CD3+ Macrophages Deliver Proinflammatory Cytokines by a CD3- and Transmembrane TNF-Dependent Pathway and Are Increased at the BCG-Infection Site

ORIGINAL RESEARCH published: 07 November 2019 doi: 10.3389/fimmu.2019.02550 CD3+ Macrophages Deliver Proinflammatory Cytokines by a CD3- and Transmembrane TNF-Dependent Pathway and Are Increased at the BCG-Infection Site Adriana Rodriguez-Cruz 1 , Dominique Vesin 2 , Lucero Ramon-Luing 3 , Joaquin Zuñiga 4 , Valérie F. J. Quesniaux 5 , Bernhard Ryffel 5 , Ricardo Lascurain 1,6 , Irene Garcia 2 and Leslie Chávez-Galán 3* Edited by: Elodie Segura, Institut Curie, France Reviewed by: Hsi-Hsien Lin, Chang Gung University, Taiwan Tina Fuchs, University of Heidelberg, Germany *Correspondence: Leslie Chávez-Galán Specialty section: This article was submitted to Antigen Presenting Cell Biology, a section of the journal Frontiers in Immunology Received: 04 May 2019 Accepted: 15 October 2019 Published: 07 November 2019 Citation: Rodriguez-Cruz A, Vesin D, Ramon-Luing L, Zuñiga J, Quesniaux VFJ, Ryffel B, Lascurain R, Garcia I and Chávez-Galán L (2019) CD3+ Macrophages Deliver Proinflammatory Cytokines by a CD3and Transmembrane TNF-Dependent Pathway and Are Increased at the BCG-Infection Site. Front. Immunol. 10:2550. doi: 10.3389/fimmu.2019.02550 1 Department of Biochemistry, Faculty of Medicine, Universidad Nacional Autónoma de México, Mexico City, Mexico, Department of Pathology and Immunology, Faculty of Medicine, Centre Medical Universitaire, University of Geneva, Geneva, Switzerland, 3 Laboratory of Integrative Immunology, Instituto Nacional de Enfermedades Respiratorias “Ismael Cosío Villegas”, Mexico City, Mexico, 4 Laboratory of Immunobiology and Genetics, Instituto Nacional de Enfermedades Respiratorias “Ismael Cosío Villegas”, Mexico City, Mexico, 5 Experimental Molecular Immunology and Neurogenetics (UMR7355), CNRS and University of Orléans, Orléans, France, 6 Hospital Nacional Homeopático, Secretaría de Salud, Mexico City, Mexico 2 Macrophages are essential cells of the innate immune response against microbial infections, and they have the ability to adapt under both pro- and anti-inflammatory conditions and develop different functions. A growing body of evidence regarding a novel macrophage subpopulation that expresses CD3 has recently emerged. Here, we explain that human circulating monocytes can be differentiated into CD3+ TCRαβ+ and CD3+ TCRαβ− macrophages. Both cell subpopulations express on their cell surface HLA family molecules, but only the CD3+ TCRαβ+ macrophage subpopulation co-express CD1 family molecules and transmembrane TNF (tmTNF). CD3+ TCRαβ+ macrophages secrete IL-1β, IL-6 IP-10, and MCP-1 by both tmTNF- and CD3-dependent pathways, while CD3+ TCRαβ− macrophages specifically produce IFN-γ, TNF, and MIP-1β by a CD3-dependent pathway. In this study, we also used a mouse model of BCG-induced pleurisy and demonstrated that CD3+ myeloid cells (TCRαβ+ and TCRαβ− cells) are increased at the infection sites during the acute phase (2 weeks post-infection). Interestingly, cell increment was mediated by tmTNF, and the soluble form of TNF was dispensable. BCG-infection also induced the expression of TNF receptor 2 on CD3+ myeloid cells, which increased after BCG-infection, suggesting that the tmTNF/TNFRs axis plays an important role in the presence or function of these cells in tuberculosis. Keywords: macrophage, CD3, TNF pathway, pro-inflammatory cytokines, BCG infection Frontiers in Immunology | www.frontiersin.org 1 November 2019 | Volume 10 | Article 2550 Activation of CD3+ Macrophages Rodriguez-Cruz et al. AUTHOR SUMMARY response is activated and a large number of leukocytes are recruited from blood vessels to the site of infection, including monocytes, which differentiate into macrophage subpopulations that in turn participate in the formation of granulomas (4). Granulomas have been assigned wide-ranging functions; on one hand, they are proposed as a vehicle to spread the pathogen, and on the other hand, as a successful manner of sequestering the pathogen together with cells producing inflammatory mediators (5). Tumor necrosis factor (TNF) is a pro-inflammatory cytokine that plays a main role in generating and maintaining the tuberculous granuloma (6). TNF is synthesized as a transmembrane form precursor (tmTNF) and then cleaved by the TNF-α-converting enzyme (TACE) upon activation stimuli to release the soluble form of TNF (solTNF). Both tmTNF and solTNF are bioactive molecules that interact with TNF receptor type 1 and 2 (TNFR1 and TNFR2) to induce cellular activation (7). T cell-derived TNF is required for the formation of granulomas, but to sustain the protective immunity against M. tuberculosis, both T cell- and macrophage-derived TNF molecules are necessary (8). Our previous studies have shown that a Mycobacterium bovis bacillus Calmette-Guérin (BCG)-induced pleural infection in TNF KO and double TNFR1 and TNFR2 (TNFR1R2) KO mice was associated with exacerbated inflammation that in turn impaired bacterial clearance and was lethal to the host (9). We also demonstrated that during BCG-induced pleurisy, myeloid-derived suppressor cells (MDSC) accumulated in the pleural cavity and tmTNF expressed by MDSC mediated immune suppression through interaction with CD4+ T cells expressing TNFR2. This interaction led to the attenuation of the excessive inflammatory response by the control of the proliferation of T cells producing IFN-γ (10). Those reports together support evidence about TNF playing a main role in controlling mycobacterial infections, both favoring an inflammatory response and activating a regulatory mechanism. A previous report noted a subpopulation of macrophages expressing the CD3/T cell receptor (TCR)αβ complex (CD3+ macrophages), generated by V(D)J recombination, accumulated in granulomas from TB patients. Using an in vitro model of BCG infection, the authors confirmed that the infection increased the frequency of CD3+ macrophages, and the presence of TNF was essential to maintain this cell subpopulation (11). Recently we demonstrated that BCG infection induces an increase of the recruitment of CD11b+ CD3+ phagocytic cells, and it is dependent of TNFR1 expression on myeloid cells, more interesting, we reported there are two CD3+ myeloid subpopulation, one more abundant and phenotypically TCRαβ− and a second TCRαβ+ (12). In this study, we used monocyte-derived macrophages (MDM) from healthy human donors to identify the frequency of CD3+ macrophages and to assess the role of TNF and CD3 in the activation and differentiation of CD3+ macrophages. Then, we used a murine model of BCG-induced pleural infection to investigate the role of tmTNF vs. solTNF to regulate the presence of CD3+ myeloid cells to the infection site. Our data corroborate the presence of human CD3+ macrophages, In response to physiologic changes or infectious challenge, macrophages play a main role in maintaining cellular homeostasis in tissues. Macrophages have high plasticity, and their differentiation process produces heterogeneous results. Recently, the eviden (...truncated)