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目的:筛选能够有效增强Epstein-Barr病毒转化的B细胞与HBsAg相关抗原肽结合能力的小分子化合物MLE(MHC loading enhancer)。方法:EB病毒转化的B细胞系与HBsAg17-31及全长在加或不加小分子化合物的情况下共孵育4 h后,流式细胞术检测平均荧光强度(MFI)。结果:FITC标记的HBsAg17-31肽段在不加小分子化合物与B细胞共孵育4 h候后(阴性对照)的45±7.87(P=0.008),对氯苯酚(pCP)为60.33±14.34(P<0.01);HBsAg全长与B细MFI为19.97±4.95,加甲醇的MFI为24.67±4.24(P=0.506),加正丁醇的则为胞共孵育4 h后,其阴性对照MFI值为26.67±1.53,加入甲醇的MFL值为31.33±2.08(P=0.078),加入正丁醇的为51.67±3.61(P<0.01),而加入pCP的MFI值为71.67±3.51(P<0.01)。结论:这些小分子化合物能够有效增强Epstein-Barr病毒转化的B细胞与FITC标记的HBsAg17-31肽段及其全长的结合能力,甲醇的催化能力较弱,正丁醇的催化能力明显强于甲醇,对氯苯酚的催化能力最强;为进一步将MLE应用于HBV免疫治疗、乙肝疫苗佐剂打下基础。
OBJECTIVE: To screen a small molecule compound MLE (MHC loading enhancer) capable of effectively binding Epstein-Barr virus-transformed B cells to HBsAg-associated antigen peptides. Methods: The average fluorescence intensity (MFI) of EB virus transformed B cell line was measured by flow cytometry after incubation with HBsAg 17-31 and full length for 4 h with or without small molecule compounds. Results: The FITC-labeled HBsAg17-31 peptide showed no significant difference between the two groups (45 ± 7.87, P = 0.008) and 60.33 ± 14.34 (PCP) P <0.01). MFI of full-length HBsAg and B-MF was 19.97 ± 4.95, with MFI of 24.67 ± 4.24 (P = 0.506) and n-butanol Was 26.67 ± 1.53. The MFL value of methanol was 31.33 ± 2.08 (P = 0.078), n-butanol was 51.67 ± 3.61 (P <0.01), while that of pCP was 71.67 ± 3.51 (P <0.01) . Conclusion: These small molecule compounds can effectively enhance the ability of Epstein-Barr virus transformed B cells to bind to the FITC-labeled HBsAg17-31 peptide and its full-length. The catalytic ability of methanol is weak and the catalytic ability of n-butanol is significantly stronger Methanol and p-chlorophenol have the strongest catalytic ability. To further apply MLE to HBV immunotherapy, hepatitis B vaccine adjuvant lays the foundation.