title: Establishment and characterization of adenosine deaminase-deficient human T cell lines.
abstract: We have established long term cell lines from a patient with adenosine deaminase (ADA)-deficient severe combined immunodeficiency by stimulation of blood and bone marrow cells with PHA and IL-2 followed by transformation of the activated cells with the human retrovirus HTLV-I. Despite the absence of detectable T cells in the patients blood, cell lines grew that carried the phenotype of mature activated T cells. TJF-2, the line established from blood, was characterized in detail. The concentration of ADA in TJF-2 cells was less than 1% of normal (3.2 U vs 413.0 U). Studies with pharmacologic inhibitors of ADA suggest that the residual adenosine deaminating activity of TJF-2 is from an enzyme distinct from true ADA, a nonspecific aminohydrolyase. Growth of TJF-2 cells was hypersensitive to inhibition by 2'-deoxyadenosine compared to normal T cells (ID50, 55 microM vs greater than 1000 microM). Analysis of 2'-deoxyadenosine-challenged cells showed that TJF-2 cells accumulated significant levels of deoxyadenosine triphosphate, whereas normal T cells did not unless they were also incubated with the ADA inhibitor deoxycoformycin. Southern and Northern blot analysis of these cells revealed a grossly intact ADA gene that produced a normal size ADA mRNA. Yet, despite ADA deficiency, cells of the TJF-2 line were otherwise indistinguishable from HTLV-I-transformed T cells derived from normal donors with respect to dependence on exogenous IL-2 for growth, clonal rearrangement patterns of TCR beta-chain genes, response to PHA, and rapid restoration of cellular volume after hypotonic challenge. The TJF-2 line thus represents a unique HTLV-I-transformed human T cell line exhibiting ADA deficiency and its expected metabolic consequences.
authors.affiliation: Metabolism Branch, National Cancer Institute, Bethesda, MD 20892.
authors.forename: D B
authors.initials: DB
authors.lastname: Kohn
authors.forename: H
authors.initials: H
authors.lastname: Mitsuya
authors.forename: M
authors.initials: M
authors.lastname: Ballow
authors.forename: J E
authors.initials: JE
authors.lastname: Selegue
authors.forename: J
authors.initials: J
authors.lastname: Barankiewicz
authors.forename: A
authors.initials: A
authors.lastname: Cohen
authors.forename: E
authors.initials: E
authors.lastname: Gelfand
authors.forename: W F
authors.initials: WF
authors.lastname: Anderson
authors.forename: R M
authors.initials: RM
authors.lastname: Blaese
mesh_terms: D000243:Adenosine Deaminase / Q000172:deficiency* / Q000235:genetics; D058892:Adenosine Deaminase Inhibitors; D002460:Cell Line; D002461:Cell Line, Transformed; D002472:Cell Transformation, Viral; D002675:Child, Preschool; D003838:Deoxyadenine Nucleotides / Q000378:metabolism; D003839:Deoxyadenosines / Q000494:pharmacology; D015368:Human T-lymphotropic virus 1; D006801:Humans; D007038:Hypotonic Solutions; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007153:Immunologic Deficiency Syndromes / Q000201:enzymology* / Q000235:genetics / Q000276:immunology; D007376:Interleukin-2 / Q000096:biosynthesis / Q000494:pharmacology; D008213:Lymphocyte Activation / Q000187:drug effects; D008297:Male; D009700:Nucleoside Deaminases / Q000172:deficiency*; D010835:Phytohemagglutinins; D012333:RNA, Messenger / Q000302:isolation & purification; D011948:Receptors, Antigen, T-Cell / Q000235:genetics; D013601:T-Lymphocytes / Q000201:enzymology* / Q000276:immunology / Q000378:metabolism
chemical_list: D058892:Adenosine Deaminase Inhibitors; D003838:Deoxyadenine Nucleotides; D003839:Deoxyadenosines; D007038:Hypotonic Solutions; D007143:Immunoglobulin Heavy Chains; D007376:Interleukin-2; D010835:Phytohemagglutinins; D012333:RNA, Messenger; D011948:Receptors, Antigen, T-Cell; D009700:Nucleoside Deaminases; D000243:Adenosine Deaminase; C026600:2'-deoxyadenosine triphosphate