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为了探讨前 S_2蛋白对慢性乙型肝炎患者(乙肝)外周血 T淋巴细胞亚群变化的关系,揭示前 S_2是否具有改变乙肝患者免疫调控能力。我们在上海有机化学研究所内,应用 Merrifield 固相化学合成法,合成了前 S_2中抗原性强的 P_(120～146)多肽段,氨基酸序列为 MQWWSTTFHQALL DPRVGL YFPAGGS,经反相高压液相 C_(18)柱纯化。测定氨基酸序列的正确性后,用0.1 mol/L PBS 溶解除菌后作为抗原。一、资料与方法选择10例乙肝患者(经 ELISA 测定 HBsAg,HBeAg,抗-HBc 和 PCR HBV DNA 全部阳性)作为研究对象,对照组为上述方法测定全部为阴性的正常人群4例。常规分离外周血中单个核细胞,在96孔培养板上每孔加入细胞悬液100μl(1×10~6细胞)。用含量为1μg、5μg、10μg(每孔含量)的前 S_2合成肽,PHA(1μg/孔)和阴性对照共5组每组均设3复孔,作用7天后制片。采用 APAAP 法观察 CD_3~+;CD_4~+;CD_8~+的百分数和 CD_4~+/
In order to investigate the relationship between the changes of T lymphocyte subsets in the peripheral blood of patients with chronic hepatitis B (hepatitis B) before S_2 protein and reveal whether the pre-S_2 has the ability to change the immune regulation of hepatitis B patients. We used the Merrifield solid-phase chemistry synthesis method to synthesize the P_ (120-146) polypeptide with strong antigenicity in the former S_2. The amino acid sequence was MQWWSTTFHQALL DPRVGL YFPAGGS. The reverse phase high pressure liquid phase C 18 ) Column. After determining the correctness of the amino acid sequence, 0.1 mol / L PBS was dissolved and sterilized as antigen. Data and Methods 10 cases of hepatitis B patients (HBsAg, HBeAg, anti-HBc and PCR HBV DNA were all tested positive by ELISA) were selected as the research object, and the control group were all 4 cases of normal population which were negative by the above method. Peripheral blood mononuclear cells were routinely isolated and 100 μl of cell suspension (1 × 10 -6 cells) was added to each well of a 96-well culture plate. Pre-S 2 synthetic peptides (1μg / well, 5μg / well), PHA (1μg / well) and negative control (5μg / well) were used to prepare 3 replicates. APAAP method was used to observe the percentage of CD_3 ~ +; CD_4 ~ +; CD_8 ~ + and CD_4 ~ + /