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The Expression of CTLA-4 in Breast Tumors and Tumor-Infiltrating Leukocytes Affects Patients’ Systemic Inflammatory Status and Varies According to Their Molecular Subtypes

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Abstract

Recent evidence has pointed out that the cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) expression is a poor prognosis factor. However, the implications of CTLA-4 expression on circulating inflammatory mediators are unclear for breast cancer. Tumor biopsies and blood samples were collected from 117 breast cancer patients. Oxidative stress parameters were evaluated in plasma samples by measuring the lipoperoxidation profile and nitric oxide metabolites (NOx). Interleukins 12 (IL-12) and 4 (IL-4) were assessed by ELISA. CTLA-4 expression was determined by immunofluorescence assessed by its labeling in tumor-infiltrating leukocytes (TILs) or breast tumors. Correlations between CTLA-4 expression in breast tumors with TCD4/TCD8 infiltrating lymphocyte and inflammation-related genes were performed using data from TIMER 2.0/TCGA databases (n = 2160). CTLA-4 expression in TILs significantly correlated to triple-negative breast tumors. Patients carrying CTLA-4-positive tumors exhibited lower plasmatic NOx levels, and those expressing CTLA-4 in TILs had reduced levels of IL-12 in plasma. No changes in either IL-4 or lipid peroxidation profiles were detected concerning any CTLA4 status. Compared to the Luminal A ones, oxidative stress parameters and cytokines were observed in patients bearing triple-negative tumors. CTLA-4 expression in all breast cancer subtypes positively correlated to TCD4/TCD8 lymphocyte infiltrates, as well as to the pro-inflammatory genes IL12A, IL4, NFKB1, NFKB2, NOS1, NOS2, and NOS3. CTLA-4 expression in both tumor and TILs can affect the systemic inflammatory status of breast cancer patients, especially antitumor molecules such as IL-12 and NOx that correlate to more aggressive disease.

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Acknowledgements

The authors are grateful for all lab and hospital personnel, funding agencies, and patients.

Funding

This work was supported by Fundação Araucária, Programa de Pesquisa Para o SUS – PPSUS, Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, Edital Universal).

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Authors and Affiliations

  1. Laboratório Biologia de Tumores, Universidade Estadual Do Oeste Do Paraná, Francisco Beltrão, PR, Brazil

    Rodrigo Kern, Janaina Carla da Silva, Mariane Okamoto Ferreira, Matheus Iago Oliveira Coletto, Stefania Tagliari de Oliveira, Fernanda Mara Alves, Thalita Basso Scandolara, Daniel Rech & Carolina Panis

  2. Programa de Pós-Graduação Em Ciências Aplicadas À Saúde, Universidade Estadual Do Oeste Do Paraná, Francisco Beltrão, PR, Brazil

    Rodrigo Kern, Janaina Carla da Silva, Mariane Okamoto Ferreira, Thalita Basso Scandolara, Daniel Rech & Carolina Panis

  3. Department of Biochemistry and Molecular Medicine, Université de Montréal, Montreal, Canada

    Janaina Carla da Silva

  4. Universidade Estadual Do Oeste Do Paraná, Campus Cascavel, PR, Brazil

    Fábio Negretti

  5. Programa de Residência Em Clínica Médica, Rede de Assistência À Saúde Metropolitana, Sarandi, PR, Brazil

    Stefania Tagliari de Oliveira

  6. Molecular Carcinogenesis Program, Brazilian National Cancer Institute (INCA), Research Coordination (CPQ), Rio de Janeiro, RJ, Brazil

    Thalita Basso Scandolara

  7. Hospital de Câncer de Francisco Beltrão, Francisco Beltrão, PR, Brazil

    Daniel Rech

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  1. Rodrigo Kern
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All authors contributed to the study conception and design, material preparation, data collection, and analysis. Rodrigo Kern and Carolina Panis wrote the first draft of the manuscript, and all authors read and approved the final manuscript.

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Correspondence to Carolina Panis.

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Kern, R., da Silva, J.C., Negretti, F. et al. The Expression of CTLA-4 in Breast Tumors and Tumor-Infiltrating Leukocytes Affects Patients’ Systemic Inflammatory Status and Varies According to Their Molecular Subtypes. Inflammation 46, 1639–1652 (2023). https://doi.org/10.1007/s10753-023-01830-5

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