Skip to main content

Archived Comments for: Intrinsic predisposition of naïve cystic fibrosis T cells to differentiate towards a Th17 phenotype

Back to article

  1. Lack of information about methodology

    Margaret Cooley, University of Tasmania

    5 December 2014

    The methods section of this paper is missing some important details that would allow appropriate assessment of the results. NO information is given about the quantitative PCR methods (primers, temperatures cycle nos etc). This is extremely important in this case, as without information about the primers and their specificity, it is impossible to evaluate why no mRNA for the mutant CFTR was detected in cftr negative cells (a DelF508 mutation still produces mRNA).

    There is little to no methodological detail about the flow cytometry analysis (what antibodies were used, what cytometer, what gating strategy) and are no labels on the Y-axis of the flow cytometry histograms in Figure 2. This makes it impossible to evaluate the gating used to identify IL-17 positive cells, and therefore to evaluate the data.

    Why were cells stimulated twice in vitro (once with anti-CD3 and anti-CD28 and then 4 days later with Ionomycin and PMA)? This would be expected to induce apoptosis/necrosis in many cells and may explain some of the results. The very small number of samples means that differences may be by chance.

     

    Competing interests

    No competing interests.

Advertisement