摘要
目的观察不同品系小鼠（Ｈ２ｂ，Ｈ２ｄ）经乙型肝炎病毒核心抗原（ＨＢｃＡｇ）基因疫苗免疫后特异性辅助性Ｔ细胞反应类型和特异性细胞毒性Ｔ细胞活性。方法Ｗｅｓｔｅｒｎｂｌｏｔ法鉴定该基因疫苗的体外表达产物；基因枪法和肌内注射法接种该基因疫苗；间接ＥＬＩＳＡ法检测小鼠血清抗ＨＢｃＩｇＧ亚类（ＩｇＧ１，ＩｇＧ２ａ）水平；５１铬释放法测定小鼠脾细胞ＨＢｃＡｇ特异性细胞毒性Ｔ细胞活性。结果两种品系小鼠经不同方法接种该基因疫苗后均产生高滴度血清抗ＨＢｃ（１∶１０９３５０～１∶３２８０５０）；抗ＨＢｃＩｇＧ亚类以ＩｇＧ２ａ占优，肌内注射法接种Ｃ５７ＢＬ／６小鼠后尤然。各组小鼠的ＨＢｃＡｇ特异性细胞毒性Ｔ细胞杀伤活性均达５０％以上。结论该基因疫苗具有良好的体液免疫和细胞免疫原性，值得进一步探索其临床应用前景。
Objective To observe the specific humoral and cellular immunogenecity of genetic vaccine based on core gene of hepatitis B virus in both C57BL/6(H 2 b)and BALB/c(H 2 d)mice. Methods The in vitro expressing product of genetic vaccine of HBV core antigen (HBcAg) was identified by Western blot. Both methods of gene gun and intramuscular immunization were used. Anti HBc(IgG)and its isotypes (IgG1, IgG2a)in mice sera were detected by indirect ELISA. HBcAg specific cytotoxic T lymphocyte(CTL)activity was measured by 51 Chromium release assay. Results The expression of HBcAg was confirmed by Western blot. High titers of serum anti HBc(1∶328 050 by gene gun method and 1∶109 350 by intramuscular method)were raised in both H 2 b and H 2 d mice after immunization with genetic vaccine of HBcAg. The levels of IgG2a isotype were generally higher than those of IgG1 in all groups of mice with both immunization methods. IgG2a was predominant in C57BL/6 mice after intramuscular immunization. These results suggested the Th1 type of immune response in mice after HBcAg genetic vaccination. HBcAg specific CTL activities were more than 50% in both species of mice with both immunization methods. Conclusion This genetic vaccine of HBcAg shows strong antigenecity in both cellular and humoral immunity. The possibility of its clinical application needs to be further investigated.
出处
《中华医学杂志》
CAS
CSCD
北大核心
1999年第6期456-459,共4页
National Medical Journal of China
基金
卫生部科学研究基金
