Oviductal antibody response to a defined recombinant sperm antigen in macques.
Macaque oviductal fluids were assayed for specific antibodies to the intra-acrosomal sperm protein SP-10 after immunizations with recombinant macaque SP-10 (re-mqSP-10), a candidate contraceptive vaccinogen. Access ports, consisting of a subcutaneous collecting reservoir and a catheter to cannulate the oviduct, were implanted into monkeys for repeated aspiration of oviductal fluid. Monkeys were inoculated i.m. once a month with an emulsion consisting of 2 mg re-mqSP-10 in a vehicle of squalene and mannin monooleate. Oviductal fluids and serum were collected during the periovulatory period for six menstrual cycles, and IgG and IgA antigen-specific antibodies in preimmune and immune fluids were compared by ELISA. Both relative and absolute concentrations of SP-10-specific immunoglobulins (Ig) were determined. Oviductal fluids from immunized animals showed significant increases in anti-SP-10 IgG at cycle 2 and at all subsequent intervals. Anti-SP-10 IgA significantly increased in oviductal fluid at cycles 4, 5, and 6. Serum anti-SP-10 IgG increased at cycle 2 and remained significantly elevated through cycle 6, while serum anti-SP-10 IgA was higher than in preimmune samples at cycle 4. Serum antibodies generated to the recombinant SP-10 recognized SP-10 extracted from macaque sperm on Western blots. Immunocytochemical staining of macaque and human sperm showed acrosomal immunofluorescence with both immune oviductal fluids and serum using both anti-IgG and anti-IgA secondary antibodies. This study demonstrates for the first time 1) IgG and IgA antibodies to a defined recombinant sperm-specific antigen in primate oviductal fluids after systemic immunization and 2) the recognition by primate oviductal fluid IgG and IgA of the endogenous contraceptive target on both human and macaque sperm.
Kurth
B E
BE
Center for Recombinant Gamete Contraceptive Vaccinogens, Department of Cell Biology, The University of Virginia, Charlottesville 22908, USA.
Weston
C
C
Reddi
P P
PP
Bryant
D
D
Bhattacharya
R
R
Flickinger
C J
CJ
Herr
J C
JC
D000177:Acrosome; D000818:Animals; D000906:Antibodies / Q000032:analysis; D000941:Antigens; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D005187:Fallopian Tubes / Q000276:immunology*; D005260:Female; D019085:Fluorescent Antibody Technique, Direct; D012739:Gonadal Steroid Hormones; D006801:Humans; D007114:Immunization; D007070:Immunoglobulin A / Q000276:immunology; D007074:Immunoglobulin G / Q000276:immunology; D066298:In Vitro Techniques; D008252:Macaca fascicularis; D008297:Male; D008565:Membrane Proteins; D011506:Proteins / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology; D013094:Spermatozoa / Q000276:immunology*
C512729:ACRV1 protein, human; D000906:Antibodies; D000941:Antigens; D012739:Gonadal Steroid Hormones; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D008565:Membrane Proteins; D011506:Proteins; D011994:Recombinant Proteins