bluuebunny/crossref_metadata_2025_split huggingface dataset:
Expression of the leukocyte common antigen CD45 by endothelium.
Authors: Forsyth K D , Chua K Y , Talbot V , Thomas W R 
Published: December 2022

Abstract
The CD45 family of Ag expressed by leukocytes play a key role in lymphocyte activation. In this study, we identify both a restricted pattern of expression of CD45 Ag by human endothelial cells and differences in m.w. of endothelial CD45 compared with lymphocyte CD45. Initially, immunoperoxidase staining of endothelial cells in culture revealed the presence of the CD45RO isoform provided the endothelial cells had been stimulated by IL-1 for several days. No other isoform of CD45 was detected by immunoperoxidase staining of endothelium. Leukocyte common antibodies, reputed to detect all isoforms of CD45, did not detect endothelial CD45RO. A polymerase chain reaction method was then used to demonstrate that the message for the CD45RO isoform was constitutively present, with increased levels after IL-1 stimulation. Western blot confirmed the presence of the RO isoform. No other CD45 isoform was detected, either by PCR amplification for message or by Western blot. There were clear differences between lymphocyte and endothelial CD45. The RO isoform expressed by endothelium has an estimated M(r) of 235,000 in contrast to lymphocyte RO that, on our gels, has an estimated M(r) of 190,000. Given the large surface area of endothelium (approximately 1 km2 in the human), the close apposition of lymphocytes and endothelium in immunologic tissues such as lymph nodes, and the pivotal role CD45 plays in activation, expression of CD45RO by endothelium may have important implications in lymphocyte-endothelial interactions. This is most likely to occur in endothelium after a few days of IL-1 stimulation, e.g., at sites of chronic inflammation, or in the draining lymph node of an inflammatory focus.
Monoclonal Ig L chain and L chain V domain fragment crystallization in myeloma-associated Fanconi's syndrome.
Authors: Aucouturier P , Bauwens M , Khamlichi A A , Denoroy L , Spinelli S , Touchard G , Preud'homme J L , Cogné M 
Published: December 2022

Abstract
Monoclonal free L chains secreted in immunoproliferative disorders are frequently involved in renal complications, including a specific proximal tubule impairment, the Fanconi's syndrome, which is generally featured by intracellular crystallization of L chain-related material. In a patient with myeloma-associated Fanconi's syndrome, hexagonal crystals (most surrounded by smooth membranes) were found in kidney proximal tubular cells and bone marrow plasma cells and phagocytes. The sequence of the patient's monoclonal kappa-chain was deduced from that of identical kappa-cDNA clones from the tumoral plasma cells. Small protein-enriched gel filtration fractions from urine yielded crystals morphologically similar and with the same 60 A periodicity on electron micrographs as those found in the cells. N-terminal sequencing and mass spectrometry studies showed that the crystals contained a 107-amino acid fragment (with a C-terminal lysine) corresponding to the V domain together with a low proportion of the entire kappa-chain. In vitro trypsin and pepsin treatment of the native entire kappa-chain yielded a homogeneous V domain fragment which, contrary to other monoclonal kappa-chains, was completely resistant to further proteolytic attack. The patient's kappa-chain also displayed an unusual self-reactivity, as demonstrated by a Western blot technique. The peculiar proneness of the V domain to resist proteolysis and to form crystals might prevent the normal cell catabolism of the L chain and lead to crystallization and renal impairment.
Origin of CD4-CD8-B220+ T cells in MRL-lpr/lpr mice. Clues from a T cell receptor beta transgenic mouse.
Authors: Zhou T , Bluethmann H , Eldridge J , Berry K , Mountz J D 
Published: December 2022

Abstract
Abnormal development of T cells in the thymus is thought to be related to autoimmune disease and the expansion of the unusual CD4-CD8-B220+ peripheral T cel subset that results in lymphadenopathy in MRL-lpr/lpr mice. Although we and others have previously shown that rearranged TCR-transgenes alter T cell development in the thymus and abrogate lymphoproliferative disease in lpr mice, the origin and developmental pathway of the LN CD4-CD8-B220+ T cells has not been fully elucidated. We therefore undertook the systematic analysis of the effect of a TCR-beta transgene on the production and differentiation of (lymph node) LN T cells and the production, differentiation, and release of thymocyte T cell populations. In nontransgenic mice, there was increased proliferation of CD4-CD8-B220+ T cells in the LN of adult MRL-lpr/lpr mice compared to MRL(-)+/+ mice, as measured by in vivo BrdU labeling. These proliferating LN T cells were greatly reduced by thymectomy of adult MRL-lpr/lpr mice 1 wk before bromodeoxyuridine labeling, indicating that recent thymic emigrants or factors were required to sustain proliferation. In the thymus, there was increased production and accumulation of CD4+CD8+TCRdull thymocytes in nontransgenic MRL-lpr/lpr compared to MRL(-)+/+ mice. As the rate of maturation from CD4+CD8+TCRdull to CD4+CD8+TCRbright was the same (6%) in both MRL-lpr/lpr and MRL(-)+/+ mice, the accumulation of the immature population in the MRL-lpr/lpr mice could not be due to a maturation defect. However, there was a decrease in apoptosis and intrathymic death of CD4+CD8+TCRdull thymocytes in MRL-lpr/lpr compared to MRL(-)+/+ mice. Introduction of the TCR-beta transgene into lpr/lpr mice normalized the proliferation of T cells in the LN. In the thymus, the TCR-beta transgene resulted in a dramatic increase in maturation efficiency and a reduction in apoptosis in MRL(-)+/+ mice. These data suggest that the TCR transgene inhibits lymphoproliferation by reducing the production of "neglected" CD4+CD8+TCRdull thymocytes that will undergo Fas Ag-mediated apoptosis. They further suggest that in lpr mice, which express a mutated Fas Ag, the "neglected" thymocytes are able to continually escape to the periphery, where they proliferate.
Abrogation of H-2-restricted CTL responses and efficient recognition of HLA-A3 molecules in DBA/2 HLA/A24 responder mice.
Authors: Barra C , Gournier H , Garcia Z , Marche P N , Jouvin-Marche E , Briand P , Fillipi P , Lemonnier F A 
Published: December 2022

Abstract
DBA/2 (H-2d) HLA-B7 x human beta 2-microglobulin transgenic and DBA/2 nontransgenic mice stimulated by DBA/2 HLA-A3 x human beta 2-microglobulin transgenic mouse spleen cells developed potent H-2Kd restricted cytolytic responses with recognition of a peptide from the second domain of the HLA-A3 H chain. These H-2Kd-restricted responses obliterated, as a rule, cytolytic responses with direct recognition of the HLA-A3 molecules, even in HLA-B7 transgenic mice. These immunodominant H-2Kd-restricted responses could be abrogated in DBA/2 HLA-A24 mice because of cross-tolerance, the HLA-A3 derived-H-2Kd presented peptide being shared by several (including A24) HLA class I H chain allelic variants. Under such experimental circumstances, strong CTL responses with exclusive direct recognition of HLA-A3 molecules constantly developed. Further analysis of these responses in six DBA/2 HLA-A24 responder mice indicated that a large fraction of the mouse V beta and V alpha genes could be used to mount such CTL responses. Thus, by combining classical HLA class I transgenesis and selective destruction of H-2K and H-2D genes, it should be possible to derive useful strains of mice for the study of HLA class I-restricted CTL responses.
Differential effect of IL-10 on dendritic cell-induced T cell proliferation and IFN-gamma production.
Authors: Macatonia S E , Doherty T M , Knight S C , O'Garra A 
Published: December 2022

Abstract
IL-10 has previously been shown to inhibit cytokine production by Th1 cells by blocking macrophage accessory cell function. In this study we demonstrate that dendritic cell-induced Th1 proliferation was unaffected by IL-10, whereas macrophage-stimulated proliferation was inhibited in the same system. In contrast dendritic cell induced IFN-gamma production by the Th1 clones was down-regulated by IL-10. Inasmuch as dendritic cells have been shown to be potent APC for T cells in primary responses we determined the effect of IL-10 on dendritic cell-induced proliferation and IFN-gamma production by CD4+ and CD8+ T cells. Dendritic cells were effective at stimulating both proliferation and IFN-gamma secretion of CD4+ or CD8+ T cells in a primary allogeneic mixed leukocyte response. In contrast, IL-10 markedly inhibited dendritic cell driven IFN-gamma production by purified CD4+ and CD8+ T cells. Down-regulation of dendritic cell-induced IFN-gamma production suggests a role for IL-10 in inhibiting the initiation of cell-mediated immune responses.
Study of activation of murine T cells with bacterial superantigens. In vitro induction of enhanced responses in CD4+ T cells and of anergy in CD8+ T cells.
Authors: Yan X J , Li X Y , Imanishi K , Kumazawa Y , Uchiyama T 
Published: December 2022

Abstract
Primary and secondary responses of murine CD4+ T cells and CD8+ T cells upon stimulation with staphylococcal enterotoxin E (SEE) bearing superantigenic properties were examined. Both isolated C57BL/6 splenic CD4+ T cells and CD8+ T cells proliferated and produced IL-2 and IFN-gamma upon stimulation with SEE in substantial levels. The amounts of IL-2 were greater in CD4+ T cells and those of IFN-gamma were somewhat greater in CD8+ T cells. SEE-induced CD4+ T lymphoblasts, larger parts of which bore the V beta 11 element in their TCR, proliferated, produced IL-2 and IFN-gamma, and showed toxin-dependent cytotoxicity in substantial levels upon restimulation with SEE. By contrast, SEE-induced CD8+ T lymphoblasts, the larger part of which bore the V beta 11 element, did not show the first two of the three responses at all upon restimulation with SEE, whereas these cells showed greater cytotoxicity. The CD8+ T lymphoblasts did not suppress the reactivity of the CD4+ T lymphoblasts. Both SEE-induced CD4+ T lymphoblasts and CD8+ T lymphoblasts proliferated and produced IL-2 and IFN-gamma in comparable levels upon stimulation with rIL-2 or mAb to CD3 or V beta 11.
The role of viral enhancer "core" motif-related sequences in regulating T cell receptor-gamma and -delta gene expression.
Authors: Hsiang Y H , Spencer D , Wang S , Speck N A , Raulet D H 
Published: December 2022

Abstract
T cells express clonally distributed alpha beta or gamma delta Ag receptor heterodimers. Transcriptional enhancers for the genes of all four subunits are active in both gamma delta and alpha beta T cells, but are less active or inactive in other cells. Conserved sequence motifs are present in all four enhancers, suggesting that common transcription factors regulate TCR gene expression. One of these motifs in the gamma 3 site of the TCR-gamma enhancer is similar to motifs found in several other lymphoid-specific and viral enhancers. This conserved "core" sequence is present in the enhancers of Moloney and SL3-3 murine leukemia viruses, important for transcription in T cells and in determining disease specificity. Here we characterize the gamma 3 site of the gamma enhancer and a corresponding homologous site, delta E3, of the TCR-delta enhancer. Our results suggest that the core site is critical for activity of the 200-bp gamma enhancer fragment and of the gamma 3 and delta E3 sites. Furthermore, we identify a nuclear factor in human T cell lines that specifically binds the core region in these and several other core-containing enhancers. This factor may be identical to or related to a purified bovine nuclear core binding factor that binds the core region of the Moloney murine leukemia virus enhancer, gamma 3 and delta E3 sites, suggesting that similar proteins regulate the TCR-gamma, delta and Moloney murine leukemia virus enhancers. Other sequences in the gamma 3 site upstream of the core sequence are also critical for activity in T cells, suggesting that at least two different factors are required for functional activity of the gamma 3 site.
Antibodies to Toxoplasma gondii major surface protein (SAG-1, P30) inhibit infection of host cells and are produced in murine intestine after peroral infection.
Authors: Mineo J R , McLeod R , Mack D , Smith J , Khan I A , Ely K H , Kasper L H 
Published: December 2022

Abstract
Monoclonal and polyclonal, monospecific antibodies to the major surface antigen of Toxoplasma gondii (SAG-1, P30) inhibit infection of human fibroblasts and murine enterocytes. Fab prepared from polyclonal, monospecific antibody to P30 also have this inhibitory effect on invasion, which indicates that this antibody directly blocks parasite infection of host cells rather agglutinating the parasite. Antibodies to another surface protein (P22) did not alter in vitro infection. If the inhibitory effect of antibody to P30 was due to steric hindrance or complexing of surface epitopes contiguous to P30, antibodies to other surface epitopes would also be inhibitory and they are not. Urea treatment of antibody (which permits discrimination of high and low avidity antibody) did not alter the effect of anti-P30 antibody. This observation indicates that the effect of the antibody to P30 was not an artifact of differences in the avidity of the antibody to P22 and P30. Heat inactivated antisera from mice infected with either RH or PTg strain T. gondii (P30+) inhibit infection of fibroblasts when challenged with autologous wild-type parasites by 87 and 40%, respectively. In contrast, these antisera have little inhibitory effect (13 and 19%, respectively) against infection of human fibroblasts by a P30-deficient mutant (PTgB). Antisera raised to the P30-deficient mutant had no significant effect on infection of cells by wild-type strains that have surface P30. The neoglycoprotein, BSA-glucosamide, competitively blocks infection of human fibroblasts by P30+ tachyzoites with surface P30 in higher level than those without surface P30. This observation indicates that there is likely to be a glycosylated host cell receptor to which T. gondii's major surface Ag SAG-1 (P30) binds. Mice infected perorally develop intestinal IgA antibody to the major 30-kDa epitope of T. gondii. Thus, the major surface epitope of T. gondii, SAG-1 (P30), has an important, functional role in infection of host cells by T. gondii and elicits an intestinal antibody response after peroral infection.
Idiotype and anti-anti-idiotype antibodies to Neisseria gonorrhoeae lipooligosaccharides with bactericidal activity but no cross-reactivity with red blood cell antigens.
Authors: Brossay L , Paradis G , Pépin A , Mourad W , Coté L , Hébert J 
Published: December 2022

Abstract
A panel of murine mAb against three different epitopes of Neisseria gonorrhoeae lipooligosaccharides (LOS) was developed. Only one of these, 3G5, displays bactericidal activity against all in vitro serum-resistant strains of N. gonorrhoeae. Evidence suggests that sialylation, which could occur in vivo, modifies some LOS epitopes in such a way that the strains become resistant to bactericidal activity and are no longer recognized by specific antibodies. The epitope recognized by our bactericidal mAb is not affected by sialylation as shown by immunoblot analysis. We also provided evidence that the 3G5 epitope is different from RBC precursor Ag, since our mAb did not induce RBC agglutination. Since LOS induce an immune response in a T cell-independent fashion and are highly toxic, they cannot be used for immunization. Use of anti-idiotypic antibodies (aId) could be a way to bypass these difficulties. Therefore, in the present study, aId were produced in rabbits and rendered idiotype-specific by appropriate adsorption. These aId specifically bind to the relevant Id but not to LOS, and inhibit only the binding of anti-LOS mAb (3G5) to LOS preparations from N. gonorrhoeae in a dose-response fashion. The specificity of our aId for the binding site of anti-LOS mAb is suggested by the binding inhibition of affinity-purified aId to Id by LOS. In addition, the capacity of aId to inhibit bactericidal activity of this anti-LOS mAb and the idiotypic cross-reactivity between rat and mouse anti-LOS antibodies support this point. Finally, the elicitation of anti-LOS activity with bactericidal activity upon immunization of naive mice with aId confirms the internal image properties of the aId. These data suggest that a bactericidal mAb suitable for immunoprotection was obtained, and the production of aId opens the door for development of a vaccine.
Cytokine expression in vivo during murine herpetic stromal keratitis. Effect of protective antibody therapy.
Authors: Staats H F , Lausch R N 
Published: December 2022

Abstract
HSV-1 topical infection on the murine cornea can induce herpetic stromal keratitis (HSK), a T cell-mediated inflammatory response that results in blindness. To begin to decipher the molecular interactions involved in this infection, extracts of infected corneas were assayed for the presence of seven different cytokines by ELISA. The most prominent cytokines detected were IL-1 alpha and IL-6. Both were elevated by day 2 after infection, reached peak levels of 82 and 538 pg/cornea, respectively, at day 10, and then diminished over the next 10 days. In contrast, TNF-alpha concentrations were not elevated over that seen in uninfected corneas at any time during the 20-day observation period. IFN-gamma and granulocyte-macrophage CSF corneal concentrations were below the sensitivity of the assay. We investigated whether passive transfer of antibody to viral glycoprotein D, which prevents HSK, influenced the production of IL-1 alpha and IL-6. It was found that corneal concentrations of IL-1 alpha were reduced threefold and IL-6 was undetectable at day 10 in the antibody-treated hosts. The levels of IL-10 and IL-4 in uninfected control and antibody-treated hosts were also monitored. Neither of these two regulatory cytokines was associated with HSK development or effective antibody therapy. Naive corneas cultured in vitro spontaneously produced IL-1 and IL-6, indicating that cells resident in the cornea had the ability to synthesize these proinflammatory cytokines. Collectively, our results imply that IL-1 alpha and IL-6 may be important contributors to the development of HSK pathogenesis.
Protein kinase C modulates the amounts of IL-1 receptor mRNA in human lung fibroblasts.
Authors: Aksamit T R , Monick M M , Hunninghake G W 
Published: December 2022

Abstract
IL-1 activates human lung fibroblasts via IL-1-specific membrane receptors. The numbers of IL-1 binding sites on fibroblasts are increased after exposure to prostaglandin E2 (PGE2) or IL-1. Increases in binding sites are associated with changes in functional responses to IL-1. In these studies, we determined if alterations in numbers of IL-1 binding sites on human lung fibroblasts were associated with parallel changes in IL-1R mRNA. In addition, since IL-1 and PGE2 can activate protein kinase C (PKC) and protein kinase A (PKA), we exposed human lung fibroblasts to 1-(5-Isoquinoline sulfonyl)-2-methylpiperafine (H7) or staurosporine, which are relatively specific inhibitors of PKC, and N-[2-(methylamino)ethyl]-5-isoquinolinesulfomamide (H8), an inhibitor of PKA, to determine whether the IL-1 and PGE2 stimulated increases in binding sites were mediated by activation of PKC or PKA. H7 decreased the base line and PGE2-stimulated increases in numbers of IL-1 binding sites. Exposure of the fibroblasts to phorbol-12-myristate-13-acetate (PMA) for 24 h, which is known to deplete PKC, also decreased the numbers of base line IL-1 binding sites. These changes were paralleled by changes in amounts of IL-1R mRNA. H7 and staurosporine also blocked IL-1 and PGE2 stimulated increases in IL-1R mRNA. In contrast, H8 had no effect on the base line or PGE2-stimulated increases in numbers of IL-1 binding sites nor did it change the amounts of IL-1R mRNA. These studies show that changes in the numbers of IL-1 binding sites on human lung fibroblasts are paralleled by changes in IL-1R mRNA. In addition, PKC activity is necessary for the expression of both base line and stimulated increases in IL-1 binding sites and IL-1R mRNA. These studies suggest that PKC activity plays an important role in the modulation of IL-1R expression.
Enhancement of tumor necrosis factor cytotoxicity by lithium chloride is associated with increased inositol phosphate accumulation.
Authors: Beyaert R , Heyninck K , De Valck D , Boeykens F , van Roy F , Fiers W 
Published: December 2022

Abstract
We have previously reported that LiCl increases considerably the cytotoxic activity of TNF towards some transformed cell lines such as L929. Here we show that treatment of these cell lines with the combination of TNF and LiCl leads to the prolonged accumulation of inositol monophosphate, inositol bisphosphate, and inositol trisphosphate, whereas treatment with TNF or LiCl alone did not. In contrast, both a LiCl-unresponsive TNF-sensitive cell line and TNF-resistant cell lines did not respond with increased accumulation of inositol phosphates (IPn) upon treatment with the combination of TNF and LiCl. Furthermore, the combination of TNF and LiCl induced a transient increase in cytidine diphosphate-diacylglycerol in L929 cells. Increased IPn and cytidine diphosphate-diacylglycerol accumulation preceded the onset of cell killing by approximately 1 h. TNF-mediated cytotoxicity and TNF-induced IPn accumulation were equally sensitive to inhibition by the phospholipase inhibitor neomycin and to stimulation by the protein kinase inhibitor staurosporine. Characterization of the inositol bisphosphate isomers by HPLC analysis revealed that the TNF + LiCl-induced increase in IPn levels was due to activation of a phospholipase C and not of a phospholipase D. In contrast to TNF, several other cytotoxic agents did not increase IPn production upon application in the presence of LiCl. The TNF + LiCl-induced increase in inositol triphosphate suggests a role for intracellular Ca2+ mobilization in TNF action. Moreover, several agents that lower the intracellular Ca2+ concentration inhibited TNF cytotoxicity. In conclusion, our data provide evidence that TNF cytotoxicity and its enhancement by LiCl are mediated by increased IPn accumulation resulting in Ca2+ mobilization.