US EPA

5372722 

Journal Article 

Differential Expression of Glutathione Transferases by Native and Cultured Human Lymphocytes 

Jones, SM; Idle, JR; Hirom, PC 

1988 

Yes 

Biochemical Pharmacology
ISSN: 0006-2952
EISSN: 1873-2968 

37 

23 

4586-4590 

English 

10.1016/0006-2952(88)90679-X 

The differential expression of glutathione transferases (GSTs) by native and cultured human lymphocytes was studied using Western immunoblot techniques. Interleukin-2 (IL2) and phytohemagglutinin (PHA) stimulated T-cell cultures and Epstein Barr virus (EBV) transformed B-cell lines were grown from lymphocytes freshly isolated from normal human venous blood. Cytosolic fractions from the cultures were analyzed by SDS polyacrylamide gel electrophoresis, and the proteins were transferred on nitrocellulose paper by electroblotting and probed using antiserum to human hepatic GSTs (mu, epsilon, and gamma). Further evaluation employed measurement of 1-chloro-2,4-dinitrobenzene (CDNB) conjugating activity and the enzyme linked immunosorbent assay (ELISA) for human GST(mu). Western blot analysis showed no immunocomplexing between human hepatic GST(epsilon) and any of the lymphocyte samples. Antiserum to human lung GST(gamma) formed complexes with mononuclear leukocytes, B-leukocytes, lymphoblastoid B-cell lines, PHA and IL2 stimulated T-cell lines but not with native T-cells. Immunocomplexing with antibodies to human hepatic GST(mu) was weakly positive for EBV transformed B-cell lines. B-cell line fractions with antibodies to GST(mu) and GST(gamma) showed increased conjugation of CDNB relative to IL2 dependent T-cell fractions. The determination of GST(mu) expression by the ELISA method yielded 36, 36, 42, and 36 percent positives for cytosolic fractions of whole blood, resting lymphocytes, IL2-dependent T-cells, and B-cell lines respectively. The authors conclude that human GSTs can be differentially expressed in cultured lymphocyte subsets.