Inhibition of human immunodeficiency virus-1 (HIV-1) by beta-chemokine analogues in mononuclear cells from HIV-1-infected patients with active tuberculosis

Abstract

Tuberculosis (TB) enhances human immunodeficiency virus-1 (HIV-1) activity in patients with dual HIV-1/TB infection. Therapies that control augmentations of HIV-1 activity at sites of Mycobacterium tuberculosis (MTB) infection may be useful in inhibition of viral expansion. Regulated upon activation, normal T-cell expressed and secreted (RANTES) analogues (AOP and NNY) are potent in inhibiting the entry of primary HIV-1 isolates into host mononuclear cells. These analogues were used to inhibit MTB-induced HIV-1 entry in blood monunuclear cells (PBMC) from patients with pulmonary TB, and pleural fluid mononuclear cells (PFMC) from patients with pleural TB. PBMC or PFMC were cultured with and without MTB in presence and absence of RANTES analogues. HIV-1 strong stop DNA was assessed by real-time polymerase chain reaction (PCR) as a measure of infection. CCR5 mRNA was assessed by real-time reverse transcription (RT)-PCR and by immunostaining and FACS analysis. HIV-1 infection was induced by MTB in vitro in PBMC from the majority (14 of 20) of HIV-1/TB subjects, and new infection was inhibited by AOP- or NNY-RANTES. HIV-1 infection was also inhibited by these reagents in MTB-induced PFMC from three of three patients with pleural TB. Expression of CCR5 mRNA was significantly induced by MTB in PBMC from patients with pulmonary TB. Further, expression of CCR5 was higher in PFMC compared to PBMC from patients with pleural TB. Also, CCR5 was fourfold higher on CD14(+) pleural mononuclear cells than on CD4(+) lymphocytes. Blocking new HIV-1 infection of mononuclear cells may be useful in control of HIV-1 during dual HIV-1/TB infection.

Fig. 1

Peripheral blood mononuclear cells (PBMC) from human immunodeficiency virus-1 (HIV-1)/tuberculosis (TB) patients were prepared and cultured in the absence (–), or presence of MTB (1 : 1; bacteria/cell). (a) HIV-1 SSDNA was measured by real-time polymerase chain reaction (PCR) and corrected for the housekeeping gene β-globin in each sample. HIV-1 SS DNA in PBMC was induced in the presence compared to the absence of Mycobacterium tuberculosis (MTB) in 14 of 20 subjects. In each subject the time-point of maximal induction of HIV-1 SSDNA by MTB was considered. In only 1 of 14 this time-point was at 120 h, in the rest the maximal induction was at 96 h. (b) HIV-1 SSDNA was assessed in the absence and presence of MTB in PBMC cultures and in the presence of MTB and AOP-RANTES or NNY-RANTES. HIV-1 SS DNA under each culture condition is expressed as a percentage of that in MTB stimulated PBMC. Percentage expression of HIV-1 DNA in MTB-induced PBMC in the presence of AOP- (▪) and NNY-RANTES (), and in PBMC cultured with medium alone (□) were compared to MTB-induced PBMC () (considered as 100%). Data presented are from 10 of 14 subjects who had inhibition by RANTES analogues. *P < 0·05.

Fig. 2

Peripheral blood mononuclear cells (PBMC) from human immunodeficiency virus-1 (HIV-1)/tuberculosis (TB) patients were prepared and cultured in the absence (–), or presence of Mycobacterium tuberculosis (MTB) (1 : 1; bacteria/cell) for up to 120 h. Expression of CCR5 mRNA was assessed by real-time reverse transcription-polymerase chain reaction (RT-PCR), and corrected to expression of the ribosomal 18 s (R18). CCR5 was induced by MTB (▪) compared to media alone (□). **P < 0·001.

Fig. 3

Fresh pleural fluid mononuclear cells (PFMC) and peripheral blood mononuclear cells (PBMC) from human immunodeficiency virus-1 (HIV-1)/tuberculosis (TB) patients with pleural TB were prepared. (a) PFMC from three donors (P1–3) were cultured with MTB (□) alone or in the presence of AOP- (▪) and NNY- RANTES (), or with media alone (−) (). HIV SS DNA was assessed and expressed as a percentage of SS DNA in MTB-stimulated PFMC (given 100%) in each individual. Inhibition in HIV-1 SS DNA in the presence of AOP and NYY-RANTES was expressed as a percentage of SS DNA in the presence of MTB alone. (b) Expression of CCR5 on CD14 (left) and CD4 (right) PFMC () and autologous PBMC (▪) from nine HIV-1/TB patients was assessed by immunostaining and FACS analysis. **P < 0·001; *P < 0·05.