肾综合征出血热病毒结构蛋白的纯化及免疫学特性分析

应用从肾综合征出血热（ＨＦＲＳ）患者血清中提取的ＩｇＧ与活化的Ｓｅｐｈａｒｏｓｅ４Ｂ偶联，制备亲和层析柱，用此亲和层析柱从ＨＦＲＳＶ感染的小白鼠乳鼠鼠脑中提取出２种分子量为６７０００和５５０００的病毒结构蛋白。经ＥＬＩＳＡ证明，此病毒结构蛋白能与ＨＦＲＳ患者血清ＩｇＧ及抗ＨＦＲＳ病毒的ＭｃＡｂ反应，效价为１６０。与６株抗ＨＦＲＳＶＭｃＡｈ试验表明，此病毒结构蛋白能与Ｈ７株反应。血凝试验证明，此病毒蛋白不能凝集鹅红细胞、鸽血球和人＂Ｏ＂型血红细胞。将纯化的病毒结构蛋白免疫家兔，证明其有较强的免疫原性，可刺激家兔产生特异性抗体；微量中和试验及乳鼠中和试验证明，中和效价为２５６，说明纯化的病毒结构蛋白具有中和抗原位点；免疫血清对感染乳鼠有一定保护作用。

Purification and Characterization of HFRS Virus Structural Protein

An immunoadsorbent column was prepared by sepharose 4B coupled with IgG isolated from HFRS patients. Using the immunoaffinity chromatography column, HFRS virus structural proteins were purified from brain tissue of sucking mice infected with the 76-118 strain of HFRS virus. Two viral structural proteins were obtained,which showed two bands on SDS-PAGE,and their molecular weights were 55K and 67K. respectively. The purified viral protein could react with positive serum and McAb against HFRS virus. The results of hemagglutination test indicate that both of the viral protein have no hemagglutination activity. Rabbits were immunized with the purified proteins.The purified viral proteins had higher immunogenicity and could induce specific antibodies against HFRSV.It was shown by neutralization test that the immune sera contained neutralizing antibodies with a titer of 1:256. The immune sera could not only neutralize HFRSV in cell culture, but also had protective effect on suckling mice infected with HFRSV. These results suggest that the two purified viral proteins are analogous to G1 and G2 glycoproteins of HFRS virus.