Journal: Science Advances

Article Title: Lung-innervating nociceptor sensory neurons promote pneumonic sepsis during carbapenem-resistant Klebsiella pneumoniae lung infection

doi: 10.1126/sciadv.adl6162

Figure Lengend Snippet: ( A ) Levels of CGRP in BALF of PBS-treated and CRKP-infected control and Trpv1 DTA mice ( n = 8 per group). ( B ) Stained sections of lungs showing β-tubulin III (TUJ1) and CGRP fibers of control and Trpv1 DTA mice. Representative images were selected from a total of nine images of three mice in each group (three images per mouse). Scale bars, 50 μm. DAPI, 4′,6-diamidino-2-phenylindole. ( C ) Left: Schematic showing organotypic culture of VG ( n = 4 VGs per well) and their stimulation with CRKP to measure CGRP release. Right: Time course of CGRP release from VG culture after stimulation with CRKP bacteria (10 6 CFU per well and 10 8 CFU per well). Unstimulated wells are VG culture with media only. Capsaicin (1.5 μM) was used as positive control. ( D ) Heatmap showing relative mRNA levels of neuropeptide receptors in CRKP-infected monocytes and neutrophils at indicated time points of infection ( n ≥ 3 per group). Un., unstimulated cells. Data in (A), (C), and (D) are the means ± SEM. Statistical analysis: Brown-Forsythe and Welch ANOVA of two-stage liner step-up procedure of Benjamini, Krieger, and Yekutieli posttests (A), one-way ANOVA with Sidak’s multiple comparison test (C), and two-way ANOVA with Sidak’s multiple comparisons posttests (D). Level of significance: * P < 0.05, ** P < 0.01, **** P < 0.0001. BioRender.com was used to create schematic in (C).

Article Snippet: Competitive CGRP inhibitor CGRP 8–37 (GenScript) (5 μg per mouse) was administered intraperitoneally into mice at 2 hours before infection.

Techniques: Infection, Control, Staining, Bacteria, Positive Control, Comparison

Journal: Science Advances

Article Title: Lung-innervating nociceptor sensory neurons promote pneumonic sepsis during carbapenem-resistant Klebsiella pneumoniae lung infection

doi: 10.1126/sciadv.adl6162

Figure Lengend Snippet: ( A ) Schematic showing monocyte and CRKP coculture in the presence and absence of neuropeptide. ( B and C ) Intracellular viable CFUs (B) and levels of ROS (C), in CRKP-infected wild-type (WT) and Ramp − / − monocytes. ( D ) Schematic of neutrophil and CRKP coculture with and without neuropeptide. ( E ) Intracellular viable CFUs in CRKP-infected WT and Ramp − / − neutrophils. ( F ) Schematic showing BMDM isolation and coculture with CRKP in the presence and absence of neuropeptides. ( G ) Intracellular viable CFUs in WT BMDMs. Data in (B), (C), (E), and (G) involve three to eight samples per group. ( H to K ) CRKP CFUs (H), proportions of myeloid subsets (I), total BAL protein (J), and panel of 13 mouse inflammatory cytokines/chemokines (K), measured in BALF of Ramp1 + / + and Ramp1 − / − mice ( n = 7 to 8 per group) at 24 hpi. ( L ) Schematic showing administration of CGRP 8–37 (5 μg per mouse) in C57BL/6 J mice and CRKP infection. ( M to P ) CFUs in whole lung (M), blood (N), liver (O), and spleen (P) of PBS-treated and CGRP 8–37 -treated mice ( n = 8 per group) after 24 hpi. Data in (B), (C), (E), (G), (H) to (K) and (M) to (P) are the means ± SEM. Statistical analysis: one-way ANOVA with Sidak’s multiple comparisons posttests [(B), (C), (E), and (G)], Mann-Whitney test [(H), (O), and (P)], unpaired t test [(J), (M), and (N)], and two-way ANOVA of two-stage liner step-up procedure of Benjamini, Krieger, and Yekutieli posttests [(I) and (K)]. Levels of significance: * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. BioRender.com was used to create schematics in [(A), (D), (F), and (L)].

Article Snippet: Competitive CGRP inhibitor CGRP 8–37 (GenScript) (5 μg per mouse) was administered intraperitoneally into mice at 2 hours before infection.

Techniques: Infection, Isolation, MANN-WHITNEY

Journal: Science Advances

Article Title: Lung-innervating nociceptor sensory neurons promote pneumonic sepsis during carbapenem-resistant Klebsiella pneumoniae lung infection

doi: 10.1126/sciadv.adl6162

Figure Lengend Snippet: Lung-innervating nociceptor neurons suppress the recruitment of neutrophils and Ly6C hi monocytes to the airspaces of lungs during CRKP lung infection. CRKP infection induces vagal TRPV1 + neurons for releasing CGRP, which in turn acts on its receptor on Ly6C hi monocytes to dampen the ROS production. This leads to an increased CRKP survival in the lungs and enhanced dissemination of bacteria to vital organs. BioRender.com was used to create model.

Article Snippet: Competitive CGRP inhibitor CGRP 8–37 (GenScript) (5 μg per mouse) was administered intraperitoneally into mice at 2 hours before infection.

Techniques: Infection, Bacteria