Description

Strain Information

Type Congenic; Mutant Strain; Targeted Mutation; Additional information on Genetically Engineered Mutant Mice. Mating System Homozygote x Homozygote         (Female x Male) Species laboratory mouse Generation N10F?+3 (25-SEP-08)   Donating Investigator Steven Shapiro,   University of Pittsburgh

Description
Homozygous mice are viable, fertile and phenotypically normal in the absence of inflammatory stress. Homozygotes do not express the targeted gene in bone marrow myeloid cells. Homozygous mice have increased susceptibility to sepsis, morbidity, and mortality following intraperitoneal injection of Gram-negative (e.g. (K. pneumoniae and E. coli), but not Gram-positive (e.g. (S. aureus), bacteria. Despite this, mutant mice are not at increased risk to spontaneous infection. Although neutrophil, T cell, and macrophage migration/recruitment to the site of infection is unaffected in homozygous mutant mice, neutrophils show impaired bactericidal activity. Further, homozygous mice treated with a broad-spectrum inflammatory stimulus (zymosan) have impaired leukocyte firm adhesion and transmigration as well as reduced pro-inflammatory cytokine production. Upon exposure to cigarette smoke, homozygous mice are protected from the accumulation of neutrophils and macrophages in the lungs, as well as the development of airspace enlargement. These mutant mice may be useful in studies of immunity and immunology, including host defense against Gram-negative bacteria, granulocyte/neutrophil function, sepsis, and cigarette smoke-induced emphysema.

Development
A targeting vector containing a neomycin phosphotransferase cDNA driven by the phosphoglycerate kinase promoter (PGK-neo) was used to replace exon 2 (which contains the histidine residue of the catalytic triad) of the endogenous gene. The construct was electroporated into 129X1/SvJ-derived RW4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6J blastocysts. The resulting chimeric animals crossed to 129X1/SvJ mice, and the strain was maintained on this background for an unknown number of generations. The donating investigator reports that mutant mice were then mated to "C57BL/6 mice from Charles River" (C57BL/6NCrl) for at least 10 generations before arrival at The Jackson Laboratory. Upon arrival, male and female mice were bred together (or may have been bred with C57BL/6NJ (Stock No. 005304) inbred mice to attempt to preserve the genetic background.

Control Information

	Control
	000664 C57BL/6J	(approximate)
	005304 C57BL/6NJ
Considerations for Choosing Controls

Additional Web Information

Congenic Nomenclature

Phenotype

Phenotype Information

View Mammalian Phenotype Terms

Mammalian Phenotype Terms

      assigned by genotype

Ela2^tm1Sds/Ela2^tm1Sds

        B6.129X1-Ela2^tm1Sds

• homeostasis/metabolism phenotype
• decreased sensitivity to xenobiotics (MGI Ref ID J:86599)
  • mice exhibit 59% protection from emphysema induced by a 6 month cigarette smoke exposure
  • following a 6 month exposure to cigarette smoke, mice exhibit fewer neutrophils and 60% less macrophages in bronchoalveolar lavage samples compared to wild-type mice

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

Ela2^tm1Sds/Ela2^tm1Sds

        129X1/SvJ-Ela2^tm1Sds

• life span-post-weaning/aging
• abnormal induced morbidity/mortality (MGI Ref ID J:47420)
  • at 16 hours following infection with Gram negative Klebsiella pneumoniae, 50% of mice are dead unlike wild-type mice that do not display mortality due to infection at 16 hours
  • at 48 hours following infection with Klebsiella pneumoniae, all mice are dead unlike wild-type mice that display 50% mortality
  • mice are less susceptible to death induced by Gram positive Staphylococcus aureus infection compared to wild-type mice
• immune system phenotype
• abnormal neutrophil physiology (MGI Ref ID J:47420)
  • despite normal neutrophil recruitment, recognition and binding, neutrophils fail to degrade phagocytosed Klebsiella pneumoniae or Escherichia coli unlike in wild-type mice
• decreased susceptibility to bacterial infection (MGI Ref ID J:47420)
  • mice are less susceptible to death induced by Gram positive Staphylococcus aureus infection compared to wild-type mice
• increased susceptibility to bacterial infection (MGI Ref ID J:47420)
  • mice appear clinically worse than wild-type mice following infection with Gram negative Klebsiella pneumoniae or Escherichia coli
  • when infected with Klebsiella pneumoniae, mice exhibit signs of severe infection including ruffled fur, hunched posture, impaired ambulation and diarrhea unlike wild-type mice
  • at 16 hours following infection with Klebsiella pneumoniae, 50% of mice are dead unlike wild-type mice that do not display mortality due to infection at 16 hours
  • at 48 hours following infection with Klebsiella pneumoniae, all mice are dead unlike wild-type mice that display 50% mortality
  • despite normal neutrophil recruitment, recognition and binding, bacterial burden is greater than in wild-type mice following infection with Klebsiella pneumoniae

Ela2^tm1Sds/Ela2^tm1Sds

        involves: 129X1/SvJ

• homeostasis/metabolism phenotype
• decreased sensitivity to xenobiotics (MGI Ref ID J:124377)
  • unlike in wild-type mice, ONO-5046 has no inhibitory effects on IL-1beta-induced leukocyte transmigration

Ela2^tm1Sds/Ela2^tm1Sds

        involves: 129X1/SvJ * C57BL/6

• immune system phenotype
• abnormal inflammatory response (MGI Ref ID J:113463)
  • following exposure to LPS then challenge with TNF at the same site to initiate a local Shwartman response, mice fail to exhibit hemorrhage or fibrin deposition, characteristic of thrombohemorrhagic vasculitis, unlike in wild-type mice despite normal neutrophil accumulation

View Research Applications

Research Applications

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

Cancer Research
Growth Factors/Receptors/Cytokines

Cardiovascular Research
Vascular Defects (defective leukocyte function)

Hematological Research
Neutrophil Defects

Immunology and Inflammation Research
CD Antigens, Antigen Receptors, and Histocompatibility Markers (genes regulating susceptibility to infectious disease and endotoxin)
Growth Factors/Receptors/Cytokines
Immunodeficiency (Neutrophil Defects)
Inflammation (Neutrophil defects)

Internal/Organ Research
Lung Defects (emphysema)

Mouse/Human Gene Homologs
leukocyte adhesion deficiency, type I

Research Tools
Cancer Research (Leukemia)
Immunology and Inflammation Research (genes regulating susceptibility to infectious disease and endotoxin)

Genes & Alleles

Gene & Allele Information

Allele Symbol		Ela2tm1Sds
Allele Name		targeted mutation 1, Steven D Shapiro
Allele Type		Targeted (knock-out)
Common Name(s)		NE-;
Mutation Made By		Steven Shapiro,   University of Pittsburgh
Strain of Origin		129X1/SvJ
ES Cell Line Name		RW-4
ES Cell Line Strain		129X1/SvJ
Gene Symbol and Name		Ela2, elastase 2, neutrophil
Chromosome		10
Gene Common Name(s)		F430011M15Rik; GE; HLE; HNE; NE; PMN-E; RIKEN cDNA F430011M15 gene; neutrophil elastase;
Molecular Note		The insertion of a neomycin selection cassette replaced exon 2, which encodes the histidine residue of the catalytic triad. Northern blot analysis of total RNA obtained from bone marrow myeloid cells showed that transcript encoded by the targeted allele was absent. Reprobing of the blot showed that the expression of the closely linked proteinase 3 was unaffected by the targeting event. An absence of elastocytic activity derived from the targeted locus was determined via kappa-elastin zymography. [MGI Ref ID J:47420]

Genotyping

Genotyping Information

Genotyping Protocols

Ela2^tm1Sds, STD PCR, vers. 1

Helpful Links

Optimizing PCR Protocols

References

References

Selected Reference(s)

Belaaouaj A; McCarthy R; Baumann M; Gao Z; Ley TJ; Abraham SN; Shapiro SD. 1998. Mice lacking neutrophil elastase reveal impaired host defense against gram negative bacterial sepsis. Nat Med 4(5):615-8. [PubMed: 9585238]  [MGI Ref ID J:47420]

Additional References

Ela2^tm1Sds related

Akk AM; Simmons PM; Chan HW; Agapov E; Holtzman MJ; Grayson MH; Pham CT. 2008. Dipeptidyl peptidase I-dependent neutrophil recruitment modulates the inflammatory response to sendai virus infection. J Immunol 180(5):3535-42. [PubMed: 18292580]  [MGI Ref ID J:131520]

Allport JR; Lim YC; Shipley JM; Senior RM; Shapiro SD; Matsuyoshi N; Vestweber D; Luscinskas FW. 2002. Neutrophils from MMP-9- or neutrophil elastase-deficient mice show no defect in transendothelial migration under flow in vitro. J Leukoc Biol 71(5):821-8. [PubMed: 11994507]  [MGI Ref ID J:76606]

Edelstein C; Shapiro SD; Klezovitch O; Scanu AM. 1999. Macrophage metalloelastase, MMP-12, cleaves human apolipoprotein(a) in the linker region between kringles IV-4 and IV-5. Potential relevance to lipoprotein(a) biology. J Biol Chem 274(15):10019-23. [PubMed: 10187779]  [MGI Ref ID J:115221]

Grenda DS; Johnson SE; Mayer JR; McLemore ML; Benson KF; Horwitz M; Link DC. 2002. Mice expressing a neutrophil elastase mutation derived from patients with severe congenital neutropenia have normal granulopoiesis. Blood 100(9):3221-8. [PubMed: 12384420]  [MGI Ref ID J:131496]

Hirahashi J; Mekala D; Van Ziffle J; Xiao L; Saffaripour S; Wagner DD; Shapiro SD; Lowell C; Mayadas TN. 2006. Mac-1 signaling via Src-family and Syk kinases results in elastase-dependent thrombohemorrhagic vasculopathy. Immunity 25(2):271-83. [PubMed: 16872848]  [MGI Ref ID J:113463]

Hirche TO; Atkinson JJ; Bahr S; Belaaouaj A. 2004. Deficiency in neutrophil elastase does not impair neutrophil recruitment to inflamed sites. Am J Respir Cell Mol Biol 30(4):576-84. [PubMed: 14565940]  [MGI Ref ID J:97295]

Hirche TO; Crouch EC; Espinola M; Brokelman TJ; Mecham RP; DeSilva N; Cooley J; Remold-O'Donnell E; Belaaouaj A. 2004. Neutrophil serine proteinases inactivate surfactant protein D by cleaving within a conserved subregion of the carbohydrate recognition domain. J Biol Chem 279(26):27688-98. [PubMed: 15078883]  [MGI Ref ID J:123985]

Hirche TO; Gaut JP; Heinecke JW; Belaaouaj A. 2005. Myeloperoxidase plays critical roles in killing Klebsiella pneumoniae and inactivating neutrophil elastase: effects on host defense. J Immunol 174(3):1557-65. [PubMed: 15661916]  [MGI Ref ID J:96406]

Lane AA; Ley TJ. 2003. Neutrophil elastase cleaves PML-RARalpha and is important for the development of acute promyelocytic leukemia in mice. Cell 115(3):305-18. [PubMed: 14636558]  [MGI Ref ID J:86458]

Lane AA; Ley TJ. 2005. Neutrophil elastase is important for PML-retinoic acid receptor alpha activities in early myeloid cells. Mol Cell Biol 25(1):23-33. [PubMed: 15601827]  [MGI Ref ID J:95151]

Levesque JP; Liu F; Simmons PJ; Betsuyaku T; Senior RM; Pham C; Link DC. 2004. Characterization of hematopoietic progenitor mobilization in protease-deficient mice. Blood 104(1):65-72. [PubMed: 15010367]  [MGI Ref ID J:90920]

Liu Z; Zhou X; Shapiro SD; Shipley JM; Twining SS; Diaz LA; Senior RM; Werb Z. 2000. The serpin alpha1-proteinase inhibitor is a critical substrate for gelatinase B/MMP-9 in vivo. Cell 102(5):647-55. [PubMed: 11007483]  [MGI Ref ID J:64322]

Raptis SZ; Shapiro SD; Simmons PM; Cheng AM; Pham CT. 2005. Serine protease cathepsin g regulates adhesion-dependent neutrophil effector functions by modulating integrin clustering. Immunity 22(6):679-91. [PubMed: 15963783]  [MGI Ref ID J:99112]

Shapiro SD; Goldstein NM; Houghton AM; Kobayashi DK; Kelley D; Belaaouaj A. 2003. Neutrophil elastase contributes to cigarette smoke-induced emphysema in mice. Am J Pathol 163(6):2329-35. [PubMed: 14633606]  [MGI Ref ID J:86599]

Wang S; Dangerfield JP; Young RE; Nourshargh S. 2005. PECAM-1, {alpha}6 integrins and neutrophil elastase cooperate in mediating neutrophil transmigration. J Cell Sci 118(Pt 9):2067-76. [PubMed: 15840647]  [MGI Ref ID J:98057]

Wang S; Voisin MB; Larbi KY; Dangerfield J; Scheiermann C; Tran M; Maxwell PH; Sorokin L; Nourshargh S. 2006. Venular basement membranes contain specific matrix protein low expression regions that act as exit points for emigrating neutrophils. J Exp Med 203(6):1519-32. [PubMed: 16754715]  [MGI Ref ID J:124377]

Young RE; Thompson RD; Larbi KY; La M; Roberts CE; Shapiro SD; Perretti M; Nourshargh S. 2004. Neutrophil elastase (NE)-deficient mice demonstrate a nonredundant role for NE in neutrophil migration, generation of proinflammatory mediators, and phagocytosis in response to zymosan particles in vivo. J Immunol 172(7):4493-502. [PubMed: 15034066]  [MGI Ref ID J:88722]

Zhao M; Trimbeger ME; Li N; Diaz LA; Shapiro SD; Liu Z. 2006. Role of FcRs in animal model of autoimmune bullous pemphigoid. J Immunol 177(5):3398-405. [PubMed: 16920981]  [MGI Ref ID J:139536]

Health & husbandry

Health & Colony Maintenance Information

Animal Health Reports

Room Number           AX11

Colony Maintenance

Breeding & Husbandry	When maintaining a live colony, these mice are bred as homozygotes. While the publications indicate mutant mice are not at increased risk to spontaneous infection, The Jackson Laboratory maintains homozygous colonies in microisolator ventilation.
Mating System	Homozygote x Homozygote         (Female x Male)
Diet Information	LabDiet® 5K52/5K67

Purchasing information

Pricing, Supply Level & Notes, Controls, General Terms & Conditions

Pricing

Supply Details

Standard Supply

Repository-Live. A collection of over 1000 strains maintained as live colonies. Individual colonies are sized to meet current customer demand. Delivery for orders of 10 mice or less ranges on average from one to eight weeks; mice are generally shipped between four to six weeks of age with a maximum shipping age of ~nine weeks. Colony sizes do not generally support stringent age specifications for large volumes of mice; however custom orders and larger quantities of mice are easily arranged. Estimated ship dates for all orders provided within 48 hours of order placement.

Supply Notes

Usually shipped between four and eight weeks of age. This strain is included in the Induced Mutant Resource Colony collection.

Control Information

	Control
	000664 C57BL/6J	(approximate)
	005304 C57BL/6NJ
Considerations for Choosing Controls
USA, Canada and Mexico - Control Pricing Information for Genetically Engineered Mutant Strains.
International - Control Pricing Information for Genetically Engineered Mutant Strains.

General Terms and Conditions

See Terms of Use

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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Ordering and Purchasing Information

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Contact Information
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Tel: 800.422.6423 or 207.288.5845
Fax: 207.288.6150
Technical Support Email Form

Terms of Use

Terms of Use

General Terms and Conditions

Contact information

General inquiries

Contracts Administration

phone:	207-288-6470
fax:	207-288-6655

JAX^® Mice & Services Conditions of Use

“Each recipient institution, including its employees and other researchers under its control (RECIPIENT), of mice or services using mice from The Jackson Laboratory (TJL) agrees that such mice, descendants of those mice derived by inbreeding or crossbreeding, including unmodified derivatives of those mice or their descendants (“MICE”) shall not be: (i) used for any purpose other than the internal research of the RECIPIENT, (ii) sold or otherwise provided to any third party for any use, or (iii) provided to any agent or other third party to provide breeding or other services with respect to MICE. Acceptance of MICE from TJL shall be deemed agreement by RECIPIENT to these conditions, and departure from these conditions requires The Jackson Laboratory’s prior written authorization.”

In case of dissatisfaction for a valid reason and claimed in writing by a purchaser within ninety (90) days of receipt of MICE, products or services, The Jackson Laboratory will, at its option, provide credit or replacement for the MICE or product received or the services provided.

No Liability

In no event shall The Jackson Laboratory, its trustees, directors, officers, employees, and affiliates be liable for any causes of action or damages, including any direct, indirect, special, or consequential damages, arising out of the provision of MICE, products or services, including economic damage or injury to property and lost profits, and including any damage arising from acts or negligence on the part of The Jackson Laboratory, its agents or employees. In purchasing or receiving MICE, products or services from The Jackson Laboratory, purchaser or recipient, or any party claiming by or through them, expressly releases and discharges The Jackson Laboratory from all such causes of action or damages, and further agrees to defend and indemnify The Jackson Laboratory from any costs or damages arising out of any third party claims.

MICE and biological materials are to be used in a safe manner and in accordance with all applicable governmental rules and regulations.

The foregoing represents the General Terms and Conditions applicable to The Jackson Laboratory’s MICE, products and services. In addition, special terms and conditions of sale of certain MICE, products and services may be set forth separately in The Jackson Laboratory web pages, catalogs, price lists, contracts, and/or other documents, and these special terms and conditions shall also govern the sale of these MICE, products and services by The Jackson Laboratory, and by its licensees and distributors.

Acceptance of delivery of MICE, products or services shall be deemed agreement to these terms and conditions. No purchase order or other document transmitted by purchaser or recipient that may modify the terms and conditions hereof, shall be in any way binding on The Jackson Laboratory, and instead the terms and conditions set forth herein, including any special terms and conditions set forth separately, shall govern the sale of MICE, products services by The Jackson Laboratory.