Measurement of lymphocyte proliferation: critical analysis of radioactive and photometric methods.
Different methods of lymphocyte proliferation are compared to identify a non-radioactive alternative to 3H-thymidine-test. The enzymatic assays evaluating the turnover of mitochondrial dehydrogenases (MTT-test) and lysosomal hexosaminidase (NAG-test) proved not sensitive enough to substitute for 3H-thymidine incorporation. The incorporation of the nucleotide analog 5-bromodeoxyuridine (BrdU) can be exploited using an ELISA-system (enzyme linked immunosorbent assay) employing a monoclonal anti-BrdU antibody to measure cell proliferation. An optimized test protocol of the BrdU-ELISA which fulfills the requirements for a sensitive and practicable non-radioactive alternative to 3H-thymidine-test is presented.
Wemme
H
H
Third Department of Internal Medicine, Johannes Gutenberg University, Mainz, Germany.
Pfeifer
S
S
Heck
R
R
Müller-Quernheim
J
J
D001345:Autoradiography / Q000379:methods* / Q000592:standards; D002455:Cell Division; D002478:Cells, Cultured; D003124:Colorimetry / Q000379:methods / Q000592:standards; D004247:DNA / Q000096:biosynthesis; D004261:DNA Replication; D006801:Humans; D007124:Immunoenzyme Techniques / Q000592:standards; D008213:Lymphocyte Activation; D010783:Photometry / Q000379:methods* / Q000592:standards
D004247:DNA