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3-Methyl-4-nitrophenol triggers nasal allergy by modulating dendritic cell properties  Xiao-Yu Liu, PhD, Yong-Jin Wu, MD, PhD, Li-Juan Song, MD, Xian-Hai.

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Presentation on theme: "3-Methyl-4-nitrophenol triggers nasal allergy by modulating dendritic cell properties  Xiao-Yu Liu, PhD, Yong-Jin Wu, MD, PhD, Li-Juan Song, MD, Xian-Hai."— Presentation transcript:

1 3-Methyl-4-nitrophenol triggers nasal allergy by modulating dendritic cell properties 
Xiao-Yu Liu, PhD, Yong-Jin Wu, MD, PhD, Li-Juan Song, MD, Xian-Hai Zeng, MD, Shuai Wang, PhD, Jiang-Qi Liu, MD, Li-Hua Mo, MSc, Xiao-Rui Geng, MD, Li-Teng Yang, MD, Rui-Di Xie, MD, Xiao-Wen Zhang, MD, PhD, Zhi-Gang Liu, MD, PhD, Ping-Chang Yang, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 143, Issue 4, Pages e7 (April 2019) DOI: /j.jaci Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 MNP increases Bcl2L12 and TH2 response–relevant factors in DCs. BMDCs were generated and exposed to MNP at the indicated concentrations in culture for 48 hours. DCs were collected and analyzed by using quantitative RT-PCR and Western blotting. The results showed levels of TIM4 (A), Bcl2L12 (B), MHC class II (C), and CD80 (D) in DCs. Data shown in bars are presented in means ± SDs. *P < .01 compared with the “0” group. Results represent 3 independent experiments. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Bcl2L12 facilitates formation of MHC class II/OVA complexes in DCs. A-C, Immunoblots show a complex of MHC class II, Bcl2L12, and OVA in extracts of WT BMDCs after exposure to OVA and MNP (Fig 2, A); a complex of MHC class II, Bcl2L12, and OVA in extracts of WT BMDCs after exposure to OVA (Fig 2, B); or a complex of MHC class II and OVA in KO BMDCs after exposure to MNP and OVA (Fig 2, C). D and E, Complex of recombinant MHC class II, rBcl2L12, and OVA in extracts of BMDCs (Fig 2, D) and a complex of recombinant MHC class II and OVA (without the presence of Bcl2L12) in extracts of BMDCs (Fig 2, E). Data are from 3 independent experiments. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig E1 Expression of Bcl2L12 in DCs of WT mice and Bcl2L12 KO mice. DCs were isolated from WT and Bcl2L12 KO mice. Total RNA was extracted from the organs and analyzed by using quantitative RT-PCR. Bars indicate mRNA levels (A) and immunoblots (B) indicate protein levels of Bcl2L12 in the main organs of WT mice and Bcl2L12 KO mice. Data are presented as means ± SDs. *P < .01 compared with WT mice. Data represent 3 independent experiments. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig E2 Bcl2L12 facilitates antigen presentation of DCs. BMDCs were generated from bone marrow of WT and Bcl2L12 KO mice. Cells were primed with MNP and OVA for 48 hours. Naive CD4+ T cells (Teff cells) were isolated from the spleens of DO11.10 mice. Teff cells were labeled with CFSE and cultured with primed BMDCs at gradient ratios for 3 days. Cells were analyzed by using flow cytometry. A, Gated histograms indicate proliferating Teff cells. B, Bars indicate summarized data of proliferating Teff cells in Fig E2, A. C-F, Bars indicate cytokine levels of CD4+ T cells in culture supernatants. Data shown in bars are presented as means ± SDs. *P < .01 compared with the “10:0” group. Data represent 3 independent experiments. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig E3 MNP/OVA-primed DCs sensitize mice. BMDCs were prepared and primed by incubating with MNP (10 μg/mL) and OVA (μg/mL), OVA alone, or saline in culture for 48 hours. BALB/c mice (6 mice per group) were adoptively transferred with primed BMDCs (106 cells/mouse) through tail vein injection, which was repeated 1 week after. Mice were killed 1 week after the second transfer. Teff cells were isolated from LMCs and splenic cells and analyzed by using the CFSE dilution assay. A, Gated histograms indicate proliferating Teff cells after treating with reagents in culture as denoted above each panel in the presence of DCs and OVA (5 μg/mL) or BSA (#; 5 μg/mL). B, Bars indicate summarized data of proliferating Teff cells in Fig E3, A. Data shown in bars are presented as means ± SDs. *P < .01 compared with the saline group. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig E4 Adoptive transfer with MNP/OVA-primed BMDCs induces allergic rhinitis in mice. BMDCs were prepared with bone marrow of WT and KO mice. Cells were primed by means of exposure to MNP/OVA in the culture for 48 hours. BMDCs (106 cells/mouse) or saline (control) were injected into BALB/c mice through tail vein puncture; injection was repeated 1 week later. One week after the second injection, mice were challenged with nasal drops containing OVA (5 mg/mL). A-C, Nasal symptoms (rhinocnesmus, sneezing, and rhinorrhea) were recorded within 1 hour after challenge with nasal drops. D-H, Mice were killed 1 hour after nasal challenge. Nasal mucosa was collected from each mouse and processed for protein extracts. Levels of specific IgE, mMCP1, IL-4, IL-5, and IFN-γ in protein extracts were determined by mean of ELISA, as shown in bars. Data shown in bars are presented as means ± SDs. *P < .01 compared with the saline group. Each group consists of 6 mice. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Fig E5 Inhibition of Bcl2L12 abolishes MNP/house mite extract (HME)–induced allergic rhinitis in mice. BALB/c (WT) mice (n = 6) and Bcl2L12 KO mice (n = 6) were treated with a mixture of MNP (10 μg/mL) and HME (10 μg/mL) by using nasal drops (50 μL per nostril per time) daily for 21 consecutive days. Control mice were treated with saline nasal drops. Mice were killed on day 21, 1 hour after treating with nasal drops. A-C, AR clinical symptoms, rhinocnesmus (Fig E5, A), sneezing (Fig E5, B), and rhinorrhea (Fig E5, C), were recorded on day 21 at 0 to 60 minutes after treating with nasal drops. D and E, Blood samples were collected from each mouse. Sera were isolated from samples and analyzed by means of ELISA. Bars indicate serum levels of specific IgE (Fig E5, D) and mMCP1 (Fig E5, E). F-H, Nasal mucosa was collected from each mouse immediately after death. Protein extracts were prepared with nasal mucosal samples and analyzed by means of ELISA. Bars indicate levels of IL-4 (Fig E5, F), IL-5 (Fig E5, G), and IFN-γ (Fig E5, H) in nasal mucosal extracts. Data shown in bars are presented as means ± SDs. Samples from individual mice were analyzed separately in triplicates. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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