Optimizing viable leukocyte sampling from the female genital tract for clinical trials: an international multi-site study

 

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dc.contributor.author McKinnon, Lyle R en_ZA
dc.contributor.author Hughes, Sean M en_ZA
dc.contributor.author De Rosa, Stephen C en_ZA
dc.contributor.author Martinson, Jeffrey A en_ZA
dc.contributor.author Plants, Jill en_ZA
dc.contributor.author Brady, Kirsten E en_ZA
dc.contributor.author Gumbi, Pamela P en_ZA
dc.contributor.author Adams, Devin J en_ZA
dc.contributor.author Vojtech, Lucia en_ZA
dc.contributor.author Galloway, Christine G en_ZA
dc.date.accessioned 2016-01-02T05:05:35Z
dc.date.available 2016-01-02T05:05:35Z
dc.date.issued 2014 en_ZA
dc.identifier.citation McKinnon, L. R., Hughes, S. M., De Rosa, S. C., Martinson, J. A., Plants, J., Brady, K. E., ... & Fialkow, M. (2013). Optimizing viable leukocyte sampling from the female genital tract for clinical trials: an international multi-site study. PloS one, 9(1), e85675. doi:10.1371/journal.pone.0085675 en_ZA
dc.identifier.uri http://hdl.handle.net/11427/16148
dc.identifier.uri http://dx.doi.org/10.1371/journal.pone.0085675
dc.description.abstract BACKGROUND: Functional analysis of mononuclear leukocytes in the female genital mucosa is essential for understanding the immunologic effects of HIV vaccines and microbicides at the site of HIV exposure. However, the best female genital tract sampling technique is unclear. Methods and FINDINGS: We enrolled women from four sites in Africa and the US to compare three genital leukocyte sampling methods: cervicovaginal lavages (CVL), endocervical cytobrushes, and ectocervical biopsies. Absolute yields of mononuclear leukocyte subpopulations were determined by flow cytometric bead-based cell counting. Of the non-invasive sampling types, two combined sequential cytobrushes yielded significantly more viable mononuclear leukocytes than a CVL (p<0.0001). In a subsequent comparison, two cytobrushes yielded as many leukocytes (∼10,000) as one biopsy, with macrophages/monocytes being more prominent in cytobrushes and T lymphocytes in biopsies. Sample yields were consistent between sites. In a subgroup analysis, we observed significant reproducibility between replicate same-day biopsies (r = 0.89, p = 0.0123). Visible red blood cells in cytobrushes increased leukocyte yields more than three-fold (p = 0.0078), but did not change their subpopulation profile, indicating that these leukocytes were still largely derived from the mucosa and not peripheral blood. We also confirmed that many CD4 + T cells in the female genital tract express the α4β7 integrin, an HIV envelope-binding mucosal homing receptor. CONCLUSIONS: CVL sampling recovered the lowest number of viable mononuclear leukocytes. Two cervical cytobrushes yielded comparable total numbers of viable leukocytes to one biopsy, but cytobrushes and biopsies were biased toward macrophages and T lymphocytes, respectively. Our study also established the feasibility of obtaining consistent flow cytometric analyses of isolated genital cells from four study sites in the US and Africa. These data represent an important step towards implementing mucosal cell sampling in international clinical trials of HIV prevention. en_ZA
dc.language.iso eng en_ZA
dc.publisher Public Library of Science en_ZA
dc.rights This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. en_ZA
dc.rights.uri http://creativecommons.org/licenses/by/4.0 en_ZA
dc.source PLoS One en_ZA
dc.source.uri http://journals.plos.org/plosone en_ZA
dc.subject.other Biopsy en_ZA
dc.subject.other White blood cells en_ZA
dc.subject.other T cells en_ZA
dc.subject.other Macrophages en_ZA
dc.subject.other Blood en_ZA
dc.subject.other Cytotoxic T cells en_ZA
dc.subject.other B cells en_ZA
dc.subject.other Cell staining en_ZA
dc.title Optimizing viable leukocyte sampling from the female genital tract for clinical trials: an international multi-site study en_ZA
dc.type Journal Article en_ZA
dc.rights.holder © 2014 McKinnon et al en_ZA
uct.type.publication Research en_ZA
uct.type.resource Article en_ZA
dc.publisher.institution University of Cape Town
dc.publisher.faculty Faculty of Health Sciences en_ZA
dc.publisher.department Institute of Infectious Disease and Molecular Medicine en_ZA
uct.type.filetype Text
uct.type.filetype Image


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This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Except where otherwise noted, this item's license is described as This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.