Rants & Raves, the new hangout for infidels on the internet, is my new favorite forum. There is (so far) no moderation, so its a strange combination of logical arguments and references, sprinkled with insults and boobs. I think its wonderful :)
I wasnt aware that it had become so popular until I started noticing the woo-ers coming out of the woodwork. First the anti-vaxer from mothering.com, now an HIV Denier. If the poster isnt being a poseur, its none other than David Rasnick.
The same David Rasnick who 'collaborated' with Matthias Rath, a professional woo-er who says vitamins can cure AIDS and cancer.
Thats another topic for another post, but what the heck is Rasnick doing on Rants & Raves? Posting on a thread about a new HIV study you might have seen on the news:
Identification of Host Proteins Required for HIV Infection Through a Functional Genomic Screen.
I dont even have to go through his whole post-- the first sentence could not be made by a virologist with a sound mind:
I was surprised to learn the Harvard study led by Professor Stephen Elledge was conducted in highly aneuploid "HeLa-derived TZM-bl cells". I've worked the past 11 years studying the consequences of aneuploidy and feel I should pass along some of my concerns about the study's experiments and the authors' conclusions.I cannot, for the life of me, figure out why this dude is 'surprised' at the use of a cell line called 'TZM-bl'. TZMs are standard in HIV research laboratories. To any HIV researcher, why these cells were used by Elledge et al is shockingly obvious. Even to someone unfamiliar with modern HIV research, like an HIV Denier might be, the reason why this cell line was used is described on the first page of the article.
HeLa cells are a cell line derived from a 1951 case of HPV induced cervical cancer (side note: some say they are no longer human). TZMs are special HeLa cells that have basically been genetically engineered for HIV research-- They express CD4 (normally only expressed on T-cells, the target of HIV-1), and the TZMs genome contains two 'reporter genes'. If the TZM is infected with HIV, a protein tat will start transcribing the two reporter genes! One reporter gene you can stain with a dye and see underneath a microscope, like I did in this post. You can also use the other reporter gene by blowing up all your cells and doing a luciferase assay.
So an HIV lab using TZMs should not be surprising to anyone supposedly as familiar with HIV research as Rasnick.
Ignoring all of that, Elledge et al also explained specifically why they used TZMs in their paper and in supporting materials. They needed the reporter genes for their initial controls and in their experimental procedures. Those reporter genes are not in patient donated PBMCs. They also gave other reasons for why TZMs were used, though they were not ideal:
TZM-bl cells were chosen due to limitations in experimental methods using more relevant T and macrophage cell lines. They proved useful for screening because they are easily transfected with siRNA, are hardy enough to survive high throughput manipulations and support a full HIV lifecycle to produce infectious virions.There are positives and negatives to all cell lines, but there are also positive and negatives to using fresh PBMCs. There are positives and negatives to using animal models or human subjects.
Im going through all of this with my own research-- I started with cell lines, then I moved to using PBMCs. Then Im going to try organotypic cell cultures, then animal models, then hopefully I will have a PhD and pass the project to someone else :) But while I want my research to be as biologically relevant as possible, Im not going to wait until vaccine trials to publish a friggen paper.
Ugh, friggen stupid comment from a stupid friggen Denier.