bluuebunny/crossref_metadata_2025_split huggingface dataset:
TGF-β Increases Leukotriene C4 Synthase Expression in the Monocyte-Like Cell Line, THP-1
Authors: Riddick Carl A. , Serio Kenneth J. , Hodulik Craig R. , Ring William L. , Regan Mark S. , Bigby Timothy D. 
Published: December 2022

AbstractThe goal of this study was to determine whether cytokines modulate leukotriene C4 (LTC4) synthase expression in mononuclear phagocytes. A panel of cytokines was surveyed for changes in LTC4 synthase mRNA in THP-1 cells. TGF-β1, -2, and -3 had significant stimulatory effects. The addition of TGF-β resulted in a time-dependent increase in LTC4 synthase mRNA at 6 h, which persisted through 48 h. Furthermore, this conditioning resulted in an increase in immunoreactive protein for LTC4 synthase through 7 days. TGF-β conditioning of cells resulted in a time- and dose-dependent increase in stimulated LTC4 synthase activity. Following transient transfection of THP-1 cells with a promoter-reporter construct containing 1.2 kb of the LTC4 synthase promoter, TGF-β treatment resulted in a 2-fold increase in reporter activity. Conditioning with TGF-β did not prolong the half-life of LTC4 synthase mRNA, as assessed by RNase protection assays in actinomycin D-treated cells. Cycloheximide exposure experiments revealed that new protein synthesis was not required for the observed stimulatory effect of TGF-β on LTC4 synthase mRNA. We conclude that LTC4 synthase expression is increased at a transcriptional level by TGF-β in mononuclear phagocytes.
One Active C1r Subunit Is Sufficient for the Activity of the Complement C1 Complex: Stabilization of C1r in the Zymogen Form by Point Mutations
Authors: Dobó József , Gál Péter , Szilágyi Katalin , Cseh Sándor , Lörincz Zsolt , Schumaker Verne N. , Závodszky Péter 
Published: December 2022

Abstract
The binding of C1 (the first component of complement) to immune complexes leads to the autoactivation of C1r through the cleavage of the Arg463-Ile464 bond in the catalytic domain. Spontaneous activation of C1r (and C1) also occurs in the fluid phase, preventing the characterization of the zymogen form of C1r. To overcome this difficulty, the zymogen form of human C1r was stabilized by mutating the Arg in the Arg463-Ile464 bond to Gln. This mutant was designated as mutant QI. Recombinant C1r (wild type (wt) or mutant) was expressed in insect cells using serum-free medium in functionally pure form; therefore, the cell culture supernatant was suitable to reconstruct C1 for the hemolytic assay. Mutant QI was a stable, nonactivable zymogen and showed no hemolytic activity in reconstituted C1. However, this stable zymogen C1r mutant could form an active mixed dimer with the wt C1r, indicating that one active C1r subunit in the C1 complex is sufficient for the full activity of the entire complex. Our experiments also showed that the exchange of C1r monomers between the C1r dimers is completed in less than 16 h even at pH 7 and 4°C. Two other mutants were also constructed by changing Arg463 to Lys, or Ile464 to Phe, and were designated as mutants KI and RF, respectively. Although these substitutions did increase the stability of the proenzyme in the cell culture supernatant, the mutant proteins retained their ability to autoactivate, and both had a wt-like hemolytic activity.
Varying Roles of E-Selectin and P-Selectin in Different Microvascular Beds in Response to Antigen
Authors: Hickey Michael J. , Kanwar Samina , McCafferty Donna-Marie , Granger D. Neil , Eppihimer Michael J. , Kubes Paul 
Published: December 2022

Abstract
Expression of E-selectin and P-selectin is critical in the effector phase of leukocyte recruitment in response to Ag. Whether their relative roles differ between tissues in response to the same Ag is unknown. In this study, a type I hypersensitivity response was elicited in C57BL/6 mice by systemic sensitization with OVA. Following local Ag challenge, endothelial selectin expression was examined in the skin and cremaster muscle microvasculature using a dual-radiolabeled mAb technique. Next, the dermal and muscle microcirculations were visualized using intravital microscopy to establish roles for P-selectin and/or E-selectin. In untreated mice, leukocyte recruitment in both skin and skeletal muscle was mediated entirely by P-selectin. Following Ag challenge, leukocyte rolling flux and adhesion were dramatically increased and leukocyte rolling velocity was unchanged in muscle. Only P-selectin expression increased in muscle, and leukocyte recruitment was entirely dependent upon this selectin. In contrast, in Ag-challenged skin, leukocyte rolling flux did not increase, but rolling velocity dropped profoundly. In skin, only E-selectin expression increased, and blockade of either E-selectin or P-selectin had minimal effect on either rolling flux or rolling velocity. Blockade of both selectins reduced rolling flux by 80% and increased rolling velocity sevenfold. These data highlight striking differences in expression of the endothelial selectins in separate microvascular beds in response to the same stimulus and demonstrate that these differences underlie very different patterns of leukocyte recruitment. The data underscore the importance of studying individual microvascular beds to understand tissue-specific leukocyte recruitment in vivo.
Antigen-Dependent and -Independent IFN-γ Modulation by Penicillins
Authors: Padovan Elisabetta , Greyerz Salome von , Pichler Werner J. , Weltzien Hans Ulrich 
Published: December 2022

AbstractThe activation of CD4+ T lymphocytes upon Ag stimulation plays a critical role in adverse immune responses including drug-specific hypersensitivity reactions. We examined the modulation of T cell phenotype induced by hapten-specific stimulation using the model of β-lactam antibiotics such as penicillin G (Pen G), Pen V, and ampicillin (Amp). When PBMC of donors suffering from hypersensitivity reactions against β-lactams were stimulated in vitro with different doses of Pen G, a preferential expansion of IL-4-producing TCRαβ+ cells was detected. A panel of T cell clones was then prepared from Pen G-specific lines after two cycles of restimulation with the hapten. For the majority of these clones, we found that high doses of Pen G induced optimal IL-4 secretion, whereas the amount of IFN-γ secreted was inversely correlated with the dose of Pen G, thus leading to a hapten-inducible shift of the fuctional phenotypes for some of the clones. Finally, Pen V and Amp were used to modulate different Ag-induced immune responses. We found that Amp had no influence on the cytokine pattern induced by specific Ag or mitogens. In contrast, Pen V inhibited the secretion of IFN-γ, but not IL-4, most likely by Ag-independent mechanisms. This last finding may open new applications for immune intervention in those diseases in which polarized Th1 responses are involved in the development of the pathology.
Increased Activation of Protein Kinase A Type I Contributes to the T Cell Deficiency in Common Variable Immunodeficiency
Authors: Aukrust Pål , Aandahl Einar Martin , Skålhegg Bjørn S. , Nordøy Ingvild , Hansson Vidar , Taskén Kjetil , Frøland Stig S. , Müller Fredrik 
Published: December 2022

AbstractThe molecular mechanisms underlying the T cell dysfunction often present in common variable immunodeficiency (CVI) are not established. cAMP-dependent protein kinase A type I (PKAI) is an important inhibitor of T cell proliferation after Ag stimulation. We therefore investigated the possibility that activation of PKAI may be involved in the development of T cell dysfunction in CVI. An exogenously added PKAI-selective antagonist (Rp-8-Br-cAMPS) induced a significant increase in anti-CD3-stimulated PBMC proliferation in 20 CVI patients compared with no effect in 15 controls. Purified T cells from 7 CVI patients with strictly defined T cell deficiency had elevated endogenous cAMP levels compared with controls. Treatment of T cells from these CVI patients with Rp-8-bromo-cAMP-phosphorothioate markedly improved anti-CD3-stimulated proliferation (up to 3.7-fold), particularly in CD4+ lymphocytes, reaching proliferation levels comparable to control values. No effect of cAMP antagonist on T cell proliferation was seen in controls. In these CVI patients, cAMP antagonist also increased IL-2 production in anti-CD3-stimulated T cells. However, exogenously added IL-2 at concentrations comparable to the achieved increase in IL-2 levels after addition of cAMP antagonist had no effect on T cell proliferation. Furthermore, the stimulatory effects of exogenously added IL-2 at higher concentrations and cAMP antagonist on T cell proliferation were additive. Our findings indicate that increased PKAI activation may be an important molecular basis for the T cell defect in CVI and suggest that the cAMP/PKAI system may be a potential molecular target for immunomodulating therapy in these patients.
Specific Vβ T Cell Subsets Are Associated with Cat and Birch Pollen Allergy in Humans
Authors: Beyer Kirsten , Häusler Tom , Kircher Moritz , Nickel Renate , Wahn Ulrich , Renz Harald 
Published: December 2022

AbstractCognate interaction between TCRs and MHC class II molecules plays an important role in initiating the allergen-specific immune response. Therefore, we analyzed the TCR distribution of human PBLs of 56 atopic and nonatopic (NA) individuals, including 4 monozygotic twin pairs, from two extended and four nuclear families. The expression of 23 Vβ and 3 Vα elements was analyzed. The blood samples of symptomatic birch pollen-sensitized individuals that were taken ≤6 wk after the birch pollen season (n = 8) showed a significantly higher frequency of Vβ16.1+ and Vβ20.1+ T cells compared with the blood samples of birch pollen-sensitized individuals that were obtained out of allergen season (n = 10) or from NA individuals (p &lt; 0.0005 and p &lt; 0.0001, respectively). Allergen-specific lymphocyte proliferation was detected in the allergic individuals, and the distribution of Vβ16.1+ and Vβ20.1+ T cells returned to normal levels after the pollen season. The frequency of these Vβ-expressing T cells correlated with the levels of allergen-specific IgE Abs. In addition, cat-sensitized individuals (n = 8) showed a significantly higher frequency of Vβ17.1-expressing T cells than did NA individuals (p &lt; 0.005). Our results indicate restricted TCR-Vβ gene usage in cat and birch pollen allergies; we suggest that both genetic and environmental factors contribute to TCR-Vβ gene expression and to the development of a specific T cell response.
Vasoactive Intestinal Peptide (VIP) and Pituitary Adenylate Cyclase-Activation Polypeptide (PACAP) Protect Mice from Lethal Endotoxemia Through the Inhibition of TNF-α and IL-6
Authors: Delgado Mario , Martinez Carmen , Pozo David , Calvo Juan R. , Leceta Javier , Ganea Doina , Gomariz Rosa P. 
Published: December 2022

AbstractThe neuropeptides vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase-activating polypeptide (PACAP) down-regulate cytokine production. Because human septic shock involves excessive cytokine production, the effect of VIP/PACAP was investigated in a high endotoxemia murine model. Both peptides protect against endotoxin-induced lethality and prevent septic shock-associated histopathological alterations. VIP/PACAP reduce serum and peritoneal TNF-α and IL-6, suggesting that the protective effect is exerted by inhibiting the production of endogenous TNF-α/IL-6. Consistent with this mechanism, VIP does not protect against septic shock induced by exogenous TNF-α. The immunomodulatory role of VIP in vivo is supported by the appearance of high levels of VIP in serum and peritoneal fluid following LPS administration. Thus, the neuropeptides VIP/PACAP protect from the lethal effect of high endotoxemia, presumably by down-regulating TNF-α and IL-6 production, and may offer an alternative in the treatment of human septic shock syndrome.
Cutting Edge: Induction of Enhanced CTL-Dependent Protective Immunity In Vivo by N-End Rule Targeting of a Model Tumor Antigen
Authors: Tobery Timothy , Siliciano Robert F. 
Published: December 2022

AbstractThere is much interest in vaccines that will enhance the induction of CTL. One mechanism to enhance Ag-specific CTL responses involves targeting Ag to undergo rapid cytoplasmic degradation by the N-end rule pathway. We have analyzed the ability of N-end rule targeting to confer protection in an immunization-challenge setting. Using the HIV-1 nef protein as a model tumor Ag, we found that in mice immunized with a vaccinia vector expressing a form of nef that is targeted for rapid cytoplasmic degradation, there was enhanced induction of nef-specific CTL and protection from a lethal challenge with the syngeneic CT26 tumor cells that had been transfected with nef. Protection from tumor challenge correlated with the magnitude of the CTL response. Thus, the targeting of tumor or viral Ags for rapid cytoplasmic degradation by the N-end rule pathway may represent a strategy for the induction of protective Ag-specific CTL responses in vivo.
The Role of the Antigen-Presenting Cell in Fas-Mediated Direct and Bystander Killing: Potential In Vivo Function of Fas in Experimental Allergic Encephalomyelitis
Authors: Thilenius Anja R. B. , Sabelko-Downes Kimberly A. , Russell John H. 
Published: December 2022

AbstractCostimulatory molecules are critical in mediating Fas-dependent direct and bystander lysis. In direct lysis, the APC is the Fas-positive target. It presents Ag to the T cell, thereby activating the T cell. The activated T cell then up-regulates FasL, allowing it to kill the APC. In bystander lysis, the APC again induces FasL expression on the T cell, but the target is a third Fas-positive cell that may lack the appropriate MHC-restricting element to activate the T cell. This study shows that ICAM-1 and B7-1 can serve as important adhesion molecules in direct killing using CD4+ T cell effectors. In bystander killing, B7-1 appears to act as a signaling molecule as well. It has been demonstrated that lpr and gld mice are less susceptible to experimental allergic encephalomyelitis than their wild-type counterparts. In this study, we show that although microglia are poor targets of direct killing, they are capable of stimulating myelin basic protein-specific T cells to kill innocent Fas-positive targets. This presents a possible mechanism for the pathogenesis of experimental allergic encephalomyelitis.
Hepatitis C Virus Envelope Glycoprotein E1 Originates in the Endoplasmic Reticulum and Requires Cytoplasmic Processing for Presentation by Class I MHC Molecules
Authors: Selby Mark , Erickson Ann , Dong Christine , Cooper Stewart , Parham Peter , Houghton Michael , Walker Christopher M. 
Published: December 2022

AbstractWe investigated whether hepatitis C virus envelope glycoprotein E1 is transported from the endoplasmic reticulum (ER) to the cytoplasm of infected cells for class I MHC processing. Target cells expressing E1 were killed by CTL lines from a hepatitis C virus-infected chimpanzee, and synthetic peptides were used to define an epitope (amino acids 233-GNASRCWVA-241) presented by the Patr-B*1601 class I MHC molecule. An unusually high concentration (&gt;100 nM) of this nonameric peptide was required for target cell lysis, but this could be reduced at least 1000-fold by replacing the asparagine at amino acid position 234 (Asn234) with aspartic acid (Asp), the anticipated anchor residue for NH2-terminal peptide binding to Patr-B*1601. Conspicuously, position 234 is part of an N-glycosylation motif (Asn-Xaa-Ser/Thr), suggesting that the Asn234 to Asp substitution might occur naturally within the cell due to deglycosylation/deamidation of this amino acid by the cytosolic enzyme peptide N-glycanase. In support of this model, we demonstrate that presentation of the epitope depended on 1) cotranslational synthesis of E1 in the ER, 2) glycosylation of the E1 molecule, and 3) a functional TAP transporter to shuttle peptide from the cytosolic to ER compartment. These results indicate for the first time that during infection of the host, viral envelope glycoproteins originating in the ER are processed in the cytoplasm for class I MHC presentation. That a posttranslational change in amino acid sequence from Asn to Asp alters the repertoire of peptides presented to CD8+ CTL has implications for the design of antiviral vaccines.
The Role of LY49 NK Cell Subsets in the Regulation of Murine Cytomegalovirus Infections
Authors: Tay Chin H. , Yu Lawrence Y. Y. , Kumar Vinay , Mason Llewelyn , Ortaldo John R. , Welsh Raymond M. 
Published: December 2022

AbstractThe distributions and functions of NK cell subsets, as defined by the expression of Ly49 NK cell receptors, were examined in murine CMV (MCMV)-infected mice. MCMV induced a reduction in NK1.1+ cell number in the spleen and an increase in the peritoneal exudate cells. Within the splenic NK1.1+ population, proportional increases in Ly49A+ and Ly49G2+ cells but decreases in Ly49C+ and Ly49D+ cells were observed 3 days post-MCMV infection, but within the peritoneal NK1.1+ cell populations there were proportional decreases in Ly49A+ cells and increases in Ly49C+, Ly49D+, and Ly49G2+ cells. Lymphocytic choriomeningitis virus did not elicit a comparable NK cell subset distribution. Lymphokine-activated killer cells were sorted into different Ly49 NK cell subsets and adoptively transferred into C57BL/6 suckling mice. Regulation of MCMV synthesis in these suckling mice was shown to be an IFN-γ-dependent, perforin- and Cmv-1-independent process, and each NK cell subset mediated anti-viral activity. In adult C57BL/6 mice, the control of MCMV in the spleen is mediated by a perforin-dependent mechanism, regulated in part by the Cmv-1 gene, which maps closely to the Ly49 family. In vivo depletions of either one or two of the Ly49 subsets in adult mice did not affect the ability of the residual NK cells to regulate MCMV synthesis. These data provide evidence of NK cell subset distribution and function in MCMV infection, but no individual subset was required for the Cmv-1-like regulation of MCMV synthesis.
Acquired Thymic Tolerance: Role of CTLA4 in the Initiation and Maintenance of Tolerance in a Clinically Relevant Autoimmune Disease Model
Authors: Issazadeh Shohreh , Zhang ManHua , Sayegh Mohamed H. , Khoury Samia J. 
Published: December 2022

AbstractInjection of Ag into the thymus of adult animals induces specific systemic tolerance. The mechanisms of acquired thymic tolerance include anergy and the deletion of Ag-specific T cells. Here, we report that anergy to nominal Ag induced via acquired thymic tolerance requires CTL-associated Ag 4 (CTLA4) engagement. The role of CTLA4 in the induction and maintenance of tolerance was then investigated in the murine experimental autoimmune encephalomyelitis model. CTLA4 blockade abrogated the induction but not the maintenance phase of acquired thymic tolerance induced by intrathymic injection of myelin Ags. In addition, CTLA4 blockade had a restricted window of action after priming with Ag, which is consistent with the expression patterns of CTLA4 in vivo. We conclude that: 1) the induction of acquired thymic tolerance requires signaling through CTLA4 and 2) CTLA4 does not appear to be required for the maintenance of acquired thymic tolerance. This is the first report documenting the role of a CTLA4 negative signaling pathway in the induction of tolerance in an autoimmune disease model.
Functional Characteristics and Survival Requirements of Memory CD4+ T Lymphocytes In Vivo
Authors: London Cheryl A. , Perez Victor L. , Abbas Abul K. 
Published: December 2022

AbstractThe phenotypic and functional characteristics of Ag-specific memory CD4+ lymphocytes are poorly defined. To examine the properties and cytokine responsiveness of these cells, we have developed an adoptive transfer system using in vitro-activated T cells expressing the DO.11 transgenic TCR specific for OVA323–339+ I-Ad. In vitro-activated DO.11 CD4+ cells exhibit comparable survival patterns at 1, 6, and 10 wk after adoptive transfer, indicating that a stable population of memory cells has been generated. In the absence of Ag, previously activated T cells survive longer than their naive counterparts in vivo, rapidly revert to a partially naive phenotype, and maintain their effector cytokine profile. The DO.11 CD4+ memory cells are capable of proliferating in response to IL-2 and IL-4, while naive DO.11 CD4+ cells exhibit no such proliferative responses.
Role of Bcl-2 in αβ T Cell Development in Mice Deficient in the Common Cytokine Receptor γ-Chain: The Requirement for Bcl-2 Differs Depending on the TCR/MHC Affinity
Authors: Nakajima Hiroshi , Leonard Warren J. 
Published: December 2022

AbstractMice lacking the common cytokine receptor γ-chain (γc) exhibit severely compromised T cell development, with diminished Bcl-2 expression in mature (CD4+ or CD8+) thymocytes and peripheral T cells. Enforced expression of Bcl-2 in these mice partially rescued αβ T cell development but not γδ T cell development. Transgenic expression of the OVA-specific DO11.10 (DO10) TCR also could modestly increase thymocyte numbers, and T cells expressing the transgenic TCR (KJ1-26+ T cells) were found in the periphery. Interestingly, the presence of KJ1-26+ T cells was dependent on the MHC background and was seen in the moderate affinity H-2d/d background but not in the higher affinity H-2d/b background in γc-deficient mice. In contrast, KJ1-26+ T cells exist in the periphery in both the H-2d/d and H-2d/b backgrounds in DO10 transgenic γc wild-type mice. These results suggest that the importance of γc-dependent signals for T cell development differs depending on the affinity of TCR for MHC. Moreover, enforced expression of Bcl-2 had a much greater effect on the development of γc-deficient T cells expressing the DO10 TCR in the high affinity H-2d/b background than in the H-2d/d background, suggesting that γc-dependent Bcl-2 expression influences T cell development in a TCR/MHC-dependent manner.
The Mechanism of Chromosome 7 Inversion in Human Lymphocytes Expressing Chimeric γβ TCR
Authors: Retière Christelle , Halary Franck , Peyrat Marie-Alix , Le Deist Françoise , Bonneville Marc , Hallet Marie-Martine 
Published: December 2022

Abstract
Functional chimeric TCR chains, encoded by VγJγCβ or VγJβCβ hybrid gene TCR, are expressed at the surface of a small fraction of αβ T lymphocytes in healthy individuals. Their frequency is dramatically increased in patients with ataxia-telangiectasia, a syndrome associated with inherited genomic instability. As the TCR γ and β loci are in an inverted orientation on chromosome 7, the generation of such hybrid genes requires at least an inversion event. Until now, neither the sequences involved in this genetic mechanism nor the number of recombinations leading to the formation of functional transcriptional units have been characterized. In this manuscript, we demonstrate that at least two rearrangements, involving classical recombination signal sequence and the V(D)J recombinase complex, lead to the formation of productive hybrid genes. A primary inversion 7 event between Dβ and Jγ genic segments generates CγVβ and CβVγ hybrid loci. Within the CγVβ locus, secondary rearrangements between Vγ and Jγ or Vγ and Jβ elements generate functional genes. Besides, our results suggest that secondary rearrangements were blocked in the CβVγ locus of normal but not ataxia-telangiectasia T lymphocytes. We also provide formal evidence that the same Dβ-3′ recombination signal sequence can be used in successive rearrangements with Jγ and Jβ genic segments, thus showing that a signal joint has been involved in a secondary recombination event.