bluuebunny/crossref_metadata_2025_split huggingface dataset:
Extracellular Signal-Regulated Kinase-2, But Not c-Jun NH2-Terminal Kinase, Activation Correlates with Surface IgM-Mediated Apoptosis in the WEHI 231 B Cell Line
Authors: Lee Jong Ran , Koretzky Gary A. 
Published: December 2022

AbstractBoth extracellular signal-regulated kinase (ERK) and c-Jun NH2-terminal kinase (JNK) have been implicated in mediating the signaling events that precede apoptosis. We studied the activation of these kinases during apoptosis of WEHI 231 B cells. Surface IgM ligation induces apoptosis of WEHI 231 cells. This effect is augmented by simultaneous engagement of CD95 and is inhibited by costimulation with either CD40 or IL-4R. We determined that surface IgM ligation activates ERK2 to a much greater level than JNK, and that IgM-mediated ERK2 activation is enhanced by costimulation with anti-CD95. Costimulation with either IL-4 or anti-CD40 interferes with anti-IgM-stimulated ERK2 activation. Transient expression of mitogen-activated protein kinase phosphatase-1 (MKP-1) inhibits both ERK2 activation and cell death following stimulation with anti-IgM and the combination of anti-IgM plus anti-CD95. CD40 engagement alone activates JNK, but IL-4 stimulation does not. N-acetyl-l-cysteine pretreatment, which blocks CD40-mediated JNK activation, does not affect the ability of CD40 to inhibit anti-IgM-mediated ERK2 activation and apoptosis. Together, these data suggest that JNK activation is not required for CD40 inhibition of surface IgM-induced cell death and that ERK2 plays an active role in mediating anti-IgM-induced apoptosis of WEHI 231 B cells.
TCR Ligation on CD8+ T Cells Creates Double-Negative Cells In Vivo
Authors: Mehal Wajahat Z. , Crispe I. Nicholas 
Published: December 2022

AbstractThe lack of CD95 in mice is associated with an accumulation of TCRαβ+ CD4−CD8− (double-negative (DN)) cells in the lymph nodes (LNs) and other organs. To test the hypothesis that these DN cells arise from TCRαβ+CD8+ cells after activation via the TCR, we have crossed an MHC class I-restricted TCR transgene (tg) onto the lpr genotype to generate two TCR-transgenic experimental groups, TCRtg+ lpr/+ (CD95-intact) and TCRtg+ lpr/lpr (CD95-deficient). Specific peptide administration resulted in peripheral deletion of TCRαβ cells from the LNs of CD95-intact and CD95-deficient mice. On day 3 after peptide administration in the CD95-deficient but not the CD95-intact mice, there was a ninefold increase in the percentage of DN cells in the LN; this increase returned to control levels by day 10. Peripheral deletion was associated with an accumulation of TCRαβ+CD8high cells in the livers of mice of both genotypes by day 3, which returned to control levels by day 10 without an increase in the percentage or total number of DN cells. Our data show that the in vivo stimulation of TCRαβ+CD8+ cells in the absence of CD95 results in an initial accumulation and an eventual loss of DN cells. This identifies a role for CD95 after TCRαβ stimulation in the efficient removal of TCRαβ+CD8+ cells after the down-regulation of CD8. CD95 is not essential for this process, because other mechanisms can compensate, but such mechanisms are less efficient in the LN.
β2-Microglobulin-Dependent T Cells Are Not Necessary for Alloantigen-Induced Th2 Responses After Neonatal Induction of Lymphoid Chimerism in Mice
Authors: Foucras Gilles , Coureau Christiane , Beijleveld Leo , Druet Philippe , Saoudi Abdelhadi , Guéry Jean-Charles 
Published: December 2022

AbstractWe have analyzed the requirement for β2-microglobulin (β2m)-dependent T cells in the generation of allogeneic Th2 responses in vivo. A neonatal injection of semiallogeneic cells in BALB/c mice induces a state of chimerism that promotes the differentiation of donor-specific CD4+ T cells toward the Th2 phenotype. Polyclonal T-B cell interactions occur in this model between host Th2 and donor B cells, resulting in the production of IgE Abs. IgE production and Th2-priming are critically dependent upon the early production of IL-4. Our data in the present paper demonstrate that: 1) IgE synthesis and the up-regulation of MHC class II and CD23 molecules on B cells are independent of β2m expression in the host, 2) no difference in the induction of CD4 alloreactive Th2 cells could be observed between β2m−/− and their wild-type control littermates when Th2-priming was measured in adult mice, and 3) the Th2 response and IgE production is induced in the complete absence of β2m-dependent T cells both in the host and in the inoculum. Therefore, using a variety of assays, we could not demonstrate diminished responses in mice with a disrupted β2m gene in this model of Th2-mediated allogeneic interaction, indicating that β2m-dependent NK1.1+ and CD8+ T cells are not required for the generation of alloreactive Th2 responses in vivo.
Granulysin-Induced Apoptosis. I. Involvement of at Least Two Distinct Pathways
Authors: Gamen Susana , Hanson Dennis A. , Kaspar Allan , Naval Javier , Krensky Alan M. , Anel Alberto 
Published: December 2022

Abstract
Granulysin is a newly described cytolytic molecule released by CTL and NK cells via granule-mediated exocytosis. It shares homology with saposin-like proteins, including NK-lysin and amoebapores, and has been implicated in the lysis of tumor cells and microbes. In the present study we show that recombinant granulysin alone induces apoptosis of Jurkat cells. This apoptosis is associated with a sixfold increase in the ceramide/sphingomyelin ratio, implicating the activation of sphingomyelinases. Granulysin- and ceramide-induced apoptosis are similar in that they both are only minimally inhibited by the more selective cysteine protease p32 (caspase 3)-like caspase inhibitor N-acetyl-Asp-Glu-Val-Asp aldehyde, while they are significantly inhibited by the more general caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone (Z-VAD-fmk). Nevertheless, while Z-VAD-fmk almost completely inhibits ceramide-induced apoptosis, a Z-VAD-fmk-resistant component was observed using granulysin. Granulysin also causes apoptosis in cells depleted of sphingomyelin by prolonged treatment with the ceramide synthase inhibitor fumonisin B1. These data indicate that granulysin induces target cell death by both ceramide- and caspase-dependent and -independent pathways.
The Invariant Chain Gene Intronic Enhancer Shows Homology to Class II Promoter Elements
Authors: Beilue Moore Bethany , Cao Zhu Alexander , McRae Theresa L. , Woo Colleen H. , Conley Sarah , Jones Patricia P. 
Published: December 2022

AbstractCoordinate expression of MHC class II proteins and the class II-associated invariant chain (Ii) is important for proper MHC class II functioning in Ag processing and presentation. The coordinate regulation of these genes results, in part, from the sharing of transcriptional regulatory regions between MHC class II and Ii genes; the Ii has previously been shown to have an upstream enhancer closely related to the essential class II promoter elements. We report here the characterization of a second enhancer in the Ii gene, located within the first intron. This intronic enhancer is contained within a 155-bp region, enhances transcription from the Ii minimal promoter, and also contains elements that are homologous to class II promoter elements X1, X2, and Y boxes.
Cutting Edge: Dendritic Cells Require Maturation via CD40 to Generate Protective Antitumor Immunity
Authors: Mackey Matthew F. , Gunn Jason R. , Maliszewski Charles , Kikutani Hitoshi , Noelle Randolph J. , Barth Richard J. 
Published: December 2022

Abstract
A critical role for CD40/CD154 interactions in the generation of protective cell-mediated tumor immunity has been demonstrated previously. Herein, we show that the failure to generate systemic tumor immunity in the absence of CD40/CD154 interactions correlates with an inhibition of Th1-type cytokine production following tumor vaccination. Furthermore, protective antitumor responses can be restored in CD40-deficient mice by the coadministration of CD40+/+ but not CD40−/− dendritic cells (DCs) with tumor Ag, suggesting that CD40 is critical for the maturation and function of DCs in vivo. Finally, we demonstrate that an IL-12-transduced but not a mock-transduced tumor vaccine induces systemic tumor immunity in anti-CD154-treated and CD154-deficient mice. These data suggest that impaired antitumor responses in the absence of CD40/CD154 interactions are the result of a lesion in APC function, namely IL-12 production, and that CD40 plays a critical role in the maturation of DCs in vivo.
β-Galactoside-Binding Protein (βGBP) Alters the Cell Cycle, Up-Regulates Expression of the α- and β-Chains of the IFN-γ Receptor, and Triggers IFN-γ-Mediated Apoptosis of Activated Human T Lymphocytes
Authors: Allione Alessandra , Wells Valerie , Forni Guido , Mallucci Livio , Novelli Francesco 
Published: December 2022

AbstractIn this paper, the effects of β-galactoside binding protein (βGBP), the LGALS1 gene product, on the cell cycle progression and expansion of activated human T lymphocytes were studied. βGBP drastically inhibits the IL-2 induced proliferation of PHA-activated T lymphocytes as well as the IL-2 independent proliferation of malignant T lymphocytes by arresting them in the S and G2/M phases of the cell cycle. In addition, βGBP up-regulates the expression of both the α- and the β-chains of the IFN-γR on activated T lymphocyte membrane. None of these effects depend on sugar binding: saturating amounts of lactose do not affect the cell cycle block nor IFN-γR up-modulation. The increased expression of both chains renders βGBP-treated T lymphoblasts sensitive to IFN-γ-induced apoptosis. Taken as a whole, these findings suggest that βGBP plays an important immunoregulatory role by switching off T lymphocyte effector functions. They also provide the first evidence of up-modulation of IFN-γR expression on T lymphocytes by a negative cell growth regulator.
Tumor-Infiltrating Lymphocytes Exhibiting High Ex Vivo Cytolytic Activity Fail to Prevent Murine Melanoma Tumor Growth In Vivo
Authors: Prévost-Blondel Armelle , Zimmermann Christine , Stemmer Christine , Kulmburg Peter , Rosenthal Felicia M. , Pircher Hanspeter 
Published: December 2022

AbstractThe identification of tumor-associated Ags recognized by CD8+ CTL and prevention of tumor outgrowth by adoptive transfer of these CTL demonstrates that CD8+ T cells play a major role in antitumor immunity. We have generated B16.F10 melanoma cells that express the glycoprotein epitope amino acid 33–41 (GP33) of the lymphocytic choriomeningitis virus (LCMV) to examine antitumor CD8+ T cell response in C57BL/6 mice immune to LCMV and in mice transgenic for the LCMV GP33-specific P14 TCR (P14 TCR mice). We find that B16.F10GP33 tumor cells grew in syngeneic C57BL/6 mice without inducing T cell tolerance. LCMV infection or adoptive transfer of LCMV-specific effector T cells delayed but did not prevent growth of preestablished tumors in these mice. However, B16.F10GP33 tumor cells were rejected in mice immune to LCMV and in mice treated with LCMV-specific effector T cells on the same day as the tumor. Surprisingly, B16.F10GP33 tumor cells grew in P14 TCR transgenic mice despite an abundance of tumor-associated Ag-specific CD8+ T cells. In these mice, freshly isolated tumor-infiltrating lymphocytes exhibited an activated phenotype and displayed high GP33-specific cytolytic activity when assessed ex vivo. Thus, B16.F10GP33 melanoma cells are able to initiate, but not to sustain, a GP33-specific CTL response sufficient to clear the tumor enduringly.
Transcription of a Minimal Promoter from the NF-IL6 Gene Is Regulated by CREB/ATF and SP1 Proteins in U937 Promonocytic Cells
Authors: Berrier Allison , Siu Gerald , Calame Kathryn 
Published: December 2022

AbstractNF-IL6 is an important transcriptional regulator of genes induced in activated monocytes/macrophages, and NF-IL6 is the only CCAAT/enhancer-binding protein (C/EBP) family member whose steady-state mRNA levels increase upon activation of monocytes (1). We show that increased transcription of the NF-IL6 gene is responsible, at least in part, for induction of NF-IL6 mRNA following activation of U937 promonocytic cells. We have identified a 104-bp minimal promoter region of the NF-IL6 gene that is sufficient for basal and activation-dependent induction of transcription in U937 cells. This region contains binding sites for the cAMP response element-binding protein/activation transcription factor (CREB/ATF) and Sp1 families of transcription factors. Each site is functionally important and contributes independently to transcription of the NF-IL6 gene in U937 cells.
Rheumatoid Factor Specificity of a VH3-Encoded Antibody Is Dependent on the Heavy Chain CDR3 Region and Is Independent of Protein A Binding
Authors: Zhang Meilin , Majid Adrian , Bardwell Philip , Vee Chris , Davidson Anne 
Published: December 2022

AbstractRheumatoid factors (RF) recognize conformational determinants located within the Fc portion of IgG. By analyzing a panel of monoclonal rheumatoid arthritis (RA)-derived RFs, we previously demonstrated that the somatically generated light chain complementarity-determining region 3 (CDR3) contributes to RF specificity. We have now generated a panel of heavy chain mutants of the B′20 Ab, a high affinity RA-derived IgM RF. B′20 also binds avidly to protein A and weakly to ssDNA and tetanus toxoid. B9601, a RF negative Ab that is highly homologous to B′20 but does not bind any of the Ags tested, and RC1, a low affinity polyreactive RF, were used to generate heavy chain mutants with framework (FR) and CDR switches. The mutated heavy chains were cotransfected into a myeloma cell line with the germline counterpart of the B′20 light chain, and the expressed Ig tested for antigenic specificity. We show that both RF specificity and polyreactivity of B′20 is dependent on its unique heavy chain CDR3 region. Replacement with a B9601 CDR3 shortened to the same length as the B′20 CDR3, and with only 5 amino acid differences, did not restore Fc binding. Conversely, absence of protein A binding of B9601 is due to the presence of a serine residue at position 82a in the B9601 heavy chain FR3 region. Together, our data suggest that Ig gene recombination events can generate B cells with autoantibody specificities in the preimmune repertoire. Abnormal release, activation, expansion, or mutation of such cells might all contribute to the generation of a high titer RF response in patients with RA.
Asymmetric Contribution to Ig Repertoire Diversity by Vκ Exons: Differences in the Utilization of Vκ10 Exons
Authors: Fitzsimmons Sean P. , Rotz Benjamin T. , Shapiro Marjorie A. 
Published: December 2022

AbstractThe mouse has approximately 140 germline Vκ genes, and functional Vκ exons are expressed at roughly equivalent levels in the preimmune repertoire. We have examined the expression of individual members of the Vκ10 family. Vκ10A and Vκ10B genes have been utilized in numerous hybridomas and myelomas, while Vκ10C has not. In this study, we have cloned the Vκ10C gene and shown that it is structurally functional, has the expected promoter elements and recombination signal sequences, and that it is capable of recombination. Vκ10C mRNA, however, is present at levels at least 1000-fold lower than Vκ10A and Vκ10B in adult spleens. While there are no sequence differences in the octamer or TATA box between Vκ10C and Vκ10A, there are three nucleotide changes in the promoter region. These promoters equally drive the expression of a reporter gene in B cells or plasma cells, but the Vκ10A promoter is able to drive expression in pre-B cell lines significantly better than the Vκ10C promoter (p &lt; 0.05). Vκ10C rearrangements can be detected in bone marrow and splenic DNA. Therefore, the lack of Vκ10C expression may reflect the inability of Vκ10C-rearranged cells to undergo positive or negative selection. Our results suggest that the available Ab repertoire is shaped not only by the number of structurally functional genes, but also by the ability of assembled genes to be expressed at critical points during B cell maturation.
The Assembly and Stability of MHC Class II-(αβ)2 Superdimers
Authors: Schafer Peter H. , Malapati Sunil , Hanfelt Kimberly K. , Pierce Susan K. 
Published: December 2022

AbstractX-ray crystallography of several MHC class II molecules revealed a structure described as a dimer of heterodimers, or a superdimer. This discovery led to the hypothesis that MHC class II molecules may interact with the TCR and CD4 as an (αβ)2 superdimer, potentially providing more stable and stimulatory interactions than can be provided by the simple αβ heterodimer alone. In this study, using chemical cross-linking, we provide evidence for the existence of the superdimers on the surface of B cells. We further characterize the superdimers and demonstrate that in lysates of B cells, I-Ek dimers and superdimers are derived from the same population of I-Ek molecules. Purified, I-Ek molecules in solution also exist as a mixture of 60-kDa dimers and 120-kDa superdimers, indicating that I-Ek has an intrinsic ability to form 120-kDa complexes in the absence of other cellular components. Peptide mapping showed that the αβ and (αβ)2 complexes are closely related and that the superdimers do not contain additional polypeptides not present in the dimers. The (αβ)2 complex displays thermal and pH stability similar to that of the αβ complex, both being denatured by SDS at temperatures above 50°C and at a pH below 5. These data support the model that MHC class II has an intrinsic ability to assume the (αβ)2 superdimeric conformation, which may be important for interactions with the TCR and CD4 coreceptor.
B and T Cells Are Required for Mouse Mammary Tumor Virus Spread Within the Mammary Gland
Authors: Golovkina Tatyana V. , Dudley Jaquelin P. , Ross Susan R. 
Published: December 2022

Abstract
Mouse mammary tumor virus (MMTV) is an infectious retrovirus transmitted through milk from mother to newborns. MMTV encodes a superantigen (SAg) whose activity is indispensable for the virus life cycle, since a genetically engineered virus with a mutation in the sag gene neither amplified in cells of the immune system of suckling pups nor infected their mammary glands. When wild-type MMTV was injected directly into the mammary glands of uninfected pubescent mice, their lymphoid as well as mammary gland cells became virus infected. To test whether this infection of lymphoid cells was dependent on SAg activity and required for virus spread within the mammary gland, we performed mammary gland injections of wild-type MMTV(C3H) into two strains of transgenic mice that lacked SAg-cognate, Vβ14+ T cells. Neither the MTV-ORF or LEL strains showed infection of their mammary glands. Moreover, no MMTV infection of their peripheral lymphocytes was detected. Similar experiments with mice lacking B cells (μ-chain knockouts) showed no detectable virus spread in the mammary glands or lymphoid tissues. These data suggest that SAg activity and MMTV-infected lymphocytes are required, not only for initial steps of viral infection, but also for virus spread within the mammary gland. Virus spread at late times in infection determines whether MMTV induces mammary tumors.
Migration Inhibitory Factor Induces Killing ofLeishmania majorby Macrophages: Dependence on Reactive Nitrogen Intermediates and Endogenous TNF-α
Authors: Jüttner Stefan , Bernhagen Jürgen , Metz Christine N. , Röllinghoff Martin , Bucala Richard , Gessner André 
Published: December 2022

AbstractMacrophage migration inhibitory factor (MIF) is a product of activated T cells, anterior pituitary cells, and macrophages. MIF plays an important role in LPS-induced shock and delayed-type hypersensitivity. Furthermore, MIF exhibits a proinflammatory spectrum of action, promoting TNF-α production by macrophages, and counter-regulates glucocorticoid suppression of cytokine production. Here, we report that purified recombinant MIF activates murine macrophages to kill Leishmania major, with maximal effects at concentrations above 1 μg/ml. This MIF-mediated activation is specific, since it can be blocked completely by anti-MIF mAb. The MIF-mediated activation is dependent on TNF-α produced endogenously by macrophages, because the administration of anti-TNF-α antiserum markedly reduced the MIF effect. No MIF-mediated activation was observed in macrophages derived from TNF receptor p55 knockout mice, thus demonstrating the requirement of the smaller TNF receptor molecule for autocrine TNF-α signaling. A highly specific inhibitor of the inducible nitric oxide synthase (iNOS), l-N6-(1-iminoethyl)lysine, dihydrochloride, also inhibited the action of MIF, suggesting an important role for iNOS in the antiparasitic properties of MIF. In line with this, no MIF-mediated activation was detected analyzing macrophages derived from iNOS-deficient mice. The effect of MIF was blocked completely by the macrophage-deactivating cytokines IL-10, IL-13, and TGF-β. Finally, the expression of MIF mRNA and protein was up-regulated in lymph nodes of mice during the first week after infection with L. major. MIF therefore represents a cytokine involved not only in the recruitment of proinflammatory cells during infection but also in the complex regulation of the antimicrobial activity of these cells.
CpG DNA Induces Sustained IL-12 Expression In Vivo and Resistance toListeria monocytogenesChallenge
Authors: Krieg Arthur M. , Love-Homan Laurie , Yi Ae-Kyung , Harty John T. 
Published: December 2022

AbstractVertebrates have evolved innate immune defense mechanisms that recognize and respond to structural patterns that are specific to microbial molecules. One such pattern recognition system is based on unmethylated CpG dinucleotides in particular sequence contexts (CpG motifs); these motifs are common in bacterial DNA but are under-represented (“CpG suppression”) and methylated in vertebrate DNA. Mice that are injected with bacterial DNA or synthetic oligodeoxynucleotides (ODNs) containing CpG motifs respond with a rapid production of IL-12 and IFN-γ. The serum levels of IL-12 were increased for at least 8 days after a single injection of CpG ODNs, but IFN-γ levels returned to baseline within 24 h. This Th1-like cytokine response to CpG motifs induces a state of resistance to infection by Listeria monocytogenes in susceptible specific pathogen-free BALB/c mice. Resistance developed within 48 h of pretreatment with CpG ODNs, persisted for at least 2 wk, and was dependent upon IFN-γ secretion. These data support the hypothesis that CpG DNA motifs are a “danger signal” that activates protective innate immune defenses and may have therapeutic potential.
Novel Regulation of Cyclooxygenase-2 Expression and Prostaglandin E2 Production by IFN-γ in Human Macrophages
Authors: Barrios-Rodiles Miriam , Chadee Kris 
Published: December 2022

AbstractCyclooxygenase-2 (COX-2) is the inducible enzyme in macrophages responsible for high output PG production during inflammation and immune responses. Although several stimuli are known to regulate COX-2, the molecular mechanisms modulating its expression by the cytokine network are poorly understood. As IFN-γ priming is essential for macrophage accessory and effector cell functions, we investigated the effect of IFN-γ on COX-2 expression in U937 human macrophages stimulated with IL-1β. A dose- and time-dependent increase in COX-2 mRNA and protein expression was evoked by IL-1β, whereas the levels of COX-1, the constitutively expressed isoform, remained unaltered. Interestingly, IFN-γ-primed cells showed 40 to 60% lower levels of COX-2 mRNA, protein expression, and PGE2 production as compared with nonprimed cells. IFN-γ-priming (50–500 U/ml) down-regulated COX-2 expression in a time- and dose-dependent fashion. Furthermore, IFN-γ inhibited COX-2 gene transcription in response to IL-1β but not to LPS. In contrast, the rate of decay of COX-2 transcripts in nonprimed and primed macrophages was similar (t1/2 = 3.2 h). The down-regulatory effect of IFN-γ on IL-1β-induced COX-2 expression was abrogated with cycloheximide. These results highlight a novel mechanism of COX-2 regulation by IFN-γ at the transcriptional level, which may affect the outcome of inflammatory and immune conditions.