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

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dc.contributor.authorMcKinnon, Lyle Ren_ZA
dc.contributor.authorHughes, Sean Men_ZA
dc.contributor.authorDe Rosa, Stephen Cen_ZA
dc.contributor.authorMartinson, Jeffrey Aen_ZA
dc.contributor.authorPlants, Jillen_ZA
dc.contributor.authorBrady, Kirsten Een_ZA
dc.contributor.authorGumbi, Pamela Pen_ZA
dc.contributor.authorAdams, Devin Jen_ZA
dc.contributor.authorVojtech, Luciaen_ZA
dc.contributor.authorGalloway, Christine Gen_ZA
dc.date.accessioned2016-01-02T05:05:35Z
dc.date.available2016-01-02T05:05:35Z
dc.date.issued2014en_ZA
dc.description.abstractBACKGROUND: 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.identifier.apacitationMcKinnon, L. R., Hughes, S. M., De Rosa, S. C., Martinson, J. A., Plants, J., Brady, K. E., ... Galloway, C. G. (2014). Optimizing viable leukocyte sampling from the female genital tract for clinical trials: an international multi-site study. <i>PLoS One</i>, http://hdl.handle.net/11427/16148en_ZA
dc.identifier.chicagocitationMcKinnon, Lyle R, Sean M Hughes, Stephen C De Rosa, Jeffrey A Martinson, Jill Plants, Kirsten E Brady, Pamela P Gumbi, Devin J Adams, Lucia Vojtech, and Christine G Galloway "Optimizing viable leukocyte sampling from the female genital tract for clinical trials: an international multi-site study." <i>PLoS One</i> (2014) http://hdl.handle.net/11427/16148en_ZA
dc.identifier.citationMcKinnon, 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.0085675en_ZA
dc.identifier.ris TY - Journal Article AU - McKinnon, Lyle R AU - Hughes, Sean M AU - De Rosa, Stephen C AU - Martinson, Jeffrey A AU - Plants, Jill AU - Brady, Kirsten E AU - Gumbi, Pamela P AU - Adams, Devin J AU - Vojtech, Lucia AU - Galloway, Christine G AB - 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. DA - 2014 DB - OpenUCT DO - 10.1371/journal.pone.0085675 DP - University of Cape Town J1 - PLoS One LK - https://open.uct.ac.za PB - University of Cape Town PY - 2014 T1 - Optimizing viable leukocyte sampling from the female genital tract for clinical trials: an international multi-site study TI - Optimizing viable leukocyte sampling from the female genital tract for clinical trials: an international multi-site study UR - http://hdl.handle.net/11427/16148 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/16148
dc.identifier.urihttp://dx.doi.org/10.1371/journal.pone.0085675
dc.identifier.vancouvercitationMcKinnon LR, Hughes SM, De Rosa SC, Martinson JA, Plants J, Brady KE, et al. Optimizing viable leukocyte sampling from the female genital tract for clinical trials: an international multi-site study. PLoS One. 2014; http://hdl.handle.net/11427/16148.en_ZA
dc.language.isoengen_ZA
dc.publisherPublic Library of Scienceen_ZA
dc.publisher.departmentInstitute of Infectious Disease and Molecular Medicineen_ZA
dc.publisher.facultyFaculty of Health Sciencesen_ZA
dc.publisher.institutionUniversity of Cape Town
dc.rightsThis 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.holder© 2014 McKinnon et alen_ZA
dc.rights.urihttp://creativecommons.org/licenses/by/4.0en_ZA
dc.sourcePLoS Oneen_ZA
dc.source.urihttp://journals.plos.org/plosoneen_ZA
dc.subject.otherBiopsyen_ZA
dc.subject.otherWhite blood cellsen_ZA
dc.subject.otherT cellsen_ZA
dc.subject.otherMacrophagesen_ZA
dc.subject.otherBlooden_ZA
dc.subject.otherCytotoxic T cellsen_ZA
dc.subject.otherB cellsen_ZA
dc.subject.otherCell stainingen_ZA
dc.titleOptimizing viable leukocyte sampling from the female genital tract for clinical trials: an international multi-site studyen_ZA
dc.typeJournal Articleen_ZA
uct.type.filetypeText
uct.type.filetypeImage
uct.type.publicationResearchen_ZA
uct.type.resourceArticleen_ZA
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