Impaired T cell function in malignant pleural effusion is caused by TGF-Œ≤ derived predominantly from macrophages
Authors: Zhen Zhang
Yu Ping
Jieyao Li
Lan Huang
Fei Wang
Yi Zhang
Dongli Yue
Xinfeng Chen
Bin Zhang
Liping Wang
Li Yang
Lifeng Li
Journal: International Journal of Cancer (2016)
Discipline: CLINICAL MEDICINE
Keywords: CD4+ T cells; CD8+ T cells; TGF-Œ≤; impaired T cell function; malignant pleural effusion (MPE)
Affiliations: Biotherapy Center, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, 450052, People's Republic of China., Department of Oncology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, 450052, People's Republic of China., Department of Oncology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, 450052, People's Republic of China. ., School of Life Sciences, Zhengzhou University, Zhengzhou, Henan, 450001, People's Republic of China., Division of Hematology/Oncology, Department of Medicine, Robert H. Lurie Comprehensive Cancer Center, Northwestern University Feinberg School of Medicine, Chicago, IL, 60611., Biotherapy Center, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, 450052, People's Republic of China. ., School of Life Sciences, Zhengzhou University, Zhengzhou, Henan, 450001, People's Republic of China. .

Abstract:  Malignant pleural effusion (MPE) is an indication of advanced cancer. Immune dysfunction often occurs in MPE. We aimed to identify the reason for impaired T cell activity in MPE from lung cancer patients and to provide clues toward potential immune therapies for MPE. The surface inhibitory molecules and cytotoxic activity of T cells in MPE and peripheral blood (PB) were analyzed using flow cytometry. Levels of inflammatory cytokines in MPE and PB were tested using ELISA. TGF-Œ≤ expression in tumor-associated macrophages (TAMs) was also analyzed. The effect of TAMs on T cells was verified in vitro. Lastly, changes in T cells were evaluated following treatment with anti-TGF-Œ≤ antibody. We found that expression levels of Tim-3, PD-1 and CTLA-4 in T cells from MPE were upregulated compared with those from PB, but levels of IFN-Œ≥ and Granzyme B were downregulated (p &lt; 0.05). The amount of TGF-Œ≤ was significantly higher in MPE than in PB (p &lt; 0.05). TGF-Œ≤ was mainly produced by TAMs in MPE. When T cells were co-cultured with TAMs, expression levels of Tim-3, PD-1 and CTLA-4 were significantly higher than controls, whereas levels of IFN-Œ≥ and Granzyme B were significantly decreased, in a dose-dependent manner (p &lt; 0.05). In vitro treatment with anti-TGF-Œ≤ antibody restored the impaired T cell cytotoxic activity in MPE. Our results indicate that macrophage-derived TGF-Œ≤ plays an important role in impaired T cell cytotoxicity. It will therefore be valuable to develop therapeutic strategies against TGF-Œ≤ pathway for MPE therapy of lung cancer. 

Retracted: No

PubPeer comments:
 Dear Dr. Xu-Feng Huang, Thank you for your attention to our research. In response to your questions, we have carefully checked the manuscript and our original data, in Fig.4c, the values of the FSC and SSC axes should be demonstrated in decimal. We made mistakes due to our unified coordinate ruler frame in log10 format. We will send the corrigendum materials to the editorial office of International Journal of Cancer for correction. This change doesnu2019t affect any conclusions of the original paper. Thanks. Best regards, Lifeng Li. , 
 #2 Dear Dr Jieyao Li, I did not make these comments. Someone has used my name to make some comments over the last several days. It does not represent my view. Kind regards Dr Xu-Feng Huang University of Wollongong, Australia , 
 Looks like someone has commented on this publication. I feel surprised by the coordinates of FSC and SSC axis in Fig.4c. They are labelled in log. But it looks that the photo is from a plot with the axis in linear, as the dots are too dispersed. In the other article from the first author (PMID:30668541), such odd markings are also seen.  Moreover, there are other issues in this publication. In Fig.4c, the lines of gates seem to drawn manually.  In the lower row of Fig.4c, there are no CD14+ cells showing at the bottom in the left panel. But the cells in the left and middle panels are from the same 7AAD-CD3- subsets, the CD14+ cells should appear in both plots.  In Fig.3, the ratios of TAM in MPE are different between panel a (62%) and b (around 50%). According to their description, these data are collected from the same subjects, the number should be the same.  In Fig.1a, the gating of CD8+ cells is not consistent. In some plots, all CD8+ cells are included (the green one). In the other plots, either the cells with dim (the blue ones) or bright (the red ones) CD8 staining are included.  , 
 15 July 2022 Expression of Concern https: onlinelibrary.wiley.com doi 10.1002 ijc.34205 Expression of Concern: Impaired T cell function in malignant pleural effusion is caused by TGF-u03b2 derived predominantly from macrophages. Lifeng Li, Li Yang, Liping Wang, Fei Wang, Zhen Zhang, Jieyao Li, Dongli Yue, Xinfeng Chen, Yu Ping, Lan Huang, Bin Zhang, Yi Zhang. Int J Cancer. 2016 Nov 15;139(10):2261-9. This Expression of Concern is for the above article, published online on 6 August, 2016 in Wiley Online Library and has been published by agreement between the journal Editor-in-Chief, the authors, and Wiley Periodicals LLC. The Expression of Concern has been agreed due to concerns about the experimental standards applied for the experiments shown in Fig. 1a and Fig. 4c. In these figures, the gating of cells was not consistent, resulting in diverse plots that include different cell populations. There were discrepancies between some images shown in the manuscript and those provided by the authors as original flow cytometry plots. Moreover, the lines of the gates for the FACS plots in Fig. 4c have been drawn manually instead of being generated consistently by the software, and in the upper left plot, the scaling of the axes is wrong. The authors explain the issues by pointing to errors in laying out the figures and an irregular gating strategy but state that the errors do not affect the conclusions of the article. Since the issues mentioned above could not be resolved finally and there is inconclusive evidence about the reliability of the flow cytometry data in this article, the journal is issuing this Expression of Concern to alert readers. ,