I got used to cytometric analysis performed on fresh blood. Recently, I read an old article, where LDA was used. Authors stimulated samples with either rIL-2 or PHA and thereafter performed cytometric analysis. They concluded:
"The present study revealed a significantly increased frequency of y6 T cells in both blood and CSF specimens of MS patients as compared with OND patients and healthy individuals. Although the difference is not statistically significant, it seems that this increased y6 T cell frequency correlates with chronic MS with a longer disease duration (>2 yr) as compared with recent-onset MS patients."
Is such methodology still relevant and trustworthy? Personally I have serious doubts.
Mentioned article: Stinissen P, Vandevyver C, Medaer R, Vandegaer L, Nies J, Tuyls L, et al. Increased frequency of gamma delta T cells in cerebrospinal fluid and peripheral blood of patients with multiple sclerosis. Reactivity, cytotoxicity, and T cell receptor V gene rearrangements. The Journal of Immunology. 1995;154(9):4883–4894.
http://www.jimmunol.org/content/154/9/4883.long
Thank you very much for help.
Hi Michal,
While it is useful It is a method where culture conditions can alter the results from what would be found if one could analyze directly ex vivo. In my case I saw a large increase in IFNg producing CD4+ T cells and reduced IL-4 and IL-10 producers, in mice treated with immunotherapy whose IgE antibody had turned to IgG2a compared to controls with a prominent IgE response. But the results can be skewed by culture conditions, thats the argument against LDA. The other difficulty in my case was that the immunotherapy receiving group had individual clones making 10-1000 fold more IFNg yet the assay was a +/- one so both a low producer and one with 1000x more were both just 1 IFNg producer.
So in your case unless you can come up with an innovative LDA I would think you are better off with an alternative method.
Randall
Thank you for the answer. Currently, I am not considering LDA in my own projects. I am preparing a review paper. I am wondering, if paper mentioned in the initial post should be trusted and therefore cited in the review or not. Ever since I've been in favour of skipping such studies, but I also feel that I have to consult this issue with someone, who has experience with LDA. Dr Randall, would you rather skip or cite such paper?
I write a lot of reviews and to be fair it should be cited since it was the technology at the time. Those reading your review can decide for themselves.
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