Distinct TLR- and NLR-Mediated Transcriptional Responses to an Intracellular Pathogen

Abstract

How the innate immune system tailors specific responses to diverse microbial infections is not well understood. Cells use a limited number of host receptors and signaling pathways to both discriminate among extracellular and intracellular microbes, and also to generate responses commensurate to each threat. Here, we have addressed these questions by using DNA microarrays to monitor the macrophage transcriptional response to the intracellular bacterial pathogen Listeria monocytogenes. By utilizing combinations of host and bacterial mutants, we have defined the host transcriptional responses to vacuolar and cytosolic bacteria. These compartment-specific host responses induced significantly different sets of target genes, despite activating similar transcription factors. Vacuolar signaling was entirely MyD88-dependent, and induced the transcription of pro-inflammatory cytokines. The IRF3-dependent cytosolic response induced a distinct set of target genes, including IFNβ. Many of these cytosolic response genes were induced by secreted cytokines, so we further identified those host genes induced independent of secondary signaling. The host response to cytosolic bacteria was reconstituted by the cytosolic delivery of L. monocytogenes genomic DNA, but we observed an amplification of this response by NOD2 signaling in response to MDP. Correspondingly, the induction of IFNβ was reduced in nod2^−/− macrophages during infection with either L. monocytogenes or Mycobacterium tuberculosis. Combinatorial control of IFNβ induction by recognition of both DNA and MDP may highlight a mechanism by which the innate immune system integrates the responses to multiple ligands presented in the cytosol by intracellular pathogens. Macrophages are critical cells of the innate immune system, contributing to immediate and robust defense against microbial infections. Macrophages detect pathogens using host receptors located on the cell surface, in phagosomal vacuoles, and in the cytosol. While fundamental to innate immunity, it is not clear if these different receptors merely provide redundant mechanisms for sensing microbial infection, or if instead they induce distinct gene expression programs that may allow for threat-specific host responses. We addressed this question by dissecting the macrophage transcriptional responses to the model intracellular bacterial pathogen Listeria monocytogenes. Using genetic and genomic approaches, we found that the macrophage response to L. monocytogenes trapped in phagosomal compartments was distinct and separable from the response to live bacteria replicating in the host cytosol. The macrophage response to cytosolic bacteria was recapitulated by bacterial nucleic acid and cell wall fragments, and induced surprisingly few primary response genes. These findings highlight a mechanism by which the innate immune system may specifically sense intracellular bacteria, as the macrophage response to Mycobacterium tuberculosis was similarly regulated.