Monoclonal antibodies as direct probes for human sperm acrosome reaction.
OBJECTIVE
To develop simple and rapid assay procedures for determining human sperm acrosome reaction under various experimental conditions.


METHODS
Specific monoclonal antibodies against human sperm acrosome were generated and utilized as probes for acrosome reaction assays by direct labelling of antibodies with fluorescein-isothiocyanate (FITC).


RESULTS
Among the generated monoclonal antibodies, HS-63 was shown to react with antigens in the acrosome content of permeabilized acrosome-intact human sperm, but not with those of the live ones. HSA-10 was found to recognize antigens on the surface of sperm inner acrosome and to react with acrosome-reacted human sperm in suspension. Following a swim-up procedure, highly motile sperm were recovered and incubated in BWW medium at 37 degrees C for 18 h. The percentages of acrosome-reacted sperm were determined at various incubation times by using FITC-labeled HS-63 or HSA-10 as the indicators in 10 min direct immunofluorescent assay. With the FITC-labeled HS-63 probe, the percentage of positively stained human sperm fixed in methanol decreased significantly after 18 h of incubation from > 90 to 70-80%. In contrast, positively stained live human sperm by HSA-10 increased significantly from < 1% to 15-30%. A high correlation was obtained between the use of monoclonal antibodies and PSA (Pissum sativum agglutinin) for direct assessment of human sperm acrosome reaction.


CONCLUSIONS
In view of the simplicity in assay procedures and evaluations, these FITC-labeled monoclonal antibodies are reliable probes for direct assessment of acrosomal status of human sperm.
Dorjee
S
S
Department of Obstetrics and Gynecology, University of British Columbia, Vancouver, Canada.
Lai
C L
CL
Lee
A
A
Lee
C Y
CY
D000177:Acrosome / Q000737:chemistry / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D019085:Fluorescent Antibody Technique, Direct; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D007118:Immunoassay; D008297:Male; D013076:Sperm Count; D013194:Staining and Labeling
D000911:Antibodies, Monoclonal