角膜不同组织免疫原性分析

Relative quantitative analysis of corneal immunogenicity

目的 了解角膜 3层组织的免疫原性相对值。方法 (1)细胞免疫 :将C57BL 6小鼠脊部皮下组织分离成囊袋 ,分别异位植入异种角膜 3层组织 ,于术后 12d用流式细胞仪检测鼠血清中抗CD2 5和抗CD71mAb标记活化的T淋巴细胞。 (2 )体液免疫 :分别用异种角膜上皮细胞和全角膜匀浆免疫动物获取抗血清 ,以酶联免疫吸附法测定含细胞的 3层角膜组织的免疫原性。结果 (1)细胞免疫 :按等组织厚度计算 ,角膜内皮、上皮及基质组织各占角膜总免疫原性的比率分别为 70 75 %、2 7 6 3%及 1 6 2 %。 (2 )体液免疫 :角膜内皮的免疫原性最强 ,角膜上皮次之 ,角膜基质最弱 ,分别约占整个角膜免疫原性的 6 2 11%、31 77%及 6 12 %。结论 3层角膜组织中角膜内皮免疫原性最强 ,基质免疫原性最弱。

Objective To quantitate the relative immunogenicity of three major corneal cell layers (epithelium, stroma, and endothelium) among the whole corneal immunogenicity respectively. Methods Cellular immunity: Three porcine major corneal cell layers were heterotopically allografted to subcutaneous layer of 40 BALB c mice respectively. Twelve days later, the peripheral white blood cells of recipients were double directly stained with anti mouse IgG fluorescein conjugate mAb and analyzed by flow cytometry. Humoral immunity: The suspension of porcine cornea or corneal epithelium was used as antigen to produce anti porcine immune sera in C57BL 6 mice. With the anti porcine immune sera, enzyme linked immunosorbent assay was performed. Results On cellular immunity detection, the immunogenicities of intact endothelium, epithelium and stroma, equal in thickness, were 70 75%, 27 63%, and 1.62% respectively. On humoral immunity detection, the immunogenicities were 62.11%, 31.77% and 6.12% in the three respective layers of equal thickness. Conclusion These findings indicate that the immunogenicity of corneal stroma is the lowest among the three corneal cell layers of equal thickness. The preliminary results provide the experimental basis for clinical application of xenogenic graft of corneal stroma.