摘要
目的建立P257-81-特异性T细胞系,在实验性自身免疫性神经炎(EAN)大鼠进行T细胞疫苗接种(TCV)的实验研究。方法P257-81多肽、IL-2和抗CD3/CD28单抗包被磁珠诱导扩增P257-81-特异性T细胞及非抗原特异性T细胞,将灭活T细胞接种大鼠,检测指标包括:瘫痪高峰期最大评分比较,淋巴细胞增殖试验、CD4+T/淋巴结单个核细胞及CD4+CD25+T/CD4+T细胞百分比测定、培养液上清IFN-γ及IL-10水平测定以及坐骨神经病理学检查。结果P257-81-特异性T细胞显著扩增,2周扩增近1000倍;P257-81-特异性TCV预防组无大鼠发病;P257-81-特异性TCV治疗组大鼠病情减轻,其坐骨神经炎性细胞浸润程度显著轻于对照组(P<0.001);P257-81-特异性TCV预防组和治疗组CD4+CD25+ T/CD4+T细胞百分比及培养液上清IL-10水平均显著高于其对照组,培养液上清IFN-γ水平均显著低于其对照组(均P<0.001)。结论抗原+IL-2+CD3/CD28单抗包被磁珠刺激是一种高效、可靠的扩增抗原特异性T细胞方法;P257-81-特异性TCV能预防EAN发生,并可治疗已发生的EAN,其机制可能为:使CD4+CD25+调节性T细胞/CD4+T细胞比例上调,诱导致病性CD4+Th1细胞向保护性CD4+Th2细胞转换。
Objective To study effect of the T cell vaccination（TCV） on the experimental autoimmune neuritis （EAN）. Methods EAN rats were immunized by injection into both hind footpads of inocula containing 200 μg of P257-81 peptide and FCA. P257-81 peptide specific-T and antigen-non-specific-T cells were isolated and expanded. Then T cells were inactivated and vaccinated into Lewis rats. Symptomatic scores were compared at the maximum of the disease. On the 14th day after immunization, we examined lymphocyte proliferation, fractions of CD4^＋ T cells among mononuclear cells in lymph nodes, frequencies of CD4^＋ CD25^＋ T ceils among CD4^- T cells, supernatant productions of IFN-γ and IL-10 secreted by lymphocytes. Histopathology of sciatic nerves was assessed. Results We applied P257-81 peptide, dynabeads coated with anti-CD3 and anti CD28 antibodies and IL-2 to induce T cells proliferation, through which we could expand P257-81 peptide specific T cells nearly 1000 fold within 2 weeks. We found that P257-81 peptide specific-T cells vaccination could both prevent and cure EAN, and antigen non- specific-T cells vaccination had no benefit. Conclusions The expansion protocol including antigen, dynabeads coated with anti-CD3 and anti CD28 antibodies and IL-2 is a kind of efficient, reliable way to expand antigen specific T cells. P257-81 peptide specific-TCV can prevent rats from EAN and therapy EAN by upregulating frequencies of CD4^＋ CD25^＋ T ceils among CD4^＋T cells, and inducing the shift of pathogenic CD4^＋Thl cells to protective CD4^＋ Th2 cells.
出处
《中国神经免疫学和神经病学杂志》
CAS
2008年第6期408-412,474,共6页
Chinese Journal of Neuroimmunology and Neurology
基金
国家自然科学基金资助项目(30470583)
