Different pathways for deoxyguanosine toxicity in T-lymphocytes of various developmental stages.
The basis of the selective cellular immunodeficiency which occurs in patients with purine nucleoside phosphorylase (PNP) deficiency still is not completely understood. We studied the mechanism of deoxyguanosine (dGuo) toxicity in proliferating lymphoid T-cells of different maturation stage, i.e. in T-cells of adult peripheral blood and cord blood and in CD3+ and CD3- subfractions of thymocytes. The mitogen-induced proliferation of T-cells from peripheral blood and cord blood and of CD3+ and CD3- subfractions of thymocytes. The mitogen-induced proliferation of T-cells from peripheral blood and cord blood and of CD3+ thymocytes, as well as the spontaneous proliferation of CD3- thymocytes, are inhibited by dGuo. CD3+ and CD3- thymocytes are significantly more sensitive to dGuo than T-cells from peripheral blood or cord blood. Among the thymocyte subfractions CD3- thymocytes appeared to be extremely sensitive. In all cell types studied, inhibition of proliferation is accompanied by intracellular increases in both guanosine triphosphate (GTP) and deoxyguanosine triphosphate (dGTP) concentrations. By use of the PNP inhibitor 8-aminoguanosine, or the metabolites hypoxanthine or deoxycytidine, the metabolism of dGuo could be selectively directed to the formation of GTP or to dGTP. Based on the pattern of rescue from dGuo intoxication under these different metabolic conditions we conclude that in CD3- thymocytes dGuo toxicity is mediated by dGTP. In all other cell types studied GTP mediates dGuo intoxication. Altogether the results show that during the maturation from immature thymocytes to mature peripheral blood T-cells a shift occurs in the pattern of dGuo toxicity since dGuo toxicity in the former is primarily caused via the dCyd kinase pathway, and in the latter mainly the degradation route is involved. Since in PNP deficiency mature T-cells do occur in the peripheral blood, we must conclude that some cells escape the stage of T-cell maturation in the thymus which is extremely sensitive to dGuo. Furthermore, the results imply that as far as T-cell development in the normal thymus is concerned, survival and death of cells might be regulated by local (deoxy) nucleoside availability.
Scharenberg
J G
JG
Department of Immunology, University Hospital for Children and Youth Het Wilhelmina Kinderziekenhuis, Utrecht, The Netherlands.
Rijkers
G T
GT
Spaapen
L J
LJ
Toebes
E A
EA
Rijksen
G
G
Duran
M
M
Staal
G E
GE
Zegers
B J
BJ
D000328:Adult; D000945:Antigens, Differentiation, T-Lymphocyte; D017252:CD3 Complex; D002648:Child; D003841:Deoxycytidine / Q000494:pharmacology; D003849:Deoxyguanosine / Q000633:toxicity*; D003854:Deoxyribonucleotides / Q000097:blood; D005312:Fetal Blood / Q000166:cytology; D006801:Humans; D019271:Hypoxanthine; D007042:Hypoxanthines / Q000494:pharmacology; D066298:In Vitro Techniques; D007231:Infant, Newborn; D008213:Lymphocyte Activation / Q000187:drug effects; D011683:Purine-Nucleoside Phosphorylase / Q000097:blood; D011948:Receptors, Antigen, T-Cell; D013601:T-Lymphocytes / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism
D000945:Antigens, Differentiation, T-Lymphocyte; D017252:CD3 Complex; D003854:Deoxyribonucleotides; D007042:Hypoxanthines; D011948:Receptors, Antigen, T-Cell; D003841:Deoxycytidine; D019271:Hypoxanthine; D011683:Purine-Nucleoside Phosphorylase; D003849:Deoxyguanosine