bluuebunny/crossref_metadata_2025_split huggingface dataset:
IL-12 exacerbates helminth-mediated corneal pathology by augmenting inflammatory cell recruitment and chemokine expression.
Authors: Pearlman E , Lass J H , Bardenstein D S , Diaconu E , Hazlett F E , Albright J , Higgins A W , Kazura J W 
Published: December 2022

Abstract
Corneal inflammation (keratitis) is a major cause of visual impairment in Onchocerca volvulus infection. Previous studies showed that onchocercal keratitis can be induced in mice following s.c. immunization and intracorneal injection with soluble O. volvulus Ags (OvAg), and that the inflammatory response is dependent on T cells and IL-4. Since recombinant IL-12 impairs IL-4-dependent, Th2-mediated responses in other parasitic infections and in models of allergic asthma, the present study was undertaken to determine the effect of IL-12 on onchocercal keratitis. Mice were injected i.p. with IL-12 or saline at the time of initial sensitization to OvAg. Surprisingly, IL-12 treatment caused significant exacerbation of corneal pathology, which was associated with increased eosinophil and mononuclear cell infiltration into the corneal stroma. Consistent with the well-documented effect of IL-12 on Th1 cell development, corneas of IL-12-treated animals had elevated expression of the Th1 cytokine IFN-gamma and diminished expression of the Th2 cytokines IL-4, IL-5, IL-10, and IL-13. However, corneas from these animals also had marked elevation of alpha- and beta-chemokines known to be active on eosinophils and mononuclear cells, including IFN-gamma-inducible protein (IP)-10, macrophage inflammatory protein-1alpha, macrophage inflammatory protein-1beta, JE/monocyte chemotactic protein-1, RANTES (regulated upon activation, normal T expressed and secreted), and eotaxin. Together, these data indicate that IL-12 exacerbates OvAg-mediated corneal pathology by enhancing chemokine expression and recruitment of inflammatory cells.
Human monocyte adhesion is modulated by endothelin B receptor-coupled nitric oxide release.
Authors: King J M , Srivastava K D , Stefano G B , Bilfinger T V , Bahou W F , Magazine H I 
Published: December 2022

Abstract
Human monocytes have the capacity to produce both endothelin 1 (ET-1) and nitric oxide (NO), yet the roles of these mediators in monocyte function remain unclear. The relationship of ET-1 and NO release to monocyte adhesion was explored using peripheral blood monocytes (PBM) and the human monocytic cell lines THP-1 and U937. Specific binding of 125I-labeled ET-1 to THP-1 was abrogated by pretreatment with the endothelin B (ET(B)) receptor antagonist, BQ-788, but not by the endothelin A (ET(A)) receptor antagonist, BQ-123, consistent with predominant ET(B) receptor expression. Direct measurement of NO with an amperometric probe demonstrated the production of nanomolar concentrations of NO by PBM and THP-1 cells upon treatment with ET-1, which was abrogated by BQ-788, but not BQ-123, pretreatment, suggesting functional coupling of ET(B) receptors to NO release. Indeed, the presence of ET(B) receptor mRNA transcripts was detected in THP-1 and is consistent with previous reports that have demonstrated functional coupling of ET(B) receptors to constitutive NO synthase activation. In contrast, U937 cells did not release NO in response to ET-1 treatment, and mRNA transcripts were not detected in these cells, consistent their failure to bind 125I-labeled ET-1, as previously determined. Exposure of PBM to ET-1 markedly reduced the adhesion of these cells to human saphenous vein, whereas PBM adhesion in the presence of BQ-788 was restored to control levels. These data demonstrate that PBM interactions with the vascular wall can be reduced by autocrine production of NO and suggest that ET(B) receptors may attenuate monocyte activity at sites of inflammation.
Mechanism of sustained E-selectin expression in cultured human dermal microvascular endothelial cells.
Authors: Kluger M S , Johnson D R , Pober J S 
Published: December 2022

Abstract
Persistent E-selectin expression has been proposed to be a unique property of dermal vascular endothelium that directs skin-specific homing of a subpopulation of circulating memory T cells. We compared the kinetics of E-selectin expression on cultured human dermal microvascular endothelial cells (HDMEC) with expression on HUVEC. Following treatment with TNF, E-selectin on HDMEC appears more slowly than on HUVEC (peak values 6-8 vs 4 h, respectively) and is sustained at significantly higher levels after 24 h. E-selectin mRNA, analyzed by S1 nuclease protection, consists of a single predominant transcript that follows a similarly transient time course in both cell types. Cell surface E-selectin is internalized more slowly on HDMEC than on HUVEC (t1/2 = 4.3 vs 1.6 h, respectively) as measured by serial FACS analyses in the presence of the protein synthesis inhibitor cycloheximide. In comparison, intercellular adhesion molecule-1 (ICAM-1) expression is not measurably reduced by either cell type under the same conditions. HDMEC are similar to HUVEC in rates of pinocytosis or receptor-mediated endocytosis. Pulse-chase analysis indicated that the degradative half-life of E-selectin protein is greater in HDMEC than in HUVEC (1.9 vs 1.5 h, respectively). E-selectin internalization in microvascular endothelial cells (EC) from lung and subcutaneous fat is slow, like HDMEC, whereas internalization in large vessel EC from saphenous vein and aorta is rapid, like HUVEC. We conclude that HDMEC sustain higher levels of expression at 24 h by slower internalization and degradation of E-selectin protein and that this may be a general property of microvascular EC.
Regulation of inducible nitric oxide synthase messenger RNA expression and nitric oxide production by lipopolysaccharide in vivo: the roles of macrophages, endogenous IFN-gamma, and TNF receptor-1-mediated signaling.
Authors: Salkowski C A , Detore G , McNally R , van Rooijen N , Vogel S N 
Published: December 2022

Abstract
To evaluate potential roles for macrophages, IFN-gamma, and TNF receptor 1 (TNFR1) in the regulation of LPS-induced inducible nitric oxide synthase (iNOS) mRNA expression, we used a model of macrophage depletion as well as IFN-gamma (GKO) and TNFR1 (TNFR1 -/-) knockout mice. LPS-induced iNOS mRNA in spleen was ablated in both macrophage-depleted and GKO mice. In livers of macrophage-depleted mice, LPS-induced iNOS mRNA was reduced by 55 to 85%, with the most profound reductions detected 6 and 8 h after LPS injection. In GKO mice, peak iNOS mRNA expression in liver (3 h) was unaffected by the loss of endogenous IFN-gamma. By 6 to 12 h after LPS challenge, however, hepatic LPS-induced iNOS mRNA and serum nitrate/nitrite levels were reduced substantially in GKO mice. Residual LPS-induced iNOS mRNA in livers of GKO mice was nearly ablated by macrophage depletion, indicating that induction of iNOS mRNA in liver requires both endogenous IFN-gamma and either macrophages and/or macrophage-derived factors. TNFR1-mediated signaling was involved in the induction of LPS-induced iNOS mRNA in liver at 3 and 6 h, but not in its maintenance at 8 h. Conversely, induction of iNOS mRNA in spleen by LPS was independent of TNFR1-mediated signaling. Our results indicate that macrophages and/or their secreted products, endogenous IFN-gamma production, and TNFR1-mediated signaling play key roles in the in vivo regulation of iNOS mRNA expression and that the extent of their involvement is both time and organ specific.
IgE regulates mouse basophil Fc epsilon RI expression in vivo.
Authors: Lantz C S , Yamaguchi M , Oettgen H C , Katona I M , Miyajima I , Kinet J P , Galli S J 
Published: December 2022

Abstract
The binding of IgE to high-affinity IgE receptors (Fc epsilon RI) on the surface of mast cells and basophils primes these cells to secrete a panel of proinflammatory mediators upon subsequent exposure to specific Ag. We now find that the level of Fc epsilon RI expression on bone marrow basophils in mice infected with the nematode Strongyloides venezuelensis exhibits a strong positive correlation with the serum concentration of IgE, as was previously reported for human blood basophils. Moreover, the administration of IgE in vivo can significantly upregulate Fc epsilon RI expression on mouse basophils, and genetically IgE-deficient (IgE -/-) mice exhibit a dramatic (approximately 81%) reduction of basophil Fc epsilon RI expression compared with the corresponding normal (IgE +/+) mice. The finding that IgE can be a major regulator of mouse basophil Fc epsilon RI expression in vivo identifies a potentially important mechanism for enhancing the expression of effector cell function in IgE-dependent allergic reactions or immunologic responses to parasites.
Development of IFN-gamma-producing CD8+ gamma delta+ T lymphocytes and IL-2-producing CD4+ alpha beta+ T lymphocytes during contact sensitivity.
Authors: Dieli F , Asherson G L , Sireci G , Dominici R , Sciré E , Salerno A 
Published: December 2022

Abstract
This paper describes the development of Ag-specific proliferation and the production of IFN-gamma and IL-2 during contact sensitivity (CS) to the hapten picryl chloride (PCl). Lymph node cells from mice immunized with PCl proliferate and produce IFN-gamma and IL-2 when re-exposed to the specific Ag in vitro. Time course experiments showed that the peak IFN-gamma production occurred at days 3 and 4 after immunization, with a sharp decline by day 6. In contrast, proliferation and IL-2 production peaked at day 3 but persisted up to day 10. Proliferation and IFN-gamma and IL-2 production displayed by immune lymph node cells were Ag-specific but required different cell populations. In fact, the production of IFN-gamma was due to a CD8+, gamma delta+ T cell, while proliferation and IL-2 production required the presence of a CD4+, alpha beta+ T cell. Furthermore, IFN-gamma production showed genetic (MHC) restriction, and finer analysis using congenic strains of mice indicated that the K molecule was the restricting element. This was confirmed by blocking the K molecule of the APC used to trigger IFN-gamma production with a specific mAb. In contrast, proliferation and IL-2 production were I-A-restricted, as demonstrated using congenic strains of mice and blocking the I-A molecule of the APCs. Further analysis using purified gamma delta+ cells revealed that these cells produced IFN-gamma in an Ag-specific and MHC (K)-restricted fashion. Injection of mice with a mAb to IL-2 blocked subsequent in vitro proliferation, as well as IL-2 and IFN-gamma production, while all three in vitro responses were unaffected by injection of a mAb to IFN-gamma given at the time of immunization. Furthermore, injection of mice with a mAb to IL-2 blocked the CS reaction when given at the time of immunization, while it had no effect when given at the time of challenge. Injection of mice with mAb to IFN-gamma at the time of challenge reduced but did not abolished CS, suggesting that IFN-gamma is important but not exclusively responsible for the CS reaction.
Nuclear Rel-A and c-Rel protein complexes are differentially distributed within human thymocytes.
Authors: Feuillard J , Dargemont C , Ferreira V , Tarantino N , Debré P , Raphael M , Körner M 
Published: December 2022

Abstract
Nuclear factor-kappa B (NF-kappa B)/Rel proteins are inducible transcriptional regulators of numerous cellular genes. They are particularly abundant in lymphoid tissues and are thought to be critical for the transcription of genes involved in immune and inflammatory responses. We have reported previously that a nuclear NF-kappa B activity was present in freshly extracted human thymocytes in the absence of in vitro treatment of these cells. In the present report, we identified NF-kappa B proteins extracted from human thymocyte nuclei as being p50/p65 and p50/c-Rel complexes. Immunochemical and immunofluorescent staining of thymus sections using specific Abs allowed visualization of nuclear NF-kappa B proteins in both thymocytes and nonthymocyte cells. This detection suggested a preferential activation of p50/c-Rel in medullary thymocytes, whereas p50/p65 was present in both cortical and medullary regions of human thymus lobules. However, the intensity of p65 labeling was much higher in several thymocytes from the medulla. p65, p50, and c-Rel activities were found in both CD4- and CD8-positive thymocytes. These observations suggest that p65 and c-Rel complexes play distinct roles in gene expression and that both forms of NF-kappa B play critical roles during late stages of the intrathymic maturation of T cells.
Syk, but not Lyn, recruitment to B cell antigen receptor and activation following stimulation of CD45- B cells.
Authors: Pao L I , Cambier J C 
Published: December 2022

Abstract
B cell Ag receptor (BCR) signaling occurs via tyrosine phosphorylation of CD79a and CD79b ITAMs, leading to recruitment and activation of Lyn and Syk tyrosine kinases and subsequent downstream events. CD45 expression is required for BCR triggering of certain of these downstream events, such as calcium mobilization and p21ras activation. However, the site in the BCR signaling cascade at which CD45 impinges is poorly defined. To address this question, we have studied CD45 function in the CD45-deficient (CD45-) and CD45-reconstituted (CD45+) J558L mu m3 plasmacytoma. In both CD45+ and CD45- cells, Ag stimulation led to CD79a and CD79b tyrosine phosphorylation as well as Syk tyrosine phosphorylation, recruitment to the receptors, and activation. In contrast to CD45+ cells, Lyn exhibited high basal tyrosine phosphorylation in the CD45- cells and was not further phosphorylated upon Ag stimulation. Mapping studies indicated that the observed constitutive phosphorylation of Lyn reflects phosphorylation of its C-terminal tyrosine, Y508, at high stoichiometry. Constitutively Y508-phosphorylated Lyn was neither recruited to the BCR nor activated upon Ag stimulation. Moreover, CD79a-ITAM phosphopeptides failed to bind Lyn from the CD45- cells. Thus, Y508 phosphorylation of Lyn occurs in the absence of cellular CD45 expression and appears to render the kinase unable to associate with the phosphorylated receptor complex via its Src homology 2 domain and to participate in signal propagation. Surprisingly, in view of previous findings implicating Src family kinases in ITAM phosphorylation, the data indicate that Ag-induced CD79a and CD79b tyrosine phosphorylation and Syk recruitment and activation can occur in the absence of CD45 expression and, hence, Src-family kinase activation.
IL-2-regulated expression of the monocyte chemotactic protein-1 receptor (CCR2) in human NK cells: characterization of a predominant 3.4-kilobase transcript containing CCR2B and CCR2A sequences.
Authors: Polentarutti N , Allavena P , Bianchi G , Giardina G , Basile A , Sozzani S , Mantovani A , Introna M 
Published: December 2022

Abstract
NK cells migrate in response to C-C chemokines, including monocyte chemotactic protein-1 (MCP-1) and MCP-3. Increased migration was observed in IL-2-activated NK cells. It was therefore of interest to define the expression in resting and activated NK cells of the MCP-1 receptor (CCR2) for which two cDNAs (A and B) have been described. Specific oligonucleotides and reverse-transcriptase PCR revealed the presence in activated NK cells and mononuclear phagocytes of the fragments expected on the basis of the reported cDNAs. In addition, amplification with a common A/B- and an A-specific oligonucleotide yielded an unexpected, abundant, 1649-bp fragment. Sequence analysis as well as Northern blotting and RNase protection with different probes revealed that the CCR2 gene is expressed in activated NK cells and mononuclear phagocytes as a predominant long transcript (3.4 kb) consisting of CCR2B, followed by a novel sequence (X), corresponding to an intron in the genome, and by a CCR2A-specific portion. The predominant long transcript is polyadenylated and present in the cytoplasm. The augmented migratory capacity of IL-2 activated vs resting NK cells was associated with increased CCR2 transcript levels.
Cloned dendritic cells can present exogenous antigens on both MHC class I and class II molecules.
Authors: Shen Z , Reznikoff G , Dranoff G , Rock K L 
Published: December 2022

Abstract
Pathways for presenting proteins from the extracellular fluids on MHC class I molecules have been described in macrophages. However, it is uncertain whether similar mechanisms exist in dendritic cells, because conventional preparations of these cells can be contaminated with macrophages. We addressed this issue by transducing granulocyte-macrophage CSF into bone marrow cultures followed by supertransfection with myc and raf oncogenes. These immortalized clones displayed dendritic morphology, and many expressed the dendritic cell-specific markers DEC-205 and 33D1 as well as high levels of MHC molecules and costimulatory molecules. Using these cloned dendritic cells, we found that exogenous OVA could be presented on both their MHC class I and class II molecules. This presentation was markedly enhanced when the Ag was particulate and internalized by phagocytosis. Presentation of particulate OVA on MHC class I molecules was insensitive to the weak base chloroquine, but was blocked by peptide aldehyde inhibitors of the proteasome, indicating that the class I-presented peptides were generated in the cytosol. Brefeldin A, which inhibits the exocytosis of newly synthesized proteins from the endoplasmic reticulum, also inhibited Ag presentation. These results establish that dendritic cells can present exogenous Ags on MHC class I molecules and appear to use a similar phagosome to cytosol pathway as macrophages. Therefore, dendritic cells are likely to play an important role in generating immune responses to tissue transplants and tumors in vivo. Furthermore, these findings provide an approach for targeting vaccine Ags into these cells to prime immune responses in vivo.
The tetraspanin protein CD82 associates with both free HLA class I heavy chain and heterodimeric beta 2-microglobulin complexes.
Authors: Lagaudrière-Gesbert C , Lebel-Binay S , Wiertz E , Ploegh H L , Fradelizi D , Conjeaud H 
Published: December 2022

Abstract
CD82 is a tetraspan transmembrane protein on NK/LAK-susceptible targets. A single highly glycosylated protein of heterogeneous molecular mass (50-90 kDa) was immunoprecipitated by anti-CD82 from Nonidet P-40 lysates of various B cell lines, Raji, Daudi, 721, and 721.134. Using the milder detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), additional proteins were coprecipitated with CD82 from surface iodinated B cell lines, including a major band at 45 kDa, identified as the HLA class I heavy chain by sequential immunoprecipitations and sequential immunoprecipitation-Western blot analysis. Cocapping experiments confirmed the molecular association of CD82 and HLA class I at the cell surface of these B cell lines. CD82 could be coprecipitated with both mature and beta 2-microglobulin (beta 2m)-free heavy chains of MHC-I from CHAPS extracts. No association between MHC-I and CD82 was found in the beta 2m-deficient Daudi cell line or after co-in vitro translation of CD82, MHC heavy chain, and beta 2m mRNA. The most likely source of free class I heavy chains at the cell surface is by dissociation of beta 2m-associated class I molecules. These results suggest that association of CD82-MHC-I takes place at the cell surface and could interfere with the capacity of the MHC-I complex to protect targets from NK-mediated cytotoxicity.
Structural C3 diversity in fish: characterization of five forms of C3 in the diploid fish Sparus aurata.
Authors: Sunyer J O , Tort L , Lambris J D 
Published: December 2022

Abstract
In virtually all species examined to date, the functionally active third component of complement (C3) is encoded by a single gene. We have recently demonstrated, however, that trout possess three structurally and functionally distinct active C3 that represent the products of at least two different C3 genes. In the present study, we provide evidence that multiple forms of functional C3 occur not only in the rainbow trout (Salmo gairdneri), a quasi-tetraploid old teleost fish, but also in the diploid gilthead sea bream (Sparus aurata), a modern teleost fish. In the gilthead sea bream, we have characterized five different forms of C3 (C3-1, C3-2, C3-3, C3-4, and C3-5); in addition, we have identified and isolated a C5-like molecule. Each of the six proteins was composed of an alpha-chain and a beta-chain; however, only the C3 isoforms contained a thioester bond in their alpha-chains. These proteins all differed in the molecular masses of their alpha- and beta-chains and in their glycosylation patterns, reactivity with various Abs, tryptic peptide maps, and NH2-terminal sequences of their chains. These observations together with the fact that each of the six proteins were also purified from a single fish suggest that the C3 isoforms represent the products of several genes. The presence of multiple forms of C3 in a modern diploid fish, very distant in evolutionary time from the trout, strongly suggests that the C3 isoforms generated once from a single C3 gene have remained functional in the genomes of these animals. These findings not only have important consequences for our understanding of the evolution of the C3 protein, but also provide evidence for the formation and generation of a new C3-related gene family.
Nature and origin of polymorphism in feline MHC class II DRA and DRB genes.
Authors: Yuhki N , O'Brien S J 
Published: December 2022

Abstract
Transcripts of the MHC class II DRA and DRB gene homologues of the domestic cat (Felis catus) were cloned and sequenced to compare the pattern and process of DR gene divergence. Homologous DRB exon 2 sequences from 36 feral domestic cats throughout the world plus from three species of Felidae (tiger cat, Iriomote cat, and Geoffroy's cat) were also determined. Limited variation in the domestic cat Feca-DRA gene was observed, but abundant variation in the Feca-DRB gene was seen comprising 61 distinct DRB alleles. Phylogenetic analyses resolved at least five monophyletic feline DRB allelic lineages (DRB*1 to *5), which are clearly distinct from those of human (HLA-DRB1 to 9 lineages), mouse (H-2Ebeta b, u, f), and dog DRB alleles. Approximately 80% of individual cats contained three to six distinct DRB sequences, indicating that feline MHC maintains two to three DRB loci. Five cats had three DRB sequences in a single allelic lineage, indicating the occurrence of recent gene duplication of feline DRB genes. DRB sequences isolated from three exotic cats demonstrated close association with a particular domestic cat DRB lineage, suggesting that these allelic lineages are derived from common ancestral alleles that existed prior to the divergence of these feline species about 10 to 15 million years ago. Patterns of synonymous and nonsynonymous nucleotide substitution rates that occurred in Ag recognition sites (ARS) and nonrecognition (NAR) sites demonstrated a strong role of natural selection--positive selection for Ag recognition sites and negative selection for nonrecognition sites of feline DRB sequences--in the process of evolution of DR molecules.