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目的建立 P2或 P0多肽诱导的实验性自身免疫性神经炎(EAN)大鼠模型,确定诱导EAN 的优选抗原和剂量,探讨 EAN 相关细胞免疫机制。方法实验组用 P2_(57-81)或 P0_(180-199)多肽加完全弗氏佐剂(FCA)免疫 Lewis 大鼠,对照组单用 FCA 免疫,致敏后每日对大鼠进行临床评分,比较高峰期最大评分,致敏第14天进行淋巴细胞增殖试验,测定 CD_4~+T/淋巴结单个核细胞(MNC)和CD_4~+CD_(25)~+T/CD_4~+T 细胞百分比,并进行坐骨神经病理检查。结果实验大鼠瘫痪高峰期最大评分,P2_(57-81)200μg组显著高于 P2_(57-81)100μg组和 P0_(180-199)200μg组(均P<0.01),P2_(57-81)100μg组与P0_(180-199)200μg组差异元统计学意义;P2_(57-81)100μg组和 P2_(57-81)200μg组对 P2_(57-81)多肽刺激反应性显著高于对照组(均 P<0.01),P0_(180-199)200μg组对 P0_(180-199)多肽刺激反应性显著高于对照组(P<0.01),P2_(57-81)100μg组和 P0_(180-199)200μg组对相应致敏多肽刺激反应性显著低于 P2_(57-81)200μg组(均P<0.05);CD_4~+T/淋巴结 MNC 百分比在各组间差异无统计学意义;CD_4~+CD_(25)~+T/CD_4~+T 细胞百分比,P2_(57-81)200μg组显著低于 P2_(57_81)100μg组、P0_(180-199)200μg组和对照组(均 P<0.01),P2_(57-81)100μg组和P0_(180-199)200μg组显著低于对照组(均P<0.01),P0_(180-199)200μg组显著低于 P2_(57-81)100μg组(P<0.01);EAN 急性期坐骨神经以炎性细胞浸润为主,P2_(57-81)200μg组重于其他实验组(均 P<0.01),P2_(57-81)200μg组慢性期无炎性细胞浸润,而表现多发性局灶性脱髓鞘和神经纤维崩解未恢复。结论200μg P2_(57-81)多肽是诱导 EAN 的优选抗原,EAN 致病与 CD_4~+T 细胞数量无关,而与致病性 T 细胞活性增强及CD_4~+CD_(25)~+T 细胞减少有关。
Objective To establish a rat model of experimental autoimmune neuritis (EAN) induced by P2 or P0 polypeptide and determine the optimal antigens and doses for inducing EAN, and to explore the mechanism of EAN-related cellular immunity. Methods Lewis rats were immunized with P2_ (57-81) or P0_ (180-199) peptides plus complete Freund’s adjuvant (FCA) in the experimental group. The control group was immunized with FCA alone. After sensitization, the rats were clinically scored The percentages of CD_4 ~ + T / lymphonocyte mononuclear cells (MNCs) and CD_4 ~ + CD_ (25) ~ + T / CD_4 ~ + T cells were measured on the 14th day after sensitization. And sciatic nerve pathology. Results The maximal score at the peak of paralysis was significantly higher in P2_ (57-81) 200μg group than in P2_ (57-81) 100μg group and P0_ (180-199) 200μg group (all P <0.01) ) Group was significantly different from that of P0_ (180-199) 200μg group; P2_ (57-81) 100μg group and P2_ (57-81) 200μg group were significantly more reactive with P2_ (57-81) (P <0.01). The reactivity of P0-1 (180-199) peptide in P0 180-199 group was significantly higher than that in control group (P0.01) (P <0.05). The percentages of CD_4 ~ + T / lymph node MNC in the 200μg group were not significantly different from those in the P2_ (57-81) 200μg group (P <0.05) The percentage of ~ (25) ~ + T / CD_4 ~ + T cells in P2_ (57-81) 200μg group was significantly lower than that in P2_ (57_81) 100μg group, P0_ (180-199) 200μg group and control group 0.01), P2_ (57-81) 100μg group and P0_ (180-199) 200μg group were significantly lower than those of the control group (P0.01). P0_ (180-199) 200μg group was significantly lower than P2_ (57-81) 100μg (P <0.01). The sciatic nerve in the acute phase of EAN was mainly infiltrated by inflammatory cells. The P2_ (57-81) 200μg group was heavier than the other experimental groups (all P <0.01) No inflammatory cell infiltration, and the performance of multiple focal demyelination and collapse of nerve fibers is not restored. CONCLUSION: 200μg P2_ (57-81) polypeptide is the optimal antigen for EAN induction. EAN pathogenicity has no relation with the number of CD_4 ~ + T cells, but increased with pathogenic T cells and CD_4 ~ + CD_ (25) ~ + T cells related.