An approach to a selection system for adenosine-deaminase-positive (ADA+) cells and detection of rat ADA+ “revertants”
✍ Scribed by Rochelle Hischhorn; Amy Ellenbogen; Frank Martiniuk
- Publisher
- John Wiley and Sons
- Year
- 1985
- Tongue
- English
- Weight
- 754 KB
- Volume
- 123
- Category
- Article
- ISSN
- 0021-9541
No coin nor oath required. For personal study only.
✦ Synopsis
We have substituted deoxyadenosine or adenosine for hypoxanthine in t h e standard HAT selection system in an attempt to select for ADA-normal (ADA+) cells. ADA-human lymphoid line cells could not utilize deoxyadenosine as an alternative to hypoxanthine as a purine source (DAT) and failed to grow but were only somewhat inhibited in growth when adenosine was substituted for hypoxanthine (AAT). In contrast, ADA+ cells utilized adenosine or deoxyadenosine as efficiently as hypoxanthine as a purine source. Growth in DAT, but not in HAT, of an artificial mixture of one ADA+ human lymphoid cells in 1,000 ADA-cells resulted in enrichment of ADA+ cells to 25-86% of total cells. When we grew a rat ADA-cell line in two variations of the DAT system, we detected at least three electrophoretically different ADA+ patterns, one of which corresponded to normal rat ADA. These could represent "revertants." Genetic deficiency of the enzyme adenosine deaminase (ADA) gives rise to a fatal infantile syndrome of severe combined immunodeficiency (SCID) . Transplantation with bone marrow from a histocompatible, related donor leads