bluuebunny/crossref_metadata_2025_split huggingface dataset:
Neonatal VH, D, and JH gene usage in rabbit B lineage cells.
Authors: Friedman M L , Tunyaplin C , Zhai S K , Knight K L 
Published: December 2022

Abstract
The neonatal antibody repertoire in both mouse and humans differs from that of the adult repertoire in that the neonatal repertoire uses a limited set of JH-proximal VH genes but the adult repertoires use many different VH genes. Rabbits are unusual in that adults use only three or four VH genes, with approximately 80% of the B cells using VH1, the 3'-most VH gene. To investigate whether the repertoire of neonatal rabbits differs from that of adults, we analyzed VH, D, and JH gene usage in B cells of neonatal rabbits. A total of 68 rearranged VDJ genes was cloned from mRNA and genomic DNA isolated from lymphoid tissues of newborn to 10-day-old rabbits. We found that 74% of the VDJ gene rearrangements utilized VH1 and 15% utilized the genes that we designated VHx or VHy. From the remaining VDJ genes we identified seven novel VH genes, one, VHz, which was found in mRNA. We conclude that the repertoire of utilized VH genes in neonates is limited and is similar to that of adult rabbits. We also found the D1, D2a, D2b, and JH4 gene segments preferentially rearranged. We suggest that the preferential usage of VH, D, and JH gene segments in VDJ genes is caused by preferential rearrangement rather than by selective expansion of B cells that utilize the gene segments.
Molecular analysis of mercury-induced antinucleolar antibodies in H-2S mice.
Authors: Monestier M , Losman M J , Novick K E , Aris J P 
Published: December 2022

Abstract
In H-2S mice, the administration of mercuric chloride results in the development of antinucleolar autoantibodies. These mice represent a valuable model to study the role of environmental factors in the development of systemic autoimmunity. We have obtained seven antinucleolar mAb from mercury-injected A.SW mice and characterized their specificities and V genes. All mAb immunoprecipitate the U3 and U8 ribonucleoprotein particles (RNP) and some (but not all) react with fibrillarin, the only currently characterized protein component of mammalian nucleolar RNP. Several VH and V kappa genes are recurrently used by these antinucleolar RNP mAb and their H chain CDR3 segments contain several acidic residues that may be important for binding to the cationic proteins composing the nucleolar RNP. Our results support the concept that in H-2S mice administration of mercury induces a specific loss of tolerance to nucleolar RNP.
Differential recognition of murine tumor-associated oncofetal transplantation antigen and individually specific tumor transplantation antigens by syngeneic cloned BALB/c and RFM mouse T cells.
Authors: Rohrer J W , Rohrer S D , Barsoum A , Coggin J H 
Published: December 2022

Abstract
We have previously demonstrated in several species that sarcomas, lymphomas, and carcinomas express a common Ag that cross-reacts with midgestation fetal cells. We also produced a mAb to that protein and characterized it as a 44-kDa glycoprotein. The cross-reactive immunity induced by immunization with tumor or fetal cells expressing the oncofetal Ag (OFA) can be adoptively transferred with cell populations containing T lymphocytes. The experiments discussed within this paper describe the establishment and characterization of two types of T lymphocytes induced by immunization with syngeneic tumor cells in two mouse strains. We find that five of the eight cloned T cells derived from spleens of BALB/c mice that had been immunized with MCA1315 fibrosarcoma cells are specific for an Ag shared by MCA1315 and MCA1321 cells. The other three clones are specific for an Ag present on MCA1315 but not on MCA1321. Also, none of the clones were reactive with the BALB/c plasmacytoma MOPC-315, which does not express OFA. We also find that 75% of the RFM T cell clones from spleens of RFM mice immune to the RFM thymoma 5T show a 5T-specific proliferative response. One of the four clones, however, responds to both 4T and 5T RFM thymoma cells. The BALB/c and RFM cross-reactive clones specifically respond to purified 44-kDa OFA derived from MCA1315 fibrosarcoma cells in the presence of syngeneic irradiated spleen cells and IL-2. All of the clones from both strains of mice, be they tumor-specific transplantation Ag specific or OFA specific, are CD4+, CD3+, alpha beta TCR+ T cells that secrete IFN-gamma on Ag stimulation.
Transforming growth factor-beta 1 inhibits inflammatory cytokine-induced C3 gene expression in astrocytes.
Authors: Barnum S R , Jones J L 
Published: December 2022

Abstract
In this report, we show that transforming growth factor-beta (TGF-beta) can significantly inhibit the capacity of IFN-gamma, IL-1 beta, and TNF-alpha to augment expression of the central component of complement C3 in the human astroglioma cell line D54-MG. Treatment of D54-MG cells with TGF-beta alone had no dose- or time-dependent effect on basal C3 protein or mRNA levels. However, TGF-beta suppressed induction of C3 expression at both the protein and mRNA level in D54-MG cells treated with inflammatory cytokines. The extent of TGF-beta-mediated suppression was cytokine-specific, and suppression of protein production did not necessarily correspond to reductions in steady-state mRNA levels for each cytokine. Similar findings were obtained at the mRNA level using primary rat astrocytes, indicating that TGF-beta can modulate C3 gene expression in nontransformed astrocytic cells. Kinetic studies demonstrated that TGF-beta mediates its suppressive effect for at least 72 h, and that pretreatment of cells with TGF-beta for as little as 2 h significantly reduced the ability of all three inflammatory cytokines to enhance C3 expression. Our results suggest that TGF-beta may play an important role in modulating the endogenous synthesis of complement by astrocytes under inflammatory conditions.
Leukotrienes in renal transplant rejection in rats. Distinct roles for leukotriene B4 and peptidoleukotrienes in the pathogenesis of allograft injury.
Authors: Spurney R F , Ibrahim S , Butterly D , Klotman P E , Sanfilippo F , Coffman T M 
Published: December 2022

Abstract
To investigate the role of leukotrienes in renal allograft rejection, we studied the effects of specific leukotriene inhibitors in a rat kidney transplant model. The enhanced renal production of leukotrienes observed in allograft recipients was reduced in a dose-dependent manner by the specific 5-lipoxygenase inhibitor MK886. This suppression of leukotriene production caused a substantial improvement in renal function. Inhibition of 5-lipoxygenase also reduced the severity of vascular inflammation and endothelial injury in allografts, and profoundly inhibited expression of donor MHC class II Ag on kidney cells. Survival of renal allograft recipients was prolonged from 10 +/- 1 days in controls to 16 +/- 1 days in animals that received a 6-day course of MK886 (p &lt; 0.05). To investigate the relative roles of LTB4 compared to peptidoleukotrienes in these processes, we treated a separate group of animals with the specific peptidoleukotriene receptor antagonist SKF106203. This agent inhibits the interaction of peptidoleukotrienes with their receptor(s) but does not affect the biologic actions of LTB4. In these studies, SKF106203 caused a modest improvement in renal allograft function that was of lesser magnitude than that seen with the 5-lipoxygenase inhibitor. SKF106203 also reduced vascular inflammation in allografts, but had no effect on expression of MHC class II Ag. We conclude that leukotrienes play a key role in the pathogenesis of renal allograft rejection. Furthermore, the detrimental effects of leukotrienes in rejection are mediated by distinct actions of LTB4 and peptidoleukotrienes.
Effects of IL-12 on in vivo cytokine gene expression and Ig isotype selection.
Authors: Morris S C , Madden K B , Adamovicz J J , Gause W C , Hubbard B R , Gately M K , Finkelman F D 
Published: December 2022

Abstract
The effects of murine rIL-12 on cytokine gene expression and Ig secretion were studied in vivo. In untreated mice IL-12 enhanced IFN-gamma and IL-10 gene expression and protein secretion, reduced base line IL-3 and IL-4 gene expression, and increased serum IgG2a concentration. In mice that had been injected with goat anti-mouse IgD antibody (G alpha M delta) to induce increases in IL-3, IL-4, and IL-10 gene expression and serum IgE, IgG1, IgG2a, and IgG3 concentrations, the simultaneous injection of IL-12 enhanced IFN-gamma and IL-10 gene expression and suppressed IL-3 and IL-4 gene expression and serum IgG and IgE responses. Anti-IFN-gamma mAb neutralized most, but not all, IFN-gamma produced by mice treated with G alpha M delta and IL-12. Anti-IFN-gamma mAb enhanced IL-3 and IL-4 gene expression, did not affect IL-10 or IFN-gamma gene expression, and increased serum IgG1, IgG2a, and IgG3 levels, but had relatively little effect on serum IgE in these mice. In contrast to its effects in G alpha M delta-treated mice, IL-12 failed to inhibit the IgE response to G alpha M epsilon antibody, which stimulates mIgE+ B cells to secrete IgE. These observations demonstrate that: 1) IL-12 may limit its own effects by inducing the production of a cytokine (IL-10) that down-regulates both IL-12 production and IL-12-induced IFN-gamma production; 2) IL-12 inhibits the production of at least one cytokine, IL-3, that is not generally regarded to be strictly Th1- or Th2-associated; 3) IL-12 inhibits switching to IgE secretion to a greater extent than it inhibits switching to other Ig isotypes; and 4) the in vivo effects of IL-12 are, to a large extent, IFN-gamma-dependent.
Peptide binding to surface class II molecules is the major pathway of formation of immunogenic class II-peptide complexes for viable antigen presenting cells.
Authors: Nygard N R , Giacoletto K S , Bono C , Gorka J , Kompelli S , Schwartz B D 
Published: December 2022

Abstract
Although studies on fixed APCs have demonstrated that peptide can bind to cell surface class II molecules, the mechanisms by which peptide-class II complexes are formed in viable cells is largely unexplored. To explore the possibility that peptide loading of class II molecules was occurring after endocytosis of peptides as well as by surface binding, we utilized an immunogenic hemagglutinin peptide (HAP 128-145) from the influenza strain A/Japan/57, and studied the appearance of surface complexes of HAP 128-145 bound to HLA-DRw11 molecules on human B-lymphoblastoid cells (BLCLs). Detection of the bound peptide was made possible by a rabbit anti-serum (alpha HAP) raised against HAP 128-145, which recognizes both the free peptide as well as peptide bound to DRw11 on living APCs. Pretreatment of the BLCLs with a variety of inhibitors of protein synthesis and intracellular trafficking failed to decrease the levels of HAP 128-145/DRw11 surface complexes. However, significant inhibition in the appearance of these complexes was caused by a decrease in the temperature at which the cells were incubated with peptide. Temperature-specific inhibition was also observed for fixed DRw11-positive APCs and purified DRw11 molecules indicating that the effect of temperature was directly on the class II molecules. We conclude that surface binding of peptide by class II molecules on human B cells is a major pathway of formation of immunogenic class II-peptide complexes for at least some soluble antigenic peptides, and that endocytosis of soluble peptides with subsequent binding of peptide by intracellular class II molecules plays little if any role in the formation of such complexes. Moreover, class II molecules have evolved to stably bind peptide optimally at physiologic temperatures, independent of cell metabolism.
Immunization with soluble hepatitis B virus surface protein elicits murine H-2 class I-restricted CD8+ cytotoxic T lymphocyte responses in vivo.
Authors: Schirmbeck R , Melber K , Kuhröber A , Janowicz Z A , Reimann J 
Published: December 2022

Abstract
Immunization with soluble proteins only rarely induces a specific response of CD8+ CTL. We describe experiments that demonstrate the efficient and specific in vivo priming of CTL in BALB/c mice immunized with soluble hepatitis B virus (HBV)-derived surface (S) protein. A single (s.c., i.p. or i.v.) injection of a low dose (30 ng to 3 micrograms per mouse) of recombinant S protein particles without adjuvants induced a CTL response. This specific cytotoxic response was read out against a panel of different S protein-expressing transfected mouse cell lines. Effector cells of this response were Ld-restricted, CD3+ CD4- CD8+ CTL. H-2d/Ld+ (BALB/c, C.B-17) mice were responders; H-2d/Ld- (dm2) mutant mice and H-2b (C57BL/6) mice were nonresponders. Injections of various dosages of a S protein-derived, immunogenic, synthetic peptide into BALB/c mice by various routes did not prime CTL. After incorporation of S protein particles into IFA or aluminum hydroxide, these protein Ag lost their ability to specifically stimulate CTL in vivo. After priming of mice with S protein emulsified in IFA or adsorbed to aluminum hydroxide boost injections with native S protein particles were inefficient in stimulating a specific CTL response. These findings are of relevance for the design of synthetic subunit vaccines for which specific stimulation of CD8+ T effector functions is desired.
A monoclonal antibody that recognizes HLA-B27 in the context of peptides.
Authors: Wang J , Yu D T , Fukazawa T , Kellner H , Wen J , Cheng X K , Roth G , Williams K M , Raybourne R B 
Published: December 2022

Abstract
The T2 mutant cell line is unable to load peptides into the MHC class I Ags inside the cells. These "empty" MHC class I Ags are not expressed on the cell surface unless the cells are cultured at low temperatures. Expression will occur at 37 degrees C only in the presence of peptides that bind to and stabilize the class I Ags. T2 cells transfected with the B*2705 gene were tested with a panel of anti-HLA-B27 mAb. Two of the antibodies, ME1 and KS3, reacted with the "empty" HLA-B27 expressed at low culture temperatures. Three antibodies, B27.M1, B27.M2, and Ye-2, were unreactive with these "empty" HLA-B27. The cells were then incubated with a panel of HLA-B27-binding peptides. One of the antibodies, Ye-2, became reactive when the cells were incubated with a peptide derived from HIV gp120 and to a less degree with a peptide derived from histone H3.3. Mouse L cells transfected with the B*2705 and the human beta 2m genes also reacted very poorly with B27.M1, B27.M2, and Ye-2. Those two peptides were also able to induce high increase in Ye-2 reactivity. Alternately, increase in Ye-2 reactivity was also observed when the L cells were incubated with IFN-gamma or TNF-alpha. These experiments indicate that the Ye-2 anti-HLA-B27 mAb recognizes HLA-B27 in the context of certain residing peptides either added exogenously or expressed endogenously. The B27.M1 and B27.M2 antibodies might share similar characteristics.
Retinoic acid inhibition of IL-1-induced IL-6 production by human lung fibroblasts.
Authors: Zitnik R J , Kotloff R M , Latifpour J , Zheng T , Whiting N L , Schwalb J , Elias J A 
Published: December 2022

Abstract
IL-6 is a multi-functional cytokine that plays an important role in normal biologic homeostasis and disease pathogenesis. Retinoids are vitamin A analogs that regulate the function of a wide variety of inflammatory and structural cells. To further understand the biology of retinoids and IL-6 we determined whether all-trans-retinoic acid (RA) and other retinoids regulate lung fibroblast IL-6 production. RA did not stimulate fibroblast IL-6 production. Instead, it inhibited the production of IL-6 by IL-1-stimulated cells. This effect was dose-dependent with an IC50 of 10(-7) M RA and significant inhibition being noted with doses of RA as low as 10(-8) M. These inhibitory effects could not be explained by cytotoxicity or a shift in the kinetics of IL-6 production. They also did not appear to involve alterations in the early events in IL-1-induced IL-6 production, because RA inhibited IL-6 production even when added 6 h after IL-1 and RA did not inhibit IL-1 binding to cell surface IL-1 receptors. RA inhibition of IL-6 protein production was associated with a comparable decrease in IL-6 mRNA accumulation and gene transcription. 13-cis-retinoic acid, retinol, retinaldehyde, all-trans etretin, Ro 13-6298, and 9-cis retinoic acid also inhibited IL-1-induced IL-6 production. However, 4-hydroxyphenyl retinamide and etretinate did not share this property. The inhibitory effects of these analogues may be mediated by nuclear retinoic acid receptors as mRNA encoding RAR-alpha, RAR-gamma, and RXR-alpha were present, and RAR-beta was induced by RA in human lung fibroblasts. These studies demonstrate that RA and other retinoid analogs inhibit IL-1-induced IL-6 production and that this effect is analog-specific and, at least partially, transcriptionally mediated.
VH and V kappa segment structure of anti-insulin IgG autoantibodies in patients with insulin-dependent diabetes mellitus. Evidence for somatic selection.
Authors: Ikematsu H , Ichiyoshi Y , Schettino E W , Nakamura M , Casali P 
Published: December 2022

Abstract
In some patients with insulin-dependent (type I) diabetes mellitus (IDDM), autoantibodies to insulin are present at diagnosis. After initiation of the treatment with not only animal but also human insulin, anti-insulin, mainly IgG, autoantibodies become a major component of the autoimmune response in virtually all IDDM patients. Their structure, however, is still relatively unknown. We analyzed the structure of the VH and V kappa segments of three human IgG mAb derived from three IDDM patients. The sequences of VH genes of two IgG, mAb13 and mAb48, were 98.3 and 96.6% identical with those of the H11 and 1.9III genes (VHIII family), respectively. The sequence of the VH gene of the third IgG, mAb49, was 98.6% identical with that of the 51p1 gene (VHI family). All three IgG mAb used V kappa III segments. The V kappa III gene sequences of mAb13 and mAb49 were 97.9 and 98.9% identical, respectively, to that of the kv3g gene; the mAb48 V kappa gene sequence was 96.5% identical to that of the kv328 gene. The VH and/or V kappa segments of these anti-insulin IgG mAb are similar to Ig V genes expressed in the fetal, and adult normal and autoimmune B cell repertoires. The nucleotide differences displayed by the three anti-insulin IgG mAb VH gene sequences, when compared with those of the closest reported germ-line genes, were concentrated in the CDR (6.2 x 10(-2) and 0.8 x 10(-2) difference/base in CDR and FR, respectively; p &lt; 0.01, chi 2 test), and yielded a significantly higher putative replacement (R) to silent (S) mutation ratio in the CDR (12.0) than in the framework (0.2). The concentration of nucleotide differences in the CDR and their high R:S putative mutation ratios were consistent with the hypothesis that these expressed VH genes underwent a process of somatic mutation and Ag-driven clonal selection. That such differences constituted somatic point-mutations was formally proved in IgG mAb13, by differentially targeted PCR amplification and Southern blot hybridization of the mAb13-producing cell line DNA. The putative germ-line gene that gave rise to the expressed VH segment was cloned using genomic DNA from PMN of the same patient whose B cells were used for the generation of this mAb. Overall, in the anti-insulin IgG mAb VH and V kappa III genes, the (putative and verified) somatic point-mutations yielded 27 amino acid replacements, of which 14 nonconserved. Four of these resulted in positively charged residues, three Arg and one His.(ABSTRACT TRUNCATED AT 400 WORDS)
Up-regulation of the MHC class II molecules on B cells by peptide ligands.
Authors: Agrawal B , Fraga E , Kane K , Singh B 
Published: December 2022

Abstract
MHC class I and class II molecules present peptide Ag to T lymphocytes. Peptides are critical in class I heavy chain folding and/or stable association with beta 2m. A recent study suggests the role of peptide Ag binding for MHC class II alpha- and beta-chain heterodimers to enter into a compact state and allow their transport to the cell surface. We have investigated the effect of peptide ligands on the expression of MHC class II I-A(d) molecules on the B cell hybridoma, TA3. These cells, when cultured in vitro, gradually lost the surface expression of I-A(d) molecules. Incubation with peptides, having high affinity for binding to intact I-A(d) molecules, significantly increased the surface expression of I-A(d) in less than 24 h. The ability of peptides to induce increased expression of I-A(d) correlated with the affinity of peptide to intact I-A(d), and an I-Ak-restricted peptide did not have an effect on I-A(d) expression. The effect could be reversed after the removal of the peptide Ag. Based on our studies with inhibitors of protein synthesis and intracellular transport, the mechanism for up-regulation of I-A(d) expression by peptides seems to involve intracellular pathway but appears not to require new protein synthesis or transport from ER. Our results suggest that the decrease in surface expression of I-A(d) on TA3 cells may result from their failure to generate or to be saturated with naturally processed peptide ligands. Thus peptide ligands are evidently important in regulating surface expression of MHC class II molecules and their recognition by T lymphocytes.
IL-6-induced calmodulin-dependent protein phosphorylation in B9 hybridoma cells.
Authors: Choi I , Na W J , Kang H S , Pyun K H 
Published: December 2022

Abstract
The addition of IL-6 to the IL-6-dependent B9-55 hybridoma cells starved for IL-6 for 16 h induced an increased phosphorylation of a 25-kda protein (p25). Phosphorylation of p25 was IL-6 dependent and maximal at 5 min after IL-6 stimulation. Phosphorylation of p25 was inhibited by N-(6-aminohexyl)-5-chloro-1-naphthalene sulfonamide (W-7), an inhibitor of calmodulin-dependent protein kinase, but not by tyrphostin nor ionomycin. When enolase was used as an exogenous substrate, similar results were obtained. Furthermore, calmodulin stimulated p25 phosphorylation in vitro. IL-6-starved B9 hybridoma cells synthesized DNA at 9 to 12 h after addition of IL-6. Preincubation of cells with W-7 before IL-6 stimulation blocked DNA synthesis. Similar effects were observed when cells were preincubated with tyrphostin or ionomycin but to a lesser extent. The concentrations of W-7 required for blocking p25 phosphorylation and DNA synthesis correlated relatively well. These results suggest that an IL-6-inducible calmodulin-dependent protein kinase is involved in IL-6 signal transduction, including p25 phosphorylation and B9 hybridoma proliferation.