bluuebunny/crossref_metadata_2025_split huggingface dataset:
Chemotaxis of Mouse Bone Marrow Neutrophils and Dendritic Cells Is Controlled by ADP-Ribose, the Major Product Generated by the CD38 Enzyme Reaction
Authors: Partida-Sanchez Santiago , Gasser Andreas , Fliegert Ralf , Siebrands Cornelia C. , Dammermann Werner , Shi Guixiu , Mousseau Betty J. , Sumoza-Toledo Adriana , Bhagat Harivadan , Walseth Timothy F. , Guse Andreas H. , Lund Frances E. 
Published: April 2014

Abstract
The ectoenzyme CD38 catalyzes the production of cyclic ADP-ribose (cADPR) and ADP-ribose (ADPR) from its substrate, NAD+. Both products of the CD38 enzyme reaction play important roles in signal transduction, as cADPR regulates calcium release from intracellular stores and ADPR controls cation entry through the plasma membrane channel TRPM2. We previously demonstrated that CD38 and the cADPR generated by CD38 regulate calcium signaling in leukocytes stimulated with some, but not all, chemokines and controls leukocyte migration to inflammatory sites. However, it is not known whether the other CD38 product, ADPR, also regulates leukocyte trafficking In this study we characterize 8-bromo (8Br)-ADPR, a novel compound that specifically inhibits ADPR-activated cation influx without affecting other key calcium release and entry pathways. Using 8Br-ADPR, we demonstrate that ADPR controls calcium influx and chemotaxis in mouse neutrophils and dendritic cells activated through chemokine receptors that rely on CD38 and cADPR for activity, including mouse FPR1, CXCR4, and CCR7. Furthermore, we show that the calcium and chemotactic responses of leukocytes are not dependent on poly-ADP-ribose polymerase 1 (PARP-1), another potential source of ADPR in some leukocytes. Finally, we demonstrate that NAD+ analogues specifically block calcium influx and migration of chemokine-stimulated neutrophils without affecting PARP-1-dependent calcium responses. Collectively, these data identify ADPR as a new and important second messenger of mouse neutrophil and dendritic cell migration, suggest that CD38, rather than PARP-1, may be an important source of ADPR in these cells, and indicate that inhibitors of ADPR-gated calcium entry, such as 8Br-ADPR, have the potential to be used as anti-inflammatory agents.
Cysteinyl Leukotrienes Are Autocrine and Paracrine Regulators of Fibrocyte Function
Authors: Vannella Kevin M. , McMillan Tracy R. , Charbeneau Ryan P. , Wilke Carol A. , Thomas Peedikayil E. , Toews Galen B. , Peters-Golden Marc , Moore Bethany B. 
Published: April 2014

Abstract
Pulmonary fibrosis is characterized by the accumulation of fibroblasts and myofibroblasts. These cells may accumulate from three potential sources: the expansion of resident lung fibroblasts, the process of epithelial-mesenchymal transition, or the recruitment and differentiation of circulating mesenchymal precursors known as fibrocytes. We have previously demonstrated that fibrocytes participate in lung fibrogenesis following administration of FITC to mice. We now demonstrate that leukotriene-deficient 5-LO−/− mice are protected from FITC-induced fibrosis. Both murine and human fibrocytes express both cysteinyl leukotriene receptor (CysLT) 1 and CysLT2. In addition, fibrocytes are capable of producing CysLTs and can be regulated via the autocrine or paracrine secretion of these lipid mediators. Exogenous administration of leukotriene (LT) D4, but not LTC4 induces proliferation of both murine and human fibrocytes in a dose-dependent manner. Consistent with this result, CysLT1 receptor antagonists are able to block the mitogenic effects of exogenous LTD4 on fibrocytes. Endogenous production of CysLTs contributes to basal fibrocyte proliferation, but does not alter fibrocyte responses to basic fibroblast growth factor. Although CysLTs can induce the migration of fibrocytes in vitro, they do not appear to be essential for fibrocyte recruitment to the lung in vivo, possibly due to compensatory chemokine-mediated recruitment signals. However, CysLTs do appear to regulate the proliferation of fibrocytes once they are recruited to the lung. These data provide mechanistic insight into the therapeutic benefit of leukotriene synthesis inhibitors and CysLT1 receptor antagonists in animal models of fibrosis.
Tissue- and Stimulus-Dependent Role of Phosphatidylinositol 3-Kinase Isoforms for Neutrophil Recruitment Induced by Chemoattractants In Vivo
Authors: Pinho Vanessa , de Castro Russo Remo , Amaral Flávio A. , de Sousa Lirlândia P. , Barsante Michele M. , de Souza Danielle G. , Alves-Filho José C. , Cara Denise C. , Hayflick Joel S. , Rommel Christian , Ruckle Thomas , Rossi Adriano G. , Teixeira Mauro M. 
Published: April 2014

AbstractPI3K plays a fundamental role in regulating neutrophil recruitment into sites of inflammation but the role of the different isoforms of PI3K remains unclear. In this study, we evaluated the role of PI3Kγ and PI3Kδ for neutrophil influx induced by the exogenous administration or the endogenous generation of the chemokine CXCL1. Administration of CXCL1 in PI3Kγ−/− or wild-type (WT) mice induced similar increases in leukocyte rolling, adhesion, and emigration in the cremaster muscle when examined by intravital microscopy. The induction of neutrophil recruitment into the pleural cavity or the tibia-femoral joint induced by the injection of CXCL1 was not significantly different in PI3Kγ−/− or WT mice. Neutrophil influx was not altered by treatment of WT mice with a specific PI3Kδ inhibitor, IC87114, or a specific PI3Kγ inhibitor, AS605240. The administration of IC87114 prevented CXCL1-induced neutrophil recruitment only in presence of the PI3Kγ inhibitor or in PI3Kγ−/− mice. Ag challenge of immunized mice induced CXCR2-dependent neutrophil recruitment that was inhibited by wortmannin or by blockade of and PI3Kδ in PI3Kγ−/− mice. Neutrophil recruitment to bronchoalveolar lavage induced by exogenously added or endogenous production of CXCL1 was prevented in PI3Kγ−/− mice. The accumulation of the neutrophils in lung tissues was significantly inhibited only in PI3Kγ−/− mice treated with IC87114. Neutrophil recruitment induced by exogenous administration of C5a or fMLP appeared to rely solely on PI3Kγ. Altogether, our data demonstrate that there is a tissue- and stimulus-dependent role of PI3Kγ and PI3Kδ for neutrophil recruitment induced by different chemoattractants in vivo.
Selective Inhibition of Inducible NO Synthase Activity In Vivo Reverses Inflammatory Abnormalities in Surfactant Protein D-Deficient Mice
Authors: Atochina-Vasserman Elena N. , Beers Michael F. , Kadire Helchem , Tomer Yaniv , Inch Adam , Scott Pamela , Guo Chang J. , Gow Andrew J. 
Published: April 2014

Abstract
Surfactant protein D (SP-D)-deficient (SP-D−/−) mice exhibit early development of emphysema. Previously we have shown that SP-D deficiency results in increased production and activity of inducible NO synthase (iNOS). In this study, we examined whether treatment with the iNOS inhibitor 1400W could inhibit the inflammatory phenotype. Mice were treated with 1400W systemically for 7 wk from 3 wk of age. Treatment reduced total lung NO synthase activity to 14.7 ± 6.1% of saline-treated 10-wk-old SP-D−/− littermates. Long-term administration of 1400W reduced lung inflammation and cellular infiltration; and significantly attenuated the increased levels of matrix metalloproteinases 2 and 9, chemokines (KC, TARC), and cytokines (IFN-γ) seen in bronchoalveolar lavage (BAL) of SP-D−/− mice. Abrogation of these levels was associated with decreasing BAL chemotactic activity for RAW cells. Two weeks of treatment with 1400W reduced total lung NO synthase (NOS) activity to 12.7 ± 6.3% of saline-treated SP-D−/− mice. Short-term iNOS inhibition resulted in attenuation of pulmonary inflammation within SP-D−/− mice as shown by decreases in total BAL cell count (63 ± 6% of SP-D−/− control), macrophage size (&gt;25 μm) within the BAL (62 ± 10% of SP-D−/− control), and a percentage of BAL macrophages producing oxidants (76 ± 9% of SP-D−/− control). These studies showed that s.c. delivery of 1400W can be achieved in vivo and can attenuate the inflammatory processes within SP-D deficiency. Our results represent the first report linking defects in the innate immune system in the lung with alterations in NO homeostasis.
Introduction of Zwitterionic Motifs into Bacterial Polysaccharides Generates TLR2 Agonists Able to Activate APCs
Authors: Gallorini Simona , Berti Francesco , Parente Pierino , Baronio Roberta , Aprea Susanna , D’Oro Ugo , Pizza Mariagrazia , Telford John L. , Wack Andreas 
Published: April 2014

Abstract
It was shown previously that bacterial polysaccharides (PS), which naturally contain both positive and negative charges, are able to activate T cells and APCs. However, the vast majority of bacterial PS are anionic and do not have these properties. In this study, we show that chemical introduction of positive charges into naturally anionic bacterial PS confers to the resulting zwitterionic PS (ZPS) the ability to activate pure human monocytes, monocyte-derived dendritic cells, and mouse bone marrow-derived dendritic cells, as do natural bacterial ZPS. Cells are induced to up-regulate MHC class II and costimulatory molecules and to produce cytokines. In mixed monocyte-T cell cocultures, ZPS induce MHC II-dependent T cell proliferation and up-regulation of activation markers. These stimulatory qualities of ZPS disappear when the positive charge is chemically removed from the molecules and thus the zwitterionic motif is destroyed. The ability of natural and chemically derived ZPS to activate APCs can be blocked by anti-TLR2 mAbs, and TLR2 transfectants show reporter gene transcription upon incubation with ZPS. In conclusion, the generation of a zwitterionic motif in bacterial PS confers the ability to activate both APCs and T cells. This finding has important implications for the design of novel polysaccharide vaccines.
Interaction between GATA-3 and the Transcriptional Coregulator Pias1 Is Important for the Regulation of Th2 Immune Responses
Authors: Zhao Xinyan , Zheng Bo , Huang Yanfang , Yang Dan , Katzman Shoshana , Chang Chawnshang , Fowell Deborah , Zeng Wei-ping 
Published: April 2014

Abstract
Th2 cytokine expression is dependent on the transcription factor GATA-3. However, the molecular interactions of GATA-3 leading to Th2 cytokine gene activation have not been well characterized. Here, we reported a number of GATA-3 associated proteins in Th2 cells, and one of such proteins Pias1 functioned as a positive transcriptional coregulator for GATA-3. When overexpressed in Th2 cells, Pias1 enhanced the expression of IL-13, and to lesser degrees, IL-4 and -5. Conversely, Pias1 siRNA down-regulated the Th2 cytokine expression. In Leishmania major infection, manipulating Pias1 expression in parasite-reactive CD4 T cells altered severity of disease caused by Th2 responses. Mechanistically, Pias1 markedly potentiated GATA-3-mediated activation of the IL-13 promoter by facilitating the recruitment of GATA-3 to the promoter. In contrast, IL-5 promoter was modestly enhanced by Pias1 and no effect was observed on IL-4 promoter. Thus, both promoter activation and additional mechanisms are responsible for regulation by Pias1.
An Effect of Parasite-Encoded Arginase on the Outcome of Murine Cutaneous Leishmaniasis
Authors: Gaur Upasna , Roberts Sigrid C. , Dalvi Rahul P. , Corraliza Inés , Ullman Buddy , Wilson Mary E. 
Published: April 2014

AbstractClassical activation of macrophages infected with Leishmania species results in expression and activation of inducible NO synthase (iNOS) leading to intracellular parasite killing. Macrophages can contrastingly undergo alternative activation with increased arginase activity, metabolism of arginine along the polyamine pathway, and consequent parasite survival. An active role for parasite-encoded arginase in host microbicidal responses has not previously been documented. To test the hypothesis that parasite-encoded arginase can influence macrophage responses to intracellular Leishmania, a comparative genetic approach featuring arginase-deficient mutants of L. mexicana lacking both alleles of the gene encoding arginase (Δarg), as well as wild-type and complemented Δarg controls (Δarg[pArg]), was implemented. The studies showed: 1) the absence of parasite arginase resulted in a significantly attenuated infection of mice (p &lt; 0.05); 2) poorer survival of Δarg in mouse macrophages than controls correlated with greater NO generation; 3) the difference between Δarg or control intracellular survival was abrogated in iNOS-deficient macrophages, suggesting iNOS activity was responsible for increased Δarg killing; 4) consistently, immunohistochemistry showed enhanced nitrotyrosine modifications in tissues of mice infected with Δarg compared with control parasites. Furthermore, 5) in the face of decreased parasite survival, lymph node cells draining cutaneous lesions of Δarg parasites produced more IFN-γ and less IL-4 and IL-10 than controls. These data intimate that parasite-encoded arginase of Leishmania mexicana subverts macrophage microbicidal activity by diverting arginine away from iNOS.
Gads−/− Mice Reveal Functionally Distinct Subsets of TCRβ+ CD4−CD8− Double-Negative Thymocytes
Authors: Zeng Ling , Dalheimer Stacy L. , Yankee Thomas M. 
Published: April 2014

Abstract
TCRβ expression in CD4−CD8− double-negative (DN) thymocytes induces signaling pathways that promote survival and proliferation, as well as differentiation into CD4+CD8+ double-positive thymocytes. The signaling pathways that regulate survival, proliferation, and differentiation remain unclear. We used Gads-deficient mice to investigate the signaling pathways that regulate these cell fates. During this investigation, we focused on TCRβ+ DN thymocytes and found that there are at least three functionally distinct subsets of TCRβ+ DN thymocytes: TCRβ+ DN3E, TCRβ+ DN3L, and TCRβ+ DN4. Survival and proliferation of TCRβ+ DN3E were independent of Gads, but survival and proliferation of TCRβ+ DN3L cells were Gads dependent. Likewise, expression of Bcl-2 in TCRβ+ DN3E cells was Gads independent, but Gads was necessary for Bcl-2 expression in TCRβ+ DN3L cells. Bcl-2 expression was not dependent on Gads in TCRβ+ DN4 cells, but proliferation of TCRβ+ DN4 cells was Gads dependent. Gads was not required for the differentiation of DN thymocytes into DP thymocytes. In fact, Gads−/− DN3E cells differentiated into DP thymocytes more readily than wild-type cells. We conclude that signaling pathways required to initiate TCRβ-induced survival and proliferation are distinct from the pathways that maintain survival and proliferation. Furthermore, signaling pathways that promote survival and proliferation may slow differentiation.
Parallel Expansion of Human Virus-Specific FoxP3− Effector Memory and De Novo-Generated FoxP3+ Regulatory CD8+ T Cells upon Antigen Recognition In Vitro
Authors: Billerbeck Eva , Blum Hubert E. , Thimme Robert 
Published: April 2014

Abstract
Although FoxP3 has been shown to be the most specific marker for regulatory CD4+ T cells, its significance in the CD8+ T cell population is not well understood. In this study, we show that the in vitro stimulation of human PBMC with hepatitis C virus or Flu virus-specific peptides gives rise to two distinct Ag-specific T cell populations: FoxP3− and FoxP3+CD8+ T cells. The FoxP3+ virus-specific CD8+ T cells share phenotypical markers of regulatory T cells, such as CTLA-4 and glucocorticoid-induced TNFR family-related gene, and do produce moderate amounts of IFN-γ but not IL-2 or IL-10. IL-2 and IL-10 are critical cytokines, however, because the expansion of virus-specific FoxP3+CD8+ T cells is blocked by IL-2- or IL-10-neutralizing mAbs. The virus-specific FoxP3+CD8+ T cells have a reduced proliferative capacity, indicating anergy, and display a cell-cell contact-dependent suppressive activity. Taken together, our results indicate that stimulation with a defined viral Ag leads to the expansion of two different cell populations: FoxP3− memory/effector as well as FoxP3+ regulatory virus-specific CD8+ T cells.
Ig Allotypic Inclusion Does Not Prevent B Cell Development or Response
Authors: Velez Maria-Gabriela , Kane Melissa , Liu Sucai , Gauld Stephen B. , Cambier John C. , Torres Raul M. , Pelanda Roberta 
Published: April 2014

Abstract
B cells expressing two different Igκ L chains (allotype included) have been occasionally observed. To determine frequency and function of these cells, we have analyzed gene-targeted mice that carry a human and a mouse Igk C region genes. Using different methodologies, we found that cells expressing two distinct κ-chains were 1.4–3% of all B cells and that they were present in the follicular, marginal zone, and B1 mature B cell subsets. When stimulated in vitro with anti-IgM, dual κ surface-positive cells underwent activation that manifested with cell proliferation and/or up-regulation of activation markers and similar to single κ-expressing B cells. Yet, when activated by divalent reagents that bound only one of the two κ-chains, dual κ B cells responded suboptimally in vitro, most likely because of reduced Ag receptor cross-linking. Nonetheless, dual κ B cells participated in a SRBC-specific immune response in vivo. Finally, we found that Ig allotype-included B cells that coexpress autoreactive and nonautoreactive Ag receptors were also capable of in vitro responses following BCR aggregation. In summary, our studies demonstrate that Igκ allotype-included B cells are present in the mouse mature B cell population and are responsive to BCR stimulation both in vitro and in vivo. Moreover, because in vitro activation in response to anti-IgM was also observed in cells coexpressing autoreactive and nonautoreactive Abs, our studies suggest a potential role of allotype-included B cells in both physiological and pathological immune responses.
Stat5 Is Essential for Early B Cell Development but Not for B Cell Maturation and Function
Authors: Dai Xuezhi , Chen Yuhong , Di Lie , Podd Andrew , Li Geqiang , Bunting Kevin D. , Hennighausen Lothar , Wen Renren , Wang Demin 
Published: April 2014

Abstract
The two closely related Stat5 (Stat5A and Stat5B) proteins are activated by a broad spectrum of cytokines. However, with the complication of the involvement of Stat5A/5B in stem cell function, the role of Stat5A/5B in the development and function of lymphocytes, especially B cells, is not fully understood. In this study, we demonstrated that Stat5A/5B−/− fetal liver cells had severe diminution of B cell progenitors but clearly had myeloid progenitors. Consistently, the mutant fetal liver cells could give rise to hemopoietic progenitors and myeloid cells but not B cells beyond pro-B cell progenitors in lethally irradiated wild-type or Jak3−/− mice. Deletion of Stat5A/5B in vitro directly impaired IL-7-mediated B cell expansion. Of note, reintroduction of Stat5A back into Stat5A/5B−/− fetal liver cells restored their abilities to develop B cells. Importantly, CD19-Cre-mediated deletion of Stat5A/5B in the B cell compartment specifically impaired early B cell development but not late B cell maturation. Moreover, the B cell-specific deletion of Stat5A/5B did not impair splenic B cell survival, proliferation, and Ig production. Taken together, these data demonstrate that Stat5A/5B directly control IL-7-mediated early B cell development but are not required for B cell maturation and Ig production.
Protection from Lethal Infection by Adoptive Transfer of CD8 T Cells Genetically Engineered to Express Virus-Specific Innate Immune Receptor
Authors: Iizuka Koho , Nakajima Chigusa , Iizuka Yoshie-Matsubayashi , Takase Mitsuyo , Kato Takako , Noda Satoshi , Tanaka Kazuo , Kanagawa Osami 
Published: April 2014

Abstract
CMV infection is one of the most common complications in immunocompromised individuals, such as organ and bone marrow transplant patients. Both innate and adaptive immune responses are required for defense against CMV infection. In murine CMV (MCMV) infection, strains harboring the MCMV-specific NK cell activation receptor, Ly49H (Klra8), are resistant. In contrast, MCMV infection of mice lacking Ly49H gene causes early mortality due to uncontrolled viral replication. In this study, we report the successful protection of mice from lethal MCMV infection with gene-transferred polyclonal CD8 T cells. CD8 T cells expressing a chimeric receptor comprising Ly49H extracellular and CD3ζ cytoplasmic domains are capable of killing target cells expressing the MCMV protein, m157. CD8 T cells expressing the chimeric receptor protect mice in vivo from lethality in the acute phase of MCMV infection, leading to the establishment of long-term protection. These data provide proof-of-principle evidence that a novel strategy for harnessing CD8 cytolytic function through TCR-independent yet pathogen-specific receptor can result in effective protection of hosts from pathogens.
Clonal Expansion of CD8+ Effector T Cells in Childhood Tuberculosis
Authors: Jacobsen Marc , Detjen Anne K. , Mueller Henrik , Gutschmidt Andrea , Leitner Sandra , Wahn Ulrich , Magdorf Klaus , Kaufmann Stefan H. E. 
Published: April 2014

Abstract
The role of CD8+ T cells in human tuberculosis (TB) remains elusive. We analyzed the T cell repertoire and phenotype in 1) children with active TB (≤4 years), 2) healthy latently Mycobacterium tuberculosis-infected children, and 3) noninfected age-matched (tuberculin skin test-negative) controls. Ex vivo phenotyping of T cell subpopulations by flow cytometry revealed a significant increase in the proportion of CD8+CD45RO−CD62L−CD28−CD27− effector T cells (TEF) in the peripheral blood of children with active TB (22.1 vs 9.5% in latently M. tuberculosis-infected children, vs 8.5% in tuberculin skin test-negative controls). Analyses of TCR variable β-chains revealed markedly skewed repertoires in CD8+ TEF and effector memory T cells. Expansions were restricted to single TCR variable β-chains in individual donors indicating clonal growth. CDR3 spectratyping and DNA sequencing verified clonal expansion as the cause for CD8+ effector T cell enrichment in individual TB patients. The most prominent enrichment of highly similar TEF clones (&gt;70% of CD8+ TEF) was found in two children with active severe TB. Therefore, clonal expansion of CD8+ TEF occurs in childhood TB with potential impact on course and severity of disease.