bluuebunny/crossref_metadata_2025_split huggingface dataset:
IL-3 alters free arachidonic acid generation in C5a-stimulated human basophils.
Authors: MacGlashan D W , Hubbard W C 
Published: January 2023

Abstract
IL-3 is known to enhance the secretion of several mediators from human basophils activated by receptor-mediated stimuli. IL-3 can cause a qualitative change in the mediator release pattern for C5a-mediated stimulation; without IL-3, C5a causes no leukotriene release whereas in the presence of IL-3, significant leukotriene occurs. This study examines the influence of a 15-min pretreatment of basophils with IL-3 (10 ng/ml) on several signal transduction events. Basophils stimulated with C5a typically displayed only a transient cytosolic Ca2+ response [Ca2+]i, attributed to the release of intracellular calcium stores. IL-3 had sporadic, statistically nonsignificant, effects on the peak of this initial response as well as inconsistent effects on the generation of a second phase in the [Ca2+]i response (i.e., that which is caused by the influx of extracellular Ca2+). IL-3 also had no effect on resting [Ca2+]i levels. Challenge of basophils in the presence of EGTA had little effect on the amount of leukotrienes generated. This maneuver did not influence the initial transient elevation of [Ca2+]i. Although basophil leukotriene release is usually slow, after exposure to IL-3 it was found that leukotriene release occurred rapidly, during the brief window of time (0 to 45 s) in which the [Ca2+]i transient occurred. These results suggested that the generation of AA was accelerated by pretreatment with IL-3. Subsequent mass measurements of free AA by gas chromatography-mass spectroscopy showed: 1) IL-3 itself caused no free AA generation; 2) without IL-3, C5a stimulated little or no free AA generation; and 3) in the presence of IL-3, C5a generated substantial levels of free AA at an accelerated rate. A similar acceleration in the rate of free AA generation, without any apparent increase in the amount, occurred in IL-3-primed basophils stimulated with FMLP. Additional studies showed that IL-3 had no effects on whole cell ATP levels and that the kinase inhibitor, staurosporine, did not inhibit the ability of IL-3 to cause enhanced responses to C5a. We conclude that the primary effect of a short period of pretreatment with IL-3 is to couple the generation of free AA to C5a-mediated stimulation in particular and to accelerate its generation after other stimuli. Enhancements of the [Ca2+]i response, while sporadic and at best modest, may have some influence on enhanced mediator release but did not clearly explain the functional effects of IL-3.
Protective effects of selectin chimeras in neutrophil-mediated lung injury.
Authors: Mulligan M S , Watson S R , Fennie C , Ward P A 
Published: January 2023

Abstract
Recombinant selectin chimeric molecules featuring the joining of the extracellular domains of L-, P-, and E-selectin to the CH2 and CH3 domains of human IgG1 have been evaluated for their ability to protect against neutrophil-dependent lung injury in rats after systemic activation of C caused by vascular infusion of cobra venom factor (CVF) or lung injury that follows intrapulmonary deposition of IgG immune complexes. Previous studies using anti-selectin antibodies have suggested that the former model is P-selectin dependent, whereas the latter is E-selectin dependent. Requirements for L-selectin have not been identified because of lack of reagents. For the current studies employing the CVF model of lung injury, infusion of P-selectin-Ig chimera reduced injury (as assessed by changes in permeability and hemorrhage) in a dose-dependent manner, with parallel reductions in lung myeloperoxidase (MPO) content. Similar results were obtained with the L-selectin-Ig chimera, whereas the E-selectin-Ig chimera was not protective and failed to alter MPO content. In contrast, in the IgG immune complex model of lung injury, the L- and E-selectin-Ig chimeras both showed dose-related protective effects and reductions in MPO content, whereas the P-selectin-Ig chimera failed to protect against injury and did not alter MPO content in this model of lung injury. In all cases of blocking of injury, this was incomplete, suggesting multi-selectin engagement or inadequate amounts of selectin-Ig chimeras employed. These data indicate that neutrophil recruitment and attendant lung injury in the CVF model are L- and P-selectin dependent and E-selectin-independent, whereas in the IgG immune complex model, neutrophil recruitment and lung injury are L- and E-selectin-dependent but independent of P-selectin. Thus, differing selectin requirements for acute inflammatory lung injury have been identified.
C33 antigen and M38 antigen recognized by monoclonal antibodies inhibitory to syncytium formation by human T cell leukemia virus type 1 are both members of the transmembrane 4 superfamily and associate with each other and with CD4 or CD8 in T cells.
Authors: Imai T , Yoshie O 
Published: January 2023

Abstract
C33 Ag and M38 Ag had been identified by mAb inhibitory to HTLV-1-induced syncytium formation. The cDNA encoding C33 Ag had revealed that it belongs to the newly defined transmembrane 4 superfamily (TM4SF). M38 Ag was detected on virtually all human cell lines and fresh leukocytes except for most granulocytes. It was also expressed on a mouse hybrid cell clone containing human chromosome 11q23-pter. Immunoprecipitation and immunoblot analyses identified a monomeric 26-kDa protein. The M38 epitope was dependent on S-S bonding. These characteristics were very similar to those reported for TAPA-1 (the target of antiproliferative antibody-1), which also belongs to TM4SF as C33 Ag. We therefore cloned the cDNA of human TAPA-1 and expressed it in COS cells. M38 indeed reacted with COS cells expressing human TAPA-1. We concluded that M38 Ag was identical to TAPA-1. To further investigate the biologic functions of C33 Ag and M38 Ag (TAPA-1) and their roles in HTLV-1-induced syncytium formation, molecules associated with these Ag were examined in T cells. Immunoprecipitation from surface-iodinated cell lysates revealed that proteins co-precipitated by C33 and M38 were mostly common including each other. Sequential immunoprecipitation-immunoblot experiments confirmed that C33 Ag and M38 Ag (TAPA-1) were associated with each other. The association was further confirmed in BHK cells doubly transfected with human cDNA for C33 Ag and TAPA-1. We extended similar analyses and found that C33 Ag and M38 Ag (TAPA-1) were regularly associated with CD4 or CD8. The association of these Ag on the cell surface was further supported by co-modulation of M38 Ag (TAPA-1), CD4 and CD8 with C33 Ag. This is the first time that a physical association between the members of TM4SF is demonstrated. Furthermore, the regular association of C33 Ag and M38 Ag (TAPA-1) with CD4 or CD8 might indicate that they play a role in expression and/or function of the CD4/CD8 co-receptor complex.
Analysis of glomeruli-eluted Gp330 autoantibodies and of Gp330 antigen of Heymann nephritis.
Authors: Makker S P 
Published: January 2023

Abstract
Quantity, charge, IgG isotype, Ag reactivity of glomerular gp330 autoantibodies (gp330Ab), and the charge of the putative Ag, gp330, were studied in active Heymann nephritis. Gp330 was anionic with an isoelectric point of 4.6. Despite variation in C3 glomerular immunofluorescence staining, C5b-9 staining was seen in all rats. Positive correlation was seen between glomerular gp330Ab and abnormal 24-h proteinuria (r = 0.637, p = 0.008), which appeared to require a certain threshold level of gp330Ab. Positive correlation was also seen between serum and glomerular gp330Ab at time of death (r = 0.55, p &lt; 0.05). No differences were seen in charge, IgG isotypes of glomerular gp330Ab, or reactivity to gp330 by Western analysis in rats with or without abnormal proteinuria. Comparison of gp330Ab IgG isotypes in immune sera and glomeruli showed no difference in IgG1 and IgG2a, but IgG2b was significantly lower in glomeruli (p &lt; 0.0001). Conclusions: 1) this is the first documentation that gp330 is highly anionic; 2) regardless of the degree of proteinuria, all immunized rats deposit in their glomeruli a qualitatively similar type of gp330Ab (charge, IgG isotype, complement activation, gp330 reactivity); 3) despite the highly anionic nature of gp330, anionic gp330Ab accumulate in glomeruli; and 4) complement activation appears to occur in every rat with gp330Ab in glomerulus and development of proteinuria is dependent on a certain threshold level of gp330Ab in the glomerulus which, in turn, correlates with the serum level of gp330Ab.
Extensive apoptosis occurring in the thymus during accelerated rejection of cardiac allografts in presensitized rats.
Authors: Tanaka K , Koga Y , Zhang X Y , Sasaki M , Wang Y , Kimura G , Nomoto K 
Published: January 2023

Abstract
Cardiac allografts of (Lewis x Brown Norway) F1 rats are rejected by Lewis recipients at 7.5 days after engrafting. When recipients are presensitized with Brown Norway skin grafts at 7 days before engrafting, the cardiac grafts are rejected between 24 and 36 h (accelerated rejection, ACR). We previously demonstrated that the number of thymocytes in recipients showing ACR is decreased to approximately one eighth at 24 h after engrafting, and the thymocytes remaining in the thymus are phenotypically and functionally more mature. In the present study, flow cytometric analysis of the thymocytes in the sensitized recipient at 24 h after engraftment demonstrated that the frequency of single positive cells (CD4+CD8- and CD4-CD8+) was increased and that of double positive cells (CD4+CD8+) was decreased, indicating that immature thymocytes preferentially disappeared during the ACR episode. Thymocytes of the recipients undergoing ACR suffered from extensive apoptosis, characterized by chromatin condensation in the nuclei, cell shrinkage, nuclear collapse, and DNA fragmentation. Serum levels of corticosterone were elevated but were within a similar range among the transplant recipients destined for acute rejection, those undergoing ACR, and isografted controls, suggesting the participation of mediator(s) other than glucocorticoid in the induction of extensive apoptosis in the transplant recipients with ACR. We propose that thymocyte apoptosis is accelerated during the allograft rejection episode.
Episialin (MUC1) inhibits cytotoxic lymphocyte-target cell interaction.
Authors: van de Wiel-van Kemenade E , Ligtenberg M J , de Boer A J , Buijs F , Vos H L , Melief C J , Hilkens J , Figdor C G 
Published: January 2023

Abstract
Episialin (MUC1) is a mucin-like glycoprotein abundantly expressed on most carcinoma cells. As a result of its extended and rigid structure, it reduces intercellular adhesion. We investigated whether this antiadhesion function allows tumor cells expressing high levels of episialin to escape from immune recognition. To test this hypothesis, we transfected episialin-negative (episialin-) melanoma cells (A375) with the MUC1 cDNA-encoding episialin. The results demonstrated that episialin-positive (episialin+) melanoma cells were significantly less susceptible to lysis than episialin- melanoma cells by both alloantigen or rIL-2-stimulated cytotoxic effector cells. In addition, cold target inhibition experiments with episialin+ and episialin- cells clearly demonstrated preferential lysis of episialin- cells. Furthermore, antibody blocking studies showed that lysis of episialin+, but not of episialin-, melanoma cells was predominantly dependent on the leukocyte function-associated Ag-1/intracellular adhesion molecule adhesion route, suggesting that episialin+ target cells adhere less efficiently to effector cells than episialin- target cells. This notion was supported by the observation that conjugate formation of the effector cells with episialin+ target cells was significantly impaired. From these results we conclude that over-expression of episialin as found on many tumor cells may indeed affect efficient lysis by cytotoxic lymphocytes and thus may contribute to escape from immune surveillance.
Antigen- and isotype-specific immune responses to a recombinant antigen-allergen chimeric (RAAC) protein.
Authors: Zhang L , Mohapatra S S 
Published: January 2023

Abstract
Ag-specific IgE and IgG antibody responses to a recombinant Ag-allergen chimeric (RAAC) protein were examined in B6D2F1 mice. The RAAC protein consisted of the truncated beta-galactosidase (beta-gal), linked at its C terminus to a polypeptide representing the conserved region of the recombinant Kentucky Bluegrass allergen encoded by the cDNA clone KBG8.3(rKBG8.3). Immunization of the mice with the RAAC protein in dextran sulfate as adjuvant led to the differential production of antibodies to the two constituents of RAAC protein with respect to their isotypic classes. Most of the antibody responses to both sets of determinants on the fusion protein were IgG1. In addition, the allergenic polypeptide of RAAC protein induced IgE antibodies, whereas the beta-gal elicited IgG2a antibodies. The same pattern of antibody isotypes was produced when the individual components, rKBG8.3 and beta-gal were separately used for immunization with dextran sulfate. On the other hand, immunization of mice with either RAAC or the beta-gal in CFA induced primarily a IgG response, and no IgE antibodies; however, under the same conditions of immunization rKBG8.3 induced IgG1 antibodies and also low levels of IgE antibodies. In contrast, the RAAC and the beta-gal induced IgG2a antibodies, whereas rKBG8.3 induced no detectable IgG2a antibodies. Furthermore, high titers of IgE antibodies were induced by the rKBG8.3 and not by the RAAC protein in dextran sulfate after the mice had been immunized twice with the same polypeptide in CFA. It is inferred from these results that the induction of isotype-specific immune responses in the animals with the same genetic background is dependent upon the Ag in question as well as the adjuvant; the latter, however, influences the magnitude but does not determine the isotype of the immune responses.
Molecular analysis of spontaneous nephrotropic anti-laminin antibodies in an autoimmune MRL-lpr/lpr mouse.
Authors: Foster M H , Sabbaga J , Line S R , Thompson K S , Barrett K J , Madaio M P 
Published: January 2023

Abstract
To explore the genetic relationship between anti-laminin and anti-DNA autoantibodies (autoAb), VH gene and gene family expression were determined among autoAb derived from an individual 6-mo-old MRL-lpr/lpr mouse. Whereas 85% of the anti-DNA Ig were identified by one of two VH family probes, 7183 and VHJ558, none of the anti-laminin antibodies (Ab) examined were recognized by these probes. Subsequent V region sequence analysis of three of the anti-laminin Ab revealed that they in fact utilized a J558 VH gene (VH50). Furthermore, FR2 and CDR2 oligonucleotide probes complementary to VH50 recognized multiple anti-laminin Ab by Northern blot analysis; the FR2 probe recognized two control anti-DNA Ab, but neither probe recognized anti-DNA Ab from the same mouse. Polymerase chain reaction amplification of MRL-lpr/lpr genomic liver DNA using primers generated from VH50 and Vk50 sequences indicated that all three anti-laminin Ig have a single replacement mutation in both their VH and Vk genes. Search of the nucleic acid databases revealed that both germline VH and Vk genes are expressed unmutated by murine lupus anti-dsDNA autoAb, previously sequenced in other laboratories. Sequence comparisons suggest that differences in anti-DNA and anti-laminin reactivity may be dependent upon somatically generated differences in the CDR3 regions of the H and L chains. The results indicate that lupus anti-laminin Ab can arise from distinct B cell populations but express the same unmutated germline V region genes as lupus anti-dsDNA autoAb. They further raise the possibility that these distinct B cell populations may be activated and expanded either: independently, by distinct Ig receptor ligands such as the Ag, laminin and DNA; or simultaneously, by a common ligand such as an anti-Id recognizing a common V region epitope.
Restrictions in the repertoire of allospecific T cells. Contribution of the alpha-helical sequence polymorphism of HLA-DR molecules.
Authors: Goronzy J J , Xie C , Hu W , Lundy S K , Weyand C M 
Published: January 2023

Abstract
We have studied the molecular diversity of the allogeneic TCR repertoire specific for the HLA-DR4 variant, DRB1*0404, by analyzing the V beta gene segment usage of CD4+ T cell clones from nine different donors and comparing the V beta repertoire in allospecific and unselected T cell populations. To investigate the forces shaping the repertoire of the alloreactive T cell response we compared the diversity of TCR specificities utilized in very similar and in disparate responder/stimulator combinations. Six of the responders shared the HLA-DRB1*0401 allele, which differs from the HLA-DRB1*0404 allele by only two amino acid substitutions at positions 71 and 86 of the HLA-DR beta 1 chain; three responders expressed HLA-DRB1 alleles with extensive polymorphisms compared with the stimulator HLA-DRB1 allele. In all nine responders, TCR specificities recruited to recognize HLA-DRB1*0404 were strongly biased toward the dominant usage of one to three V beta elements. Despite some degree of heterogeneity between different responders, the overall pattern was very similar with a hierarchy of TCR V beta elements expressed by HLA-DRB1*0404 specific T cells. A core group of V beta elements (V beta 6, 5, 2, 13.2, 18, and 7) was preferentially used, whereas other V beta elements including V beta 3, 4, and 8 were infrequently used or not used at all. Sequencing of the VDJ beta region from selected T cell clones showed no junctional bias associated with the preferential V beta gene segment usage. Surprisingly, the T cell diversity in focused and in complex alloreactive responses were equally biased. The finding that HLA-DR4+ and DR4- donors exhibited a similar bias supports the interpretation that the alpha-helical HLA-DR structure of the stimulator cell has a dominant contribution in shaping the alloreactive repertoire.
Fixation of C3 to IgG attenuates neutrophil HOCl generation and collagenase activation.
Authors: Chatham W W , Blackburn W D 
Published: January 2023

Abstract
Neutrophils stimulated with surface-associated monomeric IgG (SAIgG) release an activated collagenase in association with significant generation of hypochlorous acid (HOCl). To determine whether these neutrophil responses are modulated by IgG C fixation, neutrophils were incubated with SAIgG pretreated with normal or C-deficient sera. Treatment of SAIgG with normal sera did not attenuate neutrophil superoxide production, H2O2 generation, or extracellular release of latent collagenase and lactoferrin; however, serum treatment resulted in significant attenuation of SAIgG-induced HOCl generation (75%) and extracellular release of the azurophilic granule constituents myeloperoxidase and cathepsin G. Collagenase activity in supernatants of neutrophils incubated with SAIgG pretreated with normal sera (9.5 +/- 0.8 ng/min) was significantly less than activity in supernatants of neutrophils incubated with SAIgG not treated with sera (15.3 +/- 1.2 ng/min). Treatment of surface adherent monoclonal IgG1 and IgG2 with sera resulted in significantly greater attenuation of HOCl generation compared to serum treatment of IgG4. Attenuation of HOCl generation was not observed when SAIgG was pretreated with heat-inactivated sera, EDTA-chelated sera, or sera depleted of C3; treatment of SAIgG with C5-depleted sera yielded results comparable to treatment with intact sera. These results indicate that C3-derived ligands fixed to adherent Ig alter IgG-induced release of azurophilic granule constituents and HOCl generation by neutrophils.
In vitro maturation of neonatal human CD8 T lymphocytes into IL-4- and IL-5-producing cells.
Authors: Byun D G , Demeure C E , Yang L P , Shu U , Ishihara H , Vezzio N , Gately M K , Delespesse G 
Published: January 2023

Abstract
Naive CD8 T cells isolated from umbilical cord blood were examined for cytokine production and for surface phenotype before and after priming with anti-CD3 mAb in the presence of selected cytokines. On stimulation with anti-CD3 immobilized on CD32-B7-transfected L cells, naive and primed CD8 T cells release high levels of IFN-gamma but no IL-2, IL-4, or IL-5. IL-12-primed cells, and to a lesser extent IL-2-primed cells, have an increased capacity to produce IFN-gamma but display the same restricted pattern of cytokine production. Cells primed in the presence of IL-4 or IL-4 + IL-2 are capable of producing high levels of IL-5 but no IL-4, and their IFN-gamma-producing capacity is much reduced. Cells primed with both IL-4 + IL-12 produce high levels of IL-5 and IFN-gamma but no IL-4. IL-4-producing CD8 T cells are obtained only if IL-4- or IL-4 + IL-2-primed cells are subjected to a second cycle of activation in the presence of IL-2; restimulation of IL-4 + IL-12-primed cells under identical conditions fails to generate IL-4-producing cells but leads to the development of high IFN-gamma and IL-5 producers. Thus, unlike in CD4 T cells, IL-12 completely inhibits IL-4-induced capacity of CD8 T cells to produce IL-4. However, IL-4 and IL-12 synergize to promote the expression of IL-5. Regardless of the priming conditions, primed CD8 T cells are CD45ROhigh/RA-, CD31high and more than 50% are CD4+/CD8+; activated naive or primed CD8 T cells do not express CD40 ligand.
Reactivity of hybridomas derived from T cells activated in vivo during graft-versus-host disease.
Authors: Leibnitz R R , Lipsky P E , Thiele D L 
Published: January 2023

Abstract
To examine the specificity of T helper cells activated during murine graft-vs-host disease (GVHD), T cell hybridomas from GVHD spleens and livers were generated and analyzed. CTL-depleted C57BL/6 (B6) donor cells were injected into irradiated (B6 x bm12)F1 or (bm1 x bm12)F1 recipient mice. Five or fourteen days later, cells from livers and spleens were fused directly with the TCR-deficient (alpha beta)- BW5147 thymoma line. The in vivo-activated T cells produced hybridomas as efficiently as either T cells activated in a primary mixed lymphocyte reaction or expanded in vitro after isolation from GVHD mice. Overall, 91% (396 of 437) of hybridomas generated from GVHD animals responded to immobilized anti-CD3 and 56% (220 of 396) of these hybridomas responded specifically to APC expressing host bm1 or bm12 alloantigens. More than 80% of bm12-specific hybridomas expressed CD4; all (53 of 53) of the bm12-specific hybridomas tested reacted to homozygous bm12 APC. Of the alloreactive T hybridomas generated from B6--&gt;(bm1 x bm12)F1 GVHD mice, 7% responded to bm1 APC. Five bm1-specific hybridomas were analyzed further. One CD4+ hybridoma recognized a bm1 peptide presented by self I-Ab and was blocked by anti-Ia Ab; the other four hybridomas, two of which also expressed CD4, responded to transfected L cells expressing H-2Kbm1 and were not inhibited by anti-Ia Ab. These results indicate that a high percentage of CD4+ T hybridomas generated from freshly isolated T cells activated in vivo during GVHD are specific for host MHC class II or class I alloantigens.