Measuring turnover of SIV DNA in resting CD4+ T cells using pyrosequencing: implications for the timing of HIV eradication therapies.
Resting CD4+ T cells are a reservoir of latent HIV-1. Understanding the turnover of HIV DNA in these cells has implications for the development of eradication strategies. Most studies of viral latency focus on viral persistence under antiretroviral therapy (ART). We studied the turnover of SIV DNA resting CD4+ T cells during active infection in a cohort of 20 SIV-infected pigtail macaques. We compared SIV sequences at two Mane-A1*084:01-restricted CTL epitopes using serial plasma RNA and resting CD4+ T cell DNA samples by pyrosequencing, and used a mathematical modeling approach to estimate SIV DNA turnover. We found SIV DNA turnover in resting CD4+ T cells was slow in animals with low chronic viral loads, consistent with the long persistence of latency seen under ART. However, in animals with high levels of chronic viral replication, turnover was high. SIV DNA half-life within resting CD4 cells correleated with viral load (p = 0.0052) at the Gag KP9 CTL epitope. At a second CTL epitope in Tat (KVA10) there was a trend towards an association of SIV DNA half-life in resting CD4 cells and viral load (p = 0.0971). Further, we found that the turnover of resting CD4+ T cell SIV DNA was higher for escape during early infection than for escape later in infection (p = 0.0084). Our results suggest viral DNA within resting CD4 T cells is more labile and may be more susceptible to reactivation/eradication treatments when there are higher levels of virus replication and during early/acute infection.
Department of Microbiology and Immunology, University of Melbourne, Victoria, Australia.
Jeanette C
JC
Reece
Centre for Vascular Research, University of New South Wales, Kensington, NSW, Australia.
Alexey
A
Martyushev
Centre for Vascular Research, University of New South Wales, Kensington, NSW, Australia.
Janka
J
Petravic
Centre for Vascular Research, University of New South Wales, Kensington, NSW, Australia.
Andrew
A
Grimm
Department of Microbiology and Immunology, University of Melbourne, Victoria, Australia.
Shayarana
S
Gooneratne
Department of Microbiology and Immunology, University of Melbourne, Victoria, Australia.
Thakshila
T
Amaresena
Department of Microbiology and Immunology, University of Melbourne, Victoria, Australia.
Robert
R
De Rose
Department of Microbiology and Immunology, University of Melbourne, Victoria, Australia.
Liyen
L
Loh
Centre for Vascular Research, University of New South Wales, Kensington, NSW, Australia.
Miles P
MP
Davenport
Department of Microbiology and Immunology, University of Melbourne, Victoria, Australia.
Stephen J
SJ
Kent
D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism* / Q000821:virology; D004279:DNA, Viral / Q000097:blood*; D015658:HIV Infections / Q000097:blood / Q000628:therapy; D015497:HIV-1 / Q000378:metabolism*; D006801:Humans; D008254:Macaca nemestrina; D008954:Models, Biological; D016097:Simian Acquired Immunodeficiency Syndrome / Q000097:blood / Q000628:therapy; D015302:Simian Immunodeficiency Virus / Q000378:metabolism*
D004279:DNA, Viral