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The mode of heterosexual transmission of human immunodeficiency virus (HIV) is not yet understood. The semen of HIV-infected men contains free virus and infected cells, and it is not known which of these is more important for sexual transmission of the virus to women. Some investigators have presented in vitro studies supporting a cellular mode of transmission of HIV and have suggested that infected lymphoid cells may act as the primary source of infection. This has become known as the "Trojan Horse" hypothesis. In vivo demonstrations of such events are lacking and are not likely to be forthcoming using human subjects. To investigate the ability of normal lymphoid cells to invade the cervicovaginal mucosa in an experimental animal, we stained C3H/He (H-2Kk) mouse peritoneal lymphoid cells with bisbenzimide, a vital fluorescent DNA-binding dye, and inoculated the cells atraumatically into the vaginas of progestin-treated, BALB/c (H-2Kd) recipient mice. Donor cells were identified in recipient tissues by their bisbenzimide-fluorescent nuclei and by fluorescein staining of the membrane antigen, H-2Kk. Donor lymphoid cells were observed in histological sections of recipient cervicovaginal mucosa and also in the iliac lymph nodes of 34 of 36 recipient mice 24 h after inoculation into the vagina. The number of donor cells in the iliac lymph nodes was 8.6 +/- 1.4 (mean +/- SEM) cells per mouse with a range of 0-35 cells per mouse. Approximately 28% of the donor lymphoid cells in recipient lymph nodes expressed CD4, which in humans is the receptor for HIV. We did not detect F4/80, a marker of mature mouse macrophages in the donor cell population, on any of the migrating cells in recipient lymph nodes. However, this negative result is equivocal, because the marker might be down-regulated after transfer or the migrating macrophages might be difficult to dissociate from the recipient lymph node tissue. These observations in mice support the suggestion that HIV-containing lymphoid cells in the semen of infected men may invade the cervicovaginal mucosa after sexual intercourse and deliver the virus to a woman's internal environment. However, both the donor cells and the recipient reproductive tract of the mice in the present study differed in significant respects from their counterparts in humans that might be involved in heterosexual HIV transmission. Further studies are needed to determine whether this possible mode of virus transmission is mainly responsible for heterosexual transmission of HIV in humans.

Migration of Foreign Lymphocytes from the Mouse Vagina into the Cervicovaginal Mucosa and to the Iliac Lymph Nodes1