Horwitz B H, Zelazowski P, Shen Y, Wolcott K M, Scott M L, Baltimore D, Snapper C M
Department of Biology, Massachusetts Institute of Technology, Cambridge 02139, USA.
J Immunol. 1999 Feb 15;162(4):1941-6.
B cells lacking individual NF-kappa B/Rel family members exhibit defects in activation programs. We generated small resting B cells lacking p65 or p50 alone, or lacking both p50 and p65, then evaluated the ability of these cells to proliferate, secrete Ig, and undergo Ig class switching. B cells lacking p65 proliferated well in response to all stimuli tested. However, these cells demonstrated an isolated defect in switching to IgG3, which was associated with a decrease in gamma 3 germline CH gene expression. Whereas, previously reported, B cells lacking p50 alone had a severe proliferative defect in response to LPS, a moderate defect in response to CD40 ligand (CD40L), and normal proliferation to Ag receptor cross-linking using dextran-conjugated anti-IgD Abs (alpha delta-dex), B cells lacking both p50 and p65 exhibited severely impaired proliferation in response to LPS, alpha delta-dex, and CD40L. This defect could be overcome by simultaneous administration of alpha delta-dex and CD40L. In response to this latter combination of stimuli, B cells lacking both p50 and p65 secreted Ig and underwent isotype switching to IgG1 as efficiently as B cells lacking p50 alone. These data demonstrate a role for the p65 subunit of NF-kappa B in germline CH gene expression as well as functional redundancy between p50 and p65 during proliferative responses.
缺乏单个NF-κB/Rel家族成员的B细胞在激活程序方面存在缺陷。我们制备了单独缺乏p65或p50,或同时缺乏p50和p65的小静止B细胞,然后评估这些细胞的增殖、分泌Ig以及进行Ig类别转换的能力。缺乏p65的B细胞对所有测试刺激均能良好增殖。然而,这些细胞在转换为IgG3方面表现出单独的缺陷,这与γ3胚系CH基因表达的降低有关。此前报道,单独缺乏p50的B细胞对LPS有严重的增殖缺陷,对CD40配体(CD40L)有中度缺陷,而使用葡聚糖偶联的抗IgD抗体(αδ-葡聚糖)进行抗原受体交联时增殖正常;同时缺乏p50和p65的B细胞对LPS、αδ-葡聚糖和CD40L的增殖反应严重受损。同时给予αδ-葡聚糖和CD40L可克服这一缺陷。对于后一种刺激组合,同时缺乏p50和p65的B细胞分泌Ig并进行同型转换为IgG1的效率与单独缺乏p50的B细胞相同。这些数据证明了NF-κB的p65亚基在胚系CH基因表达中的作用,以及p50和p65在增殖反应中的功能冗余。