Complex nature of the human antisperm antibody response in SCID mice

Abstract

Human peripheral blood mononuclear (PBMs) cells were introduced into the peritoneal cavity of severely-combined immunodeficient (SCID) mice in concentrations of 2.5-4.0 x 10(7) cells per mouse. Whole mononuclear cell suspensions were used either unstimulated or following primary in vitro culture with human spermatozoa. In some experiments, immunodepletion of CD8(+) cells was carried out prior to grafting. Lymphocytes were obtained from nonsensitized (to antigen) human subjects or from individuals who were primed in vivo (vasectomized individuals in case of sperm antigens). An enzyme-linked immunosorbent assay was employed to assess total human immunoglobulin (G or M) levels as well as the specificity of the antibodies generated. We have been successful by generating primary and secondary immune responses with 'naïve' human lymphocytes, challenged with chlamydia or ovalbumin but without adjuvant or CD8(+) immunodepletion; however, we were unable to induce specific antibodies to spermatozoa under this regime in SCID male mice. We then employed female SCID mice, treated with sperm antigen extracts (glycosylated or deglycosylated) encapsulated in liposomes and human lymphocytes obtained from 'naïve' or pre-sensitized in vivo subjects. It was found that the most pronounced humoral response to sperm antigens was obtained with deglycosylated antigens and PBMs from vasectomized (in vivo pre-primed to spermatozoa) individuals. A presented SCID mice model can be helpful at understanding of antisperm antibody development and the molecular nature of generated antibodies to modified sperm antigenic entities.

Specific Fab fragments recovered by phage display technique recognizing human spermatozoa

“…Human hybridoma cell lines were obtained after fusion of PBL’s sensitized in SCID mice (Kurpisz et al. , 2004) with KR12 cell line.…”

ORIGINAL ARTICLE: In situ Reconstruction of Humoral Immune Response Against Sperm: Comparison of SCID and NOD/SCID Mouse Models

“…[9][10][11] Different approaches have been previously tried to improve the efficacy of human B lymphocytes to produce specific pre-determined antibodies. Immunocompromised mice have been applied many times to study human humoral immune responses following repopulation with human lymphoid cells.…”

“…Immunocompromised mice have been applied many times to study human humoral immune responses following repopulation with human lymphoid cells. 11 A basic goal was to obtain specific AsA which could bind only to spermatozoa; therefore, selection of the most appropriate antigenic preparation was critically important. For example, a procedure of immunodepletion of CD8 + immunosuppressive cell populations was implemented, 9,12 while following mice vaccinations, different adjuvants were applied for antibody induction regime (Freund's complete and incomplete variants).…”

“…For example, a procedure of immunodepletion of CD8 + immunosuppressive cell populations was implemented, 9,12 while following mice vaccinations, different adjuvants were applied for antibody induction regime (Freund's complete and incomplete variants). 11 Quantitative enzyme-linked immunosorbent assay (ELISA) was usually employed to assess total human immunoglobulin levels in mouse circulation as well as their specificity. 13 Enzymatic deglycosylation of the sperm antigenic extract was attempted to expose hidden behind glycocalyx protein determinants.…”

“…11 A basic goal was to obtain specific AsA which could bind only to spermatozoa; therefore, selection of the most appropriate antigenic preparation was critically important. 11 A primary aim of this study was to compare intensity and human AsA reactivities obtained in sera of SCID and NOD ⁄ SCID mice. 11 Quantitative enzyme-linked immunosorbent assay (ELISA) was usually employed to assess total human immunoglobulin levels in mouse circulation as well as their specificity.…”