食蟹猴免疫抑制模型建立初探

Establishment of Immunosuppressive Model of Cynomolgus Monkey

目的:探讨建立食蟹猴免疫抑制模型并对其进行生物学相关指标的评价。方法:以食蟹猴为造模动物,选取环磷酰胺、硫唑嘌呤、强的松、他克莫司、霉酚酸酯、环孢素等六种免疫抑制剂进行构建食蟹猴免疫抑制模型,通过物理观察和生化指标检测两方面对食蟹猴免疫抑制模型的构建进行评估,并主要选择了血液学常规、T淋巴细胞亚群、肝肾功能等指标对食蟹猴的免疫抑制状况进行观察。结果:(1)血常规检测中:在六种免疫抑制剂中,环磷酰胺、他克莫司和霉酚酸酯给药后1~3周内,白细胞计数、中性粒细胞计数均显著下降(P<0.05,P<0.01),淋巴细胞计数无明显差异,但淋巴细胞比例呈显著性升高(P<0.05,P<0.01)。环孢素A和硫唑嘌呤给药后血液学大部分指标无显著差异。而强的松给药后白细胞计数不降反升(P<0.05,P<0.01)。(2)T淋巴细胞亚群检测中:六种免疫抑制剂中,环磷酰胺和他克莫司给药后的T细胞亚群变化趋势较为稳定,均表现为T淋巴细胞比例显著升高(P<0.05,P<0.01)、Th细胞比例显著降低(P<0.05,P<0.01)、Tc细胞比例显著升高(P<0.05,P<0.01)。(3)肝肾功能检测中:他克莫司免疫抑制剂给药后1~2周内ALT水平显著升高(P<0.01),2周后ALT逐渐回落。而环磷酰胺、硫唑嘌呤、霉酚酸酯和他克莫司给药至2~3周,均导致AST水平显著升高(P<0.01)。结果说明,这几种免疫抑制剂均会导致食蟹猴肝功能出现阶段性地轻度损害。此外,检测结果表明,六种免疫抑制剂均会导致Crea和Urea水平发生变化,但Crea和Urea指标均分别在正常59~104μmol/L,3.6~9.5μmol/L范围内,表明六种免疫抑制剂对肾功能均无影响作用。结论:成功建立了食蟹猴免疫抑制模型;六种免疫抑制剂均能各自呈现出程度不等、指标不同的免疫抑制效应;环磷酰胺和他克莫司两种药物对食蟹猴进行免疫干预后,可呈现出较为确定、稳定的免疫抑制效应,是建立免疫抑制食蟹猴模型较为适宜的造模剂。

Objective: To establish cynomolgus monkey immunosuppressive model and evaluate its biological indexes. Methods: Six kinds of immunosuppressants cyclophosphamide, azathioprine, prednisone, tacrolimus, mycophenolate mofetil and cyclosporine were to be used for building cynomolgus monkey immunosuppressive model, evaluated by the two aspects of physical observation and biochemical indicator detection, and indexes of blood routine examination, T cell subset, liver and kidney function were chosen to assess the cynomolgus monkey immunosuppressive model. Results:(1) In blood routine examination, 1~3 weeks after cyclophosphamide , tacrolimus and mycophenolate administration, leukocyte and neutrophil count were significantly decreased (P<0.05, P<0.01), no significant difference in lymphocyte count, but significant elevation in the proportion of lymphocytes (P<0.05, P<0.01);After administration of cyclosporin A and azathioprine, most of the indicators of hematology showed no significant difference, and after administration of prednisone, leukocyte count elevated significantly (P<0.05, P<0.01).(2) In T cell subset detection, after administration of cyclophosphamide and tacrolimus, the change trend of T cell subset was relatively stable, showing that the proportion of T lymphocytes was significantly increased (P<0.05, P<0.01), the proportion of Th cells was significantly decreased (P<0.05, P<0.01), and the proportion of Tc cells was significantly increased (P<0.05, P<0.01).(3) In Liver and kidney function test, 1~2 weeks after administration of tacrolimus, the level of ALT was elevated remarkably, gradually fell back after two weeks. The administration of cyclophosphamide, azathioprine, mycophenolate and tacrolimus for 2 to 3 weeks resulted in significant increases in AST levels (P<0.01). The results indicated that all of these immunosuppressive agents could lead to slight stage damage of liver function in cynomolgus monkeys. In addition, the test results showed that all six immunosuppressive agents would lead to changes in Crea and Urea levels, but both Crea and Urea indexes were within the normal range of 59~104μmol/L, 3.6~9.5μmol/L respectively, indicating that all six immunosuppressive agents had no effect on renal function. Conclusion: The cynomolgus monkey immunosuppressive model was established successfully. All the six kinds of immunosuppressive agents showed different degree and indexes respectively. After the immune intervention of cyclophosphamide and tacrolimus, the immune suppression effect of cynomolgus monkeys can be determined and stable, and it is a more appropriate model builder to establish the immune suppression of cynomolgus monkey.