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IL-6 secretion of CD4+ T cells stimulated by LC3-positive extracellular vesicles in human epithelial ovarian cancer

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Abstract

Background

Ovarian cancer (OC) as the most fatal gynecological malignancy worldwide, with epithelial ovarian cancer (EOC) being the predominant and most lethal form, poses a serious threat to human health. LC3-positive extracellular vesicles (LC3+ EVs) promote tumorigenesis by educating CD4+ T cells in a murine melanoma model. However, regulation of LC3+ EVs in human EOC remains largely unknown. 

Methods

Differential analysis of Rab8a, Hsp90α and Il6 expression was performed using GEPIA2. The number of LC3+ EVs and the frequency of Heat shock protein 90α+ LC3+ EVs (HSP90α+ LC3+ EVs) in the ascites of EOC patients were tested by flow cytometry. IL-6, IL-10, IFN-γ, IL-4 and TGF-β were measured by ELISA. CD4+ T cells were isolated from peripheral blood of healthy human donors using MACS magnetic bead technology. 

Results

Higher Rab8a, Hsp90a and Il6 expression of cancer tissues compared with normal adjacent tissues in OC were found. The level of IL-6 was positively correlated with LC3+ EVs number, HSP90α+ LC3+ EVs percentage in the ascites, and ROMA index of the patient. In addition, elevated IL-6 production by CD4+ T cells induced by LC3+ EVs was observed, which was suppressed by anti-HSP90α or anti-TLR2. 

Conclusions

LC3+ EVs level and HSP90α+ LC3+ EVs percentage were associated with elevated IL-6 in the ascites of EOC patients. HSP90α on LC3+ EVs from human EOC could stimulate CD4+ T cell production of IL-6 via TLR2.

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Abbreviations

LC3+ EVs:

LC3-positive extracellular vesicles

HSP90α:

Heat shock protein 90α

EOC:

Epithelial ovarian cancer

DAMPs:

Damage-associated molecular pattern molecules

DFS:

Disease-free survival

PFS:

Progression-free survival

OS:

Overall survival

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Funding

We are grateful for the support from grants from the National Natural Science Foundation of China [NO. 8187101983], and Maternal and Child Health Research Project of Jiangsu Province [F201923].

Author information

Author notes

  1. Xiu-Lin Zhu and Hao-Jue Wang contributed to the work equally and should be regarded as the co-first authors.

Authors and Affiliations

  1. Department of Microbiology and Immunology, Medical School of Southeast University, 87 Dingjiaqiao Rd., Nanjing, 210009, Jiangsu, China

    Xiu-Lin Zhu, Xu-Ru Wang, Xuan Ji, Li Xu, Qi Yang, Jie-Ying Guo, Yan Rong & Ning Pan

  2. Department of Obstetrics and Gynecology, Zhongda Hospital, Medical School of Southeast University, 87 Dingjiaqiao Rd., Nanjing, 210009, Jiangsu, China

    Di Wu & Yun-lang Cai

  3. Department of Obstetrics and Gynecology, Xishan People’s Hospital of Wuxi City, 1128 Dacheng Rd., Wuxi, 214000, Jiangsu, China

    Hao-Jue Wang

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  1. Xiu-Lin Zhu
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Contributions

YLC, NP and HJW designed this study. XLZ and XRW performed the experiments. XLZ and HJW wrote the manuscript, performed the literature search. DW, XJ, LX, QY, JYG, and YR collected the clinical data and helped write the manuscript. YLC and NP were responsible for manuscript revision. All the authors read and approved the final manuscript.

Corresponding authors

Correspondence to Ning Pan or Yun-lang Cai.

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The authors declare that they have no conflict of interest

Ethics approval

All the human experiments were approved by the Ethics Committee for Human Studies of Southeast University and performed under protocol 2019ZDSYLL021.

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Informed consent was obtained from all the patients

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Cite this article

Zhu, XL., Wang, HJ., Wang, XR. et al. IL-6 secretion of CD4+ T cells stimulated by LC3-positive extracellular vesicles in human epithelial ovarian cancer. Clin Transl Oncol 24, 2222–2230 (2022). https://doi.org/10.1007/s12094-022-02883-y

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  • DOI: https://doi.org/10.1007/s12094-022-02883-y

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