Prostate-specific membrane antigen (PSMA) is a cell surface glycoprotein expressed predominantly in prostate secretory acinar epithelium and prostate cancer cells as well as in several extraprostatic tissues. Mouse monoclonal antibody 4G5 specific to the extracellular domain of PSMA was used to screen two phage displayed peptide libraries (9aa linear and 9aa cys library). Three 4G5-reactive phagotopes were identified. Sequence analysis of isolated clones demonstrated that the interaction motif VDPA/SK has high homology to 719-725aa on PSMA. Immunohistochemical staining of the prostate cancer sample with the PSMA-mimic phagotope (mimotope) immunized serum antibodies demonstrate that the mimotope isolated from the phage displayed peptide libraries can induce PSMA specific immune response in vivo.

Pertussis toxin (PTX) has been widely used as an adjuvant to induce Th1-mediated organ-specific autoimmune diseases in animal models. However, the cellular and molecular mechanisms remain to be defined. In this study, we showed that dendritic cells (DC) stimulated with PTX (PTX-DC) were able to substitute for PTX to promote experimental autoimmune uveitis (EAU). EAU induced by PTX-DC revealed a typical Th1 response, characterized by high uveitogenic retinal Ag interphotoreceptor retinoidbinding protein (IRBP)-specific IFN-and IL-12 production in the draining lymph nodes, as well as increased levels of anti-IRBP IgG2a and decreased levels of anti-IRBP IgG1 in the serum of IRBP-immunized mice. Furthermore, PTX-DC preferentially induced T cells to produce the Th1 cytokine, IFN-. After being stimulated with PTX, DC exhibited up-regulation of MHC class II, CD80, CD86, CD40, and DEC205. PTX-DC had also increased allostimulatory capacity and IL-12 and TNF-production. Serum IL-12 was increased in naive mice that received PTX-DC i.p. In addition, PTX activated extracellular signal-regulated kinase in DC. Following the inhibition of extracellular signal-regulated kinase signaling, the maturation of PTX-DC was reduced. Subsequently, the ability of PTX-DC to promote IFN-production by T cells in vitro and to induce EAU in vivo was blocked. The results suggest that PTX might exert an adjuvant effect on DC to promote their maturation and the production of proinflammatory cytokines, thereby eliciting a Th1 response.

目的 将小鼠前列腺癌细胞株RM-1裂解产物加载树突状细胞（DC）后，转染干扰素一γ诱导蛋白-10（IP-lO）基因构建DC瘤苗，探讨该瘤苗对小鼠抗肿瘤免疫匣应的诱导作用。方法 将RM-l细胞的裂解产物作为肿瘤抗原加载小鼠骨髓来源的Dc，并通过脂质体法转染IP-10基因，构建DC瘤苗；检测DC瘤苗的抗前列腺癌免疫治疗作用和免疫保护作用。结果 转染DC强表达IP-10，其上清对淋巴细胞有较强的趋化作用；构建的DC瘤苗能诱导特异性抗前列腺癌免疫反应，经瘤苗处理的荷瘤小鼠瘤体生长减慢，存活期延长；瘤苗还具有明显的免疫保护作用。结论 构建的前列腺癌DC瘤苗在体内能有效诱导抗肿瘤免疫反应和免疫保护功能。

目的 研究人转化生长因子βl（TGFβ1）基因修饰对大鼠骨髓树突状细胞（DC）免疫功能的影响。方法 构建TCFβ1-pcDNA3质粒并转染未成熟DC。检测转染后DC的TGFβ1表达及其功能变化，输注l周后检测TGFβ1-DC在受者睥和淋巴结的分布、T细胞捅亡水平及T绳胞端粒酶表达。结果 TGFβ1-pcDNA3船最粒转染Dc能有效抑制DC的成熟和分化，并下调DC的多种功能。TG即1基因转染不仅能降低DC对LPS的反应，同时抑制DC在MLR中刺激T细胞增殖的能力。TGFβ1-DC输注受者后，l周内可在脾和淋巴结内形成微嵌合，并有效诱导T细胞的凋亡，抑制T细胞端粒酶的活性和表达。结论 TGFβl基因能有效抑制DC的多种功能，可用于诱导机体免疫耐受。

目的 构建含小鼠次级淋巴组织趋化因子（SLC）cDNA的重组腺相关病毒载体。方法 以C57BL/6J小鼠的淋巴组织为材料抽提总RNA，用RT-PCR方法克隆小鼠SLC的成熟肽基因。PCR产物回收后经EcoRI和Xho双酶切，插人pAAv-IRES-hrTGFP质粒，用磷酸钙法，与pAAV-Rc和pHetpeR三者共同转染HEK293细胞。包装成重组的病毒颗粒，并通过绿色荧光蛋白（CFP）报告基因荧光检测及抽提病毒DNA，进行PCR扩增等进一步证实重组病毒的形成。结果 约500bp的小鼠SLC成熟肽基因被成功克隆，序列分析表明，所克隆的SLC基因与基因库注册的相同，组载体的酶切及测序鉴定表明SLC基因被定向插入。重组载体转染HEK293细胞，经荧光显微镜和病毒DNA的PCR检测，证实已完成对重组病毒的包装。并表达GEP。结论 成功构建了SLC成熟肽基因重组腺相关病毒载体。