轮状病毒P[8]G1株在KMB17细胞上的适应性及其免疫原性

Adaptability of Rotavirus P[8]G1 Strain in KMB17 Cells and Immunogenicity of Adapted Strain

目的研究轮状病毒P[8]G1株(Wa株)在人胚肺二倍体细胞KMB17上的适应性及其免疫原性。方法将人轮状病毒P[8]G1株(Wa株)在KMB17细胞上进行适应培养,连续传代10代,免疫荧光法检测病毒的增殖情况,免疫胶体金法检测病毒的抗原性,微量滴定法检测病毒的感染性滴度,并进行病毒基因组稳定性及免疫原性检测。结果轮状病毒Wa株在KMB17细胞上连续传代后,细胞病变逐渐增快;抗原性逐渐增强;至第10代达增殖高峰,病毒滴度达4.25CCID50/ml;在传代过程中,病毒基因组核酸带型保持一致;经皮下和口服2种途径免疫小鼠,血清抗体效价均达1:8192。结论轮状病毒Wa株可在KMB17细胞上稳定增殖,病毒保持了毒株的基本生物学特性,且具有较好的免疫原性。

Objective To study the adaptability of rotavirus P[8]G1 strain in KMB17 cells and the immunogenicity of adapted strain.Methods Human rotavirus P[8]G1 strain（Wa strain）was subcultured in KMB17 cells for 10 passages,then determined for proliferation by IFA,for antigenicity by immunocolloid gold method,for infectious titer by microtitration,and tested for genomic stability and immunogenicity.Results After continuous subculture in KMB17 cells,the CPE of Wa strain appeared early,while the antigenicity increased gradually.The proliferation of Wa strain of passage 10 reached the maximum,with a titer of 4.25 CCID50 /ml.The nucleic acid band pattern of viral genome showed no change during subculture.Both the serum antibody titers of mice immunized with the adapted strain by subcutaneous and oral routes reached 1：8192.Conclusion Rotavirus Wa strain was stably proliferated in KMB17 cells,which maintained the basic biological characteristics and showed good immunogenicity.