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Interaction Effects Among IFN-Γ+874, IL-2-330, IL-10-1082, IL-10-592 and IL-4-589 Polymorphisms on the Clinical Progression Of

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Interaction Effects Among IFN-Γ+874, IL-2-330, IL-10-1082, IL-10-592 and IL-4-589 Polymorphisms on the Clinical Progression Of BMJ Open: first published as 10.1136/bmjopen-2016-013279 on 23 August 2017. Downloaded from PEER REVIEW HISTORY BMJ Open publishes all reviews undertaken for accepted manuscripts. Reviewers are asked to complete a checklist review form (http://bmjopen.bmj.com/site/about/resources/checklist.pdf) and are provided with free text boxes to elaborate on their assessment. These free text comments are reproduced below. ARTICLE DETAILS TITLE (PROVISIONAL) Interaction effects among IFN-γ+874, IL-2-330, IL-10-1082, IL-10- 592, and IL-4-589 polymorphisms on the clinical progression of subjects infected with hepatitis B virus and/or hepatitis C virus: a retrospective nested case control study AUTHORS Gao, Qiu-jv; Xie, Jia-xin; Wang, Li-min; Zhou, Qiang; Zhang, Shiyong VERSION 1 - REVIEW REVIEWER Hend ibrahim Shousha endemic medicine and hepatogastroenterology department, Faculty of medicine, Cairo University REVIEW RETURNED 28-Jul-2016 GENERAL COMMENTS thank you for your effort and research idea How did you diagnose the controls who spontaneously cleared HCV or HBV? is there patients who had combined HCV and HBV infection and cleared both?? How many patients were infected with HCV, HBV or had combined http://bmjopen.bmj.com/ HBV and HCV within each group?? what is the treatment status of the patients?? Saxena R, Kaur J. Th1/Th2 cytokines and their genotypes as predictors of hepatitis B virus related hepatocellular carcinoma. World J Hepatol. 2015 Jun 18;7(11):1572-80. mentioned in their review (On associating the IL-2 (-330 T > G) genotypes with HCC progression in HBV infected individuals, we showed that both the TG on September 29, 2021 by guest. Protected copyright. and GG genotypes remained largely non-significant in HBV chronicity, among controls and carriers [54]. Similarly, the IL-2 (-330 T > G) polymorphism did not appear to modify HBV-HCC risk in the Chinese and American populations[50,77]. On the contrary, a study by Gao et al[78] reported, IL-2-330 TT genotype to be associated with an increased risk of chronic hepatitis, in case of either HBV or HCV or HBV-HCV coinfection in Chinese population (your study)) Thus it is only your study which documented the association; could this be due to the synergism or the combined population of the study with HBV and HCV or combined HBV and HCV BMJ Open: first published as 10.1136/bmjopen-2016-013279 on 23 August 2017. Downloaded from REVIEWER Sara Tatiana Moreira Human Molecular Genetics Laboratory, Parana Federal University of Technology, UTFPR, Santa Helena, Parana, Brazil. REVIEW RETURNED 15-Aug-2016 GENERAL COMMENTS 1. This research not evaluated the effect of polymorphisms on the clinical progression of the diseases because they have not studied the same individuals over time. It was studied the influence of SNPs in different clinical outcomes. The objective needs to be corrected. 2. Correct the objective of the abstract and add the correct nomenclature for the studied SNPs, including the ID (rs) of each. 5. The study protocol have been performed according to the World Medical Association Declaration of Helsinki? 8. The number of references in the introduction must be reduced. Add references to discussion, when compare with results of similar researchs. 12. Add the limitation discuss to the text. Title: Effect of interactions among IFN-γ+874, IL-2-330, IL-101082, IL-10-592, and IL-4-589 polymorphisms on the clinical progression of subjects infected with hepatitis B virus and/or hepatitis C virus Manuscript ID: bmjopen-2016-013279 In this work, the authors found that IL-10-592AC and IL-4- http://bmjopen.bmj.com/ 589CC/CT showed synergistic effect on liver inflammatory injury (P <0.01). While IFN-γ+874AA and IL-2-330TT had synergistic impact (P<0.05), IFN-γ+874AA and IL-10-1082AA had antagonistic effect (P<0.01) on the clinical progression, including asymptomatic HBV and HCV carriers, and chronic hepatitis. IL-2-330TT and IL-10- on September 29, 2021 by guest. Protected copyright. 1082AA synergistically influenced the clinical outcome (P <0.05). IFN-γ+874AA, IL-2-330TT, and IL-10-1082AA interactively affected the clinical outcome as asymptomatic HBV and HCV carriers, chronic hepatitis (P<0.05). What's more valuable in this work is the statistical method used because, as the authors said, “interleukins and the SNPs of cytokines may not play their roles alone, but interact with each other in establishing the clinical consequences of such patients”. However, the work has a large limitation also reported by the authors themselves, which corresponds to the engagement of patients infected with HBV and HCV. BMJ Open: first published as 10.1136/bmjopen-2016-013279 on 23 August 2017. Downloaded from Comments: Pg 2, line 11 - 12 “The mechanism still remains unknown”. The mechanism is not unknown, just is not fully understood. The number of references in the introduction must be reduced. Correct the objective: “evaluate the effect of interactions among the polymorphisms of IFN…on the clinical progression.” This research not evaluated the effect of polymorphisms on the clinical progression of the diseases because they have not studied the same individuals over time. It was studied the influence of SNPs in different clinical outcomes. Add the correct nomenclature for the studied SNPs, including the ID (rs) of each. The pathogenesis resulting of HCV infection is different from that derived from HBV infection. The lack of homogeneity of the sample originates vacant results, non-specific for HCV neither for HBV infection. The study protocol have been performed according to the World Medical Association Declaration of Helsinki? The sample stratification resulted in subgroups with individuals infected by HCV AND HBV, except for a subgroup, composed of individuals infected by HBV http://bmjopen.bmj.com/ AND/OR HCV. Because not follow a rule, the presence of this subgroup may have biased the results. Pg 13, line 7: herein? Eleven of the 13 references cited in the discussion refers to the biological effects of cytokines. Only two references on September 29, 2021 by guest. Protected copyright. involve comparison with other results in the literature. The authors barely discussed their results compared with others of literature, but there are several studies of polymorphisms in cytokine genes x HCV or HBV that should be cited in the discussion. Include: Discuss limitations of the study, taking into account sources of potential bias or imprecision. Discuss both direction and magnitude of any potential bias. BMJ Open: first published as 10.1136/bmjopen-2016-013279 on 23 August 2017. Downloaded from REVIEWER Sanak, Marek Department of Medicine, Jagiellonian University Medical College REVIEW RETURNED 21-Dec-2016 GENERAL COMMENTS Authors genotyped common variants in IL2, IFNG, IL10 and IL4 genes looking for their interactions with a chronic viral hepatitis B or C outcome. The comparator group were subjects who cleared the infection, chronic infection subjects were categorized according to the presence of active hepatitis symptoms. Most of my comments concern results section since numerous variants of SNP genotypes are not well presented. The main general comment is that the manuscript is disorganized. Detailed comments: 1. Study groups. What is a difference between group 1 (controls) and group (2) chronic carriers of the virus without hepatitis, if no presence of HCV genome was tested. 2. Methods. All SNPs typed should be referred to using a standard SNP database entries (rs numbers), 3. Results. It is sufficient to discuss only the risk genotype affecting elevated AST. Actually, Authors do not explain if these are the same subjects as named chronic hepatitis group 3. 4. Association with “abnormal AST level” or “abnormal amounts of ALT” does not make much sense because group 2 subjects had also intermittent elevations of transaminases. Moreover, transaminases levels tend to fluctuate between their consecutive measuruments. 5. Gene-gene interactions and the clinical outcome. Certain combinations of IFN, IL10 and IL2 variants cross-verified predicting group 2 or group 3 membership. But the number of individuals with these predicted outcomes represented only a fraction of the total group. No corrections were used for multiple comparisons. http://bmjopen.bmj.com/ 6. Results. Effect of gene-gene interactions on the liver inflammatory injury. The table 4 with results refers to all the study group, however, only in group 3 a liver biopsy was done to evaluate the inflammation. 7. Discussion. Conclusions suggest that the outcome of viral hepatitis depends on the interactions of the genetic variants studied. In fact, all these genetic variants ought to manifest by altered expression of the respective cytokines and eventual changes in the adaptive immunity response. No attempt to correlate these findings on September 29, 2021 by guest. Protected copyright. functionally with cytokine levels, lymphocyte populations or at least serological response was made. VERSION 1 – AUTHOR RESPONSE Reviewer: 1 How did you diagnose the controls who spontaneously cleared HCV or HBV? is there patients who had combined HCV and HBV infection and cleared both?? Answer: there should be under-represent. there are controls who spontaneously cleared HCV or HBV. Because there were data showing the controls were infected with HBV or HCV in plasm-donation in the late 1980s, and were tested negative to HCV or HBV in 2007. How many patients were infected with HCV, HBV or had combined HBV and HCV within each group?? what is the treatment status of the patients?? BMJ Open: first published as 10.1136/bmjopen-2016-013279 on 23 August 2017. Downloaded from Answer: There are 37(30.3%)HCV infection, 43(35.2%) HBV infection and 42(34.4%) HBV&HCV infection in ASC group, there are 18(22.2%) HCV infection, 36(44.4%) HBV infection and 27(33.3%) HBV&HCV infection in chronic hepatitis group.
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