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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
immunology and microbiology
Novel application of Ki67 to quantify antigen-specific in vitro lymphoproliferation
Journal of Immunological Methods, Volume 362, No. 1-2, Year 2010
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Description
Antigen-specific proliferation is a critical function of memory T cells that is often utilised to measure vaccine immunogenicity and T cell function. We proposed that measurement of intracellular expression of the nuclear protein, Ki67, could reliably assess specific T cell proliferation in vitro.Ki67 was expressed in CD4+ and CD8+ T cells that had undergone in vitro proliferation after 6-day culture of human whole blood or PBMC with antigens. T cells cultured with no antigen did not express Ki67. When compared to current flow cytometry based proliferation assays, Ki67 detected proliferating cells with greater sensitivity than BrdU incorporation, whereas its sensitivity was similar to dye dilution of Oregon Green (OG), a CFSE derivative. Overall, the magnitude and cytokine expression profile of proliferating T cells detected by Ki67 expression correlated strongly with T cells detected with BrdU or OG. The intra-assay variability of Ki67 proliferation was 2-3% for CD4+ T cells, and 10-16% for CD8+ T cells. Finally, we demonstrate that the Ki67 assay detects tetanus toxoid-specific CD4+ T cell proliferation after infant vaccination with tetanus toxoid (TT).Overall our data suggest that intracellular Ki67 expression provides a specific, quantitative and reproducible measure of antigen-specific T cell proliferation in vitro. © 2010 Elsevier B.V.
Authors & Co-Authors
Soares, Andreia P.
South Africa, Observatory
South African Tuberculosis Vaccine Initiative
Govender, Lerisa
South Africa, Observatory
South African Tuberculosis Vaccine Initiative
Hughes, Jane E.
South Africa, Observatory
South African Tuberculosis Vaccine Initiative
Mavakla, Wendy
South Africa, Observatory
South African Tuberculosis Vaccine Initiative
de Kock, Marwou
South Africa, Observatory
South African Tuberculosis Vaccine Initiative
Barnard, Charlene
South Africa, Observatory
South African Tuberculosis Vaccine Initiative
Pienaar, Bernadette
South Africa, Observatory
South African Tuberculosis Vaccine Initiative
Janse van Rensburg, Esme
South Africa, Observatory
South African Tuberculosis Vaccine Initiative
Jacobs, Gail
South Africa, Observatory
South African Tuberculosis Vaccine Initiative
Khomba, Gloria
South Africa, Observatory
South African Tuberculosis Vaccine Initiative
Stone, Lynnette
South Africa, Observatory
South African Tuberculosis Vaccine Initiative
Abel, Brian
South Africa, Observatory
South African Tuberculosis Vaccine Initiative
Scriba, Thomas J.
South Africa, Observatory
South African Tuberculosis Vaccine Initiative
Hanekom, Willem Albert
South Africa, Observatory
South African Tuberculosis Vaccine Initiative
Statistics
Citations: 14
Authors: 14
Affiliations: 1
Identifiers
Doi:
10.1016/j.jim.2010.08.007
ISSN:
00221759
Research Areas
Cancer
Maternal And Child Health
Study Approach
Quantitative