大鼠小肠系膜淋巴组织抗原刺激同种异体混合淋巴细胞增殖反应研究

目的探讨大鼠小肠系膜淋巴组织通过树突状细胞（DC）和外周血单核细胞（PBMC）作用后刺激混合淋巴细胞增殖强度和与DC作用后MHC类分子表达变化。方法分离培养SD大鼠DC、PBMC、混合淋巴细胞,获取Winstar大鼠肠系膜淋巴结组织悬液。CCK-8检测法测定负载肠系膜淋巴组织抗原的Dc和PBMC,及负载肠系膜淋巴组织抗原和卵白蛋白（OVA）抗原的DC对混合淋巴细胞刺激增殖能力;流式细胞仪分析肠系膜淋巴组织作用于DC后,其MHC类分子表达变化。结果获得的大鼠小肠淋巴组织悬液中,淋巴细胞占91％,其余包括少量粒细胞、浆细胞、上皮细胞等;在各组中负载肠系膜淋巴组织抗原的DC对混合淋巴细胞刺激增殖能力,均强于PBMC（P〈0．05）;在DC与混合淋巴细胞比例≥1：100各组中,负载肠系膜淋巴组织抗原的DC,对混合淋巴细胞刺激增殖能力,强于负载OVA抗原的DC（P〈0．05）;肠系膜淋巴细胞组织刺激DC后,其MHC—Ⅰ和Ⅱ类分子表达高于未加入抗原组。结论小肠系膜淋巴组织具有强抗原性;DC抗原呈递作用强于单核细胞;负载肠系膜淋巴组织抗原后的DC高表达MHC-Ⅰ、Ⅱ类分子。

Research of rat small intestinal mesentery lymphoid tissue stimulating allograft mixed lymphocyte reaction

OBJECTIVE: To evaluate the effect of the small intestinal mesenteric lymphoid tissues stimulating mixed lymphocyte reaction with dendritic cells (DC) and peripheral blood monocyte cells (PBMC), and observe the changes of the MHC molecular expression on DC. METHODS: DC, PBMC and mixed lymphocyte were separated to culture from SD rats. Lymphoid tissue suspension was adopted from small intestinal mesentery of Winstar rats. In the mixed lymphocyte reaction (MLR), the cellular proliferation of small intestinal mesenteric lymphoid tissue antigen act on DC and PBMC was detected with cell counting of CCK-8 assay, the same assay used in small intestinal mesenteric lymphoid tissue antigen and ovalbumin (OVA) acting on DC. FACS analysis was performed after lymphoid tissue suspension stimulating DC to observe the MHC molecular expression. RESULTS: In the lymphoid tissue suspension, 91% of the cells was lymphocyte, others including granulocyte, plasmocyte, epithelium. The effect of stimulating mixed lymphocyte proliferation were higher in DC groups than in PBMC groups with the small intestinal mesenteric lymphoid tissue (P < 0.05). In the proportion of DC and mixed lymphocyte >or= 1:100 groups, the mixed lymphocyte proliferation were higher in the small intestinal mesenteric lymphoid tissues groups than in the OVA groups (P < 0.05). After stimulated by the small intestinal mesenteric lymphoid tissue, DC expressed higher MHC-I and -II molecules than control groups. CONCLUSIONS: The small intestinal mesenteric lymphoid tissue has high antigenicity; the antigen presenting ability of DC was much stronger than granulocytes; DC expresses high MHC-I and MHC-II molecules after stimulated by mixed lymphoid tissue suspension.