Development of an in vitro organ culture model to study transmission of HIV-1 in the female genital tract (original) (raw)

Nature Medicine volume 6, pages 475–479 (2000)Cite this article

Human immunodeficiency virus type 1 (HIV-1) infection in adults is transmitted mainly by sexual routes, through cell-free or cell-associated virus present in the genital secretions1,2,3,4. Defining the viral and host factors of sexual transmission is important for understanding HIV-1 pathogenesis, as well as for developing a protective vaccine. One chief problem in studying the viral and host factors of HIV-1 transmission is the lack of a proper in vitro viral transmission model system that reflects the in vivo situation. There are a few reports describing HIV-1 infection of primary and transformed cells derived from genital tissues5,6,7. In addition, it was recently shown that infectious HIV-1 can be transmitted across a single monolayer of transformed epithelium cells derived from intestine (HT29, 1407 and Caco-2 cell lines) and endometrial carcinoma (HEC1 cells) by the process of transcytosis8. However, the cells used in those experiments were mostly immortalized cell lines grown as monolayers, and therefore do not mimic the stratified squamous epithelial layers present in the vagina and ectocervix of the female genital organ. The specialized multilayer mucosal tissue structure of the genital tract may serve as a barrier to HIV-1 infection and thereby in its entirety may be important in the sexual transmission of HIV-1.

Here, we describe the development of an organ culture model, derived from squamous cervical tissue, to study HIV-1 transmission across the mucosal barrier. As this model used tissues rather than a monolayer of cells (primary or transformed), it provided the natural tissue architecture, including epithelial cells, submucosa and immune cells (such as T cells, macrophages and Langerhans cells). The model allowed the evaluation of transmission of infectious virus from cell-free and cell-associated HIV-1, and identification of the major cell types in the tissue that became infected during transmission of HIV-1. Thus, this organ culture offers a model to delineate the virologic and host factors responsible for transmission of HIV-1 in the female genital tract.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Subscribe to this journal

Receive 12 print issues and online access

$209.00 per year

only $17.42 per issue

Buy this article

Prices may be subject to local taxes which are calculated during checkout

Additional access options:

References

  1. Pudney, J. & Anderson, D.J. Orchitis and human immunodeficiency virus type 1 infected cells in reproductive tissues from men with the acquired immune deficiency syndrome. Am. J. Pathol. 139, 149–160 (1991).
    CAS PubMed PubMed Central Google Scholar
  2. Vernazza, P. et al. Detection of biologic characterization of infectious HIV-1 in semen of seropositive men. AIDS 8, 1325 –1329 (1994).
    Article CAS Google Scholar
  3. Mermin, J.H., Holodiny, M., Katzenstein, D.A. & Merigan, T.C. Detection of human immunodeficiency virus DNA and RNA in semen by the polymerase chain reaction. J. Inf. Dis. 164, 769– 772 (1991).
    Article CAS Google Scholar
  4. Gupta, P. et al. High viral load in semen of HIV-1 infected men at all stages of disease and its reduction by therapy with protease and nonnucleoside RT inhibitors. J. Virol. 71, 6271– 6275 (1997).
    CAS PubMed PubMed Central Google Scholar
  5. Tan, X., Pearce-Pratt, R. & Phillips, D. Productive infection of a cervical epithelial cell line with human immunodeficiency virus: Implications for sexual transmission. J. Virol. 67, 6447–6452 (1993).
    CAS PubMed PubMed Central Google Scholar
  6. Gosselin, E. et al. Infectivity by HIV-1 of human uterine endometrial carcinoma cells. Adv. Exp. Med. Biol. 371B, 1003– 1006 (1995).
    CAS PubMed Google Scholar
  7. Howell, A. et al. Human immunodeficiency virus type 1 infection of cells and tissues from the upper and lower human female reproductive tract. J. Virol. 71, 3498–3506 ( 1997).
    CAS PubMed PubMed Central Google Scholar
  8. Bomsel, M. Transcytosis of infectious human immunodeficiency virus across a tight human epithelial cell line barrier. Nature Med. 3, 42–47 (1997).
    Article CAS Google Scholar
  9. Chackerian, B., Long, E., Luciw, P. & Overbaugh, J. HIV-1 co-receptors participates in post-entry stages of the virus replication cycle and function in SIV infection. J. Virol. 71, 3932– 3939 (1997).
    CAS PubMed PubMed Central Google Scholar
  10. Patterson, B. et al. Repertoire of chemokine receptor expression in the female genital tract. Am. J. Pathol. 153, 481– 490 (1998).
    Article CAS Google Scholar
  11. Atula, S., Grenman, R. & Syrjanen, S. Fibroblasts can modulate the phenotype of malignant epithelial cells in vitro. Exp. Cell. Res. 235 , 180–187 (1997).
    Article CAS Google Scholar
  12. Schutte, B., Tinnemans, M., Pijpers, G., Lenders, M. & Ramaekers, F. Three parameter flow cytometric analysis for simultaneous detection of cytokeratin, proliferation associated antigens and DNA content. Cytometry 21, 177–186 (1995).
    Article CAS Google Scholar
  13. Scurry, J., Beshay, V., Cohen, C. & Allen, D. Ki67 expression in lichen sclerosus of vulva in patients with and without associated squamous cell carcinoma Histopathology 32, 399– 404 (1998).
    Article CAS Google Scholar
  14. Delvenne, P., Al Saleh, W., Gilles, C., Thiry, A. & Boniver, J. Inhibition of growth of normal and human papillomavirus-transformed keratinocytes in monolayer and organotypic cultures by interferon-γ and tumor necrosis factor-α. Am. J. Pathol. 146 , 589–598 (1995).
    CAS PubMed PubMed Central Google Scholar
  15. Watson, A., Klaniecki, J. & Hanson, C. Psoralen/UV inactivation of HIV-1 infected cells for use in cytologic and immunologic procedures. AIDS Res. Hum. Retroviruses 6, 503–513 ( 1990).
    Article CAS Google Scholar
  16. Patterson, B. et al. Detection of HIV-RNA-positive monocytes in peripheral blood of HIV-positive patients by simultaneous flow cytometric analysis of intracellular HIV RNA and cellular immunophenotype. Cytometry 31, 265–327 (1998).
    Article CAS Google Scholar
  17. Levy, J. in HIV and the Pathogenesis of AIDS, 29–35 (American Society for Microbiology, Washington, DC, 1998).
    Google Scholar
  18. Miller, C. et al. Genital mucosal transmission of simian immunodeficiency virus: Animal model for heterosexual transmission of human immunodeficiency virus . J. Virol. 63, 4277–4284 (1989).
    CAS PubMed PubMed Central Google Scholar
  19. Miller, C. et al. Intravaginal inoculation of Rhesus macaques with cell-free simian immunodeficiency virus results in persistent or transient viremia. J. Virol. 68, 6391–6400 (1994).
    CAS PubMed PubMed Central Google Scholar
  20. Spira, A. et al. Cellular targets of infection and route of viral dissemination after an intravaginal inoculation of simian immunodeficiency virus into Rhesus macaques. J. Exp. Med. 183, 215– 225 (1996).
    Article CAS Google Scholar
  21. Meyers, C. Organotypic (raft) epithelial tissue culture system for the differentiation-dependent replication of papillomavirus. Meth. Cell Sci. 18, 201–210 (1996).
    Article Google Scholar
  22. Patterson, B. et al. Monitoring HIV-1 treatment in immune-cell subsets with ultrasensitive fluorescence in situ hybridization. Lancet 353 , 211–212 (1999).
    Article CAS Google Scholar

Download references

Acknowledgements

We thank C. Rinaldo for review of the manuscript; J. Andersson, K. Lore and L. Rohan for technical support; L. Mock for procuring tissues; and J. Malenka for secretarial assistance. This study was supported by Public Health Service grant RO1-HD-32256, American Foundation for AIDS Research grant #2630 and National Institutes of Health grant #2P01-A139061.

Author information

Authors and Affiliations

  1. Department of Infectious Diseases and Microbiology Graduate School of Public Health, University of Pittsburgh, Pittsburgh , 15261, Pennsylvania, USA
    Kelly B. Collins & Phalguni Gupta
  2. Department of Pediatrics, Division of Special Infectious Diseases, Children's Memorial Hospital, Northwestern University Medical School, Chicago, 60614, Illinois, USA
    Bruce K. Patterson
  3. Departments of Pathology and, Obstetrics, Gynecology and Reproductive Sciences, Magee Women's Hospital, University of Pittsburgh, Pittsburgh, 15261, Pennsylvania, USA
    Gregory J. Naus & Daniel V. Landers

Authors

  1. Kelly B. Collins
    You can also search for this author inPubMed Google Scholar
  2. Bruce K. Patterson
    You can also search for this author inPubMed Google Scholar
  3. Gregory J. Naus
    You can also search for this author inPubMed Google Scholar
  4. Daniel V. Landers
    You can also search for this author inPubMed Google Scholar
  5. Phalguni Gupta
    You can also search for this author inPubMed Google Scholar

Corresponding author

Correspondence toPhalguni Gupta.

Rights and permissions

About this article

Cite this article

Collins, K., Patterson, B., Naus, G. et al. Development of an in vitro organ culture model to study transmission of HIV-1 in the female genital tract.Nat Med 6, 475–479 (2000). https://doi.org/10.1038/74743

Download citation

This article is cited by