The short answer to the OP's question is, "yes, you can", but TZMbl cells are definitely not your best bet for producing HIV. I'll reinforce some of the comments above:

Tetherin/BST2 limits the release of viral particles, and HeLa-based cell lines like TZMbl produce very high endogenous levels of BST2 (more than any other cell line I've ever tested, including T cell lines). With TZMbl's, even if you're working with an HIV that encodes a functional Vpu, you still take a hit in virus production.

TZMbl cells of course make a lot of CD4. And depending on which papers you read/believe, CD4 either a) functions to limit release of HIV, b) prevents maturation of released viral particles, and/or c) limits the infectivity of released particles, presumably by lowering Env incorporation. So if your goal is maximize viral titers, all that CD4 is working against you, via a mechanism(s) that we don't yet understand.

However, if you are working with clinical HIV samples, you can't infect 293 cells. So, yes, TZMbl cells are convenient/easy, and can be infected by both R4- and R5-tropic HIV. It's perhaps even easier to use the Jurkat T-cell line, as they grow in suspension, and can also be infected by both R4 and R5 strains.

It depends on the RNA that packaged in the viral particles! If the full length genome of virus is used to make particles, then it will make replication competent virus (infect again and again), in case of pseudotyped (vsv-g for example) reporter virus or virus genome lacking envelope region will be replication incompetent hence only onc replication cycle possible. The rationale behind pseudotyped virus is they allow us to study the events of virus replication such as mode of virus entry, RT, nuclear entry, integration and reporter gene (luciferase for example) expression. Hence this system will not be threaten like S3 virus and so the tool can be utilized for research purpose.

Hi there,

The answer is yes. Using full-length-competent HIV viruses (such as HIV LAI) you can infect TZM-bl and these infected cells will produce progeny, typically 48 to 72 hrs post initial infection. In fact, we have a paper on the system. Pseudoviruses that are not lacking Env or other essential genes may also produce viable particles after infection in TZM-bl cells. Good luck!

Hi Nancy,

TZM, which are modified HeLa cells, are able to package and release virions, but in my experience in a very low titer compared to 293 cells. Even when obtaining significant amounts of gag antigen (p24) in the supernatant I have found very difficult to re-infect other cells. Probably one of the reasons to explain this behavior is that HeLa cells express high levels of tetherins, which retain virions on the cell surface.

Hope it helps.

Low levels of viral release could be due in some instances, vpu- viruses for example, to HeLa's constitutive expression of tetherin. Could also be related to the fact they have been transduced with CD4 and CCR5 IIRC. Perhaps CD4 expression is high enough that nef is incapable of degrading it sufficiently. This causes low infectivity as env is snared within the secretory pathway.

Thank you Ananda, I understand about the pseudo virus, they cannot do multiple rounds. The part I am thinking is real virus with HIV-1 complete genome. I don't know how efficient they could produce progeny in Hela cell.

HI, Javier. Thank you for your info. Could you please send me the link?

Hi Eduardo. Thank you!  I read a paper talking about using 7 days to produce progeny virus in TZM-bl and then infect TZM-bl cells. I don't understand why 293T is so efficient and Hela-derivative is not. It's really helpful to learn about tetherins.

Hi Jason, thanks a lot. I will read more about tetherins and CD4.

Hi,

HIV replicates pretty well in TZMbl in my hands. If you use virus that encodes Vpu, tetherin should not be a problem at all, Tetherin levels in HeLa are not particularly high. CD4 levels in TZMbl are really high because it is a transgene. So that might still slow down a bit the viral production, even when both Vpu and Nef are inhibiting CD4 presence at the cell surface.

Finally, 293T produce more viral particles because they express the SV40 large T antigen, which enhances the activity of some transcription factors including the ones controlling HIV transcription. The more viral protein expressed, the more viruses are assembled and released.

Hope that helps with your last questions.

The short answer to the OP's question is, "yes, you can", but TZMbl cells are definitely not your best bet for producing HIV. I'll reinforce some of the comments above:

Tetherin/BST2 limits the release of viral particles, and HeLa-based cell lines like TZMbl produce very high endogenous levels of BST2 (more than any other cell line I've ever tested, including T cell lines). With TZMbl's, even if you're working with an HIV that encodes a functional Vpu, you still take a hit in virus production.

TZMbl cells of course make a lot of CD4. And depending on which papers you read/believe, CD4 either a) functions to limit release of HIV, b) prevents maturation of released viral particles, and/or c) limits the infectivity of released particles, presumably by lowering Env incorporation. So if your goal is maximize viral titers, all that CD4 is working against you, via a mechanism(s) that we don't yet understand.

However, if you are working with clinical HIV samples, you can't infect 293 cells. So, yes, TZMbl cells are convenient/easy, and can be infected by both R4- and R5-tropic HIV. It's perhaps even easier to use the Jurkat T-cell line, as they grow in suspension, and can also be infected by both R4 and R5 strains.

Hi Nancy,

The real HIV-1 viruses definitely are able to replicate in the TZM-bl cells, and the progenitor viruses can infect the producing cells in turn. Usually, people use HEK293T cells to package HIV or other lentiviruses, and  use the TZM-bl cells to measure the infectivity of packaged viruses, as well as the nutralizing antibody's efficiency.

Dear Nancy,

Laboratory HIV strains, if not gene-deficient variants, replicate well in any cell culture if they match CCR5-CD4 or CXCR4-CD4 in virus strain and cell line. When you transfer virus strain from one cell line to another it can show decline in virus titre/particles replicated for 2-3 passages, then the amount usually recoveres. TZM cells is not the exclusion. Native virus replication in cell cultures is completely different story. I obtained several laboratory strains of subrype A HIV-1 (PokA-79 was sequenced and characterized) in several suitable cell cultures, it normally takes 3-4 months of cultivation and originally should be done with healthy donors PBMC-cell culture mixture, otherwise native virus does not survives in vitro. It also has one restriction, I was successfull in strain cultivation only when I had taken a mixture of infected people PBMC material for cultivation, i.g. HIV PBMC pool. Single patient's material has much less chances, though theoretically strain obtaining is possible, I had never been lucky with such attempts.

I agree with all the above comments. And in my experience, infection in tzm-bl leads very quickly to syntitia formation and will kill all the cells. Si, as stated by J. Gustin, if you want to produce virus from a molecular construct, use 293T, if it is clinical isolates, then use PBMCs (Jurkat can only be infected by X4 tropic viruses)

Thank you Corinne. 

Thank you Gregory. Did you ever test virus released from TZM-bl? I am convinced by comments that TZM-bl can produce  lots of virus.

Thank you Jean. I just got Jurkat and JLTRG. If I am using proper types of virus, will they express GFP right after infection? Normally how would people quantify?

Thank you Yi-Qun.

Thank you Bastian!