bluuebunny/crossref_metadata_2025_split huggingface dataset:
The N-Terminal Domains Target TNF Receptor-Associated Factor-2 to the Nucleus and Display Transcriptional Regulatory Activity
Authors: Min Wang , Bradley John R. , Galbraith Jennifer J. , Jones Sally J. , Ledgerwood Elizabeth C. , Pober Jordan S. 
Published: January 2023

Abstract
The subcellular localization of the TNF receptor-associated factor-2 (TRAF2) adaptor protein in human endothelial cells, which mediates proinflammatory responses of TNF, has been analyzed by confocal immunofluorescence microscopy and by Western blotting of fractionated cell extracts. Rabbit antisera reactive with either amino- or carboxyl-terminal TRAF2 peptides frequently but not uniformly stain nuclei of cultured HUVEC or the established human endothelial cell line, ECV304. However, Western blotting reveals significant heterogeneity in the reactivities of these polyclonal Abs. Transiently transfected HUVEC expressing FLAG epitope-tagged TRAF2 consistently show prominent nuclear localization, and deletion mutants of TRAF2 identify the portion of the molecule responsible for nuclear localization as the amino-terminal ring finger domain. TNF treatment does not appear to influence the localization of endogenous or transfected TRAF2 protein. Transfection of the amino-terminal half of the TRAF2 molecule, containing the ring and zinc finger domains, which localizes to the nucleus, results in activation of E-selectin but not of NF-κB promoter-reporter gene transcription or of c-Jun N-terminal kinase activation. These observations suggest that TRAF2 may reside in the nucleus and directly regulate transcription, independent of its role in cytoplasmic signal transduction.
Regulation of IL-6 Signaling by p53: STAT3- and STAT5-Masking in p53-Val135-Containing Human Hepatoma Hep3B Cell Lines
Authors: Rayanade Ravi J. , Ndubuisi MacKevin I. , Etlinger Joseph D. , Sehgal Pravin B. 
Published: January 2023

AbstractThe influence of p53 on cytokine-triggered Janus kinase-STAT signaling was investigated in human hepatoma Hep3B cell lines engineered to constitutively express the temperature-sensitive Val135 mutant of p53. In comparison to the parental p53-free Hep3B cells, these p53-Val135-containing Hep3B cell lines displayed a reduced response to IL-6 at the wild-type-like p53 temperature (32.5°C). In these cells, IL-6 induced a marked reduction in the immunologic accessibility of cytoplasmic and nuclear STAT3 and STAT5 within 20 to 30 min that lasted 2 to 4 h (STAT-masking) provided that the cells had been previously cultured at 32.5°C for at least 18 to 20 h. The onset of IL-6-induced STAT-masking required protein tyrosine kinase, protein tyrosine phosphatase, proteasomal, phospholipase C, and mitogen-activated protein kinase kinase 1 activities. The maintenance of IL-6-induced STAT-masking was dependent on continued signaling through the phosphatidylinositol-dependent phospholipase C pathway. Despite a reduction in IL-6-induced STAT3 DNA binding activity in the nuclear compartment during STAT-masking, there was increased and prolonged accumulation of tyrosine-phosphorylated STAT3 in both the cytoplasmic and nuclear compartments, indicating that the capacity of tyrosine-phosphorylated STAT3 to bind DNA was reduced during STAT-masking. Thus, IL-6-induced STAT-masking, as dramatically evident on immunomicroscopy, is a visible consequence of a novel cellular process by which a p53-Val135-induced gene product(s) regulates the association of masking protein(s) with and the DNA-binding capacity of STAT3.
Differences Between B Cell and Macrophage Transformation by the Bovine Parasite,Theileria annulata: A Clonal Approach
Authors: Sager Heinz , Bertoni Giuseppe , Jungi Thomas W. 
Published: January 2023

AbstractTheileria annulata, a tick-transmitted protozoan parasite, infects and transforms cells of the hemopoietic system, particularly those of the B cell and monocyte/macrophage lineages. Here, the effect of infection/transformation on the resulting phenotype was studied using a clonal approach. Three phenotypes of transformed cell lines could be discerned. The first is characterized by surface expression of IgM, CD21, and the B cell epitopes, B-B2 and B-B8, Ig heavy chain gene rearrangement, and mRNA expression. Such lines were obtained from fresh and cultured PBMC and at increased frequency from purified B cells, but never from fetal bone marrow cells. The second phenotype can be distinguished from the first by the absence of Ig heavy chain expression and reduced surface expression of B cell markers (CD21, B-B2, B-B8). Clones with this phenotype were obtained from transformed fetal bone marrow cells only. The third phenotype showed an absence of all of the above B cell markers, including surface IgM, and a lack of Ig heavy chain gene rearrangement. The latter clones could be maintained for several weeks after elimination of T. annulata by BW720c treatment, and they reacquired a macrophage-like phenotype. This implies that parasite-induced dedifferentiation is restricted to monocyte/macrophage, and that B cell markers are indicative of cell lineage progeny. Demonstration of surface IgM on PBMC-derived B cell clones suggests that infection of B cells with T. annulata may be an epigenetic method to immortalize ruminant B cells of a defined Ag specificity.
Human Eosinophils Produce Biologically Active IL-12: Implications for Control of T Cell Responses
Authors: Grewe Markus , Czech Wolfgang , Morita Akimichi , Werfel Thomas , Klammer Michaela , Kapp Alexander , Ruzicka Thomas , Schöpf Erwin , Krutmann Jean 
Published: January 2023

AbstractThe present study assessed the capacity of eosinophils (EOS) to synthesize the cytokine IL-12. Blood-derived, highly purified human EOS from six atopic patients and two nonatopic individuals were treated in culture with IL-4, IL-5, granulocyte-macrophage CSF, IFN-γ, TNF-α, IL-1α, RANTES, and complement 5a, respectively. The expression of both IL-12 protein and mRNAs for the p35 and p40 IL-12 subunits was strongly induced in all donors by the Th2-like cytokines IL-4 and granulocyte-macrophage CSF and was also moderately induced by TNF-α and IL-1α. IL-5 treatment resulted in IL-12 synthesis in four atopic donors and one nonatopic donor, whereas IFN-γ induced IL-12 synthesis in only two atopic donors. In contrast, RANTES exclusively induced mRNA for the p40 subunit without detectable protein release, and complement 5a had no effect on IL-12 mRNA or protein expression. EOS-derived IL-12 was biologically active, because supernatants derived from IL-4-treated EOS superinduced the Con A-induced expression of IFN-γ by a human Th1-like T cell line. This activity was neutralized by anti-IL-12 Abs. In conclusion, EOS secrete biologically active IL-12 after treatment with selected cytokines, which mainly represent the Th2-like type. Consequently, EOS may promote a switch from Th2-like to Th1-like immune responses in atopic and parasitic diseases.
SB 203580 Inhibits p38 Mitogen-Activated Protein Kinase, Nitric Oxide Production, and Inducible Nitric Oxide Synthase in Bovine Cartilage-Derived Chondrocytes
Authors: Badger Alison M. , Cook Michael N. , Lark Michael W. , Newman-Tarr Tonie M. , Swift Barbara A. , Nelson Allen H. , Barone Frank C. , Kumar Sanjay 
Published: January 2023

Abstract
Nitric oxide (NO) is implicated in a number of inflammatory processes and is an important mediator in animal models of rheumatoid arthritis and in in vitro models of cartilage degradation. The pyridinyl imidazole SB 203580 inhibits p38 mitogen-activated protein (MAP) kinase in vitro, blocks proinflammatory cytokine production in vitro and in vivo, and is effective in animal models of arthritis. The purpose of this study was to determine whether SB 203580 could inhibit p38 MAP kinase activity, NO production, and inducible NO synthase (iNOS) in IL-1 stimulated bovine articular cartilage/chondrocyte cultures. The results indicated that SB 203580 inhibited both IL-1 stimulated p38 MAP kinase activity in isolated chondrocytes and NO production in bovine chondrocytes and cartilage explants with an IC50 value of approximately 1 μM. To inhibit NO production, SB 203580 had to be present in cartilage explant cultures during the first 8 h of IL-1 stimulation, and activity was lost when it was added 24 h following IL-1. SB 203580 did not inhibit iNOS activity, as measured by the conversion of arginine to citrulline, when added directly to cultures where the enzyme had already been induced, but had to be present during the induction period. Using a 372-bp probe for bovine iNOS we demonstrated inhibition of IL-1-induced mRNA by SB 203580 at both 4 and 24 h following IL-1 treatment. The iNOS mRNA levels were consistent with NO levels in 24-h cell culture supernatants of the IL-1-stimulated bovine chondrocytes used to obtain the RNA.
Requirements for B7-CD28 Costimulation in Mucosal IgA Responses: Paradoxes Observed in CTLA4-Hγ1 Transgenic Mice
Authors: Gärdby Eva , Lane Peter , Lycke Nils Y. 
Published: January 2023

AbstractThe block in the CD80/CD86-CD28/CTLA-4 pathway in CTLA4-Hγ1 transgenic (Tg) mice results in strongly impaired systemic IgG immunity and failure to develop germinal center reactions. By contrast, here we report that mucosal immunity and IgA B cell differentiation are not affected by this block. We found abundant germinal centers and evidence of IgA switch differentiation in Peyer’s patches, normal total IgA levels, and normal numbers of IgA-labeling cells in the gut mucosa. The distribution of B-1 and B-2 cells and the relative contribution of B-1 cells to the total IgA B cells were similar in Tg and wild-type mice. Despite this, oral immunizations with keyhole limpet hemocyanin plus cholera toxin adjuvant failed to stimulate Ag-specific mucosal IgA responses in CTLA4-Hγ1 Tg mice. This was not due to a lack of adjuvant activity of cholera toxin in Tg mice, nor was this secondary to an inability to take up Ag from the gut lumen. Rather, CD4+ T cells stimulated by oral immunization in Tg mice appeared to be inappropriately primed, as evidenced by a significantly reduced level of CD40 ligand and CD44 expression and an increased expression of CD95 compared to those in wild-type mice. This study reveals a paradox in the regulation of mucosal IgA responses.
Cutting Edge: Linked Suppression of Skin Graft Rejection Can Operate Through Indirect Recognition
Authors: Wise Matt P. , Bemelman Frederike , Cobbold Steve P. , Waldmann Herman 
Published: January 2023

Abstract
Adult mice can be rendered immunologically tolerant of allogeneic tissues if transplanted under cover of mAbs to CD4 and CD8. Tolerance generated in this manner is characterized by the presence of regulatory CD4+ T cells that can recruit naive T cells to become tolerant also through “infectious tolerance.” Regulatory CD4+ T cells can also suppress rejection of third party transplant Ags provided they are expressed on the same graft as the tolerated Ags. This process of linked suppression can act across whole MHC barriers and represents a powerful mechanism with therapeutic potential. Tolerance can also be induced to reprocessed minor transplantation Ags presented through host APCs (indirect recognition). We here demonstrate that linked suppression can also be induced through the indirect pathway. This finding may be important in the development of transplantation tolerance in the clinic.
The Role of Host (Endogenous) T Cells in Chronic Graft-Versus-Host Autoimmune Disease
Authors: Chen Fangqi , Maldonado Michael A. , Madaio Michael , Eisenberg Robert A. 
Published: January 2023

AbstractChronic graft-vs-host (cGVH) disease induced by the transfer of Ia-incompatible spleen cells from one normal mouse strain (such as B6.C-H2bm12/KhEg (bm12)) to another (such as C57BL/6) causes an autoimmune syndrome resembling systemic lupus erythematosus (SLE). The role of host-derived T cells in this response is not obvious. Previous reports suggested that host T cells might serve to down-regulate the autoimmune syndrome. To address this issue more definitively, we used CD4 knockout (KO) or CD8KO C57BL/6 (B6) mice as recipients in the bm12→C57B6 cGVH model. CD4KO B6 mice injected with allogeneic bm12 spleen cells (bm12→CD4KO group) showed no evidence of cGVH disease. They made no detectable autoantibodies, including anti-chromatin, anti-dsDNA, anti-ssDNA, and rheumatoid factor. They survived at least 20 wks after induction of cGVH disease; and they did not develop nephritis, based on the absence of detectable levels of proteinuria and normal renal histology at the time of sacrifice. By contrast, CD8KO B6 mice (bm12→CD8KO group) and normal B6 mice (bm12→B6 group) injected with bm12 spleen cells generally showed similar levels of mortality, nephritis, and autoantibodies, although the autoantibody titers declined somewhat after week 8 in the bm12→CD8KO group. Control groups of recipients injected with B6 spleen cells showed no induction of autoantibodies. A surprising finding, however, was that the B6→CD8KO group developed severe histologic glomerulonephritis in the absence of autoantibodies and with decreased immune deposits. These results indicate that endogenous (host) CD4+ T cells play an essential role in the cGVH autoimmune syndrome.
Induction of Antibodies Reactive with SSA/Ro-SSB/La and Development of Congenital Heart Block in a Murine Model
Authors: Miranda-Carús M. Eugenia , Boutjdir Mohamed , Tseng Chung-E , DiDonato Francis , Chan Edward K. L. , Buyon Jill P. 
Published: January 2023

Abstract
To correlate the arrhythmogenic effects of maternal autoantibodies with the genesis of congenital heart block, female BALB/c mice were immunized with human recombinant 48-kDa SSB/La, 60-kDa SSA/Ro, 52-kDa SSA/Ro (52α), and 52β (amino acids 169–245 deleted) as well as with murine recombinant 52-kDa SSA/Ro. Control animals received β-galactosidase or a polypeptide encoded by pET-28 alone. Following primary immunization and two boosters, high titer responses to the respective Ags were established by ELISA, immunoblotting, and immunoprecipitation. Sera from mice immunized with either human 52α or 52β immunoprecipitated murine 52Ro. mRNA and protein expression of 52Ro was demonstrated in the newborn murine heart. A spectrum of atrioventricular nodal conduction abnormalities was identified by electrocardiogram. First-degree block was detected in 7% of 27 pups born to mothers immunized with 48La, 20% of 54 pups born to 60Ro-immunized mothers, 6% of 56 pups born to 52α-immunized mothers, 7% of 86 pups born to 52β-immunized mothers, and 9% of 22 pups born to mothers immunized with murine 52Ro. Advanced conduction abnormalities were only identified in offspring of 52α- or 52β-immunized mice. In the 52α group, one pup had complete block and another had second-degree block (Wenckebach type); in the 52β group, five pups had complete block. Maternal Abs to the primary immunogens were detected in the pups. No control had any conduction abnormalities. This Ab-specific animal model provides strong evidence for a pathogenic role of anti-SSA/Ro-SSB/La Abs, particularly 52Ro, in the development of congenital heart block. The range and frequency of conduction defects suggest that additional factors promote disease expression.
Requirement for Distinct Janus Kinases and STAT Proteins in T Cell Proliferation Versus IFN-γ Production Following IL-12 Stimulation
Authors: Ahn Hyun-Jong , Tomura Michio , Yu Wen-Gong , Iwasaki Masayuki , Park Woong-Ryeon , Hamaoka Toshiyuki , Fujiwara Hiromi 
Published: January 2023

Abstract
While IL-12 is known to activate JAK2 and TYK2 and induce the phosphorylation of STAT4 and STAT3, little is known regarding how the activation of these signaling molecules is related to the biologic effects of IL-12. Using an IL-12-responsive T cell clone (2D6), we investigated their requirements for proliferation and IFN-γ production of 2D6 cells. 2D6 cells could be maintained with either IL-12 or IL-2. 2D6 lines maintained with IL-12 (2D6IL-12) or IL-2 (2D6IL-2) exhibited comparable levels of proliferation, but produced large or only small amounts of IFN-γ, respectively, when restimulated with IL-12 after starvation of either cytokine. 2D6IL-12 induced TYK2 and STAT4 phosphorylation. In contrast, their phosphorylation was marginally induced in 2D6IL-2. The reduced STAT4 phosphorylation was due to a progressive decrease in the amount of STAT4 protein along with the passages in IL-2-containing medium. 2D6IL-12 and 2D6IL-2 similarly proliferating in response to IL-12 induced comparable levels of JAK2 activation and STAT5 phosphorylation. JAK2 was associated with STAT5, and IL-12-induced STAT5 phosphorylation was elicited in the absence of JAK3 activation. These results indicate that IL-12 has the capacity to induce/maintain STAT4 and STAT5 proteins, and that TYK2 and JAK2 activation correlate with STAT4 phosphorylation/IFN-γ induction and STAT5 phosphorylation/cellular proliferation, respectively.
TNF/Lymphotoxin-α Double-Mutant Mice Resist Septic Arthritis but Display Increased Mortality in Response toStaphylococcus aureus
Authors: Hultgren Olof , Eugster Hans-Pietro , Sedgwick Jonathon D. , Körner Heinrich , Tarkowski Andrzej 
Published: January 2023

AbstractTo evaluate the importance of the proinflammatory cytokines TNF and lymphotoxin-α (LTα) in an experimental model of Staphylococcus aureus sepsis and arthritis, we used TNF/LTα-double-deficient mice raised on the C57BL/6 background. Mice were i.v. inoculated with a toxic shock syndrome toxin-1 (TSST-1)-producing S. aureus strain, LS-1. Intravenous inoculation of a high dose of bacteria (1 × 107/mouse) resulted in 67% mortality in TNF/LTα-deficient mice, whereas none of the controls died (p = 0.009). Those results correlated to a significantly decreased phagocytosis in vitro and inefficient bacterial clearance in vivo in mice lacking capacity to produce TNF/LTα. Thus, at day 6 after inoculation, S. aureus could not be found in the bloodstream of controls, but bacteremia developed in all TNF/LTα-deficient mice examined (p = 0.02). Interestingly, upon infection with a lower dose of staphylococci (3 × 106/mouse) the mortality was overall low, but the frequency of arthritis was clearly higher in the wild-type group as compared with the TNF/LTα-deficient mice (40% vs 13%). Histopathologic examination revealed a lower frequency of synovitis (38% vs 90%, p &lt; 0.05) and erosivity (25% vs 60%, NS) in TNF/LTα-deficient mice as compared with wild-type counterparts. Our results show the importance of TNF/LTα in defense against systemic S. aureus infections and point out the detrimental role of these cytokines as mediators of inflammatory response in S. aureus arthritis.
MHC Class I Molecules Compete in the Endoplasmic Reticulum for Access to Transporter Associated with Antigen Processing
Authors: Knittler Michael R. , Gülow Karsten , Seelig Angela , Howard Jonathan C. 
Published: January 2023

AbstractWe have used the functionally distinct TAP alleles of the rat in cellular transfectants as tools to investigate how newly formed rat class I (RT1.A) molecules with distinct peptide requirements gain access to suitable peptides in the endoplasmic reticulum (ER). Normal maturation of RT1.Aa depends on the presence in the ER of peptides with C-terminal arginine, while restrictive TAP-B allelic group transporters fail to transport such peptides. In this situation, RT1.Aa is retained in the ER. We show that this retention is accompanied by accumulation of RT1.Aa in the ER, partly associated with TAP and partly free. In such cells, access to TAP of a second allelic product, RT1.Au, which does not require C-terminal arginine peptides, is competitively inhibited by the build-up of RT1.Aa. Nevertheless, RT1.Au loads and matures normally. Introduction of a permissive TAP-A allele competent to transport C-terminal arginine peptides releases RT1.Aa from the ER and restores RT1.Au interaction with TAP. Both class I alleles associate indiscriminately with permissive and restrictive TAP alleles. The data support the view that interaction with TAP is not a prerequisite for peptide loading by class I molecules, so long as suitable peptides are available in the ER. They further show that TAP association of a class I molecule depends on a competitive balance in the ER defined by the extent to which the peptide requirements of other class I molecules present are satisfied and not only by the intrinsic strength of the interaction with TAP.
Sequence Motifs in the Integrin α4 Cytoplasmic Tail Required for Regulation of In Vivo Expansion of Murine Lymphoma Cells
Authors: Bittner Michaela , Gosslar Uwe , Luz Arne , Holzmann Bernhard 
Published: January 2023

AbstractThe binding of integrins to cognate ligands is tightly controlled by intracellular signals. Conversely, integrin occupancy generates biochemical signals inside the cell. The present study examined whether concepts of integrin function established by in vitro analysis apply to regulation of receptor function in complex biologic settings in vivo using a mouse model of tumor metastasis. Integrin α4 subunits were truncated at amino acid Gln1014 (A4-1014), preserving the conserved GFFKR motif, and at position Glu1021 (A4-1021). In vitro adhesion assays revealed that cytoplasmic tail truncations did not affect constitutive ligand binding of α4 integrins, while agonist-induced adhesion was abolished by the A4-1014, but not by the A4-1021, mutation. Inducible ligand binding of α4 integrins was dependent on cytoskeletal function, whereas constitutive adhesion was not. In vivo metastasis formation assays demonstrated that expansion of murine T lymphoma cells in spleen is strongly inhibited by the wild-type α4 subunit and the A4-1021 mutant. In contrast, the in vivo phenotype of α4 integrin expression in lymphoma cells was completely abrogated by the A4-1014 mutation. Cross-linking of α4 integrins in vitro inhibited proliferation and induced apoptosis of LB cells expressing wild-type α4 subunits or the A4-1021 mutant, but not of LB-A4-1014 cells. In summary, these results demonstrate that sequence motifs regulating cytoskeleton-dependent α4 integrin activation in vitro are essential for the control of LB lymphoma cell expansion both in vitro and in vivo.
Recruitment of Hepatic NK Cells by IL-12 Is Dependent on IFN-γ and VCAM-1 and Is Rapidly Down-Regulated by a Mechanism Involving T Cells and Expression of Fas
Authors: Fogler William E. , Volker Kirk , Watanabe Morihiro , Wigginton Jon M. , Roessler Philip , Brunda Michael J. , Ortaldo John R. , Wiltrout Robert H. 
Published: January 2023

AbstractNK cells have been shown to be important antitumor or antiviral effector cells in the liver. In the present study we have examined the factors that regulate the initial recruitment and subsequent fate of hepatic NK and T cells in mice treated with IL-12 or IL-2. Daily administration of IL-12 caused a rapid initial increase in NK cells followed by a subsequent decrease that coincided with an accumulation of T cells. The recruitment of hepatic NK cells by IL-12, but not the subsequent T cell infiltrate, was abrogated in IFN-γ−/− mice. In contrast, daily administration of IL-2 caused a sustained increase in liver-associated NK cells that was not diminished in IFN-γ−/− mice. The IL-12-induced recruitment in both hepatic NK and T cells was abrogated by in vivo treatment with anti-VCAM-1 mAbs, while treatment with anti-ICAM-1 Abs decreased only the recruitment of T cells in the IL-12-treated mice. The rapid loss of newly recruited hepatic NK cells in IL-12-treated mice did not occur in SCID mice or in B.MRL-Faslpr (Fas−) and B6Smn.C3H-Faslgld (FasL−) mutant mice, suggesting that T cells can actively eliminate hepatic NK cells through a Fas-dependent mechanism. These findings also imply that during the endogenous innate immune response to infectious agents or tumors or in the host response induced by cytokine therapies, the biologic effects of NK cells may be limited by T cell-mediated effects.