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Article Open Access

Preliminary exploration of the role of CD8+ T cells in anti‑PD‑1 antibody‑induced myocarditis in C57BL/6 mice with Lewis lung carcinoma

  • Authors:
    • Huachun Chen
    • Fakai Li
    • Jun Song
    • Sheng Wang
    • Hanwen Xu
    • Ruoya Lyu
    • Hui Zhang
    • Guiyuan Chen
    • Xiaoyu Wu
    • Mingxing Ding
  • View Affiliations / Copyright

    Affiliations: Department of Respiratory and Critical Care Medicine, Jinhua Guangfu Tumor Hospital, Jinhua, Zhejiang 321001, P.R. China, Department of Science and Education, Jinhua Guangfu Tumor Hospital, Jinhua, Zhejiang 321001, P.R. China, Institute of Pharmacology, Jinhua Food and Drug Inspection and Testing Research Institute, Jinhua, Zhejiang 321002, P.R. China, Nursing Faculty, Medical College of Jinhua University of Vocational Technology, Jinhua, Zhejiang 321007, P.R. China, Medical Molecular Biology Laboratory, Medical College of Jinhua University of Vocational Technology, Jinhua, Zhejiang 321007, P.R. China
    Copyright: © Chen et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 56
    |
    Published online on: March 9, 2026
       https://doi.org/10.3892/br.2026.2129
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Abstract

Globally, ~33% of patients with non‑small cell lung cancer receiving anti‑PD‑1 antibody therapy may experience significant immune‑related adverse events (irAEs). Among these, myocarditis is a rare but lethal irAE. The aim of the present study was to preliminarily explore the role of CD8+ T cells in anti‑PD‑1 antibody‑induced myocarditis in C57BL/6 mice with Lewis lung carcinoma (LLC). Orthotopic transplantation models were established using wild‑type or CD8 knockout (CD8‑/‑) C57BL/6 mice. Wild‑type and CD8‑/‑ C57BL/6 mice were separately divided into three groups: Control, LLC and LLC + anti‑PD‑1. LLC cell suspensions (1x105 cells) with 50 µl Matrigel Matrix were orthotopically injected into the left lung lobe of wild‑type or CD8‑/‑ C57BL/6 mice. Following needle removal, the incision was sutured. At 10 days post‑surgery, mice in the anti‑PD‑1 groups received an intraperitoneal injection of anti‑PD‑1 antibody (200 µg). After 3 weeks, all mice were humanely euthanized via intraperitoneal injection of sodium pentobarbital (200 mg/kg). The histopathological examination of tumor, lung and heart tissue was performed by Masson's trichrome and hematoxylin and eosin staining. Reverse transcription‑quantitative PCR was carried out to determine the mRNA expression of monocyte chemotactic protein‑1, interleukin‑6, interferon‑γ and tumor necrosis factor‑α in myocardial tissue. Flow cytometry was used to analyze the ratio of CD8+ and CD4+ T cells and macrophages in myocardial tissues. Anti‑PD‑1 therapy effectively inhibited tumor growth and mitigated lung tissue damage. In wild‑type C57BL/6 mouse, treatment with anti‑PD‑1 was associated with myocardial injury, inflammatory responses and a notable increase in the ratios of CD8+ and CD4+ T cells and macrophages. However, in CD8‑/‑ C57BL/6 mice, no significant differences were observed in myocardial histopathology, inflammatory cytokine levels and the ratios of CD4+ T cells and macrophages between the control, LLC and LLC + anti‑PD‑1 groups. Anti‑PD‑1 therapy did not cause significant damage to myocardial tissue. The presence of CD8+ T cells facilitated the development of anti‑PD‑1‑induced myocarditis by activating CD4+ T cells, macrophages and inflammatory responses.
View Figures

Figure 1

Anti-PD-1 therapy improves the
histopathology of lung and tumor tissue but induces myocarditis in
wild-type mice. (A) Representative photographs of the lungs
containing the tumors. HE staining was performed to examine the
histopathology of (B) tumor (black arrow, nucleus; red arrow,
inflammatory cell infiltration), (C) lung (arrow, interstitial
congestion) and (D) heart tissue (black arrow, intercellular space;
red arrow, inflammatory cell infiltration). (E) Masson's trichrome
staining was performed to examine the histopathology of heart
tissue. Arrow, collagen fiber. Scale bar, 50 µm.
*P<0.05 vs. control, ###P<0.001 vs.
LLC. HE, hematoxylin-eosin; LLC, Lewis lung carcinoma.

Figure 2

Anti-PD-1 therapy activates
CD8+ and CD4+ T cells and macrophages and
induces secretion of inflammatory cytokines in wild-type mice. (A)
Ratios of CD68+, CD4+ and CD8+ T
cells in myocardial tissue were analyzed through flow cytometry.
(B) mRNA expression of MCP-1, IL-6, IFN-γ and TNF-α in myocardial
tissue was determined by reverse transcription-quantitative PCR.
***P<0.001, ****P<0.0001. ns, not
significant; MCP-1, monocyte chemoattractant protein-1; LLC, Lewis
lung carcinoma.

Figure 3

Effects of anti-PD-1 therapy on the
histopathology of tumor, lung and heart tissue in CD8-/-
mice. (A) Representative photographs of the lungs containing the
tumors. HE staining was performed to examine the histopathology of
(B) tumor (black arrow, nucleus; red arrow, inflammatory cell
infiltration), (C) lung (arrow, interstitial congestion) and (D)
heart tissue (black arrow, intercellular space; red arrow,
inflammatory cell infiltration). (E) Masson's trichrome staining
was performed to examine the histopathology of heart tissue. Arrow,
collagen fiber. Scale bar, 50 µm. ***P<0.001. HE,
hematoxylin-eosin; LLC, Lewis lung carcinoma; ns, not
significant.

Figure 4

Effect of anti-PD-1 therapy on
CD4+ T cell and macrophage levels and inflammatory
cytokine expression. (A) Ratios of CD68+ and
CD4+ T cells in myocardial tissue were analyzed through
flow cytometry. (B) mRNA expression of MCP-1, IL-6, IFN-γ and TNF-α
in myocardial tissue was determined via reverse
transcription-quantitative PCR. *P<0.05. ns, not
significant; LLC, Lewis lung carcinoma.
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Spandidos Publications style
Chen H, Li F, Song J, Wang S, Xu H, Lyu R, Zhang H, Chen G, Wu X, Ding M, Ding M, et al: Preliminary exploration of the role of CD8<sup>+</sup> T cells in anti‑PD‑1 antibody‑induced myocarditis in C57BL/6 mice with Lewis lung carcinoma. Biomed Rep 24: 56, 2026.
APA
Chen, H., Li, F., Song, J., Wang, S., Xu, H., Lyu, R. ... Ding, M. (2026). Preliminary exploration of the role of CD8<sup>+</sup> T cells in anti‑PD‑1 antibody‑induced myocarditis in C57BL/6 mice with Lewis lung carcinoma. Biomedical Reports, 24, 56. https://doi.org/10.3892/br.2026.2129
MLA
Chen, H., Li, F., Song, J., Wang, S., Xu, H., Lyu, R., Zhang, H., Chen, G., Wu, X., Ding, M."Preliminary exploration of the role of CD8<sup>+</sup> T cells in anti‑PD‑1 antibody‑induced myocarditis in C57BL/6 mice with Lewis lung carcinoma". Biomedical Reports 24.5 (2026): 56.
Chicago
Chen, H., Li, F., Song, J., Wang, S., Xu, H., Lyu, R., Zhang, H., Chen, G., Wu, X., Ding, M."Preliminary exploration of the role of CD8<sup>+</sup> T cells in anti‑PD‑1 antibody‑induced myocarditis in C57BL/6 mice with Lewis lung carcinoma". Biomedical Reports 24, no. 5 (2026): 56. https://doi.org/10.3892/br.2026.2129
Copy and paste a formatted citation
x
Spandidos Publications style
Chen H, Li F, Song J, Wang S, Xu H, Lyu R, Zhang H, Chen G, Wu X, Ding M, Ding M, et al: Preliminary exploration of the role of CD8<sup>+</sup> T cells in anti‑PD‑1 antibody‑induced myocarditis in C57BL/6 mice with Lewis lung carcinoma. Biomed Rep 24: 56, 2026.
APA
Chen, H., Li, F., Song, J., Wang, S., Xu, H., Lyu, R. ... Ding, M. (2026). Preliminary exploration of the role of CD8<sup>+</sup> T cells in anti‑PD‑1 antibody‑induced myocarditis in C57BL/6 mice with Lewis lung carcinoma. Biomedical Reports, 24, 56. https://doi.org/10.3892/br.2026.2129
MLA
Chen, H., Li, F., Song, J., Wang, S., Xu, H., Lyu, R., Zhang, H., Chen, G., Wu, X., Ding, M."Preliminary exploration of the role of CD8<sup>+</sup> T cells in anti‑PD‑1 antibody‑induced myocarditis in C57BL/6 mice with Lewis lung carcinoma". Biomedical Reports 24.5 (2026): 56.
Chicago
Chen, H., Li, F., Song, J., Wang, S., Xu, H., Lyu, R., Zhang, H., Chen, G., Wu, X., Ding, M."Preliminary exploration of the role of CD8<sup>+</sup> T cells in anti‑PD‑1 antibody‑induced myocarditis in C57BL/6 mice with Lewis lung carcinoma". Biomedical Reports 24, no. 5 (2026): 56. https://doi.org/10.3892/br.2026.2129
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