KSHV编码的趋化因子vMIPII在小鼠异体皮肤移植免疫排斥反应中的作用

目的　重组vMIPII在大肠杆菌中表达及其纯化。观察纯化的vMIPII蛋白对小鼠异体皮肤移植免疫排斥反应的抑制作用。方法　以 pET3 2a为vMIPII克隆基因的表达载体 ,以大肠杆菌BL2 1(DE3 ) plysS为表达菌 ,诱导表达出硫氧还蛋白 vMIPII的融合蛋白 (2 6× 10 3 D)。经金属离子亲和吸附、肠激酶消化以游离vMIPII、阳离子交换层析等步骤纯化目的蛋白。纯化的vMIPII从尾静脉连续 14d注入异体皮肤移植小鼠 (昆明鼠 /Balb/c)。结果　从 1L发酵菌液中可获得高纯度目的蛋白约 4mg。注射vMIPII的异体皮肤移植小鼠组移植皮存活时间较未注射对照组延长 7d。结论　纯化的重组vMIPII对小鼠异体皮肤移植免疫排斥反应有明显的抑制作用 ,能显著延长移植皮的存活时间

Role of KSHV-encoded chemokine vMIPII in mouse allo-skin graft rejection

Objective To obtain recombinant vMIPII expressed in E. coli, and to investigate the inhibition of vMIPII in mice allo skin transplantation rejection. Methods A thioredoxin vMIPII fusion protein(26x103) was expressed in E. coli BL21(DE3)plysS under the induction of IPTG when pET32a vMIPII was used as an expression vector. Purified vMIPII was produced through metal chelated affinity chromatography, enterokinase digestion to free vMIPII from fusion protein and cation exchange chromatography. Purified vMIPII was injected in tail vein once everyday after skin transplataton (Kunming mice to Balb/c)till day of the 14th.Results Approximately 4mg purified vMIPII were obtained from 1 liter of fermented bacteria. The allo skins of the mice which were injected in vMIPII survived 7 days more than that which were injected in salt saline. Conclusion Purified recombinant vMIPII is capable of inhibiting the allo graft rejection in mouse allo skin transplantation, markedly prolonging the survival of the allo skin.