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目的探讨酶法处理短棒状杆菌(Enzyme-digested Corynebacterium parvum product,ECPP)的效果。方法采用热灭活、超声、Pronase酶消化等方法处理短棒状杆菌(Corynebacterium parvum,CP),电镜观察其结构,并测定其蛋白、多糖及核酸含量。将BALB/c纯系小鼠、青紫蓝家兔和NIH小鼠均随机分为阴性对照组(CK,0.9%NaCl)、短棒状杆菌制剂(C.parvum product,CPP)组(1 000μg/ml CPP)、低剂量ECPP组(ECPPⅠ组,500μg/ml ECPP)和高剂量ECPP组(ECPPⅡ组,1 000μg/mlECPP),观察BALB/c纯系小鼠脾激活情况、异常毒性及血清IL-2、IFNγ及TNF-α的水平,青紫兰家兔热原反应,NIH小鼠抑瘤效果。结果电镜观察可见,与热灭活的CP相比,ECPP细胞壁外围厚度明显变薄,细菌内部结构无明显变化;ECPP的蛋白、多糖及核酸含量较CPP均明显下降,且差异均有统计学意义(P<0.01);ECPPⅡ组小鼠的脾激活指数为2.96,能抑制艾氏腹水瘤,存活率为80%,热原试验合格,异常毒性试验显示其体重增长较CPP组显著,能显著提高小鼠血清中IL-2、IFNγ及TNF-α3种细胞因子的水平。结论酶法处理短棒状杆菌制剂可降低副反应,提高机体免疫功能,具有良好的抗肿瘤作用,是一种有应用前景的新免疫调节剂。
Objective To investigate the effect of enzymatic treatment of Enzyme-digested Corynebacterium parvum product (ECPP). Methods Corynebacterium parvum (CP) was treated by heat inactivation, sonication and digestion with Pronase, and its structure was observed with electron microscope. The contents of protein, polysaccharide and nucleic acid were determined. BALB / c mice, Cyanidium blue rabbits and NIH mice were randomly divided into negative control group (CK, 0.9% NaCl), C. parvum product (CPP) group CPP group, ECPP groupⅠ, 500μg / ml ECPP group) and high-dose ECPP group (ECPPⅡgroup, 1 000μg / mlECPP) to observe the splenic activation and abnormal toxicity and the level of serum IL-2 , The levels of IFNγ and TNF-α, the pyrogen response in rabbits, the antitumor effect in NIH mice. Results Electron microscopy showed that the thickness of the outer wall of ECPP was thinner than that of heat-inactivated CP, and there was no significant change in the internal structure of the bacteria. The protein, polysaccharide and nucleic acid content of ECPP were significantly lower than those of CPP, and the differences were statistically significant (P <0.01). The spleen activation index of ECPPⅡgroup was 2.96, which could inhibit Ehrlich ascites tumor with a survival rate of 80%. The pyrogen test was passed. Abnormal toxicity test showed that the body weight gain was significantly higher than that of CPP group Levels of IL-2, IFNγ and TNF-α3 cytokines in mouse serum. Conclusion Enzymatic treatment of Corynebacterium parvum preparations can reduce side effects, improve immune function and have good anti-tumor effect. It is a promising new immunomodulator.