Transcriptome Analysis of Co-Cultures of THP-1 Human Macrophages with Inactivated Germinated Trichophyton rubrum Conidia

Bruna Aline Cantelli; Gabriela Gonzalez Segura; Tamires Aparecida Bitencourt; Mariana Heinzen de Abreu; Monise Fazolin Petrucelli; Kamila Peronni; Pablo Rodrigo Sanches; Rene Oliveira Beleboni; Wilson Araújo da Silva Junior; Nilce Maria Martinez-Rossi; Mozart Marins; Ana Lúcia Fachin

doi:10.3390/jof9050563

Transcriptome Analysis of Co-Cultures of THP-1 Human Macrophages with Inactivated Germinated Trichophyton rubrum Conidia

J Fungi (Basel). 2023 May 12;9(5):563. doi: 10.3390/jof9050563.

Authors

Bruna Aline Cantelli¹, Gabriela Gonzalez Segura¹, Tamires Aparecida Bitencourt², Mariana Heinzen de Abreu¹, Monise Fazolin Petrucelli^{1

3}, Kamila Peronni⁴, Pablo Rodrigo Sanches³, Rene Oliveira Beleboni^{1

5}, Wilson Araújo da Silva Junior³, Nilce Maria Martinez-Rossi³, Mozart Marins^{1

5}, Ana Lúcia Fachin^{1

5}

Affiliations

¹ Biotechnology Unit, University of Ribeirão Preto-UNAERP, Ribeirao Preto 14096-900, Brazil.
² Department of Biochemistry and Immunology, Ribeirão Preto Medical School, University of São Paulo, Ribeirao Preto 14049-900, Brazil.
³ Department of Genetics, Ribeirão Preto Medical School, University of São Paulo, Ribeirao Preto 14096-900, Brazil.
⁴ National Institute of Science and Technology in Stem Cell and Cell Therapy, Center for Cell-Based Therapy, Ribeirao Preto 14049-900, Brazil.
⁵ Medicine School, University of Ribeirão Preto-UNAERP, Ribeirao Preto 14096-900, Brazil.

Abstract

Although most mycoses are superficial, the dermatophyte Trichophyton rubrum can cause systemic infections in patients with a weakened immune system, resulting in serious and deep lesions. The aim of this study was to analyze the transcriptome of a human monocyte/macrophage cell line (THP-1) co-cultured with inactivated germinated T. rubrum conidia (IGC) in order to characterize deep infection. Analysis of macrophage viability by lactate dehydrogenase quantification showed the activation of the immune system after 24 h of contact with live germinated T. rubrum conidia (LGC). After standardization of the co-culture conditions, the release of the interleukins TNF-α, IL-8, and IL-12 was quantified. The greater release of IL-12 was observed during co-culturing of THP-1 with IGC, while there was no change in the other cytokines. Next-generation sequencing of the response to T. rubrum IGC identified the modulation of 83 genes; of these, 65 were induced and 18 were repressed. The categorization of the modulated genes showed their involvement in signal transduction, cell communication, and immune response pathways. In total, 16 genes were selected for validation and Pearson's correlation coefficient was 0.98, indicating a high correlation between RNA-seq and qPCR. Modulation of the expression of all genes was similar for LGC and IGC co-culture; however, the fold-change values were higher for LGC. Due to the high expression of the IL-32 gene in RNA-seq, we quantified this interleukin and observed an increased release in co-culture with T. rubrum. In conclusion, the macrophages-T. rubrum co-culture model revealed the ability of these cells to modulate the immune response, as demonstrated by the release of proinflammatory cytokines and the RNA-seq gene expression profile. The results obtained permit to identify possible molecular targets that are modulated in macrophages and that could be explored in antifungal therapies involving the activation of the immune system.

Keywords: IL-32; LPS; RNA sequencing; deep infection; dermatophytes.

Grants and funding

2019/10514-8/São Paulo Research Foundation