Sustained Egr-1 Response via p38 MAP Kinase Signaling Modulates Early Immune Responses of Dendritic Cells Parasitized by Toxoplasma gondii.

Arne L Ten Hoeve,Mohamed-Ali Hakimi,Antonio Barragan

doi:10.3389/fcimb.2019.00349

Abstract

As a response to a diverse array of external stimuli, early growth response protein 1 (Egr-1) plays important roles in the transcriptional regulation of inflammation and the cellular immune response. However, a number of intracellular pathogens colonize immune cells and the implication of Egr-1 in the host-pathogen interplay has remained elusive. Here, we have characterized the Egr-1 responses of primary murine and human dendritic cells (DCs) upon challenge with the obligate intracellular parasite Toxoplasma gondii. We report that live intracellular parasites induce a sustained high expression of Egr-1 in DCs, different from the immediate-early Egr-1 response to parasite lysates, inactivated parasites or LPS. Moreover, a distinct nuclear localization of elevated amounts of Egr-1 protein was detected in infected DCs, but not in by-stander DCs. The ERK1/2 MAPK signaling pathway mediated the canonical immediate-early Egr-1 response to soluble antigens in a MyD88/TLR-dependent fashion. In contrast, a non-canonical extended Egr-1 response that relied primarily on p38 MAPK signaling was induced by intracellular parasites and was exhibited similarly by MyD88-deficient and wildtype DCs. The extended phase Egr-1 response was dramatically reduced upon challenge of DCs with T. gondii parasites deficient in GRA24, a secreted p38-interacting protein. Further, Egr-1-silenced primary DCs maintained their migratory responses upon T. gondii challenge. Importantly, Egr-1 silencing led to elevated expression of co-stimulatory molecules (CD40, CD80) in Toxoplasma-infected DCs and in LPS-challenged immature DCs, indicating that Egr-1 responses suppressed maturation of DCs. Moreover, the IL-12 and IL-2 responses of Toxoplasma-challenged DCs were modulated in a GRA24-dependent fashion. Jointly, the data show that the Egr-1 responses of DCs to microbial external stimuli and intracellular stimuli can be selectively mediated by ERK1/2 or p38 MAPK signaling, and that Egr-1 can act as an intrinsic negative modulator of maturation in primary DCs.

Highlights

Leukocytes populate the blood and traffic the tissues, draining to the lymphatic system and back into the circulation (Friedl and Weigelin, 2008)
Highly significant differences in the cumulative fold change of early growth response protein 1 (Egr-1) mRNA expression were measured for Toxoplasma-challenged bone marrow-derived DCs (BMDCs) compared with heat-inactivated parasites, parasite lysate or LPS, while non-significant or minor differences were measured during the immediate-early phase (Figure 1B)
This indicated that the global Early growth response (Egr)-1 mRNA upregulation observed in T. gondii-challenged BMDCs was mirrored in nuclear Egr-1 protein levels in T. gondii-infected BMDCs but not in by-stander BMDCs

Summary

Introduction

Leukocytes populate the blood and traffic the tissues, draining to the lymphatic system and back into the circulation (Friedl and Weigelin, 2008). When actively invaded by T. gondii, DCs adopt a hypermigratory phenotype (reviewed in Weidner and Barragan, 2014), which mediates rapid systemic dissemination of Toxoplasma in mice (Lambert et al, 2006; Kanatani et al, 2017) This dramatic migratory activation requires the discharge of parasitic secretory organelles into the host cell cytoplasm (Weidner et al, 2013) and intracellular signaling (Fuks et al, 2012; Kanatani et al, 2017). Differences in responses have been reported for human and murine DCs and between DC subsets (Subauste and Wessendarp, 2000; Tosh et al, 2016) and the molecular mechanisms for how active invasion by the parasite modulates maturation have remained elusive

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Conclusion