Institute for Immunology and Department of Medicine, School of Medicine, University of California, Irvine, California 92697-4120, USA.
J Biol Chem. 2012 Jun 15;287(25):21520-9. doi: 10.1074/jbc.M112.366732. Epub 2012 May 3.
Immunoglobulin (Ig) class switch DNA recombination (CSR) and somatic hypermutation (SHM) are critical for the maturation of the antibody response. Activation-induced cytidine deaminase (AID) initiates CSR and SHM by deaminating deoxycytidines (dCs) in switch (S) and V(D)J region DNA, respectively, to generate deoxyuracils (dUs). Processing of dUs by uracil DNA glycosylase (UNG) yields abasic sites, which are excised by apurinic/apyrimidinic endonucleases, eventually generating double strand DNA breaks, the obligatory intermediates of CSR. Here, we found that the bivalent iron ion (Fe(2+), ferrous) suppressed CSR, leading to decreased number of switched B cells, decreased postrecombination Iμ-C(H) transcripts, and reduced titers of secreted class-switched IgG1, IgG3, and IgA antibodies, without alterations in critical CSR factors, such as AID, 14-3-3γ, or PTIP, or in general germline I(H)-S-C(H) transcription. Fe(2+) did not affect B cell proliferation or plasmacytoid differentiation. Rather, it inhibited AID-mediated dC deamination in a dose-dependent fashion. The inhibition of intrinsic AID enzymatic activity by Fe(2+) was specific, as shown by lack of inhibition of AID-mediated dC deamination by other bivalent metal ions, such as Zn(2+), Mn(2+), Mg(2+), or Ni(2+), and the inability of Fe(2+) to inhibit UNG-mediated dU excision. Overall, our findings have outlined a novel role of iron in modulating a B cell differentiation process that is critical to the generation of effective antibody responses to microbial pathogens and tumoral cells. They also suggest a possible role of iron in dampening AID-dependent autoimmunity and neoplastic transformation.
免疫球蛋白 (Ig) 类别转换 DNA 重组 (CSR) 和体细胞高频突变 (SHM) 对于抗体反应的成熟至关重要。激活诱导的胞嘧啶脱氨酶 (AID) 通过分别在转换 (S) 和 V(D)J 区 DNA 中的脱氧胞嘧啶 (dC) 脱氨来启动 CSR 和 SHM,从而产生脱氧尿嘧啶 (dU)。尿嘧啶 DNA 糖基化酶 (UNG) 对 dU 的处理产生无碱基位点,这些无碱基位点被嘌呤内切核酸酶切除,最终产生双链 DNA 断裂,这是 CSR 的必需中间体。在这里,我们发现二价铁离子 (Fe(2+),亚铁) 抑制 CSR,导致转换 B 细胞数量减少、重组后 Iμ-C(H) 转录物减少以及分泌的类别转换 IgG1、IgG3 和 IgA 抗体滴度降低,而关键 CSR 因子(如 AID、14-3-3γ 或 PTIP)或一般种系 I(H)-S-C(H) 转录没有改变。Fe(2+) 不影响 B 细胞增殖或浆细胞样分化。相反,它以剂量依赖的方式抑制 AID 介导的 dC 脱氨。Fe(2+) 对固有 AID 酶活性的抑制是特异性的,因为其他二价金属离子(如 Zn(2+)、Mn(2+)、Mg(2+) 或 Ni(2+))不能抑制 AID 介导的 dC 脱氨,并且 Fe(2+) 不能抑制 UNG 介导的 dU 切除。总体而言,我们的研究结果概述了铁在调节对微生物病原体和肿瘤细胞产生有效抗体反应至关重要的 B 细胞分化过程中的新作用。它们还表明铁在抑制 AID 依赖性自身免疫和肿瘤转化方面可能发挥作用。