title: Quantification of Residual Germinal Center Activity and HIV-1 DNA and RNA Levels Using Fine Needle Biopsies of Lymph Nodes During Antiretroviral Therapy.
abstract: 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.
authors.affiliation: 1 Kirby Institute , UNSW Sydney, Sydney, Australia .
authors.forename: William J
authors.initials: WJ
authors.lastname: Hey-Nguyen
authors.affiliation: 1 Kirby Institute , UNSW Sydney, Sydney, Australia .
authors.forename: Yin
authors.initials: Y
authors.lastname: Xu
authors.affiliation: 1 Kirby Institute , UNSW Sydney, Sydney, Australia .
authors.forename: Chester F
authors.initials: CF
authors.lastname: Pearson
authors.affiliation: 1 Kirby Institute , UNSW Sydney, Sydney, Australia .
authors.forename: Michelle
authors.initials: M
authors.lastname: Bailey
authors.affiliation: 1 Kirby Institute , UNSW Sydney, Sydney, Australia .|2 Centre for Applied Medical Research, St. Vincent's Hospital , Darlinghurst, Australia .
authors.forename: Kazuo
authors.initials: K
authors.lastname: Suzuki
authors.affiliation: 3 Department of Medical Imaging, St. Vincent's Hospital , Darlinghurst, Australia .
authors.forename: Robyn
authors.initials: R
authors.lastname: Tantau
authors.affiliation: 3 Department of Medical Imaging, St. Vincent's Hospital , Darlinghurst, Australia .
authors.forename: Solange
authors.initials: S
authors.lastname: Obeid
authors.affiliation: 3 Department of Medical Imaging, St. Vincent's Hospital , Darlinghurst, Australia .
authors.forename: Brad
authors.initials: B
authors.lastname: Milner
authors.affiliation: 4 Department of Anatomical Pathology, St. Vincent's Hospital , Darlinghurst, Australia .
authors.forename: Andrew
authors.initials: A
authors.lastname: Field
authors.affiliation: 2 Centre for Applied Medical Research, St. Vincent's Hospital , Darlinghurst, Australia .
authors.forename: Andrew
authors.initials: A
authors.lastname: Carr
authors.affiliation: 5 Holdsworth House Medical Practice , Darlinghurst, Australia .
authors.forename: Mark
authors.initials: M
authors.lastname: Bloch
authors.affiliation: 1 Kirby Institute , UNSW Sydney, Sydney, Australia .|2 Centre for Applied Medical Research, St. Vincent's Hospital , Darlinghurst, Australia .
authors.forename: David A
authors.initials: DA
authors.lastname: Cooper
authors.affiliation: 1 Kirby Institute , UNSW Sydney, Sydney, Australia .|2 Centre for Applied Medical Research, St. Vincent's Hospital , Darlinghurst, Australia .
authors.forename: Anthony D
authors.initials: AD
authors.lastname: Kelleher
authors.affiliation: 1 Kirby Institute , UNSW Sydney, Sydney, Australia .|2 Centre for Applied Medical Research, St. Vincent's Hospital , Darlinghurst, Australia .
authors.forename: John J
authors.initials: JJ
authors.lastname: Zaunders
authors.affiliation: 1 Kirby Institute , UNSW Sydney, Sydney, Australia .|2 Centre for Applied Medical Research, St. Vincent's Hospital , Darlinghurst, Australia .
authors.forename: Kersten K
authors.initials: KK
authors.lastname: Koelsch
mesh_terms: D000328:Adult; D044966:Anti-Retroviral Agents / Q000627:therapeutic use*; D044963:Biopsy, Fine-Needle; D004279:DNA, Viral / Q000097:blood; D005260:Female; D018858:Germinal Center / Q000473:pathology*; D015658:HIV Infections / Q000188:drug therapy / Q000473:pathology* / Q000821:virology*; D015497:HIV-1 / Q000302:isolation & purification*; D006801:Humans; D008198:Lymph Nodes / Q000473:pathology*; D018655:Lymphocyte Count; D008297:Male; D012367:RNA, Viral / Q000097:blood; D019562:Viral Load
chemical_list: D044966:Anti-Retroviral Agents; D004279:DNA, Viral; D012367:RNA, Viral
keywords: HIV-1 reservoir; fine needle biopsy; follicular T helper cell; germinal center; lymph node; tissue