不同免疫佐剂对寻常型天疱疮抗原EC1-2表位抗体亚型的影响

The influence of immune adjuvants on the antibody subclasses against pemphigus vulgaris antigen EC1-2 epitope

目的探讨免疫佐剂的十预对寻常型天疱疮抗原ECl-2表位抗体亚型转换的影响。方法完全弗氏佐剂(CFA)或铝[Al(OH)_3]佐剂联合重组天疱疮抗原(PVA)ECl-2融合蛋白,分别免疫C57BL/6小鼠,检测免疫后细胞因子的产生及其特异性抗体的滴度和型别,并通过间接免疫荧光(IIF)及新生鼠皮肤的体外培养,观察抗血清同抗原的亲和力及其致病作用。结果免疫后经细胞因子检测发现,CFA组小鼠IFN-γ、Th1型细胞因子增高；Al(OH)_3组则以IL-4、Th2型细胞因子增高为主。两组小鼠均有特异性的抗ECl-2 IgG型抗体产生,但其亚型不同,CFA组以IgG2a为主,而Al(OH)_3组则仅表达IgGl。CFA组小鼠血清IIF示弱的荧光沉积,体外实验示轻度棘层松解；而Al(OH)_3组则表现为明亮的荧光沉积,体外实验出现表皮内水疱。结论不同免疫佐剂直接影响寻常型天疱疮抗原ECl-2表位的抗体亚型。

Objective To study the influence of different immune adjuvants on the switch of antibody classes against pemphigus vulgaris antigen EC 1-2 epitope （ PVA EC 1-2 ）, Methods PVA EC 1-2 fusion protein, combined with complete Freund＇s adjuvant （ CFA ） or aluminum hydroxide hydrate [A1（ OH ）3], was used to immunize C57BL/6 mice. After immunization, the titers and subclasses of specific antibody as well as related cytokines were measured. The affinity of antibody subclasses with antigen and pathogenicity of antibody subclasses were studied by indiret immune-fluorescence （ IIF ） and in vitro culture of skin graft from neonatal mice, respectively. Results After immunization, the level of IFN-γ, a Th1 type cytokine, was elevated in CFA group, while the level of IL-4, a Th2 type cytokine, was increased in A1 （ OH ） 3 group. Specific antibodies were detected in both groups of mice. IgG2a subclass predominated in CFA group, but only IgGl subclass was detected in the AI （ OH ） 3 group. IIF showed a weak fluorescence between the squamous epithelial cells with the addition of antisera from CFA group, but a relatively bright fluorescence with the addition of antisera from the A1 （ OH ） 3 group. Furthermore, the in vitro cultm＇e of skin gra12 with the antisera from the AI （ OH ） 3 group showed intraepithelial blister, but that from the CFA group only showed mild acantholysis. Conclusion Different immune adjuvants may have direct influence on the antibody subclasses against PVA EC1-2.