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  • Title: Isolation and characterization of HIV-1-infected resting CD4+ T lymphocytes in breast milk.
    Author: Petitjean G, Becquart P, Tuaillon E, Al Tabaa Y, Valea D, Huguet MF, Meda N, Van de Perre P, Vendrell JP.
    Journal: J Clin Virol; 2007 May; 39(1):1-8. PubMed ID: 17387039.
    Abstract:
    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.
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