eluate antibody to 3M-1. Maximum inhibition among nearly all tested strains ranged from 46 to 56% with no discernible difference between susceptible and nonsusceptible mice. Idiotypic representation of renal eluate alpha TBM-Ab was then evaluated by competitive inhibition using a polymorphic anti-idiotypic antisera. All mice examined possessed almost identical competitive inhibition patterns, indicating surprisingly similar idiotypic representation. Thus, in susceptible or nonsusceptible mice, neither the quantitative alpha TBM-Ab response, the epitopic fine specificity of that response, nor the idiotype of eluted alpha TBM-Ab serve as distinguishing markers for susceptibility to interstitial injury. Finally, passive transfer experiments with high-titered (greater than 1:10,000) alpha TBM-Ab from SJL mice were performed to test the hypothesis that alpha TBM-Ab alone may be sufficient for the induction of alpha TBM disease. Whereas this antiserum was capable of causing typical, severe alpha TBM disease in naive susceptible SJL mice, this treatment in allotype-identical, nonsusceptible B10.S(8R) mice was completely without effect. These data demonstrate, in conclusion, that, in the absence of appropriate susceptibility genes, alpha TBM-Ab are incapable of causing alpha TBM disease. The findings support previous observations that the ability of passively transferred alpha TBM-Ab to initiate interstitial injury is dependent on the host also expressing other susceptibility genes which promote the cooperative engagement of the cell-mediated immune response.
Antigen-induced human interferon-gamma production. Differential dependence on interleukin 2 and its receptor.
Authors: Kelly C D , Welte K , Murray H W
Published: December 2022

Once stimulated with Toxoplasma gondii or cytomegalovirus (CMV) antigens, peripheral blood mononuclear cells from healthy seropositive donors secrete comparable levels of interferon-gamma (IFN-gamma). Both antigens also stimulated specific production of interleukin 2 (IL-2), a lymphokine believed to be important in IFN-gamma generation. T. gondii antigen, however, induced ninefold more IL-2 than did CMV antigen suggesting different mechanisms for antigen-stimulated IFN-gamma production. Therefore, we examined for both antigens 1) the cellular sources of IL-2 and IFN-gamma, 2) the kinetics of IL-2 production and IL-2 receptor (IL-2R) expression, and 3) the effect of antibodies to IL-2 and IL-2R on IFN-gamma secretion. For both antigens, IL-2 and IFN-gamma secretion was T4+ cell-dependent. T. gondii antigen induced high levels of IL-2 at 24 hr which increased further at 48 hr, and IFN-gamma production was strongly inhibited by antibodies to both IL-2 (90 +/- 2%) and IL-2R (80 +/- 5%). In contrast, CMV antigen stimulated low levels of IL-2 at 24 hr which declined still further by 48 hr, and CMV-stimulated IFN-gamma generation was appreciably less well inhibited by antibodies to IL-2 (47 +/- 2%) and IL-2R (31 +/- 8%). These results suggest the possibility of two mechanisms for antigen-induced IFN-gamma production--one primarily dependent on and the other largely independent of IL-2 and its receptor. Both mechanisms, however, require the activity of sensitized T4+ cells.
Characterization of serologic and cell-mediated reactivity of a 38-kDa antigen isolated from Mycobacterium tuberculosis.
Authors: Kadival G V , Chaparas S D , Hussong D
Published: December 2022

An antigen of Mycobacterium tuberculosis with an m.w. of 38,000 has been isolated by affinity chromatography using a monoclonal antibody. This antibody bound only to an antigen found in M. tuberculosis and Mycobacterium bovis BCG. The specificity of the antigen was tested in a vertical study by immunodetection on western blots reacted with hyperimmune sera against M. tuberculosis, M. bovis, and 10 other Mycobacterium species. The antigen was detected only by antisera to M. tuberculosis and M. bovis. Specificity in cell-mediated immunity was tested by skin tests in guinea pigs sensitized with M. tuberculosis, Mycobacterium intracellulare, and Mycobacterium kansasii and by lymphocyte proliferation tests. The 38-kDa antigen induced positive skin test reactions regardless of the Mycobacterium species used to sensitize the animal. The ability of the 38-kDa antigen to sensitize for cell-mediated immunity was tested by injecting mice with the 38-kDa antigen and challenging their lymphocytes in vitro with various mycobacterial antigens. Lymphocyte proliferation was observed in the presence of 38-kDa antigen, M. tuberculosis sonicate antigen, and tuberculin purified protein derivative and to M. kansasii and M. intracellulare. The 38-kDa antigen may contain a specific epitope detected by serology, but also contains epitopes that are cross-reactive for cellular immunity.
Suppressive substance produced by T cells from mice chronically infected with Trypanosoma cruzi. I. Preferential inhibition of the induction of delayed-type hypersensitivity.
Authors: Liew F Y , Scott M T , Liu D S , Croft S L
Published: December 2022

Culture supernatants of spleen cells from susceptible CBA mice chronically infected with Trypanosoma cruzi were able to inhibit the induction of delayed-type hypersensitivity (DTH) to a wide range of antigens as measured by 24-hr footpad swelling, bone marrow homing, and radioactivity accumulation assays. The suppressive activity, which was also present in the serum of these chronically infected mice, appears to be specific for the induction of DTH and had no effect on the 3-hr immediate-type hypersensitivity. It also failed to modify the expression of DTH in presensitized mice. Furthermore, it did not affect the synthesis in normal recipients of specific antibody or the induction of helper T cells or cytotoxic T cells. It also failed to induce DTH tolerance as recipient mice with markedly reduced DTH were able to develop a normal DTH response after secondary immunization. The suppressive activity was produced by an Ig- macrophage-depleted splenic T cell population, whose capacity to secrete the suppressive substance was completely abrogated by treatment in vitro with anti-L3T4 antibody and complement, but not with anti-Lyt-2 antibody and complement. These results therefore demonstrate that L3T4+ T cells from mice chronically infected with T. cruzi can produce substances which interfere with the induction of DTH. This finding may help to identify the differential antigenic stimulatory requirement for the activation of the various subsets of T cells.
Clonal heterogeneity of anti-AKR/gross leukemia virus cytotoxic T lymphocytes. Evidence for two distinct antigen systems.
Authors: Azuma H , Phillips J D , Green W R
Published: December 2022

AKR/Gross leukemia virus-induced tumor reactive cytotoxic T lymphocyte (CTL) clones were derived from C57BL/6 spleen cells. Analysis of their specificity pattern was performed by using a panel of target cells such as E male G2 and AKR.H-2bSL1 (susceptible tumors to polyclonal anti-AKR/Gross virus CTL), and cl. 18-5 and cl. 18-12 (insusceptible variant sublines derived from AKR.H-2bSL1). Several of these CTL clones were selected for further study. Lysis of Gross cell surface antigen-positive tumor cells by these clones was restricted by the H-2Kb molecule. The cell surface phenotype of these clones was Thy-1.2+, Lyt-2.2+, L3T4-, a phenotype consistent with that of polyclonal anti-AKR/Gross CTL, suggesting that they were of conventional CTL origin. According to their fine specificity pattern, the CTL clones were divided into two major groups (A and B) which were further subdivided into five and three subgroups, respectively. The specificity of group A clones was essentially the same as that of the standard polyclonal CTL population except for a variable level of natural killer-like activity by some of the CTL clones. That is, group A clones did not efficiently lyse the insusceptible variant tumors nor any of Friend-Moloney-Rauscher-positive tumors tested, but they showed strong lytic activity to susceptible tumors and iododeoxyuridine-treated insusceptible variants. Thus, their CTL activity appeared to be strictly directed to Gross cell surface antigen-positive tumors that are susceptible to polyclonal anti-AKR/Gross virus CTL. In contrast, group B clones could lyse both susceptible and insusceptible variant tumors and also a Friend virus-induced tumor (FBL3). Therefore, as defined by these CTL clones, at least two distinct antigenic systems (A and B), each with several antigenic determinants, appeared to be present. Because recent findings suggested that most of the polyclonal anti-AKR/Gross virus CTL activity appeared to be directed to N-ecotropic proviral determinants, we further investigated the nature of these two antigenic systems by use of additional target cells including lipopolysaccharide (LPS)-stimulated spleen cell blasts from AKXL recombinant inbred strains and retrovirus-infected fibroblasts. Group A clones could lyse all LPS blasts derived from AKXL recombinant inbred strains containing the AKV-1 proviral genome, but lysed only very insufficiently or did not lyse AKV-1-negative blasts containing the AKV-3 and/or AKV-4 provirus.(ABSTRACT TRUNCATED AT 400 WORDS)
Anti-NOR 90. A new autoantibody in scleroderma that recognizes a 90-kDa component of the nucleolus-organizing region of chromatin.
Authors: Rodriguez-Sanchez J L , Gelpi C , Juarez C , Hardin J A
Published: December 2022

We identified a patient (CAG) with scleroderma whose serum contained a high titer of IgG class antibodies that stained nucleoli in a pattern of independent tiny spots. When tested on isolated chromosomes, these antibodies selectively stained the nucleolus-organizing regions (NOR) of chromosomes 13, 14, 15, 21, and 22. These staining patterns were not altered when substrate cells and chromosomes were treated with RNase, 0.1 M HC1, or 4 M urea, but they were abolished by treatment with DNase and trypsin. Immunoblots performed with serum CAG on isolated nucleolar substrates identified a protein antigen of approximately 90 kDa. Antibodies affinity-purified from this protein selectively stained nucleoli and NOR chromosomal regions. Therefore, this protein is the antigen that accounts for the ability of serum CAG to recognize the NOR. In a search for the NOR 90-kDa specificity among 254 patients with various rheumatic diseases, we found nine additional patients whose sera stained metaphase chromosomes selectively at the NOR. Sera from five of them (three with scleroderma, two of unknown diagnosis) recognized a protein that electrophoretically co-migrated with the CAG antigen. Thus, scleroderma is present in at least four of six who appear to have this specificity. We conclude that autoantibodies to the NOR 90-kDa antigen have an association with scleroderma and may be useful diagnostically and as a probe for further studies of the biology of the cell nucleolus.
Lupus anticoagulant and antiplatelet properties of human hybridoma autoantibodies.
Authors: Rauch J , Meng Q H , Tannenbaum H
Published: December 2022

The clinical association of lupus anticoagulant antibodies with thrombocytopenia and thrombosis was the rationale for investigating the in vitro reactivity of these human hybridoma lupus anticoagulant antibodies with platelets. Fifty human hybridoma antibodies from 13 patients with systemic lupus erythematosus, 2 women with multiple spontaneous abortions, and 4 normal individuals were analyzed for lupus anticoagulant, antiplatelet, anti-DNA, and antiphospholipid reactivities. Of the hybridoma antibodies studied, 25 had lupus anticoagulant activity, 21 had antiplatelet reactivity, and 7 of these antibodies had both lupus anticoagulant and antiplatelet properties. No correlation was found between lupus anticoagulant antibody activity and antiplatelet, anti-denatured DNA, anticardiolipin, anti-egg phosphatidylethanolamine, antiphosphatidylserine, antiphosphatidylinositol, and antiphosphatidylcholine reactions. In contrast, antiplatelet activity was strongly correlated with antiphosphatidylethanolamine (rho = 0.761, p less than 0.001), anticardiolipin (rho = 0.748, p less than 0.001), and anti-dDNA (rho = 0.745, p less than 0.001) reactivities. Pretreatment of platelets with deoxyribonuclease, ribonuclease, trypsin, or phospholipases A2 and C resulted in different effects on the binding of individual hybridoma antibodies to platelets, suggesting that antiplatelet antibodies may recognize different epitopes on the platelet membrane. Our data demonstrate that most hybridoma lupus anticoagulant antibodies did not bind directly to platelets in vitro. This suggests that additional serum factors may be required in vivo to explain the association of these antibodies with thrombocytopenia and thrombosis.
C5a induction of human interleukin 1. Synergistic effect with endotoxin or interferon-gamma.
Authors: Okusawa S , Dinarello C A , Yancey K B , Endres S , Lawley T J , Frank M M , Burke J F , Gelfand J A
Published: December 2022

Interleukin-1 (IL-1) is a potent cytokine which possesses the ability to mediate systemic acute phase responses as well as local tissue inflammation. In these studies, we have examined the ability of C5a and C5a des Arg to induce IL-1 production in vitro. Human C5a and C5a des Arg were purified to homogeneity and were found to stimulate IL-1 release from freshly obtained human mononuclear cells into the extracellular medium. Only 2 hr of exposure to the purified complement components were necessary in order to stimulate IL-1 production. The minimal concentration of C5a required was 25 ng/ml, whereas 125 ng/ml of C5a des Arg induced comparable amounts of IL-1. This dose relationship was maintained at higher concentrations (150 ng/ml vs 750 ng/ml, respectively). That the effect was due to the anaphylatoxins themselves, and not endotoxin contamination, was shown by negative Limulus amebocyte lysate tests and employing preincubation of C5a/C5a des Arg with polymyxin B. The latter blocked a wide dose range of endotoxin-stimulated IL-1 production. However, when endotoxin was added to C5a or C5a des Arg, significant synergism in the stimulation of IL-1 production was observed, occurring at various concentrations of either agent. A similar synergism with C5a/C5a des Arg was seen with interferon-gamma. In these studies, IL-1 production was measured by bioassay employing cloned D . 10 . G4 . 1 murine T cells and by radioimmunoassay for human IL-1 beta; using C5a/C5a des Arg as stimulants, there was a high degree of correlation (r = 0.82) between the two assays. Since traumatic, infectious, and inflammatory diseases may result in the simultaneous appearance of these stimuli, the synergism described herein is likely to be clinically relevant.
Interferon-gamma, interferon-alpha/beta, and tumor necrosis factor differentially affect major histocompatibility complex class I expression in murine leukemia virus-induced tumor cell lines.
Authors: Klyczek K K , Murasko D M , Blank K J
Published: December 2022

Tumor cell lines induced by Gross murine leukemia virus were examined for cell-surface major histocompatibility complex class I expression. Three of five cell lines constitutively express H-2K and H-2D class I protein. Culturing these cells with interferon (IFN)-gamma, IFN-alpha/beta, or tumor necrosis factor increases both K and D expression in these cell lines. Two of five tumor cell lines express no class I proteins by fluorescence-activated cell sorter analysis, specific immunoprecipitation, and specific hybridization in Northern analysis. Treatment with IFN-gamma induces D, but not K protein expression in one of these cell lines. IFN-alpha/beta and tumor necrosis factor induce neither D nor K expression in this cell line. Thus, these two cytokines appear to have different mechanisms of action than IFN-gamma for altering class I expression. The other class I-negative tumor cell line does not express either K or D proteins under any conditions tested. All five cell lines express beta 2-microglobulin; this expression is increased by IFN-gamma treatment even in cell lines which do not express class I heavy chain. The results of this study demonstrate that 1) different tumor cell lines demonstrate variations in class I gene regulation, and 2) differences in regulation between class I genes may occur within a single cell line.
Immunoglobulin chain recombination among antidigoxin antibodies by hybridoma-hybridoma fusion.
Authors: Hudson N W , Mudgett-Hunter M , Panka D J , Margolies M N
Published: December 2022

Conditions necessary for in vitro chain recombination of high affinity (10(9) to 10(12) M-1) antidigoxin monoclonal antibodies resulted in decreased affinity for both intact "native" and chain recombinant molecules. Chain recombination by somatic cell fusion was used instead to study the effects on antigen specificity and idiotypy of recombinants in which an homologous light (L) chain substituted for the parental L chain. The antidigoxin antibody 26-10 utilizes a VL sequence highly homologous to that of antibody 40-20, an antidigoxin antibody which uses a different VH gene than does 26-10 and lacks significant reactivity with an anti-26-10 idiotypic serum. The drug-marked antidigoxin cell line 26-10 (gamma 2a, kappa) and a drug-marked light chain producing variant of antidigoxin hybridoma 45-20 (lambda 1) which lacks both digoxin binding and idiotypy were fused. The fusion progeny (gamma 2a, kappa, lambda 1) which binds digoxin and is idiotype-positive, was selected for kappa loss (resulting in loss of digoxin and idiotype binding) and then fused with a heavy (H) chain loss variant of antidigoxin hybridoma 40-20 (kappa, digoxin nonbinding, idiotype negative). The resultant cell line CR-57 (gamma 2a, kappa, lambda) secretes antibodies which assemble the 26-10 H chain with both the 40-20 kappa-chain and the 45-20 lambda 1-chain. The affinity purified recombinant species consisting of 26-10 H chain and 40-20 kappa-chain expresses complete 26-10 idiotypic determinants. However, this recombinant antibody binds digoxin with decreased affinity and altered specificity relative to native 26-10. The binding specificity pattern nonetheless is most similar to the H chain donor. Amino acid and nucleotide sequence analyses of the respective light chains demonstrate six variable region differences between them, two of which are in complementarity-determining regions and the remainder in the framework. Hybridoma-hybridoma fusion provides an alternative to in vitro chain recombination for studying the contribution of chain combinational diversity to antibody diversity, antigen binding, and idiotypy.
Regulation of human polymorphonuclear neutrophil (PMN) activity against Candida albicans by large granular lymphocytes via release of a PMN-activating factor.
Authors: Djeu J Y , Blanchard D K
Published: December 2022

Using a 24-hr radiolabel microassay developed in our laboratory that measures [3H]glucose uptake in residual Candida, we have identified the effector cells responsible for in vitro inhibition of Candida albicans growth as mainly polymorphonuclear neutrophils (PMN) and monocytes within the human peripheral blood cells. Highly purified T cells and large granular lymphocytes (LGL) that mediate natural killer activity which were obtained by Percoll density gradient centrifugation were found to have no innate activity against C. albicans. The LGL could not be activated by interferon-alpha, interferon-gamma or interleukin 2 to inhibit Candida growth although their K562 tumor cytotoxic activity was readily enhanced by these cytokines. Stimulation with heat-killed C. albicans also did not activate fungal growth inhibitory function in LGL and the supernatant of these activated LGL had no direct fungicidal activity. However, the activated LGL supernatant had the capability to enhance PMN function against C. albicans growth. Addition of recombinant human tumor necrosis factor, affinity-purified interferon-alpha, or interferon-gamma to PMN caused increased antifungal activity in PMN. However, antibodies to these cytokines had only a partial adverse effect on the ability of the activated LGL supernatant to stimulate PMN anti-Candida function. Therefore, the activated LGL supernatant appeared to contain a potent stimulator of PMN function which is as yet unidentified. These data indicate that LGL did not directly mediate anti-Candida activity but could indirectly influence C. albicans growth by activating PMN against the fungi through the release of a specific PMN-activating factor. Our findings therefore add another role to LGL which is the regulation of PMN function, the consequence of which is regulation of fungal immunity.
Idiotope vaccine against Streptococcus pneumoniae. A precursor study.
Authors: Ward M M , Ward R E , Huang J H , Kohler H
Published: December 2022

An analysis of nominal vs idiotope antigen-induced B cell precursors was performed in A/St mice. With the use of the splenic fragment culture system, the quantity and quality of B cell precursors responding to two anti-idiotope carrier antigens (4C11 hemocyanin and F6 hemocyanin) and nominal antigen (phosphorylcholine-hemocyanin) were compared. In addition, the effect of priming with anti-idiotope-carrier antigens on B cell precursors responding to phosphorylcholine-hemocyanin was determined. We found that one anti-idiotope-carrier antigen, 4C11-hemocyanin, and phosphorylcholine-hemocyanin stimulated similar subpopulations of primary B cells. However, the B cell population stimulated by F6-hemocyanin, the other anti-idiotype complex, was distinct. Furthermore, priming with certain idiotope antigens can direct the phosphorylcholine-hemocyanin response into the expression of idiotypes that may be the most effective in protective immunity. Our results provide essential information for the rational design of idiotype vaccines by clarifying the dynamic relationship of the B cell precursor repertoire with the in vivo antibody response in the response to nominal and idiotope antigens.
The immune response to Schistosoma mansoni infections in inbred rats. VI. Regulation by T cell subpopulations.
Authors: Phillips S M , Walker D , Abdel-Hafez S K , Linette G P , Doughty B L , Perrin P J , el Fathelbab N
Published: December 2022

These studies assess the roles of subpopulations of T lymphocytes in inducing and modulating resistance to Schistosoma mansoni. CDF rats were depleted of RT 7.1+ (anti-Pan-T), W3/25+ (anti-T helper/inducer), or OX8+ (anti-T suppressor) cells by the in vivo administration of monoclonal antibodies (mAb). The development of parasites and immunity to challenge by S. mansoni were compared with results in undepleted normal and congenitally athymic rats. Discrete subpopulations of T lymphocytes were adoptively transferred to ascertain effects upon parasite development and the protective immune response. In vitro studies, involving utilizing cocultivation of cell subpopulations +/- cyclosporin A, were utilized to dissect mechanisms. Depletion of T lymphocytes by anti-RT7.1 mAb and anti-W3/25 mAb resulted in augmented initial worm development, suboptimal resistance, and decreased antibody and delayed-type hypersensitive reactivity directed against schistosome antigens. Depletion with OX8 mAb produced opposite effects. The adoptive transfer of T cell subpopulations produced concordant results with T cell regulation expressed B cell-dependent effector mechanisms. The coadoptive transfer of cells resulted in the suppression of resistance afforded by the W3/25+ cells by OX8+ cells, which could be augmented in vitro by cyclosporin A. Thus, protective immunity to S. mansoni in rats is regulated by discrete subpopulations of T lymphocytes. The findings suggest the possibility of selective immune regulation of resistance based on the manipulation of specific T cell subpopulation.
Genetic analysis of the H-2D region using a new intra-D-region recombinant mouse strain.
Authors: Duran L W , Zeller J C , Lundy J K , Chang-Miller A , Krco C J , David C S , Pease L R
Published: December 2022

A rare D-region recombination event which gave rise to the B10.RQDB major histocompatibility complex haplotype has been examined to ascertain the nature of the crossover and to determine which class I genes are present in the new alignment of D-region genes. Serologic analysis have shown that the B10 . RQDB major histocompatibility complex recombinant mouse inherited the H-2Dd gene from the B10.T(6R) parental line and the H-2Db gene from the B10.A(2R) parental line, representing the first example of an intra-D-region crossover resulting from an intercross. Previous molecular genetic analyses of the d and b haplotypes revealed structural diversity in the organization of their D-region gene clusters. Hence, the D region is comprised of five class I genes in the d haplotype and only one in the b haplotype. Because allelic relationships among the various D-region genes are not defined, either a homologous or nonhomologous alignment of genes has generated the RQDB crossover. Therefore, the possibility that all three D-region antigen-presenting molecules (Dd, Ld, and Db) might be encoded by the RQDB haplotype was examined. Fluorescence-activated cell sorter and cytotoxic T lymphocyte analyses revealed no detectable levels of H-2Ld cell-surface expression, confirming earlier studies with antibody-mediated cytotoxicity and immunoprecipitation. Southern blot analysis localized the recombination point to within a 1-kb region at the centromeric end of the H-2Ld gene on the B10 . T(6R) chromosome in a region of high homology to the H-2Db gene on the B10 . A(2R) chromosome. Together, these studies define the D region of the RQDB haplotype as containing the five class I genes: Dd, D2d, D3d, D4d, and Db. In addition to providing insight into rare recombination events in the D region, the B10.RQDB mouse should be a useful tool for exploring the function of D-region genes.
Purification and characterization of cytostatic lymphokines produced by activated human T lymphocytes. Synergistic antiproliferative activity of transforming growth factor beta 1, interferon-gamma, and oncostatin M for human melanoma cells.
Authors: Brown T J , Lioubin M N , Marquardt H
Published: December 2022

Supernatants from activated human T lymphocytes were highly growth inhibitory for A375 human melanoma cells. Three growth inhibiting polypeptides, transforming growth factor beta 1 (TGF-beta 1), interferon-gamma (IFN-gamma), and oncostatin M, were isolated from the acid-soluble fraction of serum-free T cell-conditioned medium and purified by gel permeation chromatography and reverse-phase high performance liquid chromatography in volatile solvents at acid pH. The purification was monitored in a growth inhibition assay. The release of TGF-beta 1 biologic activity by and the purification of IFN-gamma from the medium of activated human peripheral blood T lymphocytes have been reported. We now describe the isolation of oncostatin M from the conditioned medium of activated human T cells. The concentration of oncostatin M required for half-maximal inhibition of A375 melanoma cells was approximately 4 pM when assayed in the presence of 10% fetal bovine serum. The purified oncostatin M had an apparent m.w. 28,000 and an amino-terminal sequence that was identical with the sequence of oncostatin M isolated from supernatants of macrophage-like cells. Suboptimal concentrations of TGF-beta 1 in combination with suboptimal concentrations of IFN-gamma or oncostatin M resulted in synergistic antiproliferative responses for A375 cells (1.9 and 3.1 times the expected additive responses, respectively). Combinations of oncostatin M and IFN-gamma added simultaneously to A375 cells caused an additive growth inhibitory response. These results demonstrate that oncostatin M is a novel lymphokine, and its interaction with other cytostatic polypeptide growth inhibitors may play a role in the immune regulation of tumor cell growth.
A Yersinia pseudotuberculosis protein which cross-reacts with HLA-B27.
Authors: Chen J H , Kono D H , Yong Z , Park M S , Oldstone M M , Yu D T
Published: December 2022

The most-debated question in the investigation of the spondyloarthropathies has been whether there is molecular mimicry between host HLA-B27 antigens and the arthritis-causing pathogens. We have generated a monoclonal anti-HLA-B27 antibody in our laboratory and have used a radioimmunoassay to screen a panel of bacterial species. Two strains of Yersinia pseudotuberculosis were found to be highly reactive. The cross-reactive Yersinia component was identified by Western blot to be a 19,000 component. A preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis chromatography apparatus was constructed to isolate milligram quantities of this component. To verify that the component carried the HLA-B27-specific epitope, rabbits were hyperimmunized with the purified materials. Affinity-purified antibodies from one of the immunized rabbits indeed carried anti-HLA-B27 activity. Last, antibodies generated against synthetic peptides derived from the HLA-B27.1 amino acid sequence were tested against the Yersinia component. Positive reactivity was found with antibodies generated against a peptide spanning residues 69-83 of the HLA-B27.1 protein. Since this resides in the segment responsible for the allotypic specificity of the antigen, these experiments establish the presence of molecular mimicry to a high degree of confidence.
Independent regulation of human neutrophil chemotactic receptors after activation.
Authors: Bender J G , Van Epps D E , Chenoweth D E
Published: December 2022

The fluoresceinated chemotactic factors, C5a, formyl-methionyl-leucyl-phenylalanyl-lysine (FMLPL), and casein were used in conjunction with flow cytometry to examine chemotactic factor receptor expression on polymorphonuclear leukocytes (PMN) activated with phorbol myristate acetate (PMA), C5a, or formyl-methionyl-leucyl-phenylalanine. Activation with PMA resulted in a dose-dependent increase in binding of fluorescein-labeled (FL)-casein and (FL-FMLPL) over the range of PMA concentrations from 0.5 to 50 ng/ml. In contrast, activation of PMN with PMA resulted in a dose-dependent decrease in FL-C5a binding, and activation with concentrations above 5 ng/ml resulted in a complete loss of binding. This loss of binding was not caused by inactivation of the ligand or prevented by the addition of superoxide dismutase and catalase or protease inhibitors. Furthermore, incubation of PMN with supernatants from PMN stimulated to degranulate did not reduce the availability of C5a receptors. This pattern of increased FMLPL and casein binding with decreased C5a binding was also observed with cytochalasin B-pretreated PMN that were stimulated with chemotactic factors. Parallel studies of superoxide anion generation demonstrated that PMA-treated PMN were still responsive to formyl-methionyl-leucyl-phenylalanine, but not to C5a. These data demonstrate that the activation of PMN up-regulates formyl peptide and casein receptors whereas C5a receptors are down-regulated under similar conditions.
The human beta 2-microglobulin gene. Primary structure and definition of the transcriptional unit.
Authors: Güssow D , Rein R , Ginjaar I , Hochstenbach F , Seemann G , Kottman A , Ploegh H L
Published: December 2022

The human genomic clone pb2m13 contains a functional beta 2-microglobulin (B2m) gene, which upon transfection is readily expressed in murine fibroblasts. Here we report the nucleotide sequence of the human beta 2m gene and of a nearly full length cDNA clone. A comparison with the murine beta 2m gene reveals that exon/intron boundaries are absolutely conserved. In the protein-coding regions the similarity is 70%. As far as intron sequences of the murine beta 2m gene are available, no significant similarity between human and murine genes is observed. The transcriptional start site of the human beta 2m gene was determined by S1 mapping, and comparison with the nearly full length cDNA clone now defines the transcriptional unit of the beta 2m gene. In the 5' region of the gene strong clustering of the usually underrepresented CpG dinucleotide is found resembling a similar overrepresentation in the 5' regions of the major histocompatibility complex class I genes.
The Elimination of Serum Shock
Authors: Kyes Preston , Carey Earl S.
Published: December 2022

The intravenous use of therapeutic sera in man has been seriously hampered by the severe constitutional disturbances which frequently follow the introduction of a serum directly into the blood stream. The disturbances thus produced are variously referred to as “serum shock,” “serum sickness,” “serum reaction,” “serum anaphylaxis” or more briefly as “shock.”

In the intravenous use of an antipneumococcus serum developed by one of us (1) (2) the phenomenon of such shock has been extensively observed and it is the purpose of this paper to discuss the properties of the serum which occasion the shock and to communicate a method by which were are now able to prepare a serum which provokes no shock and which retains its antibody content.

The particular serum upon which the study is based, is an anti-pneumococcus serum of high titer produced by repeated injections of Brahma cockerels with virulent pneumococci.
Cyclosporine blocks the activation of antigen-dependent cytotoxic T lymphocytes directly by an IL-2-independent mechanism.
Authors: Havele C , Paetkau V
Published: December 2022

The immunosuppressive drug Cyclosporin A (cyclosporine) inhibits the reactivation of quiescent Ag-dependent CTL in the presence of IL-2. Both proliferation and the regeneration of cytotoxicity are inhibited. The cytotoxic cells that are inhibited are Ag dependent for activation, whereas derived, Ag-independent, but still IL-2-dependent, cytotoxic cells are insensitive to cyclosporine. Cyclosporine also directly inhibits the effector phase of the cytotoxic cells, although not completely. The generation of primary CTL in mixed cultures was also blocked by cyclosporine in the presence of IL-2, in a time-dependent way that indicated that the sensitive time was early during the cultures. The CTL generated in primary cultures were significantly inhibited by cyclosporine in the assay, but this inhibition was less than for the cloned CTL lines.