C57BL/6-Tg(Scgb1a1-TFRC/OVA)1Adl/BmedJ

Stock number: 025627
Product name: B6.CC10-OVA , CC10-OVA (Thy1.2, C57BL/6) , B6.CC10-OVA (Thy1.2)
Research areas: Cancer Research, Developmental Biology Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Research Tools

Description

CC10-OVA transgenic mice express a membrane-bound exogenous antigen (ovalbumin with OT-1 and OT-11 epitopes) directed to large and small airway epithelium of the lungs by the rat Clara cell secretory protein promoter. These mice are useful for modeling antigen-specific, T-cell mediated acute bronchiolitis/pneumonitis as well as lung pathology similar to humans with acute lung transplant rejection and virus-induced lung injury.

Detailed description

CC10-OVA transgenic mice have expression of a membrane-bound exogenous antigen (ovalbumin with OT-1 and OT-11 epitopes) directed to large and small airway epithelium of the lungs (~50-60% of the airway lining) by the rat Clara cell secretory protein promoter. Transgenic carrier mice are viable and fertile with normal development and no abnormal pathology. Prior to adoptive transfer of cells the lungs remain normal. These mice are useful for modeling antigen-specific, T-cell mediated acute bronchiolitis/pneumonitis as well as lung pathology similar to humans with acute lung transplant rejection and virus-induced lung injury.
Lung-specific injury may be induced by adoptive transfer of activated OVA-specific T cells. Specifically, CC10-OVA transgenic mice adoptively transferred with 5x10⁵ activated OT-I-specific cytotoxic T lymphocytes (CTL) die within 6 days due to severe lung injury, while transfer of a lower amount of OT-I cells (1x10⁵) decreases mortality (40% survive). Transfer of 5x10⁵ OT-I cells cultured with low IL-2 resulted in significantly decreased mortality compared to the 5x10⁵ OT-I cultured under high IL-2 conditions. Transfer of 1x10⁵ OT-I cultured with low dose IL-2 was not lethal, but an acute inflammatory response was still evident in the lungs. This inflammatory phenotype resolved in 30 days (and required endogenous ICOS-expressing CD4+ bystander lymphocytes). Similarly-treated Rag1-deficient B6.CC10-OVA mice (B6.CC10-OVA.Rag1-/-) succumb to lethal lung rejection/pneumonitis (as they lack functional CD4+ lymphocytes), and this injury may be attenuated via co-transfer with T regulatory cells.
The donating investigator reports that breeding transgenic carrier mice together produces homozygous animals that are viable and fertile. In addition, those homozygous mice exhibit no difference in experimental performance compared to transgenic carrier mice produced from transgenic carrier x wildtype (noncarrier) matings.

Development

The CC10-OVA transgenic mice were created by Dr. Benjamin D. Medoff while in the laboratory of Dr. Andrew D. Luster (Massachusetts General Hospital/Harvard Medical School). The CC10-OVA transgene was designed with a 2.4 kbp promoter from rat Clara cell secretory protein (CC10 or Scgb1a1), a DNA sequence encoding the transmembrane domain (residues 2-118) of the human transferrin receptor (CD71 or TFRC) fused to a DNA sequence encoding amino acids 139-385 of mature chicken ovalbumin (the p139-387 segment including the p257-264 [SIINFEKL] and p323-339 epitopes recognized by OT-I and OT-II TCR transgenic cells, respectively). The construct was successfully injected into pronuclei of C57BL/6 eggs, which were implanted into C57BL/6 pseudopregnant females. Founder line 1, with the highest copy number (at least three copies), was identified and used. The CC10-OVA transgenic founder line 1 was subsequently bred to B6.129S7-Rag1^tm1Mom/J animals (Stock No. 002216). The colony was maintained thereafter by breeding B6.CC10-OVA.Rag1-/- animals together. An unknown breeding occurred that resulted in agouti mice. When discovered, the donating investigator proceeded to backcross the colony with C57BL/6J inbred mice for at least five generations (see SNP note below); selecting for the CC10-OVA transgene and selecting against the Rag1 null allele. The resulting B6.CC10-OVA males were sent to The Jackson Laboratory Repository in 2015. Upon arrival, sperm was cryopreserved. To establish our living colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (from Stock No. 000664).

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, 4 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.

Allele

Symbol: Tg(Scgb1a1-TFRC/OVA)1Adl
Name: transgene insetion 1, Andrew D Luster
MGI accession ID: MGI:5575977
Generation method: Transgenic
Attributes: Inserted expressed sequence
Marker symbol: Tg(Scgb1a1-TFRC/OVA)1Adl
Marker name: transgene insetion 1, Andrew D Luster
Chromosome: UN
Strain of origin: C57BL/6
Expressed genes: TFRC,transferrin receptor,human; OVA,ovalbumin,chicken
Molecular note: The CC10-OVA transgene was designed with a 2.4 kbp promoter from rat Clara cell secretory protein (CC10 or Scgb1a1), a DNA sequence encoding the transmembrane domain (residues 2-118) of the human transferrin receptor (CD71 or TFRC) fused to a DNA sequence encoding amino acids 139-385 of mature chicken ovalbumin (the p139-387 segment including the p257-264 [SIINFEKL] and p323-339 epitopes recognized by OT-I and OT-II TCR transgenic cells, respectively). The founder mouse line with the highest copy number (at least three copies) was identified and used.