The Impact of <i>TRIM67</i> Knockout on Early Intestinal Antimicrobial Capacity in Mice Infected with <i>Salmonella enterica</i> serovar Typhimurium ATCC 14028

<i>Salmonella enterica</i> serovar Typhimurium (<i>S.</i> Typhimurium) is an intracellular pathogen that survives and replicates within host cells. Macrophages, key immune cells in infection defense, play a vital role in pathogen clearance through polarization (M1/M2) and NLR...

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Main Authors: Xinyue Zhang, Qinyuan Li, Tingting Zhang, Lanlan Jia, Wentao Liu, Chao Huang, Zhengli Chen, Qihui Luo
Format: Article
Language:English
Published: MDPI AG 2025-05-01
Series:Microorganisms
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Online Access:https://www.mdpi.com/2076-2607/13/6/1267
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author Xinyue Zhang
Qinyuan Li
Tingting Zhang
Lanlan Jia
Wentao Liu
Chao Huang
Zhengli Chen
Qihui Luo
author_facet Xinyue Zhang
Qinyuan Li
Tingting Zhang
Lanlan Jia
Wentao Liu
Chao Huang
Zhengli Chen
Qihui Luo
author_sort Xinyue Zhang
collection DOAJ
description <i>Salmonella enterica</i> serovar Typhimurium (<i>S.</i> Typhimurium) is an intracellular pathogen that survives and replicates within host cells. Macrophages, key immune cells in infection defense, play a vital role in pathogen clearance through polarization (M1/M2) and NLRP3 inflammasome activation. While <i>TRIM67</i> regulates macrophage recruitment in the liver, its role in <i>S.</i> Typhimurium infection remains unclear. In this study, a <i>S.</i> Typhimurium infection model was established by orally infecting streptomycin-pretreated <i>TRIM67</i> WT and KO mice with 1 × 10<sup>9</sup> CFU of <i>S.</i> Typhimurium. <i>TRIM67</i> expression in the ileum, colon, mesenteric lymph nodes (MLNs), and peritoneal macrophages (PMs) was assessed via qRT-PCR and Western blotting. Histopathological changes were analyzed using HE and PAS staining. IHC staining, flow cytometry (FCM), qRT-PCR, and Western blotting were used to evaluate <i>TRIM67</i> knockout effects on macrophage recruitment, polarization, and NLRP3 inflammasome activation. In vitro, PMs were infected with <i>S.</i> Typhimurium (MOI 1:20), and <i>TRIM67</i>’s role in macrophage polarization and NLRP3 activation was validated. <i>S.</i> Typhimurium infection significantly upregulated <i>TRIM67</i> in the ileum, colon, and MLN. <i>TRIM67</i> knockout reduced intestinal inflammatory cell infiltration but worsened goblet cell loss and impaired digestion. Bacterial load assays revealed weakened pathogen clearance, leading to weight loss and increased mortality. <i>TRIM67</i> knockout inhibited intestinal macrophage recruitment, M1 polarization in MLN, and NLRP3 activation. In vitro, <i>TRIM67</i> knockout increased PMs’ intracellular bacterial load and suppressed NLRP3, caspase-1, and IL-1β expression. <i>TRIM67</i> knockout impairs the host’s ability to clear <i>S.</i> Typhimurium by inhibiting M1 macrophage polarization and NLRP3 inflammasome activation.
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series Microorganisms
spelling doaj-art-66a253087f8345738e6650e0729fa4b92025-08-20T03:29:44ZengMDPI AGMicroorganisms2076-26072025-05-01136126710.3390/microorganisms13061267The Impact of <i>TRIM67</i> Knockout on Early Intestinal Antimicrobial Capacity in Mice Infected with <i>Salmonella enterica</i> serovar Typhimurium ATCC 14028Xinyue Zhang0Qinyuan Li1Tingting Zhang2Lanlan Jia3Wentao Liu4Chao Huang5Zhengli Chen6Qihui Luo7Laboratory of Experimental Animal Disease Model, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, ChinaLaboratory of Experimental Animal Disease Model, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, ChinaLaboratory of Experimental Animal Disease Model, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, ChinaKey Laboratory of Animal Disease and Human Health of Sichuan Province, College of Veterinary Medicine, Chengdu 611130, ChinaKey Laboratory of Animal Disease and Human Health of Sichuan Province, College of Veterinary Medicine, Chengdu 611130, ChinaKey Laboratory of Animal Disease and Human Health of Sichuan Province, College of Veterinary Medicine, Chengdu 611130, ChinaKey Laboratory of Animal Disease and Human Health of Sichuan Province, College of Veterinary Medicine, Chengdu 611130, ChinaKey Laboratory of Animal Disease and Human Health of Sichuan Province, College of Veterinary Medicine, Chengdu 611130, China<i>Salmonella enterica</i> serovar Typhimurium (<i>S.</i> Typhimurium) is an intracellular pathogen that survives and replicates within host cells. Macrophages, key immune cells in infection defense, play a vital role in pathogen clearance through polarization (M1/M2) and NLRP3 inflammasome activation. While <i>TRIM67</i> regulates macrophage recruitment in the liver, its role in <i>S.</i> Typhimurium infection remains unclear. In this study, a <i>S.</i> Typhimurium infection model was established by orally infecting streptomycin-pretreated <i>TRIM67</i> WT and KO mice with 1 × 10<sup>9</sup> CFU of <i>S.</i> Typhimurium. <i>TRIM67</i> expression in the ileum, colon, mesenteric lymph nodes (MLNs), and peritoneal macrophages (PMs) was assessed via qRT-PCR and Western blotting. Histopathological changes were analyzed using HE and PAS staining. IHC staining, flow cytometry (FCM), qRT-PCR, and Western blotting were used to evaluate <i>TRIM67</i> knockout effects on macrophage recruitment, polarization, and NLRP3 inflammasome activation. In vitro, PMs were infected with <i>S.</i> Typhimurium (MOI 1:20), and <i>TRIM67</i>’s role in macrophage polarization and NLRP3 activation was validated. <i>S.</i> Typhimurium infection significantly upregulated <i>TRIM67</i> in the ileum, colon, and MLN. <i>TRIM67</i> knockout reduced intestinal inflammatory cell infiltration but worsened goblet cell loss and impaired digestion. Bacterial load assays revealed weakened pathogen clearance, leading to weight loss and increased mortality. <i>TRIM67</i> knockout inhibited intestinal macrophage recruitment, M1 polarization in MLN, and NLRP3 activation. In vitro, <i>TRIM67</i> knockout increased PMs’ intracellular bacterial load and suppressed NLRP3, caspase-1, and IL-1β expression. <i>TRIM67</i> knockout impairs the host’s ability to clear <i>S.</i> Typhimurium by inhibiting M1 macrophage polarization and NLRP3 inflammasome activation.https://www.mdpi.com/2076-2607/13/6/1267<i>TRIM67</i><i>Salmonella enterica</i> serovar Typhimuriummacrophage polarizationNLRP3
spellingShingle Xinyue Zhang
Qinyuan Li
Tingting Zhang
Lanlan Jia
Wentao Liu
Chao Huang
Zhengli Chen
Qihui Luo
The Impact of <i>TRIM67</i> Knockout on Early Intestinal Antimicrobial Capacity in Mice Infected with <i>Salmonella enterica</i> serovar Typhimurium ATCC 14028
Microorganisms
<i>TRIM67</i>
<i>Salmonella enterica</i> serovar Typhimurium
macrophage polarization
NLRP3
title The Impact of <i>TRIM67</i> Knockout on Early Intestinal Antimicrobial Capacity in Mice Infected with <i>Salmonella enterica</i> serovar Typhimurium ATCC 14028
title_full The Impact of <i>TRIM67</i> Knockout on Early Intestinal Antimicrobial Capacity in Mice Infected with <i>Salmonella enterica</i> serovar Typhimurium ATCC 14028
title_fullStr The Impact of <i>TRIM67</i> Knockout on Early Intestinal Antimicrobial Capacity in Mice Infected with <i>Salmonella enterica</i> serovar Typhimurium ATCC 14028
title_full_unstemmed The Impact of <i>TRIM67</i> Knockout on Early Intestinal Antimicrobial Capacity in Mice Infected with <i>Salmonella enterica</i> serovar Typhimurium ATCC 14028
title_short The Impact of <i>TRIM67</i> Knockout on Early Intestinal Antimicrobial Capacity in Mice Infected with <i>Salmonella enterica</i> serovar Typhimurium ATCC 14028
title_sort impact of i trim67 i knockout on early intestinal antimicrobial capacity in mice infected with i salmonella enterica i serovar typhimurium atcc 14028
topic <i>TRIM67</i>
<i>Salmonella enterica</i> serovar Typhimurium
macrophage polarization
NLRP3
url https://www.mdpi.com/2076-2607/13/6/1267
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