Intracellular glutathione in the peripheral blood cells of HIV-infected patients: failure to show a deficiency.
OBJECTIVE
To determine whether HIV-infected patients have a deficiency of intracellular glutathione (GSH) in peripheral blood mononuclear cells (PBMC) and erythrocytes.


METHODS
Initial experiments determining the stability of intracellular GSH preceded the measurement of GSH levels in 33 HIV-positive patients and 40 control subjects within 1 h of isolation of their blood cells. In addition, the susceptibility of erythrocytes to dapsone hydroxylamine-induced methaemoglobinaemia was evaluated.


METHODS
GSH levels were determined by an high-performance liquid chromatography method utilizing a fluorescent probe, monobromobimane. The bimane-GSH adduct formed in PBMC was also characterized by mass spectrometry. Methaemoglobin formation on exposure to dapsone hydroxylamine was determined spectrophotometrically.


RESULTS
GSH levels remained stable for only 1 h after cell isolation, thereafter showing a decrease of 20 and 60% at 4 and 24H, respectively, There was no difference in the GSH levels in PBMC and erythrocytes of the HIV-positive patients compared with controls. The GSH levels were not related to the disease stage or to CD4+ cell counts. There was no difference in GSH levels in PBMC taken from trimethoprim-sulphamethoxazole-hypersensitive and non-hypersensitive patients. Methaemoglobinaemia on exposure of erythrocytes to dapsone hydroxylamine was concentration-dependent, but there was no significant difference between patients and controls.


CONCLUSIONS
In contrast to previous studies, no deficiency of intracellular GSH in the PBMC and erythrocytes of HIV-infected patients was found. The discrepancy between studies may be methodological reflecting the instability of GSH, which requires prompt sample analysis.
Pirmohamed
M
M
Department of Pharmacology and Therapeutics, University of Liverpool, UK.
Williams
D
D
Tingle
M D
MD
Barry
M
M
Khoo
S H
SH
O'Mahony
C
C
Wilkins
E G
EG
Breckenridge
A M
AM
Park
B K
BK
D000328:Adult; D000368:Aged; D018791:CD4 Lymphocyte Count; D003622:Dapsone / Q000031:analogs & derivatives / Q000494:pharmacology; D004342:Drug Hypersensitivity; D004912:Erythrocytes / Q000737:chemistry* / Q000187:drug effects; D005978:Glutathione / Q000097:blood* / Q000172:deficiency; D015658:HIV Infections / Q000097:blood* / Q000276:immunology; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000737:chemistry*; D008297:Male; D008708:Methemoglobinemia / Q000139:chemically induced; D008875:Middle Aged; D015662:Trimethoprim, Sulfamethoxazole Drug Combination / Q000009:adverse effects
D015662:Trimethoprim, Sulfamethoxazole Drug Combination; D003622:Dapsone; D005978:Glutathione; C018209:4-amino-4'-hydroxylaminodiphenylsulfone