You can try Jurkat, CEM and SupT1 cells. As you might already know these are all T-cell lines. I have used these for transient transfections of plasmids having the HIV LTR but not for a complete HIV-1 genome as such. However, in my opinion that shouldn't be a concern.

You can use PBMCs too.

I don't understand what you mean by 'which condition is required?' If activation conditions is what you mean then you can try one of the following regularly used activation conditions depending on your further downstream requirements:

1. PMA-ionomycin activation.

2. CD3/CD28 activation.

3. TNF-a activation (potent HIV-1 LTR activator)

Thank you Rishikesh and Barbara,

We are going to discuss the possibility of having HIV transfection in our lab to investigate the role of some proteins during viral infections. These data would help us a lot. I will be in contact with you once we decided to run the experiment for more advices.

Personally we are using 293T or 293FT cells for transfection and transduction using our HIV-1/2 vectors. It actually depends on what HIV vectors you are using, being either 1st/2nd or 3rd generation vectors, HEK like 293T 293FT cells are optimized for 2nd and 3rd generation vectors, and will work fine.

hi

i personally use 293T cells to make virus by transient transfection , but they undego only single round production but you get huge amount from them. If you still want you can propagate the virus in T cell line or any other cell line but you have to change the media very often to take care of cell debris and death.

we standardized the protocol for making virus very well in our lab if you want i can send you the detailed protocol.

All the best

AB

This is an additional information. Upon DNA transfection, some cell lines produces RANTES and IFN lambda in the culture as an innate immune response (J. Immunol. 2011 Apr 15;186(8):4541-5). Thus, the transient transfection sup contains HIV particles and both cytokines. The cytokines inhibit R5 infection, thus if you try to make R5 virus stocks, you should pellet the HIV particles by ultracentrifuges with sucrose cushion, and then re-suspend the pellet in medium or PBS before storage.