Epitope mapping using the X-ray crystallographic structure of complement receptor type 2 (CR2)/CD21: Identification of a highly inhibitory monoclonal antibody that directly recognizes the CR2-C3d interface

被引:44
作者
Guthridge, JM
Young, K
Gipson, MG
Sarrias, MR
Szakonyi, G
Chen, XJS
Malaspina, A
Donoghue, E
James, JA
Lambris, JD
Moir, SA
Perkins, SJ
Holers, VM
机构
[1] Univ Colorado, Hlth Sci Ctr, Dept Med & Immunol, Denver, CO 80262 USA
[2] Univ Colorado, Hlth Sci Ctr, Dept Biochem & Mol Genet, Denver, CO 80262 USA
[3] Univ Penn, Prot Chem Lab, Philadelphia, PA 19104 USA
[4] NIAID, Immunoregulat Lab, NIH, Bethesda, MD 20892 USA
[5] Oklahoma Med Res Fdn, Oklahoma City, OK 73104 USA
[6] Univ Oklahoma, Hlth Sci Ctr, Oklahoma City, OK 73104 USA
[7] UCL, Royal Free & Univ Coll Med Sch, Dept Biochem & Mol Biol, London, England
关键词
D O I
10.4049/jimmunol.167.10.5758
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Complement receptor type 2 (CR2)/CD21 is a B lymphocyte cell membrane C3d/iC3b receptor that plays a central role in the immune response. Human CR2 is also the receptor for the EBV viral membrane glycoprotein gp350/220. Both C3d and gp350/220 bind CR2 within the first two of 15-16 repetitive domains that have been designated short consensus/complement repeats. Many mAbs react with human CR2; however, only one currently available mAb is known to block both C3d/iC3b find gp350/220 binding. We have used a recombinant form of human CR2 containing the short consensus/complement repeat 1-2 ligand-binding fragment to immunize Cr2(-/-) mice. Following fusion, we identified and further characterized four new anti-CR2 mAbs that recognize this fragment. Three of these inhibited binding of CR2 to C3d and gp350/220 in different forms. We have determined the relative inhibitory ability of the four mAbs to block ligand binding, and we have used overlapping peptide-based approaches to identify linear epitopes recognized by the inhibitory mAbs. Placement of these epitopes on the recently solved crystal structure of the CR2-C3d complex reveals that each inhibitory mAb recognizes a site either within or adjacent to the CR2-C3d contact site. One new mAb, designated 171, blocks CR2 receptor-ligand interactions with the greatest efficiency and recognizes a portion of the C3d contact site on CR2. Thus, we have created an anti-human CR2 mAb that blocks the C3d ligand by direct contact with its interaction site, and we have provided confirmatory evidence that the C3d binding site seen in its crystal structure exists in solution.
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页码:5758 / 5766
页数:9
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