The cells identified by monoclonal antibody 6B11, binds to CDR3 loop of alpha chain, are reportedly considered as iNKT cells. However, it is quite possible that a few cells may be 6B11+ but TCR beta negative. Should these population also be considered as iNKT cells?
Hi,
Do these cells also bind an alpha-GalCer loaded CD1d tetramer? If they carry the invariant alpha chain, and bind/react to the canonical CD1d-bound glycolipid, I would consider them as iNKT cells.
Pierre
I agree with Pierre. Not sure what you mean by TCR beta negative--is this Vb11 negative?
Hi
Thanks for your suggestion. I used CD3e and 6B11 for iNKT cells, but not any antibody specific to TCR beta chain (any of them including Vb11). Should I consider all CD3e+6B11+ cells as iNKT cells? What about those small population that may be TCR beta negative?
Do you think the cells lack TCR beta entirely? That would be very odd. Do they bind CD1d tetramers loaded with PBS-57?
Pierre and Mitch are right. If they are 6B11+CD3+ and other lineage negative (I.e. Not sticky B cells or myeloid cells), they should be Va24+. They need not be Vb11+ (We sorted Vb11+ and minus and found CD1d-restricted iNKT among Vb11-negative too; Exley 1997), but most are. In our hands, all 3 measures give almost identical numbers. All of these TCRa+ cells will be generic TCR beta+, most if not all, Vb11+.
Mark is absolutely correct-they need not be Vb11--most are--but they must have some type of beta chain that could be detected with a pan TCR beta antibody. A tetramer binding cell expressing a TCR composed only of alpha alpha homidimers? Has not been reported and I would suspect some type of technical problem with the beta chain staining.
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