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In 2001, the World Health Organization reported 4.3 million new human immunodeficiency virus (HIV) infections in adults globally, 41% of which were in women. During the year 2000, 27% of newly diagnosed HIV infections in the United States occurred in women. In developed countries, the perception of HIV infection has changed from an acute, lethal infection to a chronic illness; the introduction of highly active antiretroviral therapy has decreased morbidity and mortality, and new drug therapies have dramatically decreased perinatal transmission. In view of these advances, some HIV-infected individuals are considering reproduction. Following the lead of organizations in other developed countries, the American College of Obstetricians and Gynecologists has recently endorsed the use of reproductive technology in HIV-infected patients. Which patients should be offered assisted reproduction and what the optimal methods are of decreasing heterosexual and perinatal HIV transmission must be determined.

The purpose of this study was to accurately determine the T-lymphocyte subsets found in semen from healthy volunteers, to evaluate the impact of repeated ejaculation on the fiequency or type of immune cells present in semen, and to compare subset analysis in semen to that in the peripheral blood. To accomplish this, a flow cytometric method was developed to identify and count immunophenotypically distinct cells present in semen. Fresh semen samples and peripheral blood were collected over three consecutive days from nine healthy donors. Donors had normal ejaculate volume, sperm count, sperm motility, morphology, and leukocyte count. No significant inhra-donor differences were seen in these parameters over time.No siwcant differences were observed in the percentage of CD3+ cells, CD4+ cells, CD8+ cells, and the CD4:CDS ratio in semen on consecutive days.However, within the CD4+ subset, when naive and memory CD4+ cells were measured, some day to day variability was suggested. No signitlcant differences in CD3+, CD4+, CDS+, CD4/CD8 ratio, or naive and memory subsets were seen in the peripheral blood between sampling days. When semen was compared to peripheral blood some differences in immune subset values were observed, with an increase in the percentage of memory CD4+ cells in semen being the most striking. This 5nding may be relevant to HIV HIV infection (1,3,13). Although a wide range in the differential distribution of the leukocytes present in semen has been reported ( 12), some of these extremes may be due to problems inherent in traditional cell counting techniques. Little is known about the number and types of immunophenotypically distinct cells such as CD4+ and CDS+ lymphocytes and subsets of these cells that may be present in semen. A better understanding of the types of cells present in semen may be useful when studying events related to HIV transmission, other STDs, and male infertility.The primary purpose of this study was to accurately determine the T-lymphocyte subpopulations found in normal human semen and to evaluate the impact of repeated ejaculation at 24 h intervals on the frequency or type of immune cells present in semen. In order to do this, a flow cytometric method was developed to identify and count immunophenot ypically distinct cells present in semen. This approach potentially has the advantages that it allows analysis of a larger sample size than is possible using microscopic evaluation of semen preparations and reduces variation common to manual counting techniques. This study also compared subset analysis in semen to that in the peripheral blood. These values will be useful when correlating results of similar measures in HIV infected individuals currently being followed as part of a heterosexual HIV transmission study.