17 February 2021 2 1K Report

Dear All,

I am interested in studying the response of immune cells against virus-like particles (VLPs). I used the PBMC in vitro assay and treat it with the VLP using PHA as a positive ctrl while monitoring the cell proliferation using the CFSE assay. I could not get any proliferation in the VLP-treated PBMC compared to the PHA+ve Ctrl.

I am thinking to check cytokine release by FACS e.g. INF gamma (I found this in one research article but again I do not think that it is enough to see the whole picture) to identify the activated cells (followed by Multiplex cytokine assay) along with checking the expression of the co-stimulatory molecules. I am afraid that missing the first responders' innate immune cells as well as the memory cells in the PBMC resulted in misleading information regarding the immune response towards the VLP.

I am wondering if I just wait and look at the cytokine profile first or co-culture the innate/adaptive immune cells (actually I think the co-culturing system is not that easy), that's why I think it is better to shift to in vivo system.

Any suggestions to save time and resources would be appreciated. Thanks so much.

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