Evaluation of adenosine deaminase assay for analyzing T-lymphocyte density in vitro.
The proliferative capacity of T cells in response to various stimuli is commonly determined by radioactive assay based on incorporation of [3H]thymidine ([3H]TdR) into newly synthesized DNA. In order to assess techniques for application in laboratories where radioactive facilities are not present, an alternative method was tested. As an alternative, T-cell proliferation was measured by spectrophotometrically analyzing the presence of an enzyme adenosine deaminase in lymphocytes and also using a standard XTT assay. Jurkat (human) T-cell line (clone E6.1) was used for lymphocyte population. The Jurkat cell concentration was adjusted according to different cell densities and enzyme activity was determined. Cells were also seeded in complete medium up to 72 h and harvested for estimation of enzyme activity. A significant correlation between the standard cell-proliferation assay and adenosine deaminase assay was observed. The present study indicates that the assay of adenosine deaminase is a reliable and accurate method for measuring proliferation of T lymphocytes.
Kainthla
Rani Poonam
RP
Biochemistry Research Laboratory, Central India Institute of Medical Sciences, 88/2, Bajaj Nagar, Nagpur 440010, Maharashtra, India.
Kashyap
Rajpal Singh
RS
Prasad
Sweta
S
Purohit
Hemant J
HJ
Taori
Giridhar M
GM
Daginawala
Hatim F
HF
D000243:Adenosine Deaminase / Q000378:metabolism*; D001681:Biological Assay / Q000379:methods*; D002452:Cell Count; D049109:Cell Proliferation; D006801:Humans; D019169:Jurkat Cells; D013601:T-Lymphocytes / Q000166:cytology* / Q000201:enzymology*; D013997:Time Factors
D000243:Adenosine Deaminase