Production of TNFalpha and IL-6 by activated whole blood from HIV-1 infected patients detected by a one-stage procedure: relationship with the phenotype of HIV-1 isolates.
Diluted whole blood (WB) culturing may be the most appropriate milieu in which to study cytokine production in vitro. We tested TNFalpha and IL-6 production using small volumes of WB (25 microl) from HIV-1 positive patients with a one-step procedure that combines WB stimulation with LPS, PHA and cytokine measurement. We studied 49 patients without secondary infection or at distance of secondary infection staged according to the 1993 classification of the CDC and 12 healthy seronegative subjects. Heparinized blood from 5 control subjects had been collected sequentially during a period of 5 months. The individual variations of TNFalpha and IL-6 production were limited for all these individuals. In 1 out of 20 CDC group A patients, 6 out of 17 CDC group B patients and 3 out of 12 CDC group C patients, we obtained higher values of TNFalpha than the mean + 2 S.D. of the control group. In 3 out of 20 CDC group A patients, 1 out of 17 CDC group B patients without AIDS and 5 out of 12 CDC group C patients, the TNFalpha values were lower than the mean - 2 S.D. of the control group. Low IL-6 values were obtained in 1 out of 20 CDC group A patients and 1 out of 17 CDC group B patients and 3 out of 12 CDC group C patients. There was no correlation between TNFalpha production in vitro and plasma level of TNFalpha. We found no correlation between the levels of cytokines and monocyte count or between the levels of cytokines and CD4 T-cell count in peripheral blood. Our data point out a disarray in TNFalpha and IL-6 production by WB from HIV-1 infected patients. The relationship between the disarray of cytokine production and cytopathogenicity of HIV-1 isolates in the P4 cell line was investigated in this study. We found a correlation between the high level of TNFalpha produced by WB and the phenotype of HIV-1 isolates isolated from patients. The one-stage procedure used in this work is of potential value to investigate the activation status of cells for monitoring HIV-1 positive individuals and predicting HIV-1 phenotype.
Benyoucef
S
S
Laboratoire de Virologie, CHU, Lille, France.
Hober
D
D
Shen
L
L
Ajana
F
F
De Groote
D
D
Bocket-Mouton
L
L
Gérard
Y
Y
Lion
G
G
Vilain
V
V
Wattré
P
P
D001773:Blood Cells / Q000276:immunology*; D018791:CD4 Lymphocyte Count; D015496:CD4-Positive T-Lymphocytes; D018929:Cell Culture Techniques; D018920:Coculture Techniques; D003588:Cytopathogenic Effect, Viral; D005260:Female; D015658:HIV Infections / Q000276:immunology* / Q000821:virology; D015497:HIV-1 / Q000472:pathogenicity*; D006367:HeLa Cells; D006493:Heparin; D006801:Humans; D007124:Immunoenzyme Techniques; D015850:Interleukin-6 / Q000096:biosynthesis*; D007958:Leukocyte Count; D008070:Lipopolysaccharides / Q000494:pharmacology; D008297:Male; D010641:Phenotype; D010835:Phytohemagglutinins / Q000494:pharmacology; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis*
D015850:Interleukin-6; D008070:Lipopolysaccharides; D010835:Phytohemagglutinins; D014409:Tumor Necrosis Factor-alpha; D006493:Heparin