Li Zuo wei, Wen Xiao fei, Wang Ying, Luo Ming, Qiu Jian xin
Department of Urology, First People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, PR China.
Exp Clin Transplant. 2013 Dec;11(6):482-8. doi: 10.6002/ect.2012.0281. Epub 2013 Jun 6.
We sought to develop a B-cell in vitro culture system and test B cells isolated from sensitized kidney recipients and healthy controls, and assess the effectiveness of proteasome inhibitors and mycophenolic acid on antibody secretion and cell apoptosis.
CD19(+) B cells and CD19(+)CD27(+) memory B-cell subsets were detected from peripheral blood mononuclear cells obtained from 6 MICA-sensitized kidney recipients and 6 healthy controls. Peripheral blood B cells were isolated and cultured with CpG2006, PMA, MICA antigen, B-cell activating factor, CD40 ligand (CD40L), human recombinant IL-2 (rhuIL-2), rhuIL-10, rhuIL-4, and rhuIL-21. After culturing for 7 days, we tested several variables of B-cell activity including differentiation, apoptosis, and IgM production. We also assessed the effects of 2 immunosuppressive drugs (mycophenolic acid and bortezomib) on antibody secretion and cellular apoptosis.
Kidney recipients had a lower ratio of CD19+ B cells in peripheral blood mononuclear cells than did healthy controls. However, the percentage of CD19(+)CD27(+) B cells was higher in kidney recipients than in healthy controls. In the cell stimulation culture system, the ratio of CD19(+) B cells, CD19(+)CD27(+) B cells, and CD19(+)CD138(+) B cells increased after culturing for 7 days compared with unstimulated controls. In addition, the percentage of apoptotic B cells decreased, and antibody production increased in sensitized transplant patients and healthy controls. Treatment with bortezomib or mycophenolic acid induced B-cell apoptosis and inhibited secretion of antibodies.
This study describes establishment of a B-cell in vitro culture system, showing that B cells may be stimulated to secrete antibodies. The study also provides an assay for studying B cells in vitro. This study provides information suggesting that bortezomib and mycophenolic acid can inhibit B-cell antibody secretion.
我们试图建立一种B细胞体外培养系统，检测从致敏肾移植受者和健康对照者中分离出的B细胞，并评估蛋白酶体抑制剂和霉酚酸对抗体分泌和细胞凋亡的影响。
从6名对MICA致敏的肾移植受者和6名健康对照者获取的外周血单个核细胞中检测CD19(+) B细胞和CD19(+)CD27(+)记忆B细胞亚群。分离外周血B细胞，并用CpG2006、佛波酯、MICA抗原、B细胞活化因子、CD40配体(CD40L)、人重组白细胞介素-2(rhuIL-2)、rhuIL-10、rhuIL-4和rhuIL-21进行培养。培养7天后，我们检测了B细胞活性的几个变量，包括分化、凋亡和IgM产生。我们还评估了2种免疫抑制药物(霉酚酸和硼替佐米)对抗体分泌和细胞凋亡的影响。
肾移植受者外周血单个核细胞中CD19+ B细胞的比例低于健康对照者。然而，肾移植受者中CD19(+)CD27(+) B细胞的百分比高于健康对照者。在细胞刺激培养系统中，与未刺激的对照相比，培养7天后CD19(+) B细胞、CD19(+)CD27(+) B细胞和CD19(+)CD138(+) B细胞的比例增加。此外，致敏移植患者和健康对照者中凋亡B细胞的百分比降低，抗体产生增加。硼替佐米或霉酚酸治疗可诱导B细胞凋亡并抑制抗体分泌。
本研究描述了一种B细胞体外培养系统的建立，表明B细胞可被刺激分泌抗体。该研究还提供了一种体外研究B细胞的检测方法。本研究提供的信息表明，硼替佐米和霉酚酸可抑制B细胞抗体分泌。
