中文摘要：

目的建立外周血来源B细胞体外长期培养体系，观察培养过程中B细胞的增殖活化及抗原提呈功能。方法利用重组人可溶性CIM0配体（sCD40L）刺激人外周血B细胞的长期培养过程中，流式细胞技术检测B细胞DNA周期及B细胞表面CD80、CD86、主要组织相容复合物（MHC）Ⅰ类及Ⅱ分子的表达状况；观察B细胞增殖情况及B细胞活化后表面分子的表达。采用乙型肝炎病毒（HBV）核心抗原（HBcAg）-异硫氰酸荧光素（FITC）标记抗原多肽与B细胞共孵育，检测B细胞对抗原多肽的负载能力。结果人外周血B细胞在sCD40L培养体系中可长期培养达50d，在此期间，B细胞绝对计数增加近10倍，外周血B细胞比例由8．21％增高到70．67％，活化的B细胞高表达CD80、CD86、MHC—Ⅰ、MHC—Ⅱ，并可负载HBV核心抗原肽。结论sCD40L培养体系可在体外长期培养人外周血B细胞，并使B细胞增殖活化，具有抗原提呈细胞的特征。

英文摘要：

Objective To develop a method for long-term culture of human B ceils from peripheral blood mononuclear cells （PBMCs） by means of human soluble CD40 ligand （sCIMOL）, and to investigate the proliferation and antigen-presenting function of the CD40-activated B cells. Method Peripheral blood sample of 30 ml was collected from a healthy person. PBMCs were isolated and cultured in the presence of sCD40L. Flow cytometry was used to examine the proliferation, DNA cycle, and cell surface markers： CD86, CD80, major histocompatibility complex （MHC） class Ⅱ, and MHC Class Ⅰ of the B cells. The cells cultured for 45 days were divided into 2 groups ： Group 1 incubated with the fluorescein isothiocyanate （ FITC）-conjugated hepatitis-B core antigen （HBcAg-FITC） and Group 2 used as control group. Eighteen hours later cytometry and fluorescence microscopy were used to detect the peptide pulsing. Results The B ceils could be cultured up to 50 days in the sCD40L cuhure system. The ratio of B cells in the PBMCs was 8.21% at the beginning, and increased to 70.67% by day 47, and the B cell absolute count was 6.56 × 10^5 at the beginning and increased to 8.61 × 10^6 at day 50, about 10 times higher, sCD40L promoted a strong up-regulation of cell surface markers, The expression rates of CD80, CD86, MHC- Ⅱ , and MHC- Ⅰ of the sCD40L-activated B cells were 83.97% , 84.73%, 99.59% , and 98.70% respectively. Flow cytometry showed that 98.10% of the B cells co-incubated with HBcAg-FITC were loaded with HBcAg. Fluorescence microscopy showed that the HBcAg was in the cytoplasm of the B cells. Conclusion A human sCD40L culture system has been developed with the ability to culture human peripheral blood B cells, thus realizing the long-term proliferation and activation of human peripheral blood B cells that function as antigenpresenting cells.