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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
immunology and microbiology
Impact of blood processing variations on natural killer cell frequency, activation, chemokine receptor expression and function
Journal of Immunological Methods, Volume 366, No. 1-2, Year 2011
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Description
Understanding the role of natural killer (NK) cells in human disease pathogenesis is crucial and necessitates study of patient samples directly ex vivo. Manipulation of whole blood by density gradient centrifugation or delays in sample processing due to shipping, however, may lead to artifactual changes in immune response measures. Here, we assessed the impact of density gradient centrifugation and delayed processing of both whole blood and peripheral blood mononuclear cells (PBMC) at multiple timepoints (2-24. h) on flow cytometric measures of NK cell frequency, activation status, chemokine receptor expression, and effector functions. We found that density gradient centrifugation activated the NK cells and modified the chemokine receptor expression. Delays in processing beyond 8. h activated NK cells in PBMC but not in whole blood. Likewise, processing delays decreased chemokine receptor (CCR4 and CCR7) expression in both PBMC and whole blood. Finally, delays in processing PBMC were associated with a decreased ability of NK cells to degranulate (as measured by CD107a expression) or secrete cytokines (IFN-γ and TNF-α). In summary, our findings suggest that density gradient centrifugation and delayed processing of PBMC can alter measures of clinically relevant NK cell characteristics including effector functions; and therefore should be taken into account in designing clinical research studies. © 2011 Elsevier B.V.
Authors & Co-Authors
Naranbhai, Vivek
South Africa, Durban
The Nelson R. Mandela Medical School
Bartman, Patricia
South Africa, Pietermaritzburg
Edendale Non-european Hospital
Ndlovu, Dudu
South Africa, Pietermaritzburg
Edendale Non-european Hospital
Ramkalawon, Pamela
South Africa, Durban
The Nelson R. Mandela Medical School
Ndung'u, Thumbi P.
South Africa, Durban
The Nelson R. Mandela Medical School
Wilson, Douglas P.K.
South Africa, Pietermaritzburg
Edendale Non-european Hospital
Altfeld, Marcus A.
South Africa, Durban
The Nelson R. Mandela Medical School
United States, Cambridge
Harvard University
Carr, William Henry
South Africa, Durban
The Nelson R. Mandela Medical School
United States, Cambridge
Harvard University
Statistics
Citations: 39
Authors: 8
Affiliations: 3
Identifiers
Doi:
10.1016/j.jim.2011.01.001
ISSN:
00221759
Research Areas
Health System And Policy