Human rheumatoid factors with restrictive specificity for rabbit immunoglobulin G: auto- and multi-reactivity, diverse VH gene segment usage and preferential usage of V lambda IIIb.
Human rheumatoid factors with restrictive specificity for rabbit immunoglobulin G: auto- and multi-reactivity, diverse VH gene segment usage and preferential usage of V lambda IIIb.
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DOI：
10.1084/jem.179.5.1445
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发表时间：
1994-05-01
期刊：
影响因子：
--
通讯作者：
Fu SM
中科院分区：
文献类型：
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作者：
Fang Q;Kannapell CC;Gaskin F;Solomon A;Koopman WJ;Fu SM
为了确定人类风湿因子（RF）的分子和功能特性，我们建立了稳定的杂交瘤和Epstein-巴尔病毒转化的B细胞系，从三名类风湿关节炎患者和一名系统性红斑狼疮患者的滑液或外周血。获得了17个产生高滴度免疫球蛋白M（IgM）RF的细胞系，其仅与兔反应，而不与人IgG或其他哺乳动物物种的IgG反应。某些抗兔IgG RF也对其他哺乳动物抗原（Ag）具有特异性，包括HeLa细胞中发现的细胞骨架蛋白和细胞内蛋白，以及A组链球菌细胞壁提取物中存在的Ag。17个RF中的13个含有λ型轻（L）链，其中12个在血清学上被分类为λ-L链可变区（V λ）亚组的成员，命名为V λ III。在cDNA水平上确定了其中九个IgM λ RF的重链V区（VH）和V λ序列。这些抗体（Ab）使用三个VH家族中的五个VH基因，包括VH 1（dp 21/1- 4 b和dp 10 [51 p1]/hv 1051）、VH 3（dp 38/3-15和dp 77/13-21）和VH 4（dp 70/4- 4 b）。这些IgM λ RF的λ链的推导的V基因编码的氨基酸序列证实了它们的血清学分类为λ III，并且它们被进一步分类为相对不常见的V λ III亚组的成员，命名为V λ IIIb。基于cDNA分析，9个是三种不同的V λ III B种系基因的产物。从基因组DNA中克隆了两个这样的基因，命名为hsiggll 150和hsiggll 295。这些VH和V λ III B基因的独特组合可能与IgM λ RF中独特的反应模式有关。虽然这些抗体的VH和V λ区主要表达为种系编码序列，但9种多反应性抗体中有4种具有广泛的V区突变，表明Ag驱动的过程。发现λ IIIb L链优先存在于抗兔IgG RF中，并且这些Ab中的一些对其他蛋白质、细胞和细菌Ag具有特异性，这为RA和相关疾病的发病机制提供了新的见解。
To determine the molecular and functional properties of human rheumatoid factors (RF), we established stable hybridomas and Epstein- Barr virus-transformed B cell lines from the synovial fluid or peripheral blood of three patients with rheumatoid arthritis and one patient with systemic lupus erythematosus. 17 cell lines were obtained that produced high-titer immunoglobulin M (IgM) RF that reacted exclusively with rabbit but not human IgG or IgG of other mammalian species. Certain anti-rabbit IgG RF also had specificity for other mammalian antigens (Ag), including cytoskeletal proteins and intracellular proteins found in HeLa cells, as well as for Ag present in an extract prepared from the cell wall of group A streptococci. 13 of the 17 RF contained lambda-type light (L) chains, of which 12 were classified serologically as members of the lambda-L chain variable region (V lambda) subgroup, designated V lambda III. The heavy chain V region (VH) and V lambda sequences of nine of these IgM lambda RF were determined at the cDNA level. Five VH genes in three VH families were used by these antibodies (Ab), including VH1 (dp21/1-4b and dp10 [51p1]/hv1051), VH3 (dp38/3-15 and dp77/13-21), and VH4 (dp70/4-4b). The deduced V gene-encoded amino acid sequences of the lambda chains of these IgM lambda RF confirmed their serological classification as lambda III, and they were further classified as members of the relatively uncommon V lambda III subgroup, designated V lambda IIIb. Based on cDNA analyses, nine were the product of three different V lambda III b germline genes. Two such genes, designated hsiggll150 and hsiggll295, were cloned and sequenced from genomic DNA. Unique combinations of these VH and V lambda III b genes could be related to distinctive patterns of reactivity among the IgM lambda RF. Although the VH and V lambda regions of these Abs were expressed primarily as germline-encoded sequences, four of nine multireactive Abs had extensive V region mutation, indicative of an Ag-driven process. The finding that lambda IIIb L chains are preferentially found among anti- rabbit IgG RF, and that some of these Ab have specificity for other protein, cellular, and bacterial Ag, provides new insight into the pathogenesis of RA and related diseases.