I would like to design an in vitro assay in which I examine the reaction of T cells among PBMCs to the COVID-19. Due to safety issues, I can't directly culture the virus with PBMCs. On the other hand, for the stimulation of T cells and for induction of a specific response to an Ag, the presence of immunogenic antigens is sufficient. Do you suggest the same method?
What other methods are available to check the function of T lymphocytes against the virus in vitro?
Hi
Some procedures including transfection of DC cells with synthetic minigene or mRNA of corona virus , and then evaluating purified t cell activity (after incubation with transfected ĎÇ ,)using interferon gamma assay may be useful.
Hi dear alale,
I think you should use the synthetic RNA that code specific viral antigen, but I have no idea about the immunology section.
If you're just doing antigen recall... why do you need the whole virus? I mean technically, if you want ALL the potential viral antigens, just get an inactivated virus and use that.
Otherwise, there are a lot of recombinant proteins for SARS-CoV-2 and the other coronaviruses, and there are also overlapping peptides.
Now if you want to look at what T cell responses are during a natural infection and you don't have a BSL-3, there are psuedovirus systems out there that you can use by inserting your protein of interest.
Hi!
The strategy that you've thought of, seems alright to me. I'm not the expert one here though!
Mohsen Alipour, Shahrzad Shoraka Thank you so much for your response.
Given the exogenous and endogenous systems for antigen presentation by dendritic cells, doesn't this method lead to the induction of TCD8+ cells? While we do not intend to guide the response.
Shen-An Hwang Thank you so much for the information.
Assuming availability, which one do you recommend more to simulate T cell response? recombinant proteins, whole inactivated viruses, or pseudoviruses?
Could you please prioritize them?
https://www.researchgate.net/profile/Alaleh_Rezalotfi/post/In_vitro_T_cell_response_to_virus_antigenes/attachment/5ed94aea2149ed000114bc0f/AS%3A898786440257539%401591298794419/download/The-image-shows-the-differences-between-MHC-Class-I-and-MHC-Class-II.jpg
