ANALYSIS OF THE STRUCTURAL CORRELATES FOR ANTIBODY POLYREACTIVITY BY MULTIPLE REASSORTMENTS OF CHIMERIC HUMAN-IMMUNOGLOBULIN HEAVY AND LIGHT-CHAIN V-SEGMENTS

被引:144
作者
ICHIYOSHI, Y
CASALI, P
机构
[1] NYU, SCH MED, DEPT PATHOL, NEW YORK, NY 10016 USA
[2] NYU, SCH MED, KAPLAN COMPREHENS CANC CTR, NEW YORK, NY 10016 USA
关键词
D O I
10.1084/jem.180.3.885
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Polyreactive antibodies (Abs) constitute a major proportion of the early Ab repertoire and are an important component of the natural defense mechanisms against infections. They are primarily immunoglobulin M (IgM) and bind a variety of structurally dissimilar self and exogenous antigens (Ags) with moderate affinity. We analyzed the contribution of Ig polyvalency and of heavy (H) and light (L) chain variable (V) regions to polyreactivity in recombinatorial experiments involving the V-H-diversity(D)-J(H) and V(k)appa-J(k)appa gene segments of a human polyreactive IgM, monoclonal antibody 55 (mAb55), and those of a human monoreactive anti-insulin IgG, mAb13, in an in vitro C gamma l and C(k)appa human expression system. These mAbs are virtually identical in their V-H and V(k)appa gene segment sequences. First, we expressed the V-H-D-J(H) and V(k)appa-J(k)appa genes of the IgM mAb55 as V segments of an IgG molecule. The bivalent recombinant IgG Ab bound multiple Ags with an efficiency only slightly lower than that of the original decavalent IgM mAb55, suggesting that class switch to IgG does not affect the Ig polyreactivity. Second, we coexpressed the mAb55-derived H or kappa chain with the mAb13-derived kappa or H chain, respectively. The hybrid IgG Ab bearing the mAb55-derived H chain V segment paired with the mAb13-derived kappa V segment, but not that bearing the mAb13-derived H chain V segment paired with the mAb55-derived kappa V segment, bound multiple Ags, suggesting that the Ig H chain plays a major role in the Ig polyreactivity. Third, we shuffled the framework 1 (FR1) -FR3 and complementarity determining region 3 (CDR3) regions of the H and kappa chain V segments of the mAB55-derived IgG molecule with the corresponding regions of the monoreactive IgG mAb13. The mAb55-derived IgG molecule lost polyreactivity when the H chain CDR3, but not the FR1-FR3 region, was replaced by the corresponding region of mAb13, suggesting that within the H chain, the CDR3 provides the major structural correlate for multiple Ag-binding. This was formally proved by the multiple Ag-binding of the originally monoreactive mAb13-derived IgG molecule grafted with the mAb55-derived H chain CDR3. The polyreactivity of this chimeric IgG was maximized by grafting of the mAb55-derived kappa chain FR1-FRS, but not that of the kappa chain CDR3. The mAb55-derived kappa chain FR1-FR3 (an unmutated V-kappa 325 segment), however, failed to yield a polyreactive Ab when grafted onto an IgG that was in all other parts identical with mAb13. Rather, this chimeric molecule showed full specificity for insulin. Thus, the polyreactivity of the human mAb55 can be fully preserved after Ig class switch, and depends primarily on the contribution of structures that are generated through somatic rearrangement events (H chain CDR3), and structures that represent the expression of an unmutated gene (V-kappa 325 segment).
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页码:885 / 895
页数:11
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