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

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.

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Figure 1: The organ culture model with cervical tissues.
Figure 2: Hematoxylin and eosin staining of paraffin-embedded CEM-IIIB HIV-1-exposed cervical tissues from organ culture.
Figure 3: Quantification of immunostained cells of cervical tissues.
Figure 4: Transmission of HIV-1 across cervical tissues.
Figure 5: Determination of the integrity of the Transwell organ culture system.
Figure 6: Identification of cell types expressing HIV-1 RNA, by simultaneous immunophenotyping and in situ hybridization.

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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.

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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

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  1. Kelly B. Collins
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Correspondence to Phalguni Gupta.

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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

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