Quantification of Residual Germinal Center Activity and HIV-1 DNA and RNA Levels Using Fine Needle Biopsies of Lymph Nodes During Antiretroviral Therapy

• Published in: AIDS research and human retroviruses Vol. 33; no. 7; p. 648
• Main Authors: Hey-Nguyen, William J, Xu, Yin, Pearson, Chester F, Bailey, Michelle, Suzuki, Kazuo, Tantau, Robyn, Obeid, Solange, Milner, Brad, Field, Andrew, Carr, Andrew, Bloch, Mark, Cooper, David A, Kelleher, Anthony D, Zaunders, John J, Koelsch, Kersten K
• Format: Journal Article
• Language: English
• Published: United States 01.07.2017
• Subjects: Adult; Anti-Retroviral Agents - therapeutic use; Biopsy, Fine-Needle; DNA, Viral - blood; Female; Germinal Center - pathology; HIV Infections - drug therapy; HIV Infections - pathology; HIV Infections - virology; HIV-1 - isolation & purification; Humans; Lymph Nodes - pathology; Lymphocyte Count; Male; RNA, Viral - blood; Viral Load; HIV-1 reservoir; tissue; germinal center; follicular T helper cell; fine needle biopsy; lymph node
• ISSN: 1931-8405
• DOI: 10.1089/aid.2016.0171

HIV-1 reservoirs are most often studied in peripheral blood (PB), but not all lymphocytes recirculate, particularly T follicular helper (Tfh) CD4 T cells, as well as germinal center (GC) B cells, in lymph nodes (LNs). Ultrasound-guided fine needle biopsies (FNBs) from inguinal LNs and PB samples were obtained from 10 healthy controls (HCs) and 21 HIV-1-infected subjects [11 antiretroviral therapy (ART) naive and 10 on ART]. Tfh cells and GC B cells were enumerated by flow cytometry. HIV-1 DNA and cell-associated (CA) RNA levels in LNs and PB were quantified by real-time polymerase chain reaction. FNBs were obtained without adverse events. Tfh cells and GC B cells were highly elevated in ART-naive subjects, with a median GC B cell count >300-fold higher than HCs, but also remained higher in 4 out of the 10 subjects on ART. GC B cell counts and Tfh cell counts were highly correlated with each other, and also with activated T cells in LNs but not in blood. Levels of HIV-1 DNA and CA RNA viral burden in highly purified CD4 T cells from FNBs were significantly elevated compared with those in CD4 T cells from PB in the ART-naive group, but only trended toward an increase in the ART patients. FNBs enabled minimally invasive access to, and parallel measurement of residual activated T and B cells and viral burden within LNs in HIV-1-infected patients. These FNBs revealed significant GC activity that was not apparent from corresponding PB samples.