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Genetic and Functional Studies of Human Monoclonal Antibodies to HSP 60 Found in Patients of Atherosclerosis

Authors
Jang, Young-Ju; Jang, Eun-Jung
Department
Department of Microbiology
Abstract
We cloned nine monoclonal IgG Fabs against human HSP60 from PBL of atherosclerosis patients. Analysis of the variable region sequences revealed that the antibodies used diverse members of VH gene families with different DH and JH segments. However, in VL, KV3-20 gene family member along with KJ1 segment was used often. Similarities between the rearranged genes and the closest germline sequences were low. The sequences of VH were highly mutated and VH-CDR3 varied greatly in length and sequences. The ratios of R/S mutation were remarkably high in CDRs in all VH regions except one clone. Furthermore, mutations to positively charged amino acids were frequent in all VH and most VL. These results suggest that the occurrence of somatic hypermutation and antigenic selection is critical, not the usage of certain VH gene family members, in producing affinity-matured anti-HSP60 autoantibodies in atherosclerosis. However, expression of the combined germline genes of KV3-20 with KJ1 might be important for the selection at the early stage of B cell development. Two of these anti-HSP60 Fabs inhibited the uptake of human HSP60 by murine macrophage cells. One of them also reduced the release of pro-inflammatory mediators and inhibited the activation of NF-kB in HSP60-stimulated macrophages. To elucidate the functional roles of anti-HSP60 autoantibodies in atherosclerosis and the potential use of these Fabs to treat atherosclerosis, further investigation is worthy to be performed.
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