Measurement of Lymphocyte Proliferation: Critical Analysis of Radioactive and Photometric Methods

Immunobiology. 1992 Jun;185(1):78-89. doi: 10.1016/S0171-2985(11)80319-0.

Abstract

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.

Publication types

  • Comparative Study

MeSH terms

  • Autoradiography / methods*
  • Autoradiography / standards
  • Cell Division
  • Cells, Cultured
  • Colorimetry / methods
  • Colorimetry / standards
  • DNA / biosynthesis
  • DNA Replication
  • Humans
  • Immunoenzyme Techniques / standards
  • Lymphocyte Activation*
  • Photometry / methods*
  • Photometry / standards

Substances

  • DNA