体外诱导人类B细胞分泌HLA抗体的实验研究  

Experimental study on the induction of HLA antibodies by human B lymphocyte cells in vitro

作　　者：廖爱华[1] Vera Rebmann Hans Grosse-Wilde 王燕[1] 张玲[1] 

机构地区：[1]华中科技大学同济医学院生殖医学中心,湖北武汉430030 [2]埃森大学医学院免疫研究所,德国埃森45147

出　　处：《细胞与分子免疫学杂志》2007年第7期641-644,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(30571710)

摘　　要：目的:探索体外诱导人类B细胞产生特异性HLA抗体的理想培养条件。方法:以EBV转化致敏肾移植患者B淋巴细胞系(BLCL)为模型,采用ELISA和ELISPOT方法,比较3种培养体系:(1)RPMI1640+抗OPTI单克隆抗体(mAb);(2)RPMI1640+0·1mmol/L非必需氨基酸液+1mmol/L丙酮酸钠+40mg/L转铁蛋白;(3)Hybridoma无血清培养液,以及3种培养条件:(1)对照组(仅培养液),(2)培养液+rIL-4+rIL-10+鼠抗人CD40(α-CD40),(3)培养液+rIL-4+rIL-10+重组人CD40配体(CD40L),人类B细胞产生免疫球蛋白(Ig)和特异性抗HLA抗体的能力。结果:BLCL细胞在Hybridoma SFM培养体系所产生的IgG和IgM浓度分别为25·2～28·1mg/L和7·14～7·95mg/L,高于其他两种培养条件(P<0·05)。BLCL细胞在添加了IL-4、IL-10和α-CD40或CD40L的杂交瘤无血清培养液中能产生特异性抗HLA-I抗体,所获得的斑点频数分别为3·06/每104个BLCL细胞和3·55/每104个BLCL细胞,高于对照组(1·77/每104个BLCL细胞)。结论:BLCL细胞在杂交瘤无血清培养液中和添加了IL-4、IL-10和α-CD40或CD40L的培养条件下,提高分泌IgG和IgM的能力和特异性抗HLA-I类分子的抗体。AIM： To explore the optimal in vitro culture condition for producing antibodies to HLA class I molecules by human B lymphocyte cells. METHODS： By applying EBVtransformed B lymphoblast cell line （BLCL） from a sensitized kidney transplanted subject as a model, the capacity of Ig and specific HLA class I antibody production of BLCL cells was investigated by ELISA and ELISpot, under three different culture systems, RPM11640 ＋ OPTI mAb, RPM11640 ＋ 0.1 mmol/L non-essential amino acid ＋ 1 mmol/L sodium pyruvate ＋ 40 mg/L apo-transferrin,and serum free medium （SFM） for hybridoma culture, and three different culture conditions, culture medium only, culture medium supplemented with IL-4, IL-10 and α-CD40, and culture medium supplemented with IL-4, IL-10 and CD40L. RESULTS： When BLCL cells were cultured in Hybridoma SFM, the IgG and IgM concentrations in culture supernatants were 2.5.2 - 28.1 mg/L and 7. 14 - 7. 9.5 mg/L, respectively, which were higher than that of the other two culture systems （ P 〈 0.0.5 ）. Moreover, when Hybridoma SFM was supplemented with IL-4, IL-10 and α-CD40 or CD40L, the frequentcies of HLA-I antibody-secreting BLCL cells were 3.06/104 BLCL cells and 3.5.5/104 BLCL cells,which were much higher than that of SFM only group （1. 77/10^4 BLCL cells）. CONCLUSION： BLCL cells can produce both higher amount of Ig and specific HLA-I antibodies under Hybridoma SFM supplimented with IL-4, IL-10 and α-CD40 or CD40L.

关 键 词：B淋巴细胞 HLA抗体 酶联免疫斑点法 

分 类 号：R593[医药卫生—内科学]