Plasticity of Treg and imbalance of Treg/Th17 cells in patients with systemic sclerosis modified by FK506
Authors: Qian Wang
Xinjuan Liu
Xiaofeng Zeng
Jianyu Hao
Yu Wu
Mengtao Li
Journal: International Journal of Immunopathology and Pharmacology (2021)
Discipline: PHARMACOLOGY & TOXICOLOGY
Keywords: FoxP3; IL-17; Tregs; systemic sclerosis; tacrolimus.
Affiliations: Department of Gastroenterology, Beijing Chaoyang Hospital, Capital Medical University, Chaoyang District, Beijing, China, Department of Rheumatology and Clinical Immunology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Key Laboratory of Rheumatology and Clinical Immunology, Ministry of Education, Beijing, China

Abstract:  To determine the effects of Tacrolimus (FK506) on Treg cells and subpopulations in SSc patients and assess the ability of FK506 to modify the immune imbalance of Treg/Th17 cells. We analyzed PBMC from five SSc patients and six healthy control by flow cytometry after cultured with 0, 0.1, 1, or 10 ng/ml FK506 in vitro. The number of Treg cells decreased in SSc patients treated with FK506. The number of FrI cells were decreased in SSc following FK506 treatment. The drug did increase the frequency of FrII/Treg cells, but not FrII cells. However, FK506 significantly decreased FrIII in both SSc patients and controls. FK506 clearly decreased the numbers of Th17 cells and FoxP3+IL-17+ cells. The proliferation capacity of cells was also inhibited by FK506, which had a greater effect on FoxP3- cells than FoxP3+ cells. FK506 did inhibit the proliferation of FrIII cells, but not FrI or FrII cells. Our study provides that FK506 reduced the number of FoxP3low CD45RA- T cells (FrIII) by inhibiting its proliferation. Therefore, FK506 modifies Treg cells and the immune imbalance between Tregs and Th17 cells in SSc patients. 

Retracted: No

PubPeer comments:
 Figure 6A: In the bottom row, two panels appear to show a large area of overlap. Shown in bright green. ![file] , 
 Figure 6B. Two panels that represent different transfections and proteins, appear to show an overlap. Shown in yellow. ![file] , 
 Supporting Figure S3A: Several bands appear to be visible multiple times within the same blot panel. Band are sometimes a bit different and spots appear or disappear but backgrounds are remarkably similar. ![file] , 
 #3 Fig 3A is another showcase. ![file] In the CyclinD2 slice, the lower region of the first two lanes displays a different grain texture from the rest of that slice, almost as if a section of image has been stretched vertically and then smeared around a little. , 
 #4 The CyclinD1 U251 is also of interest. There is a change of texture between lanes 2 and 3, as if the two left-hand lanes had been stretched vertically from an initially-narrower slice. Also a hint of a vertical discontinuity between lanes 3 and 4. ![file] , 
 Fig 3b: ![file] , 
 #4 Fig 4A is full of surprises. Some bands have more horizontal and vertical discontinuities than a patchwork quilt. ![file] , 
 Fig 2D: ![file] ![file] , 
 Fig 1A. ![file] , 
 From PlosOne:  Journal Information **PLOS ONE is an inclusive journal community working together to advance science for the benefit of society, now and in the future.** Founded with the aim of accelerating the pace of scientific advancement and demonstrating its value, we believe all rigorous science needs to be published and discoverable, widely disseminated and freely accessible to all. Dear Editor-in-Chief Joerg Heber, if you really want to advance science for the benefit of society, now and in the future, you should do something with the articles published by Jasti S Rao and the many other problematic PlosOne articles discussed at PubPeer (871 articles at the moment). (copy of the email sent to EiC). , 
 2020 retraction. https: journals.plos.org plosone article?id=10.1371 journal.pone.0228690 After this article [1] was published, the following concerns were raised: Figure 1A SNB19, Cathepsin B: Lanes 2 and 3 appear similar. SNB19, uPAR: Lanes 2 and 4 appear similar. U251, Cathespin B: There appears to be a vertical discontinuity after lane 2, and lanes 1 and 3 appear similar. U251, uPAR: There appear to be vertical discontinuities after lanes 2 and 4. Figure 2A U251, p-p27 (ser10): There appears to be a sharp horizontal discontinuity at the top of bands in lanes 3u20135. U251, p-p27 (Thr187): The background resolution (texture) appears different in lanes 3 and 4 than in lanes 1, 2, 5. Figure 2D SNB19, p27: Lanes 4 and 5 appear similar and there are similarities in some background regions in lanes 1 and 2. U251, p27: Lanes 4 and 5 appear similar. U251, GAPDH: Lanes 3 and 4 appear similar to lanes 5 and 6, respectively. Figure 3A SNB19, Rb: There appears to be a vertical discontinuity after lane 2. SNB19, GAPDH: There appear to be similarities between upper portions of lanes 1, 2, 4, and between lanes 3, 5. U251, p-CDK2: Lanes 1 and 2 appear similar. U251, CyclinE: There appear to be similarities between image elements in lanes 1 and 2. U251, CyclinD1: The background pattern (texture) appears different between lanes 1u20132 and lanes 3u20135, and there appears to be a vertical discontinuity after lane 3. U251, CyclinD2: There appear to be similarities between image elements in lanes 4 and 5, and the background pattern (texture) for the lower portions of lanes 1, 2 appears different than that for the remainder of the panel. U251, p-Rb (Ser780): Lanes 1 and 2 appear similar, when horizontally flipped. U251, p21: Lanes 1 and 2 appear similar, and there appear to be dark gray markings below the band in lane 1. U251, Ki67: Lanes 1 and 2 appear similar, when horizontally flipped, except for an area at the top of the band in lane 1. Figure 3B U251, IP-CDK2 IB-CyclinE: There appear to be similarities between the upper halves of lanes 1 and 2. U251, u03b1Vu03b23 (native gel): There appears to be a vertical discontinuity before lane 5. Figure 4A SNB19, p-FOXO3a(Ser253): There appear to be similarities between background in lanes 1 and 2. SNB19, p-FOXO3a(Ser318): There appear to be similarities between background in lanes 1 and 2. SNB19, PI3K: There appear to be similarities between background and parts of the bands in lanes 2 and 5. SNB19, PI3K lanes 1, 4 and SNB19, p-Akt lane 2 (flipped horizontally, aspect ratio adjusted) appear similar. There appear to be similarities between SNB19, PI3K lane 3 and SNB19, p-Akt lane 3 (flipped horizontally). SNB19, GAPDH: There appear to be horizontal discontinuities in background above and below the bands, and there appears to be a vertical discontinuity after lane 2. U251, FOXO-3a: When levels are adjusted, there appear to be horizontal and vertical discontinuities around the band in lane 4. U251, p-FOXO3a(Ser253): When levels are adjusted there appears to be a vertical discontinuity after lane 2. U251, p-Akt: There appears to be a vertical discontinuity before lane 3. In Figure 6A, the U251 Control Mock and SV panels appear to show overlapping data. In Figure 6B, the SNB19 Control Mock, Cathepsin B panel (upper region) and SNB19 SV, Ki67 panel (lower region) appear similar. Figure S3A FOXO1, left panel: There appear to be similarities between lanes 1, 2, 5, and between lanes 3 and 4. p-FOXO1 (Thr24), left panel: There appear to be similarities between lanes 1 and 3, and between lanes 2 and 4. p-FOXO1 (Ser256), left panel: Areas of lanes 1 and 2 appear similar. p-FOXO1 (Ser319) FOXO4(Ser262): There appear to be similarities between lanes 1 and 2 of each panel. p-FOXO4 (Ser262): There appear to be similarities between lanes 1, 2, 3 and 5 of the left panel, and when levels are adjusted there appear to be vertical discontinuities after lanes 1, 2, and 4 of the left panel and after lane 1 of the right panel. GAPDH, left panel: There appear to be similarities between lanes 1 and 2. The above concerns call into question the integrity and reliability of the published results, and so the PLOS ONE Editors retract this article. We attempted to contact the articleu2019s authors to discuss these issues, request the underlying data, and notify them of the retraction decision, but the authors either did not reply or could not be reached. Reference 1.Gopinath S, Malla RR, Gondi CS, Alapati K, Fassett D, Klopfenstein JD, et al. (2010) Co-Depletion of Cathepsin B and uPAR Induces G0 G1 Arrest in Glioma via FOXO3a Mediated p27Kip1 Upregulation. PLoS ONE 5(7): e11668. https: doi.org 10.1371 journal.pone.0011668 pmid:20661471 ,