Petitjean G., Becquart Pierre, Tuaillon E., Al Tabaa Y., Valea D., Huguet M. F., Meda N., Van de Perre P., Vendrell J. P. (2007). Isolation and characterization of HIV-1-infected resting CD4(+) T lymphocytes in breast milk. Journal of Clinical Virology, 39 (1), p. 1-8. ISSN 1386-6532.
Titre du document
Isolation and characterization of HIV-1-infected resting CD4(+) T lymphocytes in breast milk
Petitjean G., Becquart Pierre, Tuaillon E., Al Tabaa Y., Valea D., Huguet M. F., Meda N., Van de Perre P., Vendrell J. P.
Source
Journal of Clinical Virology, 2007,
39 (1), p. 1-8 ISSN 1386-6532
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.
