Dear college,
Recently we performed ex vivo analysis of IFN-g T cell response in immunized mice. As we did not expect high frequency of T cells to our target antigen and as we wanted to see CD4+ and CD8+ T cell responses we enriched T cell populations with appropriate (CD4 or CD8)T cell separation columns and performed IFN+ Elispots. The purity of separation was 85% and we did not mind that because we reasoned that non-T cells can serve as antigen-presenting cells in our in vitro stimulation. My question is can those non-T cells seriously affect T cell response in a way that that compasion of of the results in immunized and non immunized will be not correct. Will be very greatful for your suggestions.
NK and NKT cells can produce a lot of IFN-g. These innate cells can respond independent of T-cell specific antigens. Moreover, they are the most frequent contaminant in the CD8 or CD4 positive selection.
Thank you very much for your suggestion. It will certainly help to shape our reply to a reviewer.
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