Indirect mixed antiglobulin reaction (MAR) as a screening procedure for antisperm antibodies. I. Methodological...

Area of Science:

• Immunology
• Reproductive Medicine
• Clinical Chemistry

Background:

• Direct mixed antiglobulin reaction (MAR) testing requires motile sperm, limiting its application.
• Antisperm antibodies, particularly IgG and IgA, can impact male fertility.
• A need exists for a practical screening method to characterize the immunoglobulin class of antisperm antibodies.

Purpose of the Study:

• To develop and standardize an indirect MAR assay for detecting IgG and IgA antisperm antibodies.
• To establish a practical screening procedure for identifying the Ig-class of antisperm antibodies.
• To overcome limitations of the direct MAR test by utilizing donor spermatozoa.

Main Methods:

• Investigated indirect MAR using donor spermatozoa sensitized with patient serum or seminal plasma.
• Optimized immunoglobulin removal from sensitized donor spermatozoa using a swim-up method.
• Standardized indirect MAR with glutaraldehyde-fixed sheep erythrocytes coated with IgG or IgA and specific anti-immunoglobulin antibodies.

Main Results:

• The swim-up method proved optimal for reducing nonbound immunoglobulin.
• The indirect MAR assay was standardized for sensitivity and ease of reading.
• The method allows for the comparison of IgG and IgA antibody reactions on an approximately equimolecular basis.

Conclusions:

• The indirect MAR is a viable alternative to direct MAR for detecting and characterizing antisperm antibodies.
• This standardized assay provides a sensitive and practical tool for clinical application in male infertility investigations.
• The method facilitates the differentiation of IgG and IgA antisperm antibody involvement.