title: Cellular HIV-1 DNA levels in patients receiving antiretroviral therapy strongly correlate with therapy initiation timing but not with therapy duration.
abstract: BACKGROUND
  Viral reservoir size refers to cellular human immunodeficiency virus-1 (HIV-1) DNA levels in CD4(+) T lymphocytes of peripheral blood obtained from patients with plasma HIV-1-RNA levels (viral load, VL) maintained below the detection limit by antiretroviral therapy (ART). We measured HIV-1 DNA levels in CD4(+) lymphocytes in such patients to investigate their clinical significance.
  
  
  METHODS
  CD4(+) T lymphocytes were isolated from the peripheral blood of 61 patients with a VL maintained at less than 50 copies/ml for at least 4 months by ART and total DNA was purified. HIV-1 DNA was quantified by nested PCR to calculate the copy number per 1 million CD4(+) lymphocytes (relative amount) and the copy number in 1 ml of blood (absolute amount). For statistical analysis, the Spearman rank or Wilcoxon signed-rank test was used, with a significance level of 5%.
  
  
  RESULTS
  CD4 cell counts at the time of sampling negatively correlated with the relative amount of HIV-1 DNA (median = 33 copies/million CD4(+) lymphocytes; interquartile range [IQR] = 7-123 copies/million CD4(+) lymphocytes), but were not correlated with the absolute amounts (median = 17 copies/ml; IQR = 5-67 copies/ml). Both absolute and relative amounts of HIV-1 DNA were significantly lower in six patients in whom ART was initiated before positive seroconversion than in 55 patients in whom ART was initiated in the chronic phase, as shown by Western blotting. CD4 cell counts before ART introduction were also negatively correlated with both the relative and absolute amounts of HIV-1 DNA. Only the relative amounts of HIV-1 DNA negatively correlated with the duration of VL maintenance below the detection limit, while the absolute amounts were not significantly correlated with this period.
  
  
  CONCLUSIONS
  The amounts of cellular HIV-1 DNA in patients with VLs maintained below the detection limit by the introduction of ART correlated with the timing of ART initiation but not with the duration of ART. In addition, CD4(+) T lymphocytes, which were newly generated by ART, diluted latently infected cells, indicating that measurements of the relative amounts of cellular HIV-1 DNA might be underestimated.
authors: AIDS Medical Center, National Hospital Organization Osaka National Hospital, Osaka, Japan. dai@onh.go.jp
authors: Dai
authors: D
authors: Watanabe
authors: Shiro
authors: S
authors: Ibe
authors: Tomoko
authors: T
authors: Uehira
authors: Rumi
authors: R
authors: Minami
authors: Atsushi
authors: A
authors: Sasakawa
authors: Keishiro
authors: K
authors: Yajima
authors: Hitoshi
authors: H
authors: Yonemoto
authors: Hiroki
authors: H
authors: Bando
authors: Yoshihiko
authors: Y
authors: Ogawa
authors: Tomohiro
authors: T
authors: Taniguchi
authors: Daisuke
authors: D
authors: Kasai
authors: Yasuharu
authors: Y
authors: Nishida
authors: Masahiro
authors: M
authors: Yamamoto
authors: Tsuguhiro
authors: T
authors: Kaneda
authors: Takuma
authors: T
authors: Shirasaka
references: Proc Natl Acad Sci U S A. 1998 Jul 21;95(15):8869-73
references: 9671771
references: J Infect Dis. 2005 Jul 1;192(1):46-55
references: 15942893
references: J Clin Microbiol. 2005 Dec;43(12):6183-5
references: 16333125
references: J Infect Dis. 2008 Feb 1;197(3):411-9
references: 18248304
references: Science. 1997 Nov 14;278(5341):1295-300
references: 9360927
references: Curr HIV Res. 2008 Jan;6(1):43-8
references: 18288974
references: Nat Med. 2010 Apr;16(4):460-5
references: 20228817
references: AIDS. 2000 Jan 7;14(1):23-9
references: 10714564
references: J Acquir Immune Defic Syndr Hum Retrovirol. 1998 Dec 15;19(5):478-83
references: 9859961
references: AIDS. 1999 Jun 18;13(9):1045-9
references: 10397534
references: Microbiol Immunol. 2004;48(10):767-72
references: 15502410
references: AIDS. 2004 Jan 2;18(1):45-9
references: 15090828
references: J Virol Methods. 2009 May;157(2):141-6
references: 19126416
references: J Leukoc Biol. 2010 Apr;87(4):621-6
references: 20130221
references: AIDS Res Hum Retroviruses. 2000 Dec 10;16(18):1939-47
references: 11153076
references: AIDS Res Hum Retroviruses. 2001 Nov 20;17(17):1599-604
references: 11779347
references: N Engl J Med. 1999 May 27;340(21):1614-22
references: 10341273
references: AIDS. 2006 Aug 22;20(13):1727-36
references: 16931937
references: J Virol Methods. 2005 Mar;124(1-2):157-65
references: 15664064
references: BMC Infect Dis. 2005 Mar 21;5:15
references: 15780144
references: J Infect Dis. 2002 May 15;185(10):1409-16
references: 11992275
references: J Infect Dis. 2006 Jul 1;194(1):29-37
references: 16741879
references: J Infect Dis. 1999 Aug;180(2):320-9
references: 10395845
references: Curr HIV Res. 2008 May;6(3):261-6
references: 18473790
references: AIDS. 2010 Jul 17;24(11):1641-9
references: 20543660
references: J Virol Methods. 2010 Mar;164(1-2):135-8
references: 19963009
references: AIDS. 2001 Apr 13;15(6):665-73
references: 11371680
references: Retrovirology. 2009 Feb 12;6:15
references: 19216757
references: Nat Med. 2009 Aug;15(8):893-900
references: 19543283
references: Nat Med. 2003 Jun;9(6):727-8
references: 12754504
references: J Clin Microbiol. 2007 Apr;45(4):1288-97
references: 17314225
references: Nature. 1997 May 8;387(6629):183-8
references: 9144289
references: J Leukoc Biol. 2010 Apr;87(4):575-88
references: 19801499
references: J Acquir Immune Defic Syndr. 2008 Jul 1;48(3):350-4
references: 18545149
references: J Infect Dis. 2000 Jan;181(1):354-7
references: 10608788