HMGB1 blockade attenuates experimental autoimmune myocarditis and suppresses Th17-cell expansion
Authors: Tianqing Peng
Dong Zheng
Zhaoliang Su
Huaxi Xu
Shengjun Wang
Zhijun Jiao
Chenglin Zhou
Haitao Zhu
Caixia Sun
Siamak Sandoghchian
Yu Zhang
Yanfang Liu
Journal: European Journal of Immunology (2011)
Discipline: IMMUNOLOGY
Keywords: HMGB1; Dilated cardiomyopathy; Th17 cells; autoimmune myocarditis; monoclonal antibody
Affiliations: Department of Immunology, Jiangsu University, Zhenjiang, PR China.

Abstract:  High-mobility group box 1 (HMGB1), a non-histone nuclear protein, has been implicated in cardiovascular diseases. Dilated cardiomyopathy (DCM), one of the leading causes of heart failure, is often caused by coxsackievirus B3-triggered myocarditis and promoted by the post-infectious autoimmune process. Th17 cells, a novel CD4(+) T subset, may be important in the pathogenesis of autoimmune myocarditis. In the present study, we attempted to block HMGB1 function with a monoclonal antibody specific for HMGB1 B box and investigated the effects of the blockade on Th17 cells and experimental autoimmune myocarditis (EAM). After induction of EAM, HMGB1 protein levels were significantly elevated both in the heart and blood. Administration of an anti-HMGB1 B box mAb attenuated cardiac pathological changes and reduced the number of infiltrating inflammatory cells in the heart during EAM. These protective effects of HMGB1 blockade correlated with a reduced number of Th17 cells in local tissues and lower levels of IL-17 in the serum. Furthermore, in vitro, studies demonstrated that HMGB1 promoted Th17-cell expansion. Therefore, we speculate that HMGB1 blockade ameliorates cardiac pathological changes in EAM by suppressing Th17 cells. 

Retracted: No

PubPeer comments:
 Figure 5A * Red boxes: The uncorrelated protein control and 72h panels look more similar than expected. The gated percentages and top parts of the plots are different, however. * Blue boxes: The 24h and 96h panels look more similar than expected. The gated percentages are different, however. ![file] , 
 #1 #1 Thank you for raising this problem. The determinants of naive CD4+T cells differentiation into Th17 is the combination of anti-CD3 mAb, anti-CD28 mAb, TGF-b, and IL-6. The anti-CD3 mAb, and anti-CD28 mAb provide conditions for the activation and proliferation of naive CD4+ T, and TGF-b and IL-6 provide conditions for polarization into Th17 cells. In the present work, all the groups, the purified naive CD4+T cells undergoes the same processing, therefore, the CD4+ T cells only were divided into IL-17 positive or negative, the images should be similar, and our intervention factors (HMGB1 or unassociated protein) can only weakly affect the number of IL-17 positive or negative. Furthermore, our system for inducing Th17 cells polarization under anti-CD3 mAb, anti-CD28 mAb, TGF-b, and IL-6 was not working well, the percentage of Th17 cells after anti-CD3 mAb, anti-CD28 mAb, TGF-b, and IL-6 treatment was up to 30-40%u3002 I also hope you can give more suggestions for our research , 
 Dear Dr. Su, thank you for your answer, which was long, but did not address my concern that the areas in the red boxes looked very, very, very similar. More similar than expected, even if one would run the same sample twice through the FACS machine. In fact, all the dots appear to be on the exact same position, if one compares those two regions. Can you address that concern, please? Maybe by providing the original, high resolution images, or the raw files? , 
 #3 Dear Elisabeth Thank you for your suggestion. We definitely say that the results are obtained from repeated experiments and reliable. Since the last time you pointed out this problem, we have tried to find the raw data, but it has been too long time past for us to find more detailed raw data. We just found a word document from an old computer disk recovery, which is labelled HMGB1 treated CD4+ T cells, but there is neither grouping information nor information about which experiment it is. To confirm that we did not lie, we also uploaded this raw data. Regarding why the graphics you mentioned are so similar, we truly donu2019t know. In fact, we also feel that the images we upload are also very similar. Best wishes Zhaoliang SU ![file] ![file] , 
 #4 Dear Zhaoliang Su, Maybe you can upload the original FSC files for review and post the link for download? Same for the other publication with similar problems: https: pubpeer.com publications 8B4EA7033E16E3D613B197BE221F88#5 Thanks! , 
 Dear Dr. Su, thank you for posting images from other experiments. I have no concerns about these images, but i was concerned about the published images, for which you unfortunately could not find the originals. So, my concerns remain. ,