Rapid flow cytometry based cytotoxicity assay for evaluation of NK cell function.

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dc.contributor.authorMhatre, Snehal
dc.contributor.authorMadkaikar, Manisha
dc.contributor.authorGhosh, Kanjaksha
dc.contributor.authorDesai, Mukesh
dc.contributor.authorPujari, Vaishali
dc.contributor.authorGupta, Maya
dc.date.accessioned2014-12-12T10:30:57Z
dc.date.available2014-12-12T10:30:57Z
dc.date.issued2014-10
dc.description.abstractAssessment of natural killer cells (NK-cell) cytotoxicity is used not only in research settings but is also important in diagnosis of various diseases. NK-cell cytotoxicity assays are based on measurement of target cells killed by cytotoxic cells analyzed either by chromium (51Cr) release assay or flow cytometry. Both these methods use peripheral blood mononuclear cells (PBMC) or pure NK-cell population and hence require large volume of blood sample which is difficult to obtain in pediatric patients and patients with cytopenia. Hence, a flow cytometric assay was designed to determine NK cell activity using whole blood, eliminating the need for isolation of PBMCs or pure NK cells. This assay is based on a dual fluorescent staining of target cells (K562 cell line). The DIOC18 dye labeled K562 cells are incubated with whole blood and then counterstained with 7-AAD enabling the measurement of dead target cell and then percent cytotoxicity is calculated. This study compared the NK cell cytotoxicity using PBMC and whole blood in clinically relevant samples. There was no significant difference between two assays in the measurement of lytic activity or in reproducibility in the repeated samplings of healthy individuals. The whole blood assay required less volume of blood and also less processing time as compared to PBMC assay. It was also validated by testing patients diagnosed with familial hemophagocytic lymphohistiocytosis expected to have low NK-cell activity. This assay is rapid, sensitive and reproducible and requires significantly less volume of blood which is important for clinical evaluation of NK-cell function.en_US
dc.identifier.citationMhatre Snehal, Madkaikar Manisha, Ghosh Kanjaksha, Desai Mukesh, Pujari Vaishali, Gupta Maya. Rapid flow cytometry based cytotoxicity assay for evaluation of NK cell function. Indian Journal of Experimental Biology. 2014 Oct; 52(10): 983-988.en_US
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/153792
dc.language.isoenen_US
dc.source.urihttps://nopr.niscair.res.in/handle/123456789/29442en_US
dc.subjectFlow cytometryen_US
dc.subjectNatural Killer cell cytotoxicityen_US
dc.subjectWhole blooden_US
dc.subject.meshAdult
dc.subject.meshCell Survival --physiology
dc.subject.meshFemale
dc.subject.meshFlow Cytometry --methods
dc.subject.meshHumans
dc.subject.meshK562 Cells
dc.subject.meshKiller Cells, Natural --cytology
dc.subject.meshKiller Cells, Natural --physiology
dc.subject.meshLeukocytes, Mononuclear --cytology
dc.subject.meshLeukocytes, Mononuclear --physiology
dc.subject.meshMale
dc.subject.meshMiddle Aged
dc.subject.meshReproducibility of Results
dc.subject.meshYoung Adult
dc.titleRapid flow cytometry based cytotoxicity assay for evaluation of NK cell function.en_US
dc.typeArticleen_US
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