bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cutting Edge: B Cell-Deficient Mice Develop Experimental Allergic Encephalomyelitis with Demyelination After Myelin Oligodendrocyte Glycoprotein Sensitization
Authors: Hjelmström Peter , Juedes Amy E. , Fjell Jenny , Ruddle Nancy H. 
Published: December 2022

AbstractMyelin oligodendrocyte glycoprotein (MOG) induced experimental allergic encephalomyelitis (EAE) is an animal model for the central nervous system disease multiple sclerosis (MS). The roles of individual components of the immune system have not been completely defined in the mouse model, and to determine the role of B cells and Abs in the induction of EAE and demyelination, B cell-deficient μMT (H-2b) mice were immunized with MOG peptide 35–55. The μMT mice were susceptible to MOG-induced EAE and developed a chronic sustained disease, with inflammatory lesions and primary demyelination in the spinal cord, brain, and optic nerves, similar to that seen in wild-type C57BL/6 mice. The inflammatory cells in the central nervous system of μMT mice included both activated and memory T cells and macrophages. The data suggest that B cells and Abs are not necessary for primary demyelination in MOG-induced EAE in mice.
Immunodominance in the CTL Response Against Minor Histocompatibility Antigens: Interference Between Responding T Cells, Rather than with Presentation of Epitopes
Authors: Wolpert Elisabeth Z. , Grufman Per , Sandberg Johan K. , Tegnesjö Anna , Kärre Klas 
Published: December 2022

AbstractWe have investigated mechanisms involved in immunodominance of the CTL response of C57BL/6 (B6) mice against cells of BALB.B origin. This transplantation barrier consists of at least 40 minor histocompatibility (H) Ags. Insufficient presentation of nondominant epitopes in the presence of dominant epitopes was investigated as a possible mechanism for immunodominance. Ag presentation was assessed by recognition of dendritic cells of BALB.B origin, MLC restimulatory capacity, and quantification of cell surface presentation by peptide elution from intact cells. Cells from BALB.B mice, which fail to elicit CTL against nondominant epitopes, presented nondominant epitopes to a similar extent as cells from minor H congenic mice; the latter do elicit CTL against nondominant minor H Ags. Nevertheless, presentation of nondominant and dominant epitopes by the same APC appeared to be an important factor for immunodominance to occur, since simultaneous immunization with the epitopes on separate cells elicited CTL against both types of epitopes. This suggested that immunodominance is determined in the interaction between different responding T cells and the APC. Support for this was obtained in an in vitro model in which the CTL response against a nondominant epitope was inhibited by the concomitant response against a dominant epitope. This study suggests that immunodominance in the CTL response against certain minor H Ags results from interference between T cell responses and not from insufficient presentation of peptide epitopes. The study also provides an in vitro model for further investigations of the immunodominance phenomenon.
Expression and Function of Fas on Cells Damaged by γ-Irradiation in B6 and B6/lpr Mice
Authors: Booker Jessica K. , Reap Elizabeth A. , Cohen Philip L. 
Published: December 2022

Abstract
Fas (CD95) is a cell surface protein that mediates apoptosis. lpr is a mutation of the Fas gene caused by a retroviral insertion resulting in premature termination of transcription and aberrant splicing of Fas mRNA. Mice homozygous for the lpr gene develop lymphoproliferation and produce autoantibodies closely resembling those of human systemic lupus erythematosus. While lpr mice have been reported to express low levels of normally spliced Fas mRNA, it is unknown whether they express functional Fas protein. Here we show that splenocytes from lpr mice that have been damaged by γ-irradiation expressed Fas protein. Fas was up-regulated on irradiated B6 cells and could be detected on B6/lpr cells undergoing apoptosis following in vitro culture. Detection of Fas on live lpr cells was demonstrable when apoptosis was blocked by zinc. In a short term chimera system, Fas was shown to play a role, in vivo, in the disposition of radiation-injured cells from both normal and lpr mice. The addition of anti-Fas Ab to in vitro cultures resulted in an increase in apoptosis in both B6 and B6/lpr cells. Detection of intact Fas message and low levels of Fas protein in lpr mice has led to the consideration of lpr as a leaky mutation. This study demonstrates that lpr mice can produce functional Fas protein. This system is also appropriate for identifying the in vivo role of Fas/FasL in apoptosis following other cell manipulations.
CD40 Ligand/Trimer DNA Enhances Both Humoral and Cellular Immune Responses and Induces Protective Immunity to Infectious and Tumor Challenge
Authors: Gurunathan Sanjay , Irvine Kari R. , Wu Chang-You , Cohen Jeffrey I. , Thomas Elaine , Prussin Calman , Restifo Nicholas P. , Seder Robert A. 
Published: December 2022

AbstractCD40/CD40 ligand interactions have a central role in the induction of both humoral and cellular immunity. In this study, we examined whether a plasmid expressing CD40 ligand/trimer (CD40LT) could enhance immune responses in vivo. BALB/c mice were injected with plasmid expressing β-galactosidase DNA with or without CD40LT DNA or IL-12 DNA, and immune responses were assessed. Mice vaccinated with β-gal DNA plus CD40LT DNA or IL-12 DNA had a striking increase in Ag-specific production of IFN-γ, cytolytic T cell activity, and IgG2a Ab. The mechanism by which CD40LT DNA enhanced these responses was further assessed by treating vaccinated mice with anti-IL-12 mAb or CTLA-4 Ig (CTLA4Ig). Production of IFN-γ and CTL activity was abrogated by these treatments, suggesting that CD40LT DNA was mediating its effects on IFN-γ and CTL activity through induction of IL-12 and enhancement of B7 expression, respectively. Physiologic relevance for the ability of CD40LT DNA to enhance immune responses by the aforementioned pathways was shown in two in vivo models. First, with regard to CTL activity, mice vaccinated with CD40LT DNA did not develop metastatic tumor following challenge with lethal dose of tumor. Moreover, in a mouse model requiring IL-12-dependent production of IFN-γ, mice vaccinated with soluble Leishmania Ag and CD40LT DNA were able to control infection with Leishmania major. These data suggest that CD40LT DNA could be a useful vaccine adjuvant for diseases requiring cellular and/or humoral immunity.
Adenovirus-Mediated Expression of a Dominant Negative Mutant of p65/RelA Inhibits Proinflammatory Gene Expression in Endothelial Cells Without Sensitizing to Apoptosis
Authors: Soares Miguel P. , Muniappan Ashok , Kaczmarek Elzbieta , Koziak Katarzyna , Wrighton Christopher J. , Steinhäuslin Florence , Ferran Christiane , Winkler Hans , Bach Fritz H. , Anrather Josef 
Published: December 2022

Abstract
We hypothesized that blocking the induction of proinflammatory genes associated with endothelial cell (EC) activation, by inhibiting the transcription factor nuclear factor κB (NF-κB), would prolong survival of vascularized xenografts. Our previous studies have shown that inhibition of NF-κB by adenovirus-mediated overexpression of IκBα suppresses the induction of proinflammatory genes in EC. However, IκBα sensitizes EC to TNF-α-mediated apoptosis, presumably by suppressing the induction of the NF-κB-dependent anti-apoptotic genes A20, A1, manganese superoxide dismutase (MnSOD), and cellular inhibitor of apoptosis 2. We report here that adenovirus mediated expression of a dominant negative C-terminal truncation mutant of p65/RelA (p65RHD) inhibits the induction of proinflammatory genes, such as E-selectin, ICAM-1, VCAM-1, IL-8, and inducible nitric oxide synthase, in EC as efficiently as does IκBα. However, contrary to IκBα, p65RHD does not sensitize EC to TNF-α-mediated apoptosis although both inhibitors suppressed the induction of the anti-apoptotic genes A20, A1, and MnSOD equally well. We present evidence that this difference in sensitization of EC to apoptosis is due to the ability of p65RHD, but not IκBα, to inhibit the constitutive expression of c-myc, a gene involved in the regulation of TNF-α-mediated apoptosis. These data demonstrate that it is possible to block the expression of proinflammatory genes during EC activation by targeting NF-κB, without sensitizing EC to apoptosis and establishes the role of c-myc in controlling induction of apoptosis during EC activation. Finally, these data provide the basis for a potential approach to suppress EC activation in vivo in transgenic pigs to be used as donors for xenotransplantation.
Characterization of C3dg Binding to a Recess Formed Between Short Consensus Repeats 1 and 2 of Complement Receptor Type 2 (CR2; CD21)
Authors: Prodinger Wolfgang M. , Schwendinger Michael G. , Schoch Jürgen , Köchle Maria , Larcher Clara , Dierich Manfred P. 
Published: December 2022

AbstractTo allow for a better characterization of the ligand binding structures of human complement receptor type 2 (CR2; CD21), we have established an IgG1 κ mouse mAb, FE8, that interferes efficiently with binding of C3dg and EBV to CR2. In contrast to mAb OKB7, the only well-characterized mAb with similar specificity, mAb FE8 blocked binding of soluble C3dg or particles carrying multiple copies of surface-bound C3dg to CR2 or induced complete removal of these ligands from the receptor. In vitro EBV infection of B lymphocytes, on the other hand, was abrogated by mAbs FE8 and OKB7 with similar dose-response characteristics. As FE8 was shown to recognize a discontinuous epitope, a series of overlapping peptides derived from SCR1 and -2 and immobilized on cellulose was screened with FE8. The results suggest that up to five discontinuous sequences contributed to the epitope. The sequence 63-EYFNKYS-69, located between the two SCR units, reacted most intensively. Two other sequences, 16-YYSTPI-21 and 105-NGNKSVWCQANN-116, are located between Cys1 and Cys2 of SCR1 and around Cys3 of SCR2, respectively. Based on the solution structure for two factor H SCRs, a three-dimensional model of SCR1 and -2 was generated. The FE8 binding peptide sequences were located in relative proximity to each other, bounding the recess formed between SCR1 and -2. This potential of mAb FE8 is currently unique and may be exploited for interfering with conditions of unwanted recognition of C3dg-coated structures by the immune system.
IL-7 Reconstitutes Multiple Aspects of v-Abl-Mediated Signaling
Authors: Banerjee Arnob , Rothman Paul 
Published: December 2022

AbstractThe mechanism by which early lymphoid cells are selectively transformed by v-Abl is currently unknown. Previous studies have shown constitutive activation of IL-4 and IL-7 signaling pathways, as measured by activation of Janus protein kinase (JAK)1, JAK3, STAT5, and STAT6, in pre-B cells transformed by v-Abl. To determine whether activation of these cytokine signaling pathways by v-Abl is important in the cellular events induced by the Abelson murine leukemia virus, the effects of IL-4 and IL-7 on pre-B cells transformed with a temperature-sensitive v-Abl mutant were examined. Whereas IL-4 had little or no effect, IL-7 delayed both the apoptosis and cell cycle arrest that occur upon v-Abl kinase inactivation. IL-7 also delayed the decreases in the levels of c-Myc, Bcl-2, and Bcl-xL that occur upon loss of v-Abl kinase activity. IL-7 did not maintain v-Abl-mediated differentiation arrest of the pre-B cells, as activation of NF-κB and RAG gene transcription was unaffected by IL-7. These results identify a potential role for IL-7 signaling pathways in transformation by v-Abl while demonstrating that a combination of IL-4 and IL-7 signaling cannot substitute for an active v-Abl kinase in transformed pre-B cells.
B Cell Responses to a Peptide Epitope. VI. The Kinetics of Antigen Recognition Modulates B Cell-Mediated Recruitment of T Helper Subsets
Authors: Vijayakrishnan Lalitha , Manivel Venkatasamy , Rao Kanury V. S. 
Published: December 2022

AbstractThe ability of Ag-primed B cells to recruit distinct Th subsets was examined using two analogous synthetic peptides, G41CT3 and G28CT3, as model Ags. With sequence differences at only two positions, these peptides were identical both with respect to fine specificity of Abs induced and ability to prime T cells. Lymph node cell populations primed with peptide G41CT3, when challenged with the homologous Ag, yielded predominantly Th2 cytokines. In contrast, a challenge with the heterologous Ag, G28CT3, resulted in a markedly increased production of Th1 cytokines. These distinctions derived from altered APC function of Ag-primed B cells due to differential kinetics of recognition of the two Ags by surface Ig receptors, as confirmed by binding studies with a panel of anti-G41CT3 mAbs. A concentration-dependent circular dichroism study revealed differences in the nature of intermolecular associations for these two peptides. Furthermore, the on-rate of peptide G28CT3 binding to Ab also increased with increasing peptide concentration, implying a dependence on intermolecular interactions. This, in turn, correlated well with the ability of peptide G28CT3 to preferentially activate either Th1 or Th2 cells. Thus, the relative proportion of Th1 vs Th2 cells recruited by Ag-primed B cells is governed by the on-rate of Ag binding to surface Ig receptors, with higher on-rates promoting Th1 recruitment. Further, even subtle changes in solution behavior of an Ag can markedly influence the kinetics of recognition by B cells.
The Fas/Fas Ligand System Is Involved in the Pathogenesis of Autoimmune Myocarditis in Rats
Authors: Ishiyama Shigeru , Hiroe Michiaki , Nishikawa Toshio , Shimojo Takashi , Abe Shinji , Fujisaki Hiroyuki , Ito Hiroshi , Yamakawa Katsutoshi , Kobayashi Nobuyuki , Kasajima Takeshi , Marumo Fumiaki 
Published: December 2022

Abstract
The mechanisms responsible for myocardial injury and cell death in myocarditis are still unclear. We examined whether myocardial cell death occurs via apoptosis in myosin-induced autoimmune myocarditis in rats and whether the Fas/Fas ligand (FasL) system plays a role in this apoptosis. On days 14, 17, 21, and 35 after immunization with porcine heart myosin, some cardiomyocytes and infiltrating lymphocytes were found to be apoptotic on in situ terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end-labeling (TUNEL) assay, but none was on day 60 and in control rats. Apoptotic indices peaked at day 17, and laddering of genomic DNA from the affected myocardium was observed on days 17 and 21 on agarose gel electrophoresis. The expression of Fas mRNA and protein was detected on days 17 and 21 in some cardiomyocytes and infiltrating lymphocytes by Northern blot analysis and immunohistochemistry, respectively. In addition, FasL was detected in some infiltrating lymphocytes on days 14, 17, and 21 by both in situ hybridization and immunostaining, and FasL-positive lymphocytes were mainly CD4+ cells. Some rats were injected with anti-Fas Ab (0.1 mg/kg) or anti-FasL Ab (0.1 mg/kg), and subsequently, inflammatory lesions exhibited less severe than did untreated rats with myocarditis. These findings suggest that cell death via apoptosis of cardiomyocytes and lymphocytes is one of the mechanisms of myocardial injury in autoimmune myocarditis, and that the Fas/FasL system might play a role in the induction of this apoptosis.
NF-κB Activation in CD27 Signaling: Involvement of TNF Receptor-Associated Factors in Its Signaling and Identification of Functional Region of CD27
Authors: Yamamoto Hiroyasu , Kishimoto Tadamitsu , Minamoto Seijiro 
Published: December 2022

AbstractCD27 belongs to TNF receptor family, and it is unique in this family for its disulfide-linked homodimerization of 55-kDa monomers. In the present study we demonstrate that overexpression of CD27 in 293 cells induces a low level of NF-κB activation, and the ligation of the receptor by its corresponding ligand (CD70) augments this signal dramatically. Either TNF receptor-associated factor-2 (TRAF2) or TRAF3 binds to the CD27 molecule from the coimmunoprecipitation experiment. This NF-κB activation signal is inhibited by dominant negative TRAF2 or intact TRAF3, indicating that TRAF2 and TRAF3 works as a mediator and an inhibitor, respectively. The activated NF-κB complex contains at least two components, p50 and p65, but not p52. All these phenomena have also been observed in the TNF receptor type II, CD30 and CD40 signaling system, indicating that this receptor family uses the common or similar molecules for this signal. Finally, we identified the 13-amino acid alignment in the cytoplasmic region of the CD27 molecule (residues 238–250 amino acids), which is critical for the NF-κB activation signal and also for its association with TRAFs. This amino acid alignment contains the EEEG sequence, which is essential for interaction of CD30 or CD40 with TRAFs (TRAF1 and TRAF2, but not TRAF3), and also contains the PIQED sequence, which is similar to PXQXT that is known to be necessary for interaction of TNF receptor II and CD30 with TRAFs (TRAF1, 2, and 3).
Autocrine/Paracrine IFN-αβ Mediates the Lipopolysaccharide-Induced Activation of Transcription Factor Stat1α in Mouse Macrophages: Pivotal Role of Stat1α in Induction of the Inducible Nitric Oxide Synthase Gene
Authors: Gao Jian Jun , Filla Michael B. , Fultz Marion J. , Vogel Stefanie N. , Russell Stephen W. , Murphy William J. 
Published: December 2022

AbstractWe have examined the role of Stat1α in the induction by LPS of the mouse inducible nitric oxide synthase (EC 1.14.13.39) gene. LPS induced both the tyrosine phosphorylation of Stat1α and the production of nitric oxide in a time- and dose-dependent manner. The phosphorylation of Stat1α elicited by LPS differed from that observed using IFN-γ or IFN-β, in that LPS induced less phosphorylated protein and the time course of induction was much delayed (2–4 h compared with 30 min). Cycloheximide inhibited LPS-mediated Stat1α phosphorylation. In addition, cell culture supernatants derived from macrophages treated with LPS for 4 h could be transferred to naive macrophage cultures resulting in rapid (30 min), rather than delayed (4 h), phosphorylation of Stat1α. Together, these results implicated an autocrine/paracrine effector protein(s) in the phosphorylation process. LPS stimulated phosphorylation of Stat1α in peritoneal macrophages derived from IFN-γ-knockout mice, negating any possibility that IFN-γ was the mediator. By contrast, neutralizing Ig raised against mouse IFN-αβ inhibited both the delayed LPS-mediated phosphorylation of Stat1α and the rapid induction of phosphorylation induced by supernatants from LPS-stimulated cultures. Collectively, these results show that LPS-induced IFN-αβ production, Stat1α activation, and nitrite accumulation closely parallel one another, suggesting that indirect activation of transcription factor Stat1α by IFN-αβ is a critical determinant of LPS-mediated inducible nitric oxide synthase gene expression.
Recognition of Two Overlapping CTL Epitopes in HIV-1 p17 by CTL from a Long-Term Nonprogressing HIV-1-Infected Individual
Authors: Harrer Thomas , Harrer Ellen , Barbosa Peter , Kaufmann Friedemann , Wagner Ralf , Brüggemann Susanne , Kalden Joachim R. , Feinberg Mark , Johnson R. Paul , Buchbinder Susan , Walker Bruce D. 
Published: December 2022

AbstractHIV-1 infection has been shown to elicit strong CTL responses in some infected persons, but few data are available regarding the relationship between targeted epitopes and in vivo viral quasispecies. In this study, we examined the CTL response in a person infected for 15 yr with a CD4 count persistently &gt;500 cells/μl. The dominant in vivo activated CTL response was directed against two overlapping Gag CTL epitopes in an area of p17 known to be essential for viral replication. The 9-mer SLYNTVATL (amino acids 77–85) was recognized in conjunction with HLA-A2, whereas the overlapping 8-mer TLYCVHQR (amino acids 83–91) was recognized by HLA-A11-restricted CTL. Analysis of in vivo virus sequences both in PBMC and plasma revealed the existence of sequence variation in this region, which did not affect viral replication in vitro, but decreased recognition by the A11-restricted CTL response, with maintenance of the A2-restricted response. These results indicate that an essential region of the p17 protein can be simultaneously targeted by CTL through two different HLA molecules, and that immune escape from CTL recognition can occur without impairing viral replication. In addition, they demonstrate that Ag processing can allow for presentation of overlapping epitopes in the same infected cell, which can be affected quite differently by sequence variation.
Attenuation of HLA-DR Expression by Mononuclear Phagocytes Infected withMycobacterium tuberculosisIs Related to Intracellular Sequestration of Immature Class II Heterodimers
Authors: Hmama Zakaria , Gabathuler Reinhard , Jefferies Wilfred A. , de Jong Gary , Reiner Neil E. 
Published: December 2022

AbstractMHC class II expression was examined in macrophages infected with Mycobacterium tuberculosis. IFN-γ increased the surface expression of class II molecules in THP-1 cells and this was markedly reduced in cells infected with M. tuberculosis. Despite this effect, steady state levels of HLA-DRα, HLA-DRβ, and invariant (Ii) chains were equivalent in control and infected cells. Metabolic labeling combined with pulse-chase experiments and biochemical analysis showed that the majority of class II molecules in infected cells became resistant to endoglycosidase H, consistent with normal Golgi processing. However, results of intracellular staining and dual color confocal microscopy revealed a significant defect in transport of newly synthesized class II molecules through the endocytic compartment. Thus, compared with findings in control cells, class II molecules in infected cells colocalized to a minimal extent with a lysosomal-associated membrane protein-1+ endosomal compartment. In addition, in contrast to control cells, class II molecules in infected cells failed to colocalize with endocytosed BSA under conditions where this marker is known to label late endosomes, lysosomes, and the MHC class II compartment. Consistent with defective transport along the endocytic pathway, the maturation of SDS-stable class II αβ dimers—dependent upon removal of Ii chain and peptide loading of class II dimers in the MHC class II compartment—was markedly impaired in M. tuberculosis-infected cells. These findings indicate that defective transport and processing of class II molecules through the endosomal/lysosomal system is responsible for diminished cell surface expression of MHC class II molecules in cells infected with M. tuberculosis.