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目的研究外周血单个核细胞(PBMC)中乙型肝炎病毒(HBV)包膜抗原基因变异的意义。方法收集19例慢性乙肝患者的PBMC和配对血清,用PCR扩增PBMC及血清中HBV包膜基因片段(S,preS1,preS2),并对其中各5例PBMC及配对血清中的preS1和preS2基因片段,分别进行核苷酸序列分析。结果S、preS1和preS2片段在血清和PBMC中的检出率,分别为100%、947%、100%和0、263%、263%。与HBVADR序列比较,10份标本HBVDNA核苷酸序列分析,发现2份preS1和3份preS2的核苷酸序列,没有发生改变;8份preS1和7份preS2的核苷酸序列,均发生了一个至多个位点的点突变。结论PBMC中存在的包膜基因不完整,因而不会形成完整的HBV病毒颗粒,故不支持HBV可潜伏于PBMC并发展成为体内HBV再感染的来源。preS1点突变编码的氨基酸位点,集中在aa21~47区域,可能会影响HBV的吸附和穿入,及组织亲嗜性的改变。HBV亚基因片段在PBMC中,是否会影响细胞的功能,有待进一步研究。
Objective To investigate the significance of hepatitis B virus (HBV) envelope antigen gene mutation in peripheral blood mononuclear cells (PBMCs). Methods PBMCs and matched sera from 19 chronic hepatitis B patients were collected. The HBV envelope gene fragments (S, preS1, preS2) in PBMC and serum were amplified by PCR, and the preS1 and preS2 genes in 5 PBMCs and matched serum Fragment, respectively, for nucleotide sequence analysis. Results The detection rates of S, preS1 and preS2 in serum and PBMC were 100%, 947%, 100% and 0,263%, 263% respectively. Compared with the HBVADR sequence, the nucleotide sequence of the HBVDNA in 10 samples showed no change in the nucleotide sequence of 2 copies of preS1 and 3 copies of preS2. One nucleotide sequence of 8 copies of preS1 and 7 copies of preS2 Point mutations to multiple sites. Conclusion The envelope genes in PBMC are incomplete and therefore do not form a complete HBV virion. Therefore, they do not support that HBV can be latent in PBMC and develop into a source of HBV re-infection in vivo. PreS1 point mutation encoded amino acid sites, concentrated in aa21 ~ 47 area, may affect the adsorption and penetration of HBV, and tissue changes in the tropism. HBV subunit fragment in PBMC, will affect the function of cells, pending further study.