GIF regulation of B cells and CD4 cells

GIF 对 B 细胞和 CD4 细胞的调节

• 批准号: 7173448
• 负责人: KATSUJI SUGIE
• 金额: $ 31.49万
• 依托单位: LA JOLLA INSTITUTE FOR IMMUNOLOGY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-02-01 至 2009-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7173448
• 关键词: Address; Adoptive Transfer; Antibodies; Antibody Formation; Antigen Presentation; Antigen-Presenting Cells; Antigens; Asthma; B-Cell Activation; B-Lymphocytes; Binding; Biological; Biological Assay; Bone Marrow; CD4 Positive T Lymphocytes; Cell Surface Proteins; Cell division; Cells; Chemicals; Chimera organism; Cloning; Collaborations; Complementary DNA; Cysteine; Cytokine Receptors; Cytoplasm; Cytosol; Endosomes; Enzyme-Linked Immunosorbent Assay; Event; Facility Construction Funding Category; Flow Cytometry; Gene Targeting; Generations; Hematopoietic; Immune response; Immunization; In Vitro; Infection; Inflammation; Interleukin-4; Knowledge; Leishmania major; Lysosomes; Mass Spectrum Analysis; Measurement; Measures; Mediating; Messenger RNA; Migration Inhibitory Factor; Modification; Mus; Ovum; Phenotype; Phosphorylation; Post-Translational Protein Processing; Process; Production; Protein Sequence Analysis; Proteins; Receptors, Antigen, B-Cell; Regulation; Reporting; Rest; Role; Sampling; Somatic Cell; Source; T-Cell Activation; T-Independent Antigens; T-Lymphocyte; Testing; Time; Tissues; Work; allergic airway inflammation; aluminum sulfate; cDNA Library; cell mediated immune response; cell type; crosslink; cytokine; disulfide bond; expression cloning; glycosylation inhibiting factor; in vivo; mouse model; phenylpyruvate tautomerase; programs; receptor; receptor expression; research study; response; trafficking; transcription factor; uptake

DESCRIPTION (provided by applicant): The function of a cytokine called GIF or MIF has been unclear for a long time. However, recent studies have demonstrated that a posttranslational modification of this cytokine at C60 is required for the binding to the receptor and for the capability to inhibit B cell Ig switch, BCR-mediated antigen uptake, and IL-4 secretion from CD4 cells. GIF-/- mice displayed enhanced antibody responses to a T-dependent antigen regardless of Ig isotype but normal responses to a T-independent antigen. CD4 cells from GIF-/- mice differentiated toward a Th2 phenotype as compared with wild type cells. GIF receptor was undetectable in resting mouse T and B cells, but after in vitro stimulation the receptor was induced on these cells. GIF receptor appeared to be a 50-52 kDa cell surface protein. To determine whether the biological effects of GIF is mediated through the interaction of GIF and its receptor, anti-receptor mAb will be generated and the cDNA encoding the receptor will be cloned. To define the target cells for this cytokine, the expression of the receptor will be delineated using cells and mRNA samples from various hematopoietic and nonhematopoietic tissues. To clarify the role of GIF in regulating B cell activation in T-B collaboration, HEL-specific B cells from MD4 BCR Tg mice and Ova-specific CD4 cells from DO11.10 TCR Tg mice, either on GIF+/+ or -/- background, will be stimulated in vitro with HEL-Ova conjugate. The secretion of GIF, the induction of its receptor, and the events associated with B cell activation will be analyzed in the course of the cognate T-B interaction. The effects of GIF on BCR-dependent antigen uptake, trafficking, and processing will be analyzed. To determine the role of GIF on cytokine production and Th differentiation of CD4 cells, the expression of GIF receptor and the secretion of GIF upon T cell activation will be analyzed. The source of GIF that regulates Th differentiation will be determined by adoptive transfer experiments using CD4 cells from GIF+/+ and -/- DO11.10 TCR Tg mice and bone marrow chimera constructions. The mechanism by which GIF inhibits Th2 differentiation and/or induces Th1 differentiation will be analyzed with regard to Stat phosphorylation and transcription factors. The role of GIF in regulating Th2-type inflammation will be evaluated in the mouse model of antigen-induced bronchial asthma. These experiments will establish the role of this cytokine in CD4 T cell-mediated immune responses.

描述（由申请人提供）：一种称为GIF或MIF的细胞因子的功能长期以来一直不清楚。然而，最近的研究表明，该细胞因子需要在C60处进行翻译后修饰才能与受体结合，并能够抑制B细胞Ig开关、bcr介导的抗原摄取和CD4细胞分泌IL-4。GIF-/-小鼠对t依赖性抗原表现出增强的抗体反应，与Ig同型无关，但对t非依赖性抗原表现出正常反应。与野生型细胞相比，来自GIF-/-小鼠的CD4细胞向Th2表型分化。静止小鼠T细胞和B细胞中检测不到GIF受体，但在体外刺激后，这些细胞被诱导出该受体。GIF受体是一个50-52 kDa的细胞表面蛋白。为了确定GIF的生物学效应是否通过GIF与其受体的相互作用介导，我们将生成抗受体mAb，并克隆编码受体的cDNA。为了确定这种细胞因子的靶细胞，受体的表达将使用来自各种造血和非造血组织的细胞和mRNA样本来描述。为了阐明GIF在T-B协同作用中调节B细胞活化的作用，我们在体外用HEL-Ova偶联物刺激GIF+/+或-/-背景下MD4 BCR Tg小鼠的hell特异性B细胞和DO11.10 TCR Tg小鼠的ova特异性CD4细胞。GIF的分泌、受体的诱导以及与B细胞活化相关的事件将在同源T-B相互作用的过程中进行分析。GIF对bcr依赖性抗原摄取、运输和加工的影响将被分析。为了确定GIF对CD4细胞细胞因子产生和Th分化的作用，我们将分析在T细胞活化时GIF受体的表达和GIF的分泌。通过采用来自GIF+/+和-/- DO11.10 TCR Tg小鼠的CD4细胞和骨髓嵌合体构建的过继性转移实验，确定调节Th分化的GIF来源。GIF抑制Th2分化和/或诱导Th1分化的机制将根据Stat磷酸化和转录因子进行分析。我们将在小鼠抗原诱导支气管哮喘模型中评估GIF对th2型炎症的调节作用。这些实验将确定这种细胞因子在CD4 T细胞介导的免疫应答中的作用。