Choosing the Appropriate Control

Abstract

To the Editor:We recently read the article by Kobayashi and colleagues,“ Expression of Inducible Nitric Oxide Synthase and Inflammatory Cytokines in Alveolar Macrophages of ARDS Following Sepsis” (June 1998).1Kobayashi A Hashimoto S Kooguchi K et al.Expression of inducible nitric oxide synthase and inflammatory cytokines in alveolar macrophages of ARDS following sepsis.Chest. 1998; : 1632-1639Abstract Full Text Full Text PDF PubMed Scopus (91) Google Scholar The article concerns the fluorometric determination of intracellular alveolar macrophage inducible nitric oxide synthase (iNOS), interleukin-1β, interleukin-6, and interleukin-8.Dr. Kobayashi and colleagues1Kobayashi A Hashimoto S Kooguchi K et al.Expression of inducible nitric oxide synthase and inflammatory cytokines in alveolar macrophages of ARDS following sepsis.Chest. 1998; : 1632-1639Abstract Full Text Full Text PDF PubMed Scopus (91) Google Scholar conclude from their data that there is strong expression of iNOS in alveolar macrophages from ARDS patients. It is our opinion that in their report, the increase in immunostaining of alveolar macrophages in the ARDS group might represent nonspecific staining of their anti-iNOS antibody. It is true that the application of the anti-iNOS antibody was associated with an increase in staining in the ARDS group as compared to the control group. This result could be true of any antibody. The investigators should demonstrate that the increase holds when comparing the specific anti-iNOS antibody to a nonspecific antibody within the ARDS group. We respectfully suggest that further experiments be performed with a nonspecific antibody of the same subclass as the anti-iNOS antibody included. With these control experiments, a conclusion could be made that references the nature of the specific anti-iNOS antibody's pattern of staining in ARDS alveolar macrophages as compared to the pattern of the nonspecific-isotype antibody with a similar fluorochrome. To the Editor: We recently read the article by Kobayashi and colleagues,“ Expression of Inducible Nitric Oxide Synthase and Inflammatory Cytokines in Alveolar Macrophages of ARDS Following Sepsis” (June 1998).1Kobayashi A Hashimoto S Kooguchi K et al.Expression of inducible nitric oxide synthase and inflammatory cytokines in alveolar macrophages of ARDS following sepsis.Chest. 1998; : 1632-1639Abstract Full Text Full Text PDF PubMed Scopus (91) Google Scholar The article concerns the fluorometric determination of intracellular alveolar macrophage inducible nitric oxide synthase (iNOS), interleukin-1β, interleukin-6, and interleukin-8. Dr. Kobayashi and colleagues1Kobayashi A Hashimoto S Kooguchi K et al.Expression of inducible nitric oxide synthase and inflammatory cytokines in alveolar macrophages of ARDS following sepsis.Chest. 1998; : 1632-1639Abstract Full Text Full Text PDF PubMed Scopus (91) Google Scholar conclude from their data that there is strong expression of iNOS in alveolar macrophages from ARDS patients. It is our opinion that in their report, the increase in immunostaining of alveolar macrophages in the ARDS group might represent nonspecific staining of their anti-iNOS antibody. It is true that the application of the anti-iNOS antibody was associated with an increase in staining in the ARDS group as compared to the control group. This result could be true of any antibody. The investigators should demonstrate that the increase holds when comparing the specific anti-iNOS antibody to a nonspecific antibody within the ARDS group. We respectfully suggest that further experiments be performed with a nonspecific antibody of the same subclass as the anti-iNOS antibody included. With these control experiments, a conclusion could be made that references the nature of the specific anti-iNOS antibody's pattern of staining in ARDS alveolar macrophages as compared to the pattern of the nonspecific-isotype antibody with a similar fluorochrome. Choosing the Appropriate ControlCHESTVol. 115Issue 4PreviewTo the Editor: Full-Text PDF