I was analyzing CNS cells for any infiltration of T cells in one of our disease models and found dual staining of TCRgd and TCRb, has anyone found this or maybe its an articraft? any thoughts?
Dear Abbas,
TCRgd and TCRab are separate populations, so what you observed was likely an artifact. This could be an interference generated by the anti-CD3 antibodies as these Abs may recognize close epitopes with the TCRgd. BD recommend aTCRgd clone 11F2 instead of clone B1 for double staining with aCD3 (it worked well in our hands with the 11F2, no more double positive populations).
Good luck,
Mohamed
Although, I agree that an artifact is the most likely explanation (especially with cells derived after extensive purification/digestion, which require careful gating - you have live/dead discrimination, dumb channels etc?), one might add that some weird ab/gd- chimeric TCRs do exist. The most recent publication I am aware of e.g. is PMID: 25452463.
i have proper gating strategy, no doublets/live dead staining. That paper seemed interesting, let me have a look. thanks Gerhard
@Ewa: do you see separate gamma delta T cells as well or all are just double positives alpha, beta, gamma, delta?
this is exciting, i am going to run the flow one more time to see this population.
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