The quality of sperm chromatin is an important factor in fertilization and is especially critical where one spermatozoon is artificially selected for fertilizing an egg (as in intracytoplasmic sperm injection). In this study, flow cytometry after staining of human spermatozoa with Acridine Orange was used to study chromatin structure. A method is described for estimating the percentage of cells in a human sperm sample that have completed epididymal maturation in regard to chromatin condensation. Of the 121 samples of the semen that were examined, nine contained a higher percentage of hypocondensed spermatozoa and six samples contained elevated amounts of hypercondensed spermatozoa. In addition to aberrancies in chromatin condensation other defects showed up as satellite populations of spermatozoa with higher than normal ratios of red/green fluorescence after Acridine Orange staining. Such defects were found in 15 semen samples. The use of swim-up and Percoll gradient centrifugation methods was shown to improve the percentage of spermatozoa with normal chromatin structure in some samples with poor initial quality.
Reaginic humoral antibodies to human seminal plasma were present in two women with systemic reactions. In one patient, IgE antibodies were demonstrated by direct skin tests, leukocyte histamine release, passive transfer to a nonallergic human recipient, the radioallergosorbent test, inhibition of the radioallergosorbent test and neutralization of passive transfer antibodies. A similar mechanism was established in the other case by direct skin tests and antigen-induced leukocyte histamine release. Sephadex G-100 fraction 2, derived from human seminal plasma, showed greater reaginic activity than other chromatographic fractions. Two other patients with histories of localized reactions did not develop humoral antibodies but did evince cell-mediated immunity to seminal fluid antigens. One patient with localized reactions also had significant titers of IgM and IgG sperm agglutinating antibodies to seminal plasma. Both reaginic and cell-mediated sensitization to human sperm extract were also observed in this series of patients. Histocompatibility leukocyte antigen (HLA) typing of all patients and their sexual partners showed a marked degree of shared histocompatibility focus antigens in members of each of two couples.
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