Sequence Analysis of Monoclonal Antibodies Derived from a Patient with Idiopathic Thrombocytopenic Purpura

Abstract

Autoantibodies directed at the platelet membrane glycoprotein lb (GPIb) can mediate a severe form of idiopathic thrombocytopenic purpura (ITP). The platelet-specific antibody from plasma of one patient (DM) with this form of ITP displays a public idiotype termed DMId. The DMId idiotype has been found in the plasma of several patients with ITP, usually in association with GPIb-specific autoantibodies. As a step in the understanding of the molecular genetics of this form of ITP we have determined the nucleotide sequences of expressed V region genes selected from a panel of five human lymphoblastoid cell lines derived from patient DM. Two of the lines secreted antibodies that bound GPIb, and three of the lines secreted antibodies that expressed DMId. The H chain sequences of the DMId-positive antibodies and of one of the GPIb-binding antibodies belong to the V_H4 family. The second GPIb-binding antibody belongs to the V_H 1 family. All have multiple substitutions from previously published sequences giving these antibodies the appearance of having been antigen driven. These results are consistent with the hypothesis that autoantibodies in ITP arise from a “normal” immune response inappropriately directed at platelet antigens. Further, our results suggest that V_H4 gene segments may be recruited preferentially into the DMId-positive, GPIb-specific autoantibody response.

Key Words:

• Anti-platelet autoantibodies
• immunoglobulin heavy chain variable region genes
• idiopathic thrombocytopenic purpura
• platelet glycoprotein lb