Please use this identifier to cite or link to this item:
Title: Sensitive quantification of the HIV-1 reservoir in gut-associated lymphoid tissue
Author: Morón López, Sara
Puertas, Maria C.
Gálvez, Cristina
Navarro, Jordi
Carrasco, Anna
Esteve i Comas, Maria
Manyé, Josep
Crespo, Manel
Salgado, Maria
Martinez Picado, Javier
Keywords: VIH (Virus)
Cèl·lules T
HIV (Viruses)
T cells
Issue Date: 17-Apr-2017
Publisher: Public Library of Science (PLoS)
Abstract: Background: The implementation of successful strategies to achieve an HIV cure has become a priority in HIV research. However, the current location and size of HIV reservoirs is still unknown since there are limited tools to evaluate HIV latency in viral sanctuaries such as gut-associated lymphoid tissue (GALT). As reported in the so called "Boston Patients", despite undetectable levels of proviral HIV-1 DNA in blood and GALT, viral rebound happens in just few months after ART interruption. This fact might imply that current methods are not sensitive enough to detect residual reservoirs. Showing that, it is imperative to improve the detection and quantification of HIV-1 reservoir in tissue samples. Herein, we propose a novel non-enzymatic protocol for purification of Lamina Propria Leukocytes (LPL) from gut biopsies combined to viral HIV DNA (vDNA) quantification by droplet digital PCR (ddPCR) to improve the sensitivity and accuracy of viral reservoir measurements (LPL-vDNA assay). Methods: Endoscopic ileum biopsies were sampled from 12 HIV-1-infected cART-suppressed subjects. We performed a DTT/EDTA-based treatment for epithelial layer removal followed by non-enzymatic disruption of the tissue to obtain lamina propria cell suspension (LP). CD45+ cells were subsequently purified by flow sorting and vDNA was determined by ddPCR. Results: vDNA quantification levels were significantly higher in purified LPLs (CD45+) than in bulk LPs (p<0.01). The levels of vDNA were higher in ileum samples than in concurrent PBMC from the same individuals (p = 0.002). As a result of the increased sensitivity of this purification method, the Poisson 95% confidence intervals of the vDNA quantification data from LPLs were narrower than that from bulk LPs. Of note, vDNA was unambiguously quantified above the detection limit in 100% of LPL samples, while only in 58% of bulk LPs. Conclusion: We propose an innovative combined protocol for a more sensitive detection of the HIV reservoir in gut-associated viral sanctuaries, which might be used to evaluate any proposed eradication strategy.
Note: Reproducció del document publicat a:
It is part of: PLoS One, 2017, vol. 12, num. 4, p. e0175899
Related resource:
ISSN: 1932-6203
Appears in Collections:Articles publicats en revistes (Medicina)

Files in This Item:
File Description SizeFormat 
691997.pdf2.2 MBAdobe PDFView/Open

This item is licensed under a Creative Commons License Creative Commons