The critical role of arginine residues in the binding of human monoclonal antibodies to cardiolipin

Previously we reported that the variable heavy chain region (V(H)) of a human beta(2 )glycoprotein I-dependent monoclonal antiphospholipid antibody (IS4) was dominant in conferring the ability to bind cardiolipin (CL). In contrast, the identity of the paired variable light chain region (V(L)) determined the strength of CL binding. In the present study, we examine the importance of specific arginine residues in IS4V(H )and paired V(L )in CL binding. The distribution of arginine residues in complementarity determining regions (CDRs) of V(H )and V(L )sequences was altered by site-directed mutagenesis or by CDR exchange. Ten different 2a2 germline gene-derived V(L )sequences were expressed with IS4V(H )and the V(H )of an anti-dsDNA antibody, B3. Six variants of IS4V(H), containing different patterns of arginine residues in CDR3, were paired with B3V(L )and IS4V(L). The ability of the 32 expressed heavy chain/light chain combinations to bind CL was determined by ELISA. Of four arginine residues in IS4V(H )CDR3 substituted to serines, two residues at positions 100 and 100 g had a major influence on the strength of CL binding while the two residues at positions 96 and 97 had no effect. In CDR exchange studies, V(L )containing B3V(L )CDR1 were associated with elevated CL binding, which was reduced significantly by substitution of a CDR1 arginine residue at position 27a with serine. In contrast, arginine residues in V(L )CDR2 or V(L )CDR3 did not enhance CL binding, and in one case may have contributed to inhibition of this binding. Subsets of arginine residues at specific locations in the CDRs of heavy chains and light chains of pathogenic antiphospholipid antibodies are important in determining their ability to bind CL

Anti-phospholipid human monoclonal antibodies inhibit CCR5-tropic HIV-1 and induce β-chemokines

Traditional antibody-mediated neutralization of HIV-1 infection is thought to result from the binding of antibodies to virions, thus preventing virus entry. However, antibodies that broadly neutralize HIV-1 are rare and are not induced by current vaccines. We report that four human anti-phospholipid monoclonal antibodies (mAbs) (PGN632, P1, IS4, and CL1) inhibit HIV-1 CCR5-tropic (R5) primary isolate infection of peripheral blood mononuclear cells (PBMCs) with 80% inhibitory concentrations of <0.02 to ∼10 µg/ml. Anti-phospholipid mAbs inhibited PBMC HIV-1 infection in vitro by mechanisms involving binding to monocytes and triggering the release of MIP-1α and MIP-1β. The release of these β-chemokines explains both the specificity for R5 HIV-1 and the activity of these mAbs in PBMC cultures containing both primary lymphocytes and monocytes

Perspectives New Roles for Rheumatoid Factor

In the 1940s, anti-IgG autoantibodies were discovered in the sera of patients with rheumatoid arthritis and given the term rheumatoid factors (RF). &apos; The measurement of agglutinating IgM-RF is still the most useful serological test for the diagnosis of rheumatoid arthritis. High titers of IgM-RF are also present in primary Sjogren&apos;s syndrome, mixed cryoglobulinemia, and in several different chronic infections (1). Recent advances in molecular and cellular immunology are gradually revealing the genetic and environmental factors that control RF production. Two sets of results were largely unexpected. First, genes encoding RF are present in most normal people, and their expression is carefully regulated during the development of the immune system. Second, lymphocytes with RF receptors on their plasma membranes are remarkably abundant in normal people, who have only low levels of circulatin