T lymphocyte activation results in an increased expression of beta-1,4-galactosyltransferase: phorbol ester induces a similar enhancement in the absence of mitosis.
We previously showed that in vitro activated human T lymphocytes expressed increased amounts of beta-1,6-branched N-linked oligosaccharides (Lemaire S etal. (1994) J Biol Chem269: 8069-74), which have been proposed to participate in the regulation of the immune process. In the present paper, we compared the activity and expression of beta-1,4-galactosyltransferase (GalT), one of the glycosyltransferases involved in the biosynthesis of these beta-1,6-branched N-linked oligosaccharides, before and after in vitro activation of T lymphocytes after a 40h treatment with a mixture of phorbol 12-myristate 13-acetate and Phaseolus vulgaris lectin. After treatment, the enzymatic activity of the GalT was significantly increased and immunoblot experiments performed with a monoclonal antibody to human GalT showed an increased intensity of the GalT band at 49 kDa, attributable to an enhancement of GalT mRNA level, as shown by Northern blots. However, treatment of the same T-lymphocytes by phorbol ester alone, which is unable to induce mitosis, resulted in a comparable increase of the expression of GalT. Moreover, these phorbol ester-treated T lymphocytes, analysed by flow cytometry exhibited a two-fold increase in the expression of GalT. Finally, confocal fluorescence microscopy performed on all T lymphocytes (treated or not) showed that the flow cytometric signal of GalT originates from intracellular, Golgi-associated antigen only since no surface GalT was detected.
Lemaire
S
S
UFR Biomédicale des Saints-Péres, Paris, France.
Derappe
C
C
Pasqualetto
V
V
Mrkoci
K
K
Berger
E G
EG
Aubéry
M
M
Néel
D
D
D005434:Flow Cytometry; D006801:Humans; D008213:Lymphocyte Activation / Q000187:drug effects*; D008214:Lymphocytes / Q000187:drug effects / Q000201:enzymology*; D008938:Mitosis / Q000187:drug effects*; D009237:N-Acetyllactosamine Synthase / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D010835:Phytohemagglutinins / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014158:Transcription, Genetic
D010835:Phytohemagglutinins; C100156:leukoagglutinins, plants; D009237:N-Acetyllactosamine Synthase; D013755:Tetradecanoylphorbol Acetate