26 September 2010

NKTs? (Natural Killer T cells)


Here's a practical question. I want to clone out human CD8+ T cells, starting with frozen PBMCs. Before I stimulated them with CMV peptide (and an artificial APC), I used magnetic beads to produce a somewhat more purified population. I used anti-CD56 beads to remove NK cells, and anti-CD19 beads to remove B cells. After a few rounds of stimulation/proliferation followed by rest, I sorted only the CD8+ cells also expressing early activation markers (CD69 and CD137). I then cultured these through several more cycles of stimulation and rest.
I then stained the cells with certain markers I was interested in, and performed flow cytometry. I found 14% of the population of one of the cultures to be CD8+, CD4+, CD3+ triple positive cells. Unfortunately, I did not stain with anti-CD56. My guess is that these peripheral double positive T cells are NKT cells. Does anyone have any ideas?

1 comment:

  1. I'm glad this wasn't something I should've picked up in class. Waaaay over my head!

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