Comments on: Zinc and rhinovirus replication

By: reverse phone lookup

reverse phone lookup — Thu, 17 Nov 2011 21:19:00 +0000

nice theme. but it takes a while to load

By: Nadine

Nadine — Wed, 16 Feb 2011 11:18:00 +0000

hi
I intend to do plaque assay for adenovirus carrying specific target gene, can anybody suggest any negative control, in addition to vector without gene?

By: profvrr

profvrr — Thu, 17 Dec 2009 01:44:36 +0000

You make very good points, and I do plan to look at the effects of
ZnCl2 in a respiratory cell line. If the sole target of ZnCl2 is a
viral protein or RNA, then it might not matter what cell types the
study is done in. For this reason I decided to get the project started
in a very easy cell line to work with, HeLa cells, and then compare
the findings in a respiratory line. Thanks for reading and listening,
and good luck with HIV.

By: Guin

Guin — Thu, 17 Dec 2009 01:38:17 +0000

In counting the PFU and assaying for viral titre, HeLa cells sounds like a great choice. However, when the goal is to try to identify biologically relevant mutants to find out zinc's mechanism of action, I am prone to think respiratory cells would be a better choice? Again, I don't know much about cell cultures and so there may be factors that I have not considered (for instance, how much will a virus be affected when different cell lines are used). Perhaps you can test the growth of some respiratory epithelial cells under a range of ZnCl2 concentrations and see whether this model is viable… Good luck! I'll keep reading! By the way, I love your blog, TwiV and TwiP. I'm a PhD student in HIV.

By: profvrr

profvrr — Thu, 17 Dec 2009 00:34:13 +0000

It's a perfectly reasonable question. Clearly HeLa cells, being of
cervical origin, are not relevant to the biology of rhinovirus. They
are very convenient for studying aspects of viral replication and
genetics that depend upon efficient viral replication. In the case of
these experiments, it is easy to plaque rhinoviruses in HeLa cells and
to select for drug resistant mutants. But your comment will provoke me
to pull some respiratory cells out of the freezer and see if I can get
rhinovirus to form plaques on them.

By: Guin

Guin — Thu, 17 Dec 2009 00:25:52 +0000

I've been following this Rhinovirus experiment you're performing. Please excuse my lack of knowledge: I thought HeLa cells are derived from a cervical cancer cell line. Rhinovirus causes common cold so I'm assuming it infects cells in the airway? What's the rationale of using HeLa cells? (I also noticed the original publication used HeLa cells as well.)

By: profvrr

profvrr — Wed, 16 Dec 2009 17:44:36 +0000

By: Guin

Guin — Wed, 16 Dec 2009 17:38:17 +0000

By: profvrr

profvrr — Wed, 16 Dec 2009 16:34:13 +0000

By: Guin

Guin — Wed, 16 Dec 2009 16:25:52 +0000

By: profvrr

profvrr — Tue, 01 Dec 2009 10:03:39 +0000

That's what I'll be doing today – trying a range of ZnCl2 concentrations.

By: EC

EC — Fri, 27 Nov 2009 22:31:06 +0000

The ZnCl2 results seem consistent with other potential metal-ion based therapeutics, e.g. cobalt(III) hexammine, which also has an IC50 in the < 0.1 mM region against several types of viruses. Maybe that concentration level is something common to broad-spectrum metal-ion antivirals. You might be interested in running some tests in parallel.

By: BN

BN — Wed, 25 Nov 2009 17:59:32 +0000

How about first determining maximum Zn concentration that cells can withstand, then using that concentration for your plaque assay?

By: profvrr

profvrr — Wed, 25 Nov 2009 08:12:26 +0000

I haven't tried mixing virus with ZnCl2 before plating, but it's worth
a try. I'll do it next week.

By: chrisupton

chrisupton — Tue, 24 Nov 2009 21:43:10 +0000

Does anything happen if the virus is held in the ZnCl2 before infection? ie stocks of viruses diluted out of ZnCl2 before infection.

By: uberVU - social comments

uberVU - social comments — Mon, 23 Nov 2009 19:41:33 +0000

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