Recombinant Soluble Human Complement Receptor Type 1 Inhibits Antisperm Antibody- and Neutrophil-Mediated Injury to Human Sperm1

Abstract

The pathogenesis of antisperm antibody (ASA)-mediated infertility is postulated to be related in part to complement (C)-dependent sperm dysfunction in the female genital tract. We have previously demonstrated that C can be involved in ASA-mediated sperm injury by the deposition of activated C3 fragments and the assembly of terminal membrane attack complex (C5b-9) leading to C3-mediated sperm binding to neutrophils or C5b-9-mediated sperm motility loss. This study evaluated the protective effect of recombinant soluble C receptor type 1 (sCR1) on ASA-and C-mediated neutrophil/sperm interaction, neutrophil aggregation, and sperm motility loss. Motile sperm with or without neutrophils were incubated in the presence of 10% C-fixing ASA+ serum or ASA- control sera in the presence or absence of sCR1. After defined incubation periods, the following neutrophil and sperm parameters were evaluated: 1) neutrophil aggregation (by the flow cytometric pulse processing method), 2) sperm phagocytosis (by light microscopy), 3) the deposition of C3 cleavage fragments (C3b, iC3b, and C3d) on motile sperm (by immunofluorescence flow cytometry), and 4) the relation between sperm motility loss and sperm-bound C3d. Only the coincubation of neutrophils with sperm in the presence of C-fixing ASA+ sera resulted in marked neutrophil aggregation (20.5 +/- 0.26% vs. 2.4% +/- 1.6; p < 0.0001) and a concomitant increase in neutrophils containing ingested sperm (71 +/- 5.8% vs. 3.5%; p < 0.0001). Soluble CR1 inhibited ASA- and C-mediated neutrophil aggregation by 46% and sperm phagocytosis by 57%. Motile sperm incubated with C-fixing ASA- sera showed a time-dependent increase in the binding of C3 fragments as detected by flow cytometry using anti-iC3b neoantigen, anti-C3c, and anti-C3d monoclonal antibodies (mAbs). A negative correlation (r2 = -0.930; p < 0.001) was found between the increase in sperm-associated C3d fluorescence and the percentage motile sperm in the presence of ASA- sera. Soluble CR1 (200 micrograms/ml) maximally inhibited the binding of anti-C3b, anti-C3c, and anti-C3d mAbs to sperm by 96%, 83%, and 72%, respectively. Thus, sCR1 abrogated the binding of C3 fragments to human sperm and fully protected sperm from C5b-9-mediated sperm immobilization. These findings suggested the therapeutic potential of sCR1 as an intravaginal pharmacophore to prevent C-dependent sperm dysfunction and related inflammatory events in the female genital tract.

Disruption of Mouse CD46 Causes an Accelerated Spontaneous Acrosome Reaction in Sperm

“…The female reproductive tract is abundant in complement, partly because it requires a periodic removal mechanism for millions of allogeneic spermatozoa. One reported detrimental effect of complement on sperm in the female reproductive tract is that females who produce antisperm antibody become infertile when the antibody inactivates sperm with complement (8). It is also reported that the estrogen stimulates the synthesis of C3 in rat uterus (38).…”