6533b854fe1ef96bd12aeb03

RESEARCH PRODUCT

Measurement of T Cell Activation After 16‐hr In Vitro Stimulation with Concanavalin A

Borros M. Arneth

subject

CD4-Positive T-LymphocytesCell ExtractsErythrocytesTime FactorsHistologyLymphocyteCD3T cellCD8-Positive T-LymphocytesLymphocyte ActivationBiochemistryImmunophenotypingFlow cytometryConcanavalin AmedicineHumansCytotoxic T cellWhole bloodbiologymedicine.diagnostic_testAntibodies MonoclonalCD28General MedicineFlow CytometryMolecular biologyLymphocyte SubsetsMedical Laboratory Technologymedicine.anatomical_structureConcanavalin Abiology.protein

description

A flow cytometry assay that can be used to directly determine the proportion of activated T lymphocytes in human whole blood samples after stimulation with concanavalin A is presented here. Human whole blood is incubated with fluorescently labeled antibodies (against CD3, CD4, CD8, and CD69), erythrocytes are then lysed, and the samples are analyzed using a flow cytometer. The assay presented is able to differentiate between CD4+ and CD8+ T lymphocytes. Thus, it is possible to quantify both lymphocyte populations in parallel, as well as the respective proportions of activated T lymphocytes, all from one sample. An additional advantage of this assay is that it was developed to assay whole blood, thereby reducing the likelihood of introducing pre-analytic error and facilitating the calculation of quantitative ratios. Thus, the method can be used for both in vitro and ex vivo experiments.

yearjournalcountryeditionlanguage
2010-01-14Current Protocols in Cytometry
https://doi.org/10.1002/0471142956.cy0628s51