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.
Hello, Emy!
You look at this research articles:
Article Noninfectious papilloma virus-like particles inhibit HIV-1 r...
Figure 1
VLPs inhibit HIV-1 replication. (A-B) PHA-stimulated PBMCs were infected with (A) X4 virus (HIV-1NL4.3) or (B) R5 virus (HIV-1Ad8) as described in “Materials and methods.” The infected cells were cultured for 7 days in the presence of various concentrations of VLPs. (C) PHA-stimulated PBMCs were infected with X4 virus for 2 hours and then cultured for 7 days in the absence or the presence of 1 μg/mL baculovirus extract or VLPs. (D) PHA-stimulated PBMCs were infected with X4 virus for 2 hours and then cultured for 7 days in the absence or the presence of 1 μg/mL VLPs, SV40-VLPs, or BVLPs.
Article Direct stimulation of T lymphocytes by immunosomes: Virus-li...
Fig. 4.
Accessory cell-independent activation of T lymphocytes by immunosomes. (A) PBMC, highly purified T cells, or T cells supplemented with 10% monocytes were stimulated with medium, undecorated VL
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