Paradoxical expression of adenosine deaminase in T cells cultured from a patient with adenosine deaminase deficiency and combine immunodeficiency.

Abstract

T lymphocytes cultured from a patient (T.D.) with adenosine deaminase (ADA) deficiency expressed ADA activity in the normal range, inconsistent with her severe immunodeficiency, metabolic abnormalities, and with the absence of ADA activity in her B lymphocytes and other nucleated hematopoietic cells.

Expression pattern of CD2 locus control region containing retroviral vectors in hemopoietic cells in vitro and in vivo

“…34 Unexpectedly, the average expression levels of most CD2-LCR containing retroviruses were quite similar to that of LgAL(⌬Mo+PyF101). The N8F vector, however, exhibited significantly lower hADA expression levels.…”

Tandem mass spectrometry, but not T-cell receptor excision circle analysis, identifies newborns with late-onset adenosine deaminase deficiency

“…Red blood cells from patient P4 were found to have no ADA enzyme activity, and white blood cells were found to have levels of activity that were about 3% to 4% of normal values. According to previous reports, 8,18,20 healthy subjects with partial ADA deficiency have levels of ADA activity in white cells that are 5% to 70% of those of control subjects, whereas patients with SCID have levels of ADA activity that are less than 1% to 2% of those of control subjects.…”

“…17 Method C was a radiochemical thin-layer chromatographic assay. 18 Normal values for ADA activity in erythrocytes and white blood cells are 63.0 6 41.4 and 1197 6 516 nmol/h/mg protein, respectively. Untreated patients with new diagnoses of early-onset ADA-SCID and those with delayedand late-onset ADA-SCID usually have less than 1% of the normal activity of ADA in red and white blood cells.…”

Clustered Charged Amino Acids of Human Adenosine Deaminase Comprise a Functional Epitope for Binding the Adenosine Deaminase Complexing Protein CD26/Dipeptidyl Peptidase IV

“…1) were available from a previous study (23). Alanine substitutions were introduced into the wild type human ADA cDNA by PCR mutagenesis essentially as described (28,29). All final ADA cDNA PCR products were cloned into pBluescript II KS and fully sequenced using the ABI 377 PRISM DNA Sequencing Instrument (Applied Biosystems, Foster City, CA).…”

“…[ 35 S]Methionine-labeled ADA was generated in vitro from ADA cDNAs in pBluescript, using the TNT Coupled Wheat Germ Extract System (Promega, Madison, WI). Translation products were analyzed by SDS-PAGE and fluorography, and they were also electrophoresed on cellulose acetate and stained for ADA activity in situ as described previously (28).…”