Is it normal that I had large amount of IFN-gamma with balb/c splenocytes (after 7 days incubation)? First with addition of Con A and then with stimulation with anti-cd3, anti-cd3/anti-cd28. All the ressource I saw said that balb/c are used for Th2 helper T cells. Do you have any suggestions?
As first impression, I think no; I don't say precisely, but I would suppose that these cells have been stimulated or activated in some way, for example number of passages in culture or number of incubation days with the same culture medium, or the presence of something in the composition medium. I would consider also the serum: do you use the de-complemented or complemented serum? Usually, the balb/c cells are very slowly in their growth, with a body cell very big: when the cells are faster and with a changed morphology, it needs restart.
Assuming the mice are naive, it is not surprising.
If you were to analyze spleen T-cells for intracellular Th1/Th2/Th17 cytokines before culture, you will find that there are significantly more Th1 cells. After stimulation these cells will release IFNg, which in turn will block any Th2 differentiation.
I still don't understand what you are trying to achieve by stiimulating spleen cells with ConA and then anti-CD3/anti-CD28. There is no need for both.
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