Hi I`m trying to detect immune response in vitro with my nanoparticle (virus-like particles) incorporating SARS-CoV-2 Spike protein. I inoculated quite a lot of nanoparticle in PBMC and inoculated them for 24 hours, but could not detect any cytokine or complement with ELISA or intracellular cytokine staining. Does anyone know the reason?
Do I have to supplement PBMC with GM-CSF or IL-4 first to get them reacting to my antigen?
Thanks in advance.
Agree with Lyudmila. Check if PBMCs are viable. Check LPS and lectin activation of cytokine synthesis. Also screen spike protein nanoparticle conjugate stimulated cell supernatants using a cytokine array to get a more holistic picture. You may have screened for cytokines not affected, while missing the ones modulated by treatment.
Edmund Pool Lyudmila Akhmaltdinova
Thank you for the reply. At least the cell viability looked ok when counting the cells. IL-2 and IFNy were the two cytokines I checked, but I understand that different cytokines are expressed at different timepoints. I'll try the cytokine array at different infection time points.
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