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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.
D B Kohn, and
H Mitsuya, and
M Ballow, and
J E Selegue, and
J Barankiewicz, and
A Cohen, and
E Gelfand, and
W F Anderson, and
R M Blaese
September 1984,
Molecular and cellular biology,
D B Kohn, and
H Mitsuya, and
M Ballow, and
J E Selegue, and
J Barankiewicz, and
A Cohen, and
E Gelfand, and
W F Anderson, and
R M Blaese
March 1982,
The Journal of biological chemistry,
D B Kohn, and
H Mitsuya, and
M Ballow, and
J E Selegue, and
J Barankiewicz, and
A Cohen, and
E Gelfand, and
W F Anderson, and
R M Blaese
January 1984,
Advances in experimental medicine and biology,
D B Kohn, and
H Mitsuya, and
M Ballow, and
J E Selegue, and
J Barankiewicz, and
A Cohen, and
E Gelfand, and
W F Anderson, and
R M Blaese
June 1980,
The Journal of biological chemistry,
D B Kohn, and
H Mitsuya, and
M Ballow, and
J E Selegue, and
J Barankiewicz, and
A Cohen, and
E Gelfand, and
W F Anderson, and
R M Blaese
January 1977,
Ciba Foundation symposium,
D B Kohn, and
H Mitsuya, and
M Ballow, and
J E Selegue, and
J Barankiewicz, and
A Cohen, and
E Gelfand, and
W F Anderson, and
R M Blaese
September 1976,
Cancer research,
D B Kohn, and
H Mitsuya, and
M Ballow, and
J E Selegue, and
J Barankiewicz, and
A Cohen, and
E Gelfand, and
W F Anderson, and
R M Blaese
September 1975,
Experimental cell research,
D B Kohn, and
H Mitsuya, and
M Ballow, and
J E Selegue, and
J Barankiewicz, and
A Cohen, and
E Gelfand, and
W F Anderson, and
R M Blaese
September 1989,
Somatic cell and molecular genetics,
D B Kohn, and
H Mitsuya, and
M Ballow, and
J E Selegue, and
J Barankiewicz, and
A Cohen, and
E Gelfand, and
W F Anderson, and
R M Blaese
February 1984,
The Journal of biological chemistry,
D B Kohn, and
H Mitsuya, and
M Ballow, and
J E Selegue, and
J Barankiewicz, and
A Cohen, and
E Gelfand, and
W F Anderson, and
R M Blaese
August 2001,
The Journal of pharmacology and experimental therapeutics,
