羧基荧光素二醋酸盐琥珀酰亚胺酯标记淋巴细胞评估粘膜耐受对大鼠实验性结肠炎模型免疫状态的影响

ims: To investigate the influence of mucosal tolerance on the immunologic status of the experimental colitis. Methods: Rat models of experimental colitis were established by enema with 2,4,6-trinitrobenzenesulphonic acid (TNBS). Ovalbumin(OVA) was used as inducing antigen and lipopolysaccharide (LPS) as adjuvant. Rat models of experimental colitis were grouped according to different therapies as following: OVA oral administration, OVA nasal administration, OVA and LPS oral administration, OVA and LPS nasal administration, LPS oral administration. PBS was orally or nasally administrated as controls. Splenic lymphocytes were extracted and labeled with 5,6-Carboxyfluorescein diacetate succinimidyl ester , and splenic mononuclear cells proliferation test was developed with OVA as stimulant. Fluorescence intensity of cells was tested by flow cytometry after 72 hours and proliferation index was calculated. Results: Colonic macroscopical scores and histological scores decreased significantly after OVA and LPS oral administration, OVA and LPS nasal administration or LPS oral administration. Among them OVA and LPS nasal administration reduced colonic scores most significantly. CFSE labeled lymphocyte proliferation test showed that experimental colitis interfered the induction of tolerance by oral administration, but it has no influence on induction of tolerance via nasal administration and induction of mucosal tolerance via nasal administration of antigen and adjuvant. Conclusions: 5,6-Carboxyfluorescein diacetate succinimidyl ester labeled lymphocyte proliferation test could be employed to evaluate therapeutic effect of mucosal tolerance. Nasal tolerance combined with adjuvant had therapeutic effect on TNBS induced experimental colitis, which could have potential clinical applications.