Kulik Liudmila, Hewitt Finnegan B, Willis Van C, Rodriguez Rosa, Tomlinson Stephen, Holers V Michael
Division of Rheumatology, Department of Medicine, University of Colorado School of Medicine, Aurora, CO 80045, USA.
Mol Immunol. 2015 Feb;63(2):479-88. doi: 10.1016/j.molimm.2014.10.005.
Although reagents are available to block mouse complement receptor type 2 and/or type 1 (CR2/CR1, CD21/CD35) function in acute or short term models of human disease, a mouse anti-rat antibody response limits their use in chronic models. We have addressed this problem by generating in Cr2−/− mice a mouse monoclonal antibody (mAb 4B2) to mouse CR2/CR1. The binding of murine mAb 4B2 to CR2/CR1 directly blocked C3dg (C3d) ligand binding. In vivo injection of mAb 4B2 induced substantial down regulation of CR2 and CR1 from the B cell surface, an effect that lasted six weeks after a single injection of 2 mg of mAb. The 4B2 mAb was studied in vivo for the capability to affect immunological responses to model antigens. Pre-injection of mAb 4B2 before immunization of C57BL/6 mice reduced the IgG1 antibody response to the T-dependent antigen sheep red blood cells (SRBC) to a level comparable to that found in Cr2−/− mice. We also used the collagen-induced arthritis (CIA) model, a CR2/CR1-dependent autoimmune disease model, and found that mice pre-injected with mAb 4B2 demonstrated substantially reduced levels of pathogenic IgG2a antibodies to both the bovine type II collagen (CII) used to induce arthritis and to endogenous mouse CII. Consistent with this result, mice pre-injected with mAb 4B2 demonstrated only very mild arthritis. This reduction in disease, together with published data in CII-immunized Cr2−/− mice, confirm both that the arthritis development depends on CR2/CR1 receptors and that mAb 4B2 can be used to induce biologically relevant receptor blockade. Thus mAb 4B2 is an excellent candidate for use in chronic murine models to determine how receptor blockage at different points modifies disease activity and autoantibody responses.
尽管有试剂可在人类疾病的急性或短期模型中阻断小鼠补体受体2型和/或1型(CR2/CR1,CD21/CD35)的功能,但小鼠抗大鼠抗体反应限制了它们在慢性模型中的应用。我们通过在Cr2−/−小鼠中产生针对小鼠CR2/CR1的小鼠单克隆抗体(mAb 4B2)解决了这个问题。鼠源mAb 4B2与CR2/CR1的结合直接阻断了C3dg(C3d)配体的结合。体内注射mAb 4B2可诱导B细胞表面CR2和CR1的大量下调,单次注射2 mg mAb后,这种作用可持续六周。对4B2 mAb在体内影响对模型抗原免疫反应的能力进行了研究。在C57BL/6小鼠免疫前预先注射mAb 4B2,可将对T细胞依赖性抗原绵羊红细胞(SRBC)的IgG1抗体反应降低到与Cr2−/−小鼠相当的水平。我们还使用了胶原诱导性关节炎(CIA)模型,这是一种CR2/CR1依赖性自身免疫性疾病模型,发现预先注射mAb 4B2的小鼠对用于诱导关节炎的牛II型胶原(CII)和内源性小鼠CII的致病性IgG2a抗体水平大幅降低。与此结果一致,预先注射mAb 4B2的小鼠仅表现出非常轻微的关节炎。疾病的这种减轻,连同在CII免疫的Cr2−/−小鼠中已发表的数据,证实了关节炎的发展依赖于CR2/CR1受体,并且mAb 4B2可用于诱导生物学相关的受体阻断。因此,mAb 4B2是用于慢性小鼠模型以确定不同时间点的受体阻断如何改变疾病活动和自身抗体反应的极佳候选物。