肾综合征出血热纯化疫苗候选毒株在Vero细胞上的适应传代及其免疫原性研究

目的 将肾综合征出血热纯化疫苗候选毒株适应于Vero细胞，并对其抗原性和免疫原性进行研究。方法 将汉滩病毒H8207株和汉城病毒Y86013株在Vero细胞上进行连续终末稀释传代，采用间接免疫荧光法、酶联免疫吸附试验和空斑减少中和试验，研究传代后毒株的繁殖特性、病毒滴度、抗原量及免疫原性。结果 经过5次终末稀释传代，两株病毒在Vero细胞上均显示出良好的适应性，从第六代开始病毒滴度稳定在7．0Lg TCID50/m1以上，第八代两毒株抗原量均已达到1：64，H8207株抗原量继续升高，最高时达1：256。用两株病毒不同培养代次上清液制备的单价原液灭活疫苗免疫家兔二针，免疫血清对同型毒株的中和效价均达到1：10。结论 两株病毒已适应于Vero细胞，且具有病毒滴度高和免疫原性良好的特性，适合用作Vero细胞肾综合征出血热灭活纯化疫苗的候选毒株。

Passage adaptability of candidate strains for hemorrhagic fever with renal syndrome purified vaccine in Vero cells and their immunogenicity

OBJECTIVE: To adapt the candidate strains of hemorrhagic fever with renal syndrome (HFRS) purified vaccine to Vero cells and to study their antigenicity and immunogenicity. METHODS: The viral strains H8207 (Hantaan virus, HTN) and Y86013 (Seoul virus, SEO) were continuously propagated in Vero cell by the terminal dilution method and studied the characteristics of virus multiplication, viral titers and the amounts of virus antigen after serial passages. Three batches of crude monovalent inactivated vaccine were developed using the different passages of these 2 viral strains. RESULTS: The strains H8207 and Y86013 adapted to Vero cells and stably grew on the cells with high titers. Rabbits immunized with the crude vaccines of H8207 and Y86013 showed 100% sero-conversion and the neutralizing antibody titers of the rabbit immune sera reached 1?10 at 4 weeks after 2 times of immunization. CONCLUSIONS: The results suggest that these 2 candidate strains had adapted to Vero cells, possessed high titers and good immunogenicity and be feasible to prepare the HFRS purified vaccine in Vero cells.