Description

Strain Information

Type Congenic; Targeted Mutation; Additional information on Genetically Engineered and Mutant Mice. Visit our online Nomenclature tutorial. Additional information on Congenic nomenclature. Mating System Homozygote x Homozygote         (Female x Male)   12-AUG-10 Species laboratory mouse Generation N10+F10 (22-MAY-13) Generation Definitions   Donating Investigator Dr. Richard A. Flavell,   Yale University School of Medicine

Description
The toll-like receptor gene targeted in this strain is a sensor for monomeric flagellin, a component of bacterial flagella known to be a virulence factor. These mice have been used to investigate the mechanism of flagellin detection and signalling in antibacterial immune responses to Salmonella typhimurium and Pseudomonas aeruginosa. The gene has been found to be essential for the recognition of bacterial flagellin both in vivo and ex vivo. Mice that are homozygous for this targeted deletion are viable and fertile. Transcripts of this gene are absent from bone marrow-derived macrophages in homozygotes.

Relative to controls, homozygotes have increased fat mass throughout the body, particularly in visceral fat, a substantial increase in serum triglyceride and cholesterol levels, increased blood pressure, and higher than normal production of pro-inflammatory cytokines interferon gamma and interleukin 1 beta in adipose tissue. Basal insulin levels and insulin production in response to a glucose challenge are significantly elevated, and the response to exogenous insulin is reduced compared with that of controls. The ability to restore blood glucose to baseline levels after administration of a bolus of glucose is impaired, and a mild elevation in blood glucose levels is found after an overnight fast. After 8 weeks on a high fat diet fasting glucose concentrations exceed 120 mg/dL, inflammatory infiltrates are found in the pancreatic islets, and hepatic steatosis is observed.

Homozygotes offer a model of metabolic syndrome for which the hyperphagia, obesity, hyperglycemia, insulin resistance, colomegaly, and increased pro-inflammatory cytokines can be transferred from diseased homozygotes to wild-type hosts by tranplanting gut microbiota. Treatment of homozygotes with broad spectrum antibiotics for 12 weeks corrects the metabolic syndrome. Histerectomy rederivation to updgrade the microbial flora of homozygotes results in diminished severity of colitis and a more uniform phenotype of mild inflammation and obesity, including a 20% increase in body mass at 20 weeks of age.

Development
A targeting vector was used to replace the coding region with a loxP-flanked neomycin resistance cassette inserted in the opposite transcriptional orientation. 129S1/Sv Oca2⁺ Tyr⁺ Kitl⁺-derived W9.5 embryonic stem (ES) cells were used to create the mutation. Resultant male chimeric mice were mated to C57BL/6 females. The donating investigator reported that this strain was backcrossed ten times to C57BL/6/Ncr mice (see SNP note below).

A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. While the 27 markers throughout the genome suggested a C57BL/6 genetic background, at least 1 of 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6J ; C57BL/6N genetic background.

Control Information

	Control
	000664 C57BL/6J
Considerations for Choosing Controls

Phenotype

Phenotype Information

View Related Disease (OMIM) Terms

Related Disease (OMIM) Terms provided by MGI

- Potential model based on gene homology relationships. Phenotypic similarity to the human disease has not been tested. Legionnaire Disease, Susceptibility to   (TLR5) Systemic Lupus Erythematosus, Susceptibility to, 1; SLEB1   (TLR5)

View Mammalian Phenotype Terms

Mammalian Phenotype Terms provided by MGI

      assigned by genotype

The following phenotype information may relate to a genetic background differing from this JAX^® Mice strain.

Tlr5^tm1Flv/Tlr5^tm1Flv

        involves: 129S1/Sv * C57BL/6

• immune system phenotype
• abnormal cytokine secretion
  • after i.p. injection with flagellin, wild-type mice show elevated serum levels of Il-6 at 2 hours and Il-12 at 2 and 4 hours after challenge but mutants are unresponsive   (MGI Ref ID J:111778)
  • elevated Il-6 and Il-12 mRNA levels are detected in cultured wild-type dendritic cells in response to flagellin; mutant DCs show little response although response to LPS is similar to wild-type   (MGI Ref ID J:111778)
  • increases in Il-6 and keratinocyte chemoattractant in BALFs after challenge are present in wild-type but absent in mutants   (MGI Ref ID J:111778)
• abnormal dendritic cell physiology
  • when mutants are injected i.p. with flagellin, splenic cells show no response after 6 hours, whereas cells from wild-type mice display maturation markers in response to flagellin   (MGI Ref ID J:111778)
  • mutant cells show normal responses to LPS injection   (MGI Ref ID J:111778)
• abnormal response to transplant
  • when wild-type bone marrow is transplanted into Tlr5 mutants, significant amounts of Il-6 are found in sera upon flagellin challenge; when Tlr5 -null bone marrow is put into wild-type mice, flagellin challenge only results in a small Il-6 response   (MGI Ref ID J:111778)
• decreased neutrophil cell number
  • neutrophils are absent from BALFs from challenged mutants but wild-type show significant accumulation   (MGI Ref ID J:111778)
• increased susceptibility to autoimmune diabetes   (MGI Ref ID J:158055)
• increased susceptibility to bacterial infection
  • when infected by i.p. with S. typhimurium (SB300) and a non-flagellated form (SB762) wild-type and mutants show similar survival time and bacterial load in spleens and livers   (MGI Ref ID J:111778)
  • infection with P. aeruginosa (1 x 10⁶ cfu) results in similar survival rates to wild-type at 7 weeks of age (~85-90%)   (MGI Ref ID J:111778)
  • at dose of 6 x 10⁶ cfu, mutants succumb to infection in 2 days   (MGI Ref ID J:111778)
• increased susceptibility to induced colitis
  • while 10% of homozygotes have severe colitis and and additional 30% have less severe colitis, the severity of colitis is greatly attenuated in homozygotes rederived into a cleaner environment   (MGI Ref ID J:158055)
• increased susceptibility to induced pancreatitis
  • when homozygotes are fed a high-fat diet they develop inflammatory infiltrates in the pancreatic islets, have fasting glucose concentrations greater than 120 mg/dL, and display hepatic steatosis   (MGI Ref ID J:158055)
• lung inflammation
  • after intranasal (i.n.) challenge with flagellin, at 10 and 24 hours after challenge bronchoalveolar lavage fluids (BALFs) from mutants show no cell infiltration while wild-type mice have elevated cell content and infiltration   (MGI Ref ID J:111778)
• homeostasis/metabolism phenotype
• abnormal glucose homeostasis
  • when homozygotes are fed a high-fat diet they develop inflammatory infiltrates in the pancreatic islets, have fasting glucose concentrations greater than 120 mg/dL, and display hepatic steatosis   (MGI Ref ID J:158055)
  • after a 15 hour fast homozygotes on a normal diet have slightly elevated glucose levels and when non-fasting homozygotes are administered a bolus of glucose they display an impaired ability to restore blood glucose to baseline levels   (MGI Ref ID J:158055)
• • increased circulating insulin level
    • basal serum insulin levels are significantly higher than normal, with an average over .55 nm/mL   (MGI Ref ID J:158055)
  • insulin resistance
    • the amount of insulin produced in response to a glucose challenge is significantly increased to approximately 1.2 nm/mL, serum levels of lipocalin 2 are elevated, and the number and size of pancreatic islets that stain positive for insulin is increased   (MGI Ref ID J:158055)
    • although food restriction prevents increased body and fat pad mass, increased serum glucose, lipids, and insulin, food restricted homozygotes still have a decreased response to exogenous insulin   (MGI Ref ID J:158055)
• abnormal physiological response to xenobiotic
  • decimation of the gut microbiota by treatment with broad spectrum antibiotics for 12 weeks beginning at wean age corrects the metabolic syndrome that develops in the absence of antibiotics   (MGI Ref ID J:158055)
• increased circulating cholesterol level
  • significantly higher serum cholesterol levels   (MGI Ref ID J:158055)
• increased circulating triglyceride level
  • significantly higher serum triglyceride levels   (MGI Ref ID J:158055)
• increased susceptibility to diet-induced obesity
  • when homozygotes are fed a high-fat diet they develop inflammatory infiltrates in the pancreatic islets, have fasting glucose concentrations greater than 120 mg/dL, and display hepatic steatosis   (MGI Ref ID J:158055)
• adipose tissue phenotype
• abnormal adipose tissue physiology
  • adipose tissue has a higher than normal production of interferon gamma and interleukin 1 beta   (MGI Ref ID J:158055)
• increased epididymal fat pad weight
  • at 20 weeks of age the epididymal fat pads are approximately 2 fold greater than normal   (MGI Ref ID J:158055)
• increased total body fat amount
  • magnetic resonance imaging shows increased fat mass throughout the body, especially the visceral fat   (MGI Ref ID J:158055)
• growth/size phenotype
• increased body weight   (MGI Ref ID J:158055)
• • obese
    • by 4 weeks of age the body mass is an average of 15% above that of normal controls and in homozygotes rederived into a clean facility the body mass at 20 weeks of age is 20% greater than normal controls in both males and females   (MGI Ref ID J:158055)
• increased susceptibility to diet-induced obesity
  • when homozygotes are fed a high-fat diet they develop inflammatory infiltrates in the pancreatic islets, have fasting glucose concentrations greater than 120 mg/dL, and display hepatic steatosis   (MGI Ref ID J:158055)
• digestive/alimentary phenotype
• abnormal gut flora balance
  • the gut microbiotica shows enrichment or reduction of 116 bacterial phylotypes relative to wild-type controls and transplanting gut microbiota from homozygotes to germ-free control hosts confers many aspects of the metabolic disease phenotype   (MGI Ref ID J:158055)
• increased susceptibility to induced colitis
  • while 10% of homozygotes have severe colitis and and additional 30% have less severe colitis, the severity of colitis is greatly attenuated in homozygotes rederived into a cleaner environment   (MGI Ref ID J:158055)
• hematopoietic system phenotype
• decreased neutrophil cell number
  • neutrophils are absent from BALFs from challenged mutants but wild-type show significant accumulation   (MGI Ref ID J:111778)
• cardiovascular system phenotype
• increased systemic arterial blood pressure   (MGI Ref ID J:158055)
• • hypertension   (MGI Ref ID J:158055)
  • increased systemic arterial diastolic blood pressure
    • average is raised to approximately 110mm Hg   (MGI Ref ID J:158055)
  • increased systemic arterial systolic blood pressure
    • average is increased to appoximately 130mm Hg   (MGI Ref ID J:158055)
• behavior/neurological phenotype
• increased eating behavior   (MGI Ref ID J:158055)
• polyphagia
  • homozygotes consume approximately 10% more food than wild-type controls and have greater stool output   (MGI Ref ID J:158055)
• endocrine/exocrine gland phenotype
• increased susceptibility to induced pancreatitis
  • when homozygotes are fed a high-fat diet they develop inflammatory infiltrates in the pancreatic islets, have fasting glucose concentrations greater than 120 mg/dL, and display hepatic steatosis   (MGI Ref ID J:158055)
• liver/biliary system phenotype
• hepatic steatosis
  • when homozygotes are fed a high-fat diet they develop inflammatory infiltrates in the pancreatic islets, have fasting glucose concentrations greater than 120 mg/dL, and display hepatic steatosis   (MGI Ref ID J:158055)
• respiratory system phenotype
• lung inflammation
  • after intranasal (i.n.) challenge with flagellin, at 10 and 24 hours after challenge bronchoalveolar lavage fluids (BALFs) from mutants show no cell infiltration while wild-type mice have elevated cell content and infiltration   (MGI Ref ID J:111778)

Tlr5^tm1Flv/Tlr5^tm1Flv

        involves: 129S1/Sv

• immune system phenotype
• abnormal leukocyte migration
  • flagellin-treated mice exhibit reduced Ly6C^hi monocyte emigration from the bone marrow compared with similarly treated wild-type mice   (MGI Ref ID J:171379)
• cellular phenotype
• abnormal leukocyte migration
  • flagellin-treated mice exhibit reduced Ly6C^hi monocyte emigration from the bone marrow compared with similarly treated wild-type mice   (MGI Ref ID J:171379)

View Research Applications

Research Applications

This mouse can be used to support research in many areas including:

Immunology, Inflammation and Autoimmunity Research
CD Antigens, Antigen Receptors, and Histocompatibility Markers
      Tlr deficiency
      genes regulating susceptibility to infectious disease and endotoxin
Immunodeficiency
      Tlr deficiency
Inflammation
      Tlr deficiency

Tlr5^tm1Flv related

Diabetes and Obesity Research
Hyperglycemia
Insulin Resistance

Immunology, Inflammation and Autoimmunity Research
Inflammation

Internal/Organ Research
Gastrointestinal Defects
      colitis

Genes & Alleles

Gene & Allele Information provided by MGI

Allele Symbol		Tlr5tm1Flv
Allele Name		targeted mutation 1, Richard Flavell
Allele Type		Targeted (knock-out)
Mutation Made By		Dr. Richard Flavell,   Yale University School of Medicine
Strain of Origin		129S1/Sv-Oca2<+> Tyr<+> Kitl<+>
ES Cell Line Name		W9.5/W95
ES Cell Line Strain		129S1/Sv-Oca2<+> Tyr<+> Kitl<+>
Gene Symbol and Name		Tlr5, toll-like receptor 5
Chromosome		1
Gene Common Name(s)		SLEB1; TIL3;
Molecular Note		A loxP-flanked neomycin selection cassette replaced the coding exon for the gene. mRNA expression in mutant and WT BM-derived macrophages was assessed by RT-PCR. Transcripts were absent in mutant but not in WT macrophages. [MGI Ref ID J:111778]

Genotyping

Genotyping Information

Genotyping Protocols

Tlr5{tm1Flv}, Standard PCR

Helpful Links

Genotyping resources and troubleshooting

References

References provided by MGI

Selected Reference(s)

Feuillet V; Medjane S; Mondor I; Demaria O; Pagni PP; Galan JE; Flavell RA; Alexopoulou L. 2006. Involvement of Toll-like receptor 5 in the recognition of flagellated bacteria. Proc Natl Acad Sci U S A 103(33):12487-12492. [PubMed: 16891416]  [MGI Ref ID J:111778]

Vijay-Kumar M; Aitken JD; Carvalho FA; Cullender TC; Mwangi S; Srinivasan S; Sitaraman SV; Knight R; Ley RE; Gewirtz AT. 2010. Metabolic Syndrome and Altered Gut Microbiota in Mice Lacking Toll-Like Receptor 5. Science 328(5975):228-31. [PubMed: 20203013]  [MGI Ref ID J:158055]

Additional References

Tlr5^tm1Flv related

Feng T; Cong Y; Alexander K; Elson CO. 2012. Regulation of Toll-like receptor 5 gene expression and function on mucosal dendritic cells. PLoS One 7(4):e35918. [PubMed: 22545147]  [MGI Ref ID J:187190]

Garaude J; Kent A; van Rooijen N; Blander JM. 2012. Simultaneous targeting of toll- and nod-like receptors induces effective tumor-specific immune responses. Sci Transl Med 4(120):120ra16. [PubMed: 22323829]  [MGI Ref ID J:183811]

Henao-Mejia J; Elinav E; Jin C; Hao L; Mehal WZ; Strowig T; Thaiss CA; Kau AL; Eisenbarth SC; Jurczak MJ; Camporez JP; Shulman GI; Gordon JI; Hoffman HM; Flavell RA. 2012. Inflammasome-mediated dysbiosis regulates progression of NAFLD and obesity. Nature 482(7384):179-85. [PubMed: 22297845]  [MGI Ref ID J:181354]

Huber S; Gagliani N; Zenewicz LA; Huber FJ; Bosurgi L; Hu B; Hedl M; Zhang W; O'Connor W Jr; Murphy AJ; Valenzuela DM; Yancopoulos GD; Booth CJ; Cho JH; Ouyang W; Abraham C; Flavell RA. 2012. IL-22BP is regulated by the inflammasome and modulates tumorigenesis in the intestine. Nature 491(7423):259-63. [PubMed: 23075849]  [MGI Ref ID J:189217]

Jarchum I; Liu M; Lipuma L; Pamer EG. 2011. Toll-Like Receptor 5 Stimulation Protects Mice from Acute Clostridium difficile Colitis. Infect Immun 79(4):1498-503. [PubMed: 21245274]  [MGI Ref ID J:170621]

Kinnebrew MA; Buffie CG; Diehl GE; Zenewicz LA; Leiner I; Hohl TM; Flavell RA; Littman DR; Pamer EG. 2012. Interleukin 23 Production by Intestinal CD103(+)CD11b(+) Dendritic Cells in Response to Bacterial Flagellin Enhances Mucosal Innate Immune Defense. Immunity 36(2):276-87. [PubMed: 22306017]  [MGI Ref ID J:181332]

Kirkland D; Benson A; Mirpuri J; Pifer R; Hou B; Defranco AL; Yarovinsky F. 2012. B Cell-Intrinsic MyD88 Signaling Prevents the Lethal Dissemination of Commensal Bacteria during Colonic Damage. Immunity 36(2):228-38. [PubMed: 22306056]  [MGI Ref ID J:181331]

Rivas MN; Koh YT; Chen A; Nguyen A; Lee YH; Lawson G; Chatila TA. 2012. MyD88 is critically involved in immune tolerance breakdown at environmental interfaces of Foxp3-deficient mice. J Clin Invest 122(5):1933-47. [PubMed: 22466646]  [MGI Ref ID J:184544]

Shi C; Jia T; Mendez-Ferrer S; Hohl TM; Serbina NV; Lipuma L; Leiner I; Li MO; Frenette PS; Pamer EG. 2011. Bone Marrow Mesenchymal Stem and Progenitor Cells Induce Monocyte Emigration in Response to Circulating Toll-like Receptor Ligands. Immunity 34(4):590-601. [PubMed: 21458307]  [MGI Ref ID J:171379]

Shi Z; Cai Z; Yu J; Zhang T; Zhao S; Smeds E; Zhang Q; Wang F; Zhao C; Fu S; Ghosh S; Zhang D. 2012. Toll-like receptor 11 (TLR11) prevents Salmonella penetration into the murine Peyer patches. J Biol Chem 287(52):43417-23. [PubMed: 23135279]  [MGI Ref ID J:193751]

Sun Y; Karmakar M; Taylor PR; Rietsch A; Pearlman E. 2012. ExoS and ExoT ADP ribosyltransferase activities mediate Pseudomonas aeruginosa keratitis by promoting neutrophil apoptosis and bacterial survival. J Immunol 188(4):1884-95. [PubMed: 22250085]  [MGI Ref ID J:181171]

Ubeda C; Lipuma L; Gobourne A; Viale A; Leiner I; Equinda M; Khanin R; Pamer EG. 2012. Familial transmission rather than defective innate immunity shapes the distinct intestinal microbiota of TLR-deficient mice. J Exp Med 209(8):1445-56. [PubMed: 22826298]  [MGI Ref ID J:189145]

Zhang X; Wang T; Deng T; Xiong W; Lui P; Li N; Chen Y; Han D. 2013. Damaged spermatogenic cells induce inflammatory gene expression in mouse Sertoli cells through the activation of Toll-like receptors 2 and 4. Mol Cell Endocrinol 365(2):162-73. [PubMed: 23123736]  [MGI Ref ID J:193757]

van 't Veer C; van den Pangaart PS; Kruijswijk D; Florquin S; de Vos AF; van der Poll T. 2011. Delineation of the Role of Toll-like Receptor Signaling during Peritonitis by a Gradually Growing Pathogenic Escherichia coli. J Biol Chem 286(42):36603-18. [PubMed: 21690093]  [MGI Ref ID J:177619]

Health & husbandry

Health & Colony Maintenance Information

Animal Health Reports

Room Number           MGL375

Colony Maintenance

Breeding & Husbandry	When maintained as a live colony, homozygotes may be bred.
Mating System	Homozygote x Homozygote         (Female x Male)   12-AUG-10
Diet Information	LabDiet® 5K52/5K67

Pricing and Purchasing

Pricing, Supply Level & Notes, Controls

Control Information

	Control
	000664 C57BL/6J
Considerations for Choosing Controls
Control Pricing Information for Genetically Engineered Mutant Strains.

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The Jackson Laboratory's Genotype Promise

The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX^® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX^® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX^® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project.

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