Evaluation of T‐lymphocyte subsets present in semen and peripheral blood of healthy donors: A report from the heterosexual transmission study

Abstract

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.

Associations between Virologic and Immunologic Dynamics in Blood and in the Male Genital Tract

“…As previously described (19,57), our HIV-infected participants had fewer CD4 ϩ T cells in semen than the HIV-uninfected group, which is consistent with findings at other mucosal sites (10,54). In the male genital tract, low numbers of CD4 ϩ T cells, which are the primary target cells for HIV replication, may explain the inefficient rate of transmission seen for HIV compared to other STI (62,74).…”

Lymphocyte immunoregulatory cells present in semen from human immunodeficiency virus (HIV)-infected individuals: A report from the HIV heterosexual transmission study

“…The enriched mononuclear cell fraction was harvested and the cells were resuspended in HBSS containing 2% FBS and 0.1% sodium aide. Cells were aliquoted in each tube for three-color immunofluorescence labeling with monoclonal antibodies directed to specific surface antigens as previously described (8).…”

“…We have recently reported a flow cytometric method for identifying and counting immunophenot ypically distinct cells present in normal semen (8). This application allows analysis of a greater sample size compared to traditional microscopic evaluation of semen preparations.…”

Murine Sperm Expresses Toll-Like Receptor (TLR) Family that Responds to the Pathogens Released from Virus, and Decreases Fertilization Ability by the Stimuli

“…In pig, porcine reproductive and respiratory syndrome virus (PRRSV), Aujeszky's disease virus and porcine parvovirus (PPV) decrease sperm fertilization activity, which results in economic losses in domestic animals [23][24][25]. The reduction of sperm functions due to virus infections has been thought to be caused by the immune function accumulating leukocytes in semen [26]. Leukocytes express TLR3, TLR7, TLR8 and TLR9 that recognize virus-released pathogens, ssRNA, double strand RNA or unmethylated CpG DNA, then secrete cytokines or chemokine families [27].…”