摘要
目的观察转染FcγRⅡb1基因能否纠正SLE患者B细胞的过度活化。方法构建人FcγRⅡb1基因真核载体,通过电穿孔转染SLE患者B细胞。分别采用抗人μ链的F(ab’)2片段[F(ab’)2anti-μ]和抗人μ链的完整IgG(IgG anti-μ)2种抗体刺激B细胞,同时以正常人B细胞作对照。采用分光光度计检测激活B细胞胞内[Ca2+]i反应,3H-TdR掺入法检测B细胞的增殖能力,ELISA法检测B细胞分泌抗体的功能。结果分别以F(ab’)2anti-μ和IgG anti-μ激活SLE患者B细胞后,细胞内[Ca2+]i反应、cmp值及IgG分泌量的比值均显著低于正常人(P<0.01),通过转染FcγRⅡb1基因后,这些比值均显著提高(P<0.01或P<0.05)。结论转染FcγRⅡb1基因能有效纠正SLE患者B细胞过度的活化、增殖及抗体分泌。
Inhibitory signaling abnormality of FcγRⅡb1 possibly contributes to the mechanism of hyperactivity of B lymphocytes in patients with systemic lupus erythematosus (SLE). This study was designed to investigate whether FcγRⅡb1 gene transfection could correct the hyperactivity of B lymphocytes of human SLE. Firstly, normal human FcγRⅡb1 gene eukaryotic expression vectors were constructed and transfected into human B lymphocytes from SLE by electroporation method. Then, the B lymphocytes from SLE and normal controls were stimulated with F(ab')2 anti-μ and IgG anti-μrespectively. The fluxes of intracytoplasmic calcium ([Ca2+]i) of B lymphocytes activated by different activators were measured by fluorescence spectrophotometric method; the proliferative capacity of B lymphocytes was detected with 3H-TdR incorporation method; and the IgG production by B lymphocytes cultured with activators was assayed by ELISA. We found that the ratios of [Ca2+]i response, cmp value, and IgG production of F(ab')2 anti-ix- or IgG anti-ix-stimulated SLE B lymphocytes were significant lower, as compared with normal group (P〈 0.01), but these indexes would significantly increase (P〈 0.01 or P〈 0.05 ) after FcγRⅡb1 gene transfection. These data suggest that FcγRⅡb1 bl gene transfection is able to effectively correct the excessive activation, proliferative capacity, and antibody production of B lymphocytes from patients with SLE.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2014年第1期45-48,52,共5页
Immunological Journal
基金
四川省科技厅基金项目(2008JY0080-1)