Journal
INNATE IMMUNITY
Volume 26, Issue 6, Pages 459-472Publisher
SAGE PUBLICATIONS LTD
DOI: 10.1177/1753425920915507
Keywords
TLR; TLR3; AhR; innate immunity; genetic screen
Categories
Funding
- Institut Curie, INSERM, CNRS
- INCA [40832]
- LNCC PARIS [RS19/75-1]
- ARC [PJA 20181208045]
- Sidaction [2018-1-AEQ-11984, VIH2016126002]
- ANRS
- ANRS [ECTZ25472, ECTZ36691]
- E ' cole normale superieure de Lyon
- Association de la Recherche contre le Cancer (ARC)
- [ANR-10-IDEX-0001-02 PSL]
- [ANR-11-LABX-0043]
- [ANR 15-CE130009-01]
- [ANR-14-CE14-0004-02]
- [ANR-17-CE15-0025-01]
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A subset of TLRs is specialised in the detection of incoming pathogens by sampling endosomes for nucleic acid contents. Among them, TLR3 senses the abnormal presence of double-stranded RNA in the endosomes and initiates a potent innate immune response via activation of NF-kappa B and IRF3. Nevertheless, mechanisms governing TLR3 regulation remain poorly defined. To identify new molecular players involved in the TLR3 pathway, we performed a genome-wide screen using CRISPR/Cas9 technology. We generated TLR3(+) reporter cells carrying a NF-kappa B-responsive promoter that controls GFP expression. Cells were next transduced with a single-guide RNA (sgRNA) library, subjected to sequential rounds of stimulation with poly(I:C) and sorting of the GFP-negative cells. Enrichments in sgRNA estimated by deep sequencing identified genes required for TLR3-induced activation of NF-kappa B. Among the hits, five genes known to be critically involved in the TLR3 pathway, including TLR3 itself and the chaperone UNC93B1, were identified by the screen, thus validating our strategy. We further studied the top 40 hits and focused on the transcription factor aryl hydrocarbon receptor (AhR). Depletion of AhR had a dual effect on the TLR3 response, abrogating IL-8 production and enhancing IP-10 release. Moreover, in primary human macrophages exposed to poly(I:C), AhR activation enhanced IL-8 and diminished IP-10 release. Overall, these results reveal AhR plays a role in the TLR3 cellular innate immune response.
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