I have been trying to infect HaCaT cells using 3 different HRV strains for two months but I can't have any CPE even using high MOI and 4 weeks infection
Hey Fedy,
I have to begin by admitting that I have no experience of Rhinos.. But I would suggest that you try mixing trypsinized. Maybe it is better to "quench" the trypsin by making suspension of the detached cells with medium with FCS, followed by pelleting and resuspension of the cells to fresh medium. Or directly with the rhinovirus inoculum. This way I at least get very nice infection rate with some viruses. Useful for some applications/analyses :)
BTW, maybe this is trivial, but do they surely cause CPE in cell lines?
BR,
Jussi
Hey Jussi
Thanks for your answer.
Yes HRV causes CPE in HeLa cells and fibroblasts. I use fibroblastic cells (HEL-299) as positive control and i get very nice and clear CPE.
I tried your method today, I'll let you know if it works ^^.
Thank you for the tip ;)
Best regards
Fedy
I have used only Hela cells for rhinovirus infections. There is BEAS-2B cells which you can try. There are several literature showing infection in this cell line.
One of the most common method to make the cell line infectious is to adapt the cells with the virus by blindly passaging the cell-line with your virus several times (most literature cite 10-15 passages).
Or depending on the HRV group (Major or minor group) you can try cell lines with ICAM-1 or LDR recpetors. These are the highly reported receptors for HRVs. On the other hand you can try to transfect your experimental cell-line with these receptors and try to see if you get any CPE.
All the best.
Hi Lav
Thank you for the answer
I am keeping on blind passages on HaCaT cells but this procedure is like lottery and need several months.
I already tested my HaCaT cells and they do express HRV receptors (IF staining). So HaCaT cells are supposed to be sensitive to HRV expression, but they are probably not permissive. RT-PCR results are negative on infected HaCaT lysate.
BR
Strange. Hope you have the information about the major or minor group HRVs and you have tested for the receptor of HRV of your interest.
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