We read that NK cells are CD 3- and CD 56+/CD 16+.
Recently I and across two IVD approved kits from two manufacturer for lymphocyte subset assay. Kit from BD (cat no. 644611) have CD16/CD56+ selection for NK cells while kit from Beckman Coulter (cat.no. 6607073) has only CD 56 marker for NK cells.
Why there is discrepancy in it?
What should we follow only cd56 or CD16/CD 56+??
Both are correct, because NK cells are not a homogenous population.
Article Polyfunctionality of natural killer cell in healthy donors
If you are interested in cytotoxic NK cells, I would use CD3-CD56dimCD16+ as a way to isolate your cells.
If you are interested in cytokine production, CD3-CD56high is the populations with higher cytokine secretion.
If you wish to look at differential effects on your cells, you can also use markers as CD69, CD25 or NKG2D to asses levels of activations in human NK cells, as well as degranulation marker, CD107a.
Peripheral blood NK cells are CD16 + but NK cells in lymph nodes are CD16-. So it depends on the sample that are isolating them from. If blood use the kit that selects for CD16+ cells.
You should confirm that the cells that you isolate are CD3 negative.
CD3-CD56+ isolate NK cells. CE3+CD56+ isolates NKT cells. Bright vs dim CD56 are largely the same as CD56+/- CD16. Not absolutely correct statement. See
CD56bright natural killer (NK) cells: an important NK cell subset Article CD56bright natural killer (NK) cells: An important NK cell subset
Indeed, NK cells are mostly double positive for CD16/CD56 markers. But in reality it is a heterogeneous population composed mainly of two large subsets. precisely based on those two markers. There is a subpopulation that for some authors are immature and for others they are called secretors that are CD56bright / CD16 low or negative, (about 10% of the total NK). There is another subpopulation called cytotoxic (about 90 %) that expresses CD16 constitutively. Both kits are useful but it depends on the purpose of your assay and the bias you are willing to tolerate. If you do not want to identify the subpopulations of NK, you can use either of the two kits interchangeably, since with CD56 alone, you lose a few NKs that are CD16 +/CD56- in healthy individuals since they are only elevated in chronic viral infections . If you want to identify the subpopulations then, I would recommend you using a cocktail of CD3 / CD16 / CD56, with different fluorochromes.
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