6533b82afe1ef96bd128b970

RESEARCH PRODUCT

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

J. Müller-quernheimSibylle PfeiferRosario HeckHeike Wemme

subject

DNA ReplicationLymphocyteImmunologyDehydrogenaseLymphocyte proliferationLymphocyte ActivationImmunoenzyme TechniquesPhotometrymedicineHumansImmunology and AllergyHexosaminidaseCells Culturedchemistry.chemical_classificationbiologyChemistryCell growthDNAHematologyMolecular biologymedicine.anatomical_structureEnzymeBiochemistryMonoclonalbiology.proteinAutoradiographyColorimetryAntibodyCell Division

description

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.

https://doi.org/10.1016/s0171-2985(11)80319-0