bluuebunny/crossref_metadata_2025_split huggingface dataset:
IgE-binding factors from mouse T lymphocytes. III. Role of antigen-specific suppressor T cells in the formation of IgE-suppressive factor.
Authors: Jardieu P , Uede T , Ishizaka K 
Published: January 2023

Abstract
BDF1 mice were given three i.v. injections of ovalbumin (OA) to induce antigen-specific suppressor T cells. Incubation of spleen cells of OA-treated mice with homologous antigen resulted in the formation of IgE-suppressive factor. This factor was not derived from antigen-specific suppressor T cells, but suppressor T cells were essential for determining the nature of IgE-binding factors formed. In the spleen cells of OA-treated mice, antigenic stimulation of antigen-primed Lyt-1+ (helper) T cells resulted in the formation of inducers of IgE-binding factor, whereas Lyt-2+, I-J+ T cells released glycosylation-inhibiting factor (GIF), and these two factors, in combination, induced unprimed Lyt-1+ T cells to form IgE-suppressive factor. The role of GIF is to inhibit the assembly of N-linked oligosaccharides on IgE-binding factors during their biosynthesis, and thereby provide them with a biologic activity: suppression of the IgE response. Under the experimental conditions employed, GIF was released spontaneously from antigen-specific suppressor T cells. However, antigenic stimulation of the cells enhanced the release of the factor. GIF from antigen-specific suppressor T cells has a m.w. of 25,000 to 30,000, as estimated by using gel filtration, binds to anti-I-J alloantibodies and to a monoclonal antibody specific for lipomodulin, and has affinity for specific antigen. The possible relationship between antigen-specific GIF and antigen-specific suppressor factors is discussed.
Inhibition of class I and class II MHC-restricted antigen presentation by cytotoxic T lymphocytes specific for an exogenous antigen.
Authors: Rock K L , Rothstein L , Fleischacker C , Gamble S 
Published: January 2023

Abstract
Ag in the extracellular fluids can be internalized, processed, and presented in association with class I MHC molecules on specialized APC in normal spleen. We examine the fate of these APC after they present Ag to a CTL. When splenocytes present exogenous OVA to CTL, their ability to subsequently present native Ag in association with both class I and class II molecules is inhibited. CTL do not inhibit the ability of splenocytes to present processing independent peptides with class I or class II molecules. Inhibition of Ag presentation is only observed in the presence of the specific Ag recognized by the CTL. This inhibition is MHC-restricted. In the presence of specific Ag, CTL inhibit the ability of APC to present unrelated Ag. However, bystander APC are not affected by activated CTL. Taken together these results indicate that when APC present exogenous Ag to CTL, they are inhibited or killed. The CTL that mediates this activity has a conventional CD4-CD8+ phenotype and utilizes a TCR-alpha beta. The potential significance of these findings and their possible relationship to phenomena associated with Ts cells are discussed.
IL-12 receptor. I. Characterization of the receptor on phytohemagglutinin-activated human lymphoblasts.
Authors: Chizzonite R , Truitt T , Desai B B , Nunes P , Podlaski F J , Stern A S , Gately M K 
Published: January 2023

Abstract
IL-12 is a 75-kDa heterodimeric cytokine composed of disulfide-bonded 35-kDa and 40-kDa subunits. Included among the biologic activities mediated by IL-12 is induction of proliferation of PHA-activated human PBL. The concentration of IL-12 required to stimulate maximum proliferation of PHA-activated lymphoblasts is 50 to 100 pM. In this study, highly purified 125I-labeled IL-12 (7 to 15 microCi/microgram; 50 to 100% bioactive) was used to characterize the receptor for IL-12 on 4-day PHA-activated lymphoblasts. The binding of 125I-labeled IL-12 to PHA-activated lymphoblasts was saturable and specific because the binding of radiolabeled ligand was only inhibited by IL-12 and not by other cytokines. The kinetics of [125I]IL-12 binding to PHA-activated lymphoblasts was rapid at both 4 degrees C and 22 degrees C; reaching equilibrium within 60 min. At 22 degrees C, the rate of dissociation of [125I]IL-12 was slow in the absence of competing IL-12 (t1/2 = 5.9 h) and more rapid in the presence of 25 nM competing IL-12 (t1/2 = 2.5 h). The kinetically derived equilibrium dissociation constant ranged from 10 to 83 pM. Analysis of steady state binding data by the method of Scatchard identified a single binding site with an apparent equilibrium dissociation constant of 100 to 600 pM and 1000 to 9000 sites/lymphoblast. The equilibrium dissociation constant for competing ligands and sites per cell calculated from unlabeled IL-12 competition experiments ranged from 164 to 315 pM and 1067 to 3336, respectively, which is in good agreement with the values determined from steady state binding. The variations in KD and sites per cell were dependent on the individual preparations of lymphoblasts. Although the steady state binding data were consistent with a single class of high affinity binding sites, the kinetic dissociation data indicates a cooperative interaction between receptors on PHA-activated lymphoblasts. Affinity cross-linking of surface bound [125I]IL-12 to PHA-activated lymphoblasts at 4 degrees C identified a major complex of approximately 210 to 280 kDa. Anti-IL-12 antibodies also immunoprecipitated a complex of approximately 210 to 280 kDa that was produced by cross-linking unlabeled IL-12 to 125I-labeled lymphoblast cell-surface proteins. Cleavage of this complex with reducing agent identified one radiolabeled protein of approximately 110 kDa. These data suggest that the IL-12 binding site on PHA-activated lymphoblasts may be composed of a single protein of approximately 110 kDa.(ABSTRACT TRUNCATED AT 400 WORDS)
IL-2 dependence of IL-9 expression in human T lymphocytes.
Authors: Houssiau F A , Renauld J C , Fibbe W E , Van Snick J 
Published: January 2023

Abstract
Human IL-9 is a T cell-derived lymphokine that is abundantly expressed upon activation with mitogens. The observation that IL-9 induction peaks as late as 28 h after stimulation suggested the involvement of secondary signals in this process. The finding reported here that IL-9 expression is blocked by cycloheximide strongly supports this hypothesis. Moreover, we identify IL-2 as the critical element controlling IL-9 expression in T cells. We show (i) that anti-IL-2R antibodies block IL-9 expression in T cells stimulated with PMA and anti-CD3 and (ii) that IL-2, of a panel of cytokines, is the only molecule that synergizes with PMA for IL-9 induction. The latter finding is confirmed in a T cell leukemia line. Finally, we demonstrate that IL-2 plays a regulatory role in the induction of other cytokines, such as IL-4, IL-5, IL-6, and granulocyte/macrophage-CSF, in fresh peripheral T cells.
Identification of two type IIa IgG-binding proteins expressed by a single group A streptococcus.
Authors: Otten R A , Raeder R , Heath D G , Lottenberg R , Cleary P P , Boyle M D 
Published: January 2023

Abstract
Functional heterogeneity associated with Ig-binding proteins expressed by group A streptococci is well documented. In this study we have demonstrated that treatment of group A streptococcal isolate 64/14 with CNBr resulted in the solubilization of two different sized proteins that displayed identical functional reactivity with human IgG1, IgG2, and IgG4 (characteristics of a type IIa binding protein). Monospecific polyclonal antibodies to each form of type IIa molecule were prepared and no antigenic cross-reactivity between the two m.w. forms of type IIa binding protein could be detected. The smaller m.w. protein was shown to be identical or closely related to the recombinant type IIa protein cloned from strain CS110. These studies provide further evidence for the heterogeneity of type II Ig-binding proteins expressed by pathogenic group A streptococci.
Cytotoxic T lymphocytes from HIV-1 seropositive individuals recognize immunodominant epitopes in Gp160 and reverse transcriptase.
Authors: Lieberman J , Fabry J A , Kuo M C , Earl P , Moss B , Skolnik P R 
Published: January 2023

Abstract
The CTL response to HIV-1 is more vigorous than for any known human pathogen and may be a significant factor in preventing the progression to symptomatic disease. T cell lines, generated by non-specific stimulation with PHA and IL-2, may be reproducibly used to identify HIV-1 isolate-invariant epitopes recognized by the CTL of infected individuals. The CTL response in each of 12 infected individuals to envelope and reverse transcriptase (RT) is dominated by the recognition of one or two viral isolate-invariant epitopes. Seven subjects respond to a single gp160 epitope; three subjects recognize 2 gp160 epitopes. There is a significant increase in recognition of epitopes in the C terminal 104 amino acids of gp41 (p less than 0.002); in fact 40% of the subjects that respond to gp160 recognize the C terminal 20-mer. The CTL-mediated lysis of gp160-expressing targets is MHC restricted, but not all individuals that share the same serologically defined class I-restricting element respond to the same epitope. Recognition of the terminal 20mer is restricted by both A30 and B8. The response to RT in six subjects is distributed over the RT protein. The six subjects recognize four separate regions defined by truncated RT-vaccinia recombinants, but none of the subjects' CTL demonstrate significant recognition of the RT epitope identified in H-2k mice and some humans.
Mhc-DQB repertoire variation in hominoid and Old World primate species.
Authors: Otting N , Kenter M , van Weeren P , Jonker M , Bontrop R E 
Published: January 2023

Abstract
Comparison of 87 distinct Mhc-DQB sequences, obtained from 13 primate species, demonstrates that five out of eight trans-species Mhc-DQB allele lineages are at least 30 million years old and predate divergence of hominoid and Old World primate species. One lineage may be much older because its members are not only traced back in higher primates, but also are present in a New World primate species. Comparing Mhc-DQB repertoire variation in distinct species, allows one to pinpoint when certain polymorphisms were lost or gained in primate evolution. Heterogeneity observed among members of trans-species Mhc-DQB allele lineages can be explained in major part by point mutations, whereas intraexonic crossing-over is a potent mechanism in generating new allele lineages. The stability of Mhc-DQB polymorphisms is influenced by selective forces because distinct allele lineages appear to have accumulated nucleotide substitutions and amino acid replacements at different rates.
Evidence for the expression of non-HLA-A,-B,-C class I genes in the human fetal liver.
Authors: Houlihan J M , Biro P A , Fergar-Payne A , Simpson K L , Holmes C H 
Published: January 2023

Abstract
HLA class I Ag expression was investigated in the human fetal liver. By immunostaining, mAb to monomorphic class I Ag showed widespread reactivity with both epithelial and hemopoietic cells. By contrast, mAb to polymorphic determinants showed more restricted reactivity that was confined to a proportion of hemopoietic cells: the hepatic epithelium was essentially unreactive. This suggested that the developing liver might express nonclassical HLA class I. Class I Ag were examined in membrane and cytosol fractions of mid-trimester fetal liver. Because of its broad reactivity with HLA-A,-B, mAb Q1/28 was selected to identify classical class I Ag in these studies. Immunoprecipitations were carried out against radiolabeled glycoprotein extracts of fetal liver membranes. W6/32 detected a 40-kDa product characteristic of nonclassical class I proteins, as well as a 43-kDa product, in lysates immunodepleted with Q1/28. By immunoblotting, an anti-H chain antiserum (HC) identified a Q1/28- 40-kDa component and a 43-kDa Q1/28+ component in fetal liver membrane glycoproteins. The fetal liver cytosol fraction was found to contain a 42- to 43-kDa product by W6/32-chromatography. This component partitioned to the aqueous phase upon condensation in TritonX-114 detergent and by immunoblotting was reactive with monomorphic mAb HC10 but not with Q1/28. Total RNA and polymerase chain reaction amplified class I transcripts of fetal liver were probed using oligonucleotides specific for HLA-E, -F, and -G. HLA-F, was readily detected in total RNAs by Northern analysis. HLA-E, HLA-F, and HLA-G were all detected in fetal liver by polymerase chain reaction. Differential expression of these genes may occur between the first and second trimester of liver development. Overall therefore, the human fetal liver expresses multiple class I protein products and contains transcripts for non-classical class I genes; particularly HLA-F.
IL-7 up-regulates the expression of IL-8 from resting and stimulated human blood monocytes.
Authors: Standiford T J , Strieter R M , Allen R M , Burdick M D , Kunkel S L 
Published: January 2023

Abstract
Blood monocytes are important cellular sources of a vast array of bioactive substances, including regulatory and chemotactic cytokines. The regulation of these cytokines is of critical importance to the expression of acute and chronic inflammatory responses. IL-7, a T and B cell-activating cytokine, has recently been shown to have stimulatory effects on the expression of several monocyte-derived proinflammatory cytokines. We now describe the induction of IL-8 mRNA and extracellular protein from human blood monocytes by IL-7. The up-regulation of IL-8 mRNA by IL-7 was not altered by concomitant treatment with cycloheximide, suggesting that the direct stimulatory effects of IL-7 were not dependent upon de novo protein synthesis. In addition, IL-7 significantly potentiated the production of IL-8 from LPS-, TNF-, and IL-1-treated peripheral blood monocytes. Our findings suggest that IL-7 may play a critical role in the modulation of macrophage-derived cytokine expression and may function in vivo as an important proinflammatory cytokine.
Diphtheria toxin- and Pseudomonas A toxin-mediated apoptosis. ADP ribosylation of elongation factor-2 is required for DNA fragmentation and cell lysis and synergy with tumor necrosis factor-alpha.
Authors: Morimoto H , Bonavida B 
Published: January 2023

Abstract
We have reported that diphtheria toxin (DTX) mediates target cell lysis and intranucleosomal DNA fragmentation (apoptosis) and also synergizes with TNF-alpha. In this paper, we examined which step in the pathway of DTX-mediated inhibition of protein synthesis was important for induction of cytolytic activity and for synergy. Using a DTX-sensitive tumor cell line, we first examined the activity of the mutant CRM 197, which does not catalyze the ADP ribosylation of elongation factor-2 (EF-2). CRM 197 was not cytolytic for target cells and did not mediate intranucleosomal DNA fragmentation of viable cells. The failure of CRM 197 to mediate target cell lysis suggested that the catalytic activity of DTX is prerequisite for target cell lysis. This was corroborated by demonstrating that MeSAdo, which blocks the biosynthesis of diphthamide, inhibited DTX-mediated protein synthesis inhibition and also blocked target cell lysis. Furthermore, the addition of nicotinamide, which competes with NAD+ on the DTX action site of EF-2, also blocked DTX-mediated lysis. These findings suggest that ADP-ribosylation of EF-2 may be a necessary step in the pathway leading to target cell lysis. In contrast to the sensitive line, the SKOV-3 tumor cell line is sensitive to protein synthesis inhibition by DTX but is not susceptible to cytolysis and apoptosis by DTX. Thus, protein synthesis inhibition by DTX is not sufficient to mediate target cell lysis. The synergy in cytotoxicity obtained with the combination of DTX and TNF-alpha was examined in order to determine the pathway mediated by DTX in synergy. Like the direct lysis by DTX, synergy was significantly reduced by MeSAdo and by nicotinamide. Furthermore, synergy was not observed with combination of CRM 197 and TNF-alpha. These results demonstrate that, in synergy, DTX may utilize the same pathway required for its cytolytic activity. Pseudomonas aeruginosa exotoxin shared most the properties shown for DTX. Altogether, these findings demonstrate that DTX-mediated apoptosis is initiated at a step beyond the ADP ribosylation of EF-2.
Mapping the major human T helper epitopes of tetanus toxin. The emerging picture.
Authors: Reece J C , Geysen H M , Rodda S J 
Published: January 2023

Abstract
Progress on the mapping of Th epitopes of tetanus toxin (tt) has been slow due to reliance on studies of clones. In this paper, human Th cell epitopes of tt were mapped using proliferation tests on PBMC in response to synthetic peptides. PBMC from nine donors were tested over the entire set of tt homologous overlapping dodecapeptides. The 1304 peptides were initially tested as 66 pools, each containing an average of 20 peptides. PBMC from individual donors responded to as few as 1 and as many as 17 of the 66 peptide pools. The sequences responsible for proliferation were identified for the two most frequently recognized pools, and for another two pools within a major immunodominant region. Three new epitope sequences were mapped in detail and based on their recognition by most individuals are likely to be promiscuous. A cocktail of peptides including the newly identified Th cell epitopes was able to induce proliferation in PBMC from 24 of 31 tetanus toxoid (TT)-responsive donors. This cocktail is a chemically defined reagent that can be used to quantitate in vitro Ag-specific Th cells in PBMC from most subjects, and may thus be useful for serial measurements of specific immunity such as in subjects undergoing immunotherapy or immunosuppressive treatment.
Inducible nitric oxide synthase from a rat alveolar macrophage cell line is inhibited by nitric oxide.
Authors: Griscavage J M , Rogers N E , Sherman M P , Ignarro L J 
Published: January 2023

Abstract
The objective of this study was to determine whether inducible nitric oxide (NO) synthase from a rat alveolar macrophage cell line (NR8383) activated by LPS plus IFN-gamma could be regulated by NO, one of the two products of the enzymatic reaction. This study was based on previous observations in this laboratory that NO is a negative feedback modulator of constitutive NO synthase from rat cerebellum. NO synthase activity was determined by monitoring the formation of 3H-L-citrulline from 3H-L-arginine in the presence of added cofactors. NO synthase catalyzed the conversion of L-arginine to equimolar quantities of NO and L-citrulline. NO and S-nitrosothiols inhibited NO synthase activity and this effect was enhanced by superoxide dismutase and attenuated by oxyhemoglobin. Nitrite and nitrate, the oxidation products of NO, as well as L-citrulline, the amino acid end-product, produced no significant effects on NO synthase activity. The inhibitory effect of NO on NO synthase appeared to be partially reversible upon addition of oxyhemoglobin. The inhibitory effect of NO was mimicked by other heme ligands including carbon monoxide, cyanide, and manganese-protoporphyrin IX. These observations indicate that (1) enzyme-bound heme plays a mechanistic role in the catalytic conversion of L-arginine to NO plus L-citrulline; (2) NO may function as a negative feedback modulator of inducible NO synthase by interacting with enzyme-bound heme; and (3) negative feedback modulation by NO may represent a mechanism by which the potentially toxic L-arginine-NO pathway in activated alveolar macrophages is turned off.
Tyrosine phosphorylation of a gamma-chain homodimer associated with Fc gamma RIII (CD16) in cultured human monocytes.
Authors: Masuda M , Verhoeven A J , Roos D 
Published: January 2023

Abstract
The efficient expression of transmembrane-anchored Fc gamma RIIIa requires the presence of other peptides, such as the gamma-chain of the IgE receptor I or the zeta-chain of the TCR. We found that Fc gamma RIIIa in cultured human monocytes is specifically associated with the gamma-chain homodimer, and that the gamma-chains in this complex are phosphorylated on tyrosine residues. Anti-Fc gamma RIII immunoprecipitates, which were prepared from 1% digitonin lysates of cultured human monocytes, incorporated phosphate into a homodimer consisting of two 14-kDa polypeptides when incubated with [gamma-32P]ATP. Identity of this co-associated structure of Fc gamma RIIIa as the gamma-chain dimer was confirmed by elution of the protein from the anti-Fc gamma RIII immunoabsorbent with 1% Nonidet P-40 detergent and reimmunoprecipitation with anti-gamma-chain antibody. Phosphoamino acid analysis showed that the gamma-chain exclusively contained phosphotyrosine. The gamma-chain was also phosphorylated when electropermeabilized cells were activated by cross-linking Fc gamma RIIIa. The gamma-chain may play an important role in signal transduction via Fc gamma RIIIa in human macrophages.
Heterogeneity of human sperm surface antigens identified by indirect immunoprecipitation of antisperm antibody bound to biotinylated sperm.
Authors: D'Cruz O J , Haas G G , Lambert H 
Published: January 2023

Abstract
Target Ag recognized by clinically important C-fixing anti-sperm antibodies (ASA) were identified by indirect immunoprecipitation after incubation of biotinylated, motile sperm with ASA-positive sera from autoimmune, isoimmune, and vasectomized patients. This method assessed native protein thus permitting analysis of conformation-dependent membrane Ag that would be missed by previously reported Ag identification techniques. Sera from 35 ASA-positive and 10 ASA-negative patients were exposed to capacitated, surface-biotinylated motile sperm. After detergent-extraction of the membrane fraction, the immune complexes were precipitated with protein G-agarose. The Ag were eluted under reducing conditions, electrophoresed on SDS-10% PAGE, electroblotted onto nitrocellulose membranes, and visualized by the biotin/avidin-peroxidase detection method. By this method, only high titered ASA-positive sera demonstrated variable reactivity restricted to a total of eight to nine protein bands. Individual sera revealed a differential reactivity to a constant set of protein bands (63/61, 58/56, 29, and 21/19 kDa) and a preferential reactivity to either 45-, 43-, or 41-kDa protein Ag that were patient specific. None of these bands were precipitable by ASA-negative sera. ASA directed against the 63/61-kDa and 58/56-kDa polypeptides were found in 85% of immunoprecipitable sera; the 29-kDa was detectable with 70% of sera. Immunoreactivity to the 45-kDa to 41-kDa bands was detectable with 30% of the immunoprecipitable sera. Although the pattern of Ag recognized by ASA from autoimmune, isoimmune, and vasectomized patients was similar, our results indicate that in addition to the recognition of common human sperm surface determinants, distinct sperm surface epitopes are also recognized as foreign by the patient's immune system. Sera that immunoprecipitated the predominant monomeric 29-kDa or the dimeric 58/56-kDa Ag when preincubated with capacitated sperm markedly inhibited sperm binding and penetration to the human zona when compared with ASA-negative serum controls. It is suggested that C-fixing ASA directed to these surface determinants may account for some of the clinical manifestations of ASA-mediated infertility.