In vivo overexpression of Core2 N-acetylglucosaminyltransferase prevents repopulation of the bone marrow with colony forming cells but fails to affect normal T cell development.
UDP-GlcNAc:Galbet1 --> 3GalNAc-R beta1 --> 6N-acetylglucosaminyltransferase (Core2 N-acetyl-glucosaminyltransferase, C2GnT; EC 2.4.1.102) forms beta1 --> 6N-acetyl-glucosaminyl linkages in O-glycoproteins and creates branches for the addition of N-acetyl-lactosamine antennae. Changes in C2GnT activity have been associated with immune disorders, malignancies, and T-cell ontogeny. In this study, we used SCID (severe combined immune deficiency) mice to determine the effects of C2GnT overexpression on hemopoiesis, and in particular, on thymocyte development. BALB/c bone marrow cells transfected with C2GnT using the retroviral murine stem cell vector were used to repopulate SCID mice. Mice were analysed 3 weeks to 3 months after bone marrow transfer. Elevated levels of C2GnT activity in bone marrow, spleen, and thymus from mice repopulated with C2GnT transfected bone marrow cells indicated that C2GnT was overexpressed in recipient mice. In C2GnT repopulated mice, up to 50% of T cells showed an increase in CD43 130-kDa expression, compared with T cells from control animals, indicative of an elevated C2GnT activity in these cells. Furthermore, T-cell subset numbers appeared to be normal, suggesting that C2GnT overexpression did not alter T-cell ontogeny. Interestingly, C2GnT overexpression negatively affected the repopulation of myeloid cells. Only insignificant numbers of interleukin-3/granulocyte-macrophage colony stimulating factor (IL-3/GM-CSF) responsive bone marrow cells were found to be retrovirally transfected in C2GnT repopulated mice, whereas up to 50% of IL-3/GM-CSF responsive bone marrow cells were found to be retrovirally transfected in corresponding controls. These data indicate that in vivo overexpression of C2GnT negatively interferes with the myeloid differentiation pathway but does not affect T-cell development.
Fellinger
W J
WJ
The Biomedical Research Centre, University of British Columbia, Vancouver, Canada.
Barran
P
P
Merkens
H
H
Corbel
S Y
SY
Ziltener
H J
HJ
D000818:Animals; D015703:Antigens, CD; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D015971:Gene Expression Regulation, Enzymologic / Q000276:immunology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D007377:Interleukin-3 / Q000494:pharmacology; D051917:Leukosialin; D008198:Lymph Nodes / Q000166:cytology / Q000201:enzymology / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D016513:Mice, SCID; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics*; D009355:Neomycin / Q000494:pharmacology; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D012190:Retroviridae / Q000235:genetics; D012795:Sialoglycoproteins / Q000235:genetics; D013154:Spleen / Q000166:cytology / Q000201:enzymology / Q000276:immunology; D016176:T-Lymphocyte Subsets / Q000166:cytology* / Q000502:physiology; D013950:Thymus Gland / Q000166:cytology / Q000201:enzymology / Q000276:immunology; D014162:Transfection
D015703:Antigens, CD; D007377:Interleukin-3; D051917:Leukosialin; D011500:Protein Synthesis Inhibitors; D012795:Sialoglycoproteins; C497863:Spn protein, mouse; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D017351:N-Acetylglucosaminyltransferases; C053024:beta-1,3-galactosyl-O-glycosyl-glycoprotein beta-1,6-acetylglucosaminyl transferase; D009355:Neomycin