期刊
JOURNAL OF CLINICAL VIROLOGY
卷 39, 期 1, 页码 1-8出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jcv.2007.02.004
关键词
breast milk cells; CD4(+) T lymphocytes isolation; ELISpot assay; flow cytometry; HIV-1 reservoir
类别
An HIV-1 reservoir comprised primarily of latently infected resting CD4(+) T lymphocytes that can be stimulated in vivo to produce virus may play a critical role in mother-to-child postnatal transmission of HIV-1 by breastfeeding. Here, we describe an experimental protocol for the detection of resting CD4(+) T cell HIV-1 reservoir from breast milk. We adapted a method for the purification of resting CD4(+) T lymphocytes in blood to isolate resting CD4 T+ cells in breast milk from HIV-1-infected-lactating women (n = 18) and from controls (n = 3). Purified resting CD4 T+ cells from blood and breast milk samples of HIV-1-infected-lactating women were polyclonally stimulated to characterize and enumerate HIV-1-antigen-secreting cells (HIV-1-Ag-SCs) by an enzyme-linked immunospot (ELISpot) assay. Resting CD4 T+ cells represented more than 90% of purified viable breast milk cells. CD4(+) T cell polyclonal stimulation combined with the ELISpot assay led to the characterization of a breast milk T cell HIV-1 reservoir greater than the blood reservoir (median 400 and 57.14 HIV-1-Ag-SCs/10(6) resting CD4(+) T cells, respectively,p < 0.001). our strategy could be adapted to other body fluids and be useful for characterizing new HIV-1 reservoirs. (c) 2007 Elsevier B.V. All rights reserved.
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