I am working on a project developing an ovine vaccine and I am interested in looking for differences in the release of some cytokines before and after administrating the vaccine to the sheep.
I have already established the protocol for IFN-gamma Elispot and I am trying now with Interleukin 8 using ConA as a positive control and is not working. The reagents that I am using for IL8 Elispot are ovine and designed for ELISA, I haven't found Ovine reagents for Elispot available. There is not a lot of literature published either.
Any suggestions? Thanks a lot!
IL8 is mainly secreted by activated monocytes, which express the CD208 (DC-Lamp).
In sheep vaccination tends to significantly increase the level of circulating monocytes DC Lamp +.
So you could follow vaccination on blood samples by FACS analysis intracyto with a CD208 antibody, certain monoclonal antibodies crossing with all animal and human species.
In sheep the monocytes are easily isolated by FACS FSC-SSC, and so the percentage of CD208 + cells in the window of monocytes can be correlated with their activation rate and their ability to produce Il8.
This may be simpler than an Elispot.
Thanks Jean-Jacques, I would like to try by FACS, not on this trial but maybe on the following one.
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