Long-term culture and T cell receptor analysis of T cell clones isolated from a patient with adenosine deaminase deficiency and type I diabetes.
We performed limiting dilution culture of T cells from a patient affected by primary immunodeficiency as a result of complete lack of adenosine deaminase (ADA) activity and also affected by insulin-dependent diabetes mellitus (type I diabetes). Despite the occurrence of immunodeficiency, we were able to raise and grow T cell clones derived from this patient in long-term culture. These T cells displayed ADA enzymatic activity and produced interleukin-2 after engagement of their T cell receptor (TCR)/CD3 complex. We analyzed the TCR repertoire of such clones by nucleotide sequencing of TCR beta chains. The results show that the T cell clones express different V beta but similar J regions. However, the CDR3 regions which are implicated in antigen recognition were found to be heterogeneous.
De Berardinis
P
P
Institute of Protein Biochemistry and Enzymology, CNR, Naples, Italy.
Ombra
M N
MN
Buono
C
C
Toraldo
R
R
Vetrano
F
F
Guardiola
J
J
D000243:Adenosine Deaminase / Q000172:deficiency*; D000595:Amino Acid Sequence; D001483:Base Sequence; D002478:Cells, Cultured; D017931:DNA Primers / Q000737:chemistry; D003922:Diabetes Mellitus, Type 1 / Q000276:immunology*; D006801:Humans; D007153:Immunologic Deficiency Syndromes / Q000276:immunology*; D066298:In Vitro Techniques; D007376:Interleukin-2 / Q000378:metabolism; D008969:Molecular Sequence Data; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics*; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology*; D013997:Time Factors
D017931:DNA Primers; D007376:Interleukin-2; D016693:Receptors, Antigen, T-Cell, alpha-beta; D000243:Adenosine Deaminase