Hi Dinesh,

I have never worked with chicken cells but what does 'activate chicken cells' mean? Also, I immortalized some embryonic chicken liver cells while I was at Usask. Not sure if they will be of any use to you. I immortalized them using SV40T antigen.

Regards,

Arinjay.

Arinjay Banerjee

Thank you for the response Jay. Activates means more macrophage like functioning. This cell line is derived from Bone marrow cells of galine transformed by avian leukemia virus. I used Phorbol myristate acetate (PMA/synthetic mitogen) to activates it.That definitely increased the intracellular level of reactive oxygen species level (i measured it).. PMA which is also a protein kinase C activator may help to increase the phagocytosis. I think i should test both activated and unactivated HD11 cells and check the difference.

I have used PMA before and it definitely stimulates macrophages. The answer you will get for your infection studies in these macrophages will be different. I guess it depends what your question is. Do you want to test the effects of a primed macrophage response on virus replication or do you simply want to test how the macrophage would respond to infection? Are you interested in quantifying ROS responses or antiviral cytokine responses?

Arinjay Banerjee Yes! I am going to infect them with wild-type Salmonella and some mutants strains. What concentration of of PMA did you use Jay? Do you find any toxic effect to the macrophages? T

Yes. There are toxic effects. I would titrate the amount of PMA on your cell type to determine the minimum dose with highest efficacy.

Arinjay Banerjee Thank you Jay.I will titrate and see.