CD13hi Neutrophil-like myeloid-derived suppressor cells exert immune suppression through Arginase 1 expression in pancreatic ductal adenocarcinoma
Authors: Guiling Ding
Chenyan Zhao
Yan Gu
Ying Chen
Biao Yang
Minghua Zhu
Jing Zhang
Xiongfei Xu
Chin-Lee Wu
Gang Jin
Shiwei Guo
Danghui Yu
Min Shi
Journal: OncoImmunology (2017)
Discipline: BIOLOGY & BIOCHEMISTRY
Keywords: Immune escape; myeloid-derived suppressor cells; pancreatic ductal adenocarcinoma; tumor microenvironment.
Affiliations: Department of Pathology, Changhai Hospital, Second Military Medical University , Shanghai, China., Department of Pathophysiology, Second Military Medical University , Shanghai, China., National Key Laboratory of Medical Immunology & Institute of Immunology, Second Military Medical University , Shanghai, China., Department of General Surgery, Changhai Hospital, Second Military Medical University , Shanghai, China., Department of Pathology, Massachusetts General Hospital, Harvard Medical University , Boston, MA, USA.

Abstract:  Perineural invasion and immunosuppressive tumor microenvironment are the distinct features of pancreatic ductal adenocarcinoma (PDAC). Heterogeneous myeloid-derived suppressor cells (MDSCs) are potent suppressors of antitumor immunity, posing obstacles for cancer immunotherapy. Increasing evidences have demonstrated the accumulation of MDSCs in PDAC patients. However, the role of MDSCs in perineural invasion of PDAC and the existence of novel MDSC subsets during PDAC remain unclear. This study found that lymphocytic perineural cuffs were frequently present in chronic pancreatitis (CP) tissues and adjacent non-neoplastic pancreatic tissues (ANPTs), but not in PDAC with perineural invasion. Meanwhile, we found that neutrophil-like MDSCs (nMDSCs), but not monocyte-like MDSCs (mMDSCs), were significantly increased in PBMCs and tumor tissues of PDAC patients. Further observation identified two distinct subsets of nMDSCs, CD13hi and CD13low nMDSCs in PDAC patients, which have not been reported previously. Despite a similar morphology, CD13hi nMDSCs expressed higher levels of CD11b, CD33, CD16 and arginase 1 but lower levels of CD66b than CD13low nMDSCs. Importantly, CD13hi MDSCs, compared with CD13low nMDSCs, more effectively suppressed alloreactive T cell responses via an arginase-1-related mechanism. After tumor resection, the circulating CD13hi nMDSCs were decreased markedly. PDAC patients with more CD13hi nMDSCs had a shorter overall survival than those with less CD13hi nMDSCs. To conclude, we identified two novel MDSC subsets with different characteristics and functions in PDAC, demonstrated the association of the two MDSC subsets with cancer progression, and explored their roles in perineural invasion and immune escape of PDAC. 

Retracted: No

PubPeer comments:
 CD33 and CD11b is mainly expressed on MDSCs membrane and they have been shown to be a marker of MDSCs. According to Figure2 A in this manuscript, CD33 and CD11b were mainly expressed in nucleus because they colocalated with DAPI. Could authors give a reasonable explanation? ![file] , 
 Could authors re-check the source of IL12 CBA reagentuff1f I CANNOT find this reagent on website of BD Bioscience. , 
 Could authors re-check the source of IL12 CBA reagentuff1f I CANNOT find this reagent on website of BD Bioscience. ![file] , 
 Dear Meligethes Kraatzi, If you remove the pseudo-color and subsequently adjust lightness and contrast of Fig 2A, you will find CD11b image and CD33 image shared a lot of similarities. See figure below: ![file] , 
 ![file] Figure S2 B in this paper reuse of the same figures from First author s doctoral thesis (http: www.wanfangdata.com.cn details detail.do?_type=degree id=D151385)uff0cbut marked with different molecules. In his her doctoral thesis , the author declared green cells were CD13+ and red cells were CD16+. In this paper, he she declared green cells were CD11b+ and red cells were CD33+. Could the author give an explaination? , 
 Unstained control reused in Fig3 C. ![file] , 
 Dear Elaeocarpus Foveolatusuff0cI noticed the reused unstained controls have different ratio (red box). ![file] , 
 #1 In the manuscriptuff0cwe just show the pictures of low power microscope. The comment of Meligethes Kraatziu00a0is not accurate because he she maybe didnu2019t see the picture clearly. The figures below are the original pictures of my experiment at the same time. Under high power microscope (X 400 times), We were see the fluorescence located of positive cells are mostly in the cell membranes and a few in the cytoplasm clearly! There is no doubt of the membrane located of CD33 and CD11b in the manuscript. ![file] , 
 #3 We used IL-12 p70 CBA reagent from BD bioscience and you surely could search and find it in the homepage of BD bioscience. The catalog number is 558283. ![file] , 
 #4 1. The commentator thinks that the figures are the result after adjusting the lightness and contrast of the pictures. The figures are the original pictures, and the manual photoshop technology mentioned by the commentator seems very professional, but we really don t know how to do that. 2. To the Polyscias Sessiliflorau2019s commentuff0cI want ask she he a questionuff1fDoes he she have done any immunofluorescence experiment beforeuff1fThere are different non-specific stains in the immunofluorescence experiment. Non-specific staining is a common problem in immunofluorescence experiments. If the fluorescence located of the positive cells are accurate, there is no doubt of the experiment pictures. There is a non-specific staining background in the pictures, because the author does not photoshop the pictures and fake as the commentator thought. , 
 #5 To Helina Flavisquamau2019s comment, I think you donu2019t know the long story of my studies of MDSCs. The doctoral thesis figures and the manuscript figure S2B are all true, The similarly of those because the stain positive cells are the same subtype of immune cells in continuous sections of the same case. In 2009, We found a new subtype immune cells are increased in the peripheral blood of pancreatic cancer . My doctoral thesis is finished in the year of 2011. We all guess the new subtype immune cells are MDSCs, but there are no uniform anthropogenic biomarkers of MDSCs. The commentator can consult the literature of MDSC before the year of 2011, the uniform markers of MDSCs in all literatures were Gr-1 and CD11b of mice. We waiting for a long time, and restart the experiment after 3 years later. There many data vertified CD33 and CD11b are recognized as uniform marker antibodies of MDSCs, so we restart the situ immunofluorescence experiment in the continuous sections of the same case, and the conclusion supports the pre experiment in the period of doctor. The doctoral thesis is only the preliminary experimental results of this article, and has not been published in any journal. I want ask Helina Flavisquama a question? How do you get the co-author doctoral thesis? The link you sent is not real. If you have any questions, welcome to sent a email to me but not this anonymous way. , 
 #6 As you showed, the histograms of CD66b and CD124 used actually the same unstained control because both anti-human CD66b and anti-human CD124 were PE-conjugated. So we stained the cells with these two antibodies individually and other required markers and then overlay PE channels in different populations using unstained control PE channel. , 
 #7 Really the numbers of unstained controls in these two histograms are not shown correctly due to typing error during figure preparation. Thanks for your careful reading and critical checking. If it is required, we will contact with the editor of the journal and revise this typing error in this figure. ,