Impact of Notch1 Deletion in Macrophages on Proinflammatory Cytokine Production and the Outcome of Experimental Autoimmune Encephalomyelitis.
Notch signaling is involved in regulating TLR-mediated responses in activated macrophages. In this study, we investigated the impact of Notch signaling in macrophages in an experimental autoimmune encephalomyelitis (EAE) model. To examine the impact of deficiency in Notch signaling in activated macrophages in EAE, an adoptive transfer of activated macrophages derived from Notch1(fl/fl) × Mx1cre(+/-) (Notch1 knockout [N1KO]) or CSL/Rbp-jκ(fl/fl) × Mx1cre(+/-) (CSL/RBP-Jκ KO) mice was performed prior to induction of EAE. Mice receiving activated N1KO macrophages showed decreased severity of EAE compared with mice receiving wild-type or CSL/RBP-Jκ KO macrophages. In vitro restimulation of splenocytes by myelin oligodendrocyte glycoprotein 35-55 peptide from these mice revealed that cells from mice receiving N1KO macrophages produced significantly less IL-17 compared with the control mice, whereas IFN-γ production was similar in both groups. We found that activated N1KO, but not CSL/RBP-Jκ KO, macrophages produced less IL-6 and had lower CD80 expression compared with wild-type and did not exhibit any defect in IL-12p40/70 production, whereas activated macrophages from CSL/RBP-Jκ KO mice phenocopied γ-secretase inhibitor treatment for reduced IL-12p40/70 production. Furthermore, the nuclear translocation of the NF-κB subunit c-Rel was compromised in γ-secretase inhibitor-treated and CSL/RBP-Jκ KO but not N1KO macrophages. These results suggest that Notch1 and CSL/RBP-Jκ in macrophages may affect the severity of EAE differently, possibly through modulating IL-6 and CD80 expression, which is involved in the Th17 but not Th1 response.
Graduate Program in Biotechnology, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand;
Wipawee
W
Wongchana
Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, MA 01003;
Rebecca G
RG
Lawlor
Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, MA 01003; Molecular and Cellular Biology Program, University of Massachusetts, Amherst, MA 01003;
Barbara A
http://orcid.org/0000-0001-8255-0556
BA
Osborne
Department of Microbiology, Faculty of Science, Omics Sciences and Bioinformatics Center, Chulalongkorn University, Bangkok 10330, Thailand; and Center of Excellence in Immunology and Immune-mediated Diseases, Chulalongkorn University, Bangkok 10330, Thailand tanapat.p@chula.ac.th.
Tanapat
T
Palaga
D019264:Adoptive Transfer; D053829:Amyloid Precursor Protein Secretases / Q000037:antagonists & inhibitors; D000818:Animals; D018122:B7-1 Antigen / Q000096:biosynthesis; D002478:Cells, Cultured; D018920:Coculture Techniques; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000276:immunology*; D005260:Female; D017353:Gene Deletion; D051819:Immunoglobulin J Recombination Signal Sequence-Binding Protein / Q000235:genetics*; D007371:Interferon-gamma / Q000096:biosynthesis; D053762:Interleukin-12 Subunit p40 / Q000096:biosynthesis; D020381:Interleukin-17 / Q000096:biosynthesis; D015850:Interleukin-6 / Q000096:biosynthesis; D008264:Macrophages / Q000276:immunology* / Q000637:transplantation; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D020569:Proto-Oncogene Proteins c-rel / Q000378:metabolism; D051881:Receptor, Notch1 / Q000235:genetics*; D015398:Signal Transduction / Q000276:immunology; D058504:Th17 Cells / Q000276:immunology*
D018122:B7-1 Antigen; D051819:Immunoglobulin J Recombination Signal Sequence-Binding Protein; D053762:Interleukin-12 Subunit p40; D020381:Interleukin-17; D015850:Interleukin-6; C497762:Notch1 protein, mouse; D020569:Proto-Oncogene Proteins c-rel; C497571:Rbpj protein, mouse; D051881:Receptor, Notch1; D007371:Interferon-gamma; D053829:Amyloid Precursor Protein Secretases