C.Cg-Tg(Tcra6C2,Tcrb6C2)16Chi/J

Stock number: 027510
Product name: Env28 TCR transgenic line 6C2.16
Research areas: Cancer Research, Immunology, Inflammation and Autoimmunity Research, Internal/Organ Research, Research Tools, Virology Research

Description

The Env28 TCR transgenic line 6C2.16 has a CD8⁺ T cell repertoire that is specific for the L^d-restricted ENV28 epitope (hepatitis B virus envelope/surface antigen residues 28-39 [IPQSLDSWWTSL]). These mice may be useful in studying cellular mechanisms of CD8⁺ T cell immunity, memory and protection from HBV; especially early events in the host-virus relationship that determine the outcome of hepatotropic human-virus infections.

Detailed description

In humans, the hepatitis B virus (HBV) causes acute and chronic hepatitis and hepatocellular carcinoma. Clearance of HBV requires functional virus-specific CD8⁺ T cell responses.
The Env28 TCR transgenic line 6C2.16 is a BALB/c-congenic (H-2^d) mouse strain carrying T cell receptor α and β transgenes that recognize an L^d-restricted ENV28 epitope (residues 28-39 [IPQSLDSWWTSL] of the HBV envelope/surface antigen [HBsAg]). These mice have a CD8⁺ T cell repertoire (greater than 83% of the splenic CD8⁺ T cells) that is specific for the ENV28 epitope. The ENV28-specific CD8⁺ T cells are immunologically naive with respect to their cognate antigen.
Hemizygous mice are viable and fertile. To date (July 2015), it has not been attempted to make this strain homozygous.

Development

The Env28 TCR transgenic line 6C2.16 was designed by Dr. Francis V. Chisari (The Scripps Research Institute). To generate the T cell clones, the H-2^d mouse P815 mastocytoma target cell line was incubated with the major envelope polypeptide amino acids 21 to 40 from hepatitis B virus (HBV) envelope/surface antigen (HBsAg). The resulting L^d-restricted CD3⁺ CD4^- CD8⁺ cytotoxic T lymphocytes (CTL) clone "6C2" was found to recognize an epitope located between residues 28-39 of HBsAg (ENV28; IPQSLDSWWTSL). The identity of the T cell receptor (TCR) Vα and Vβ chains on the surface of the "6C2" T cells was found to be Vα4.1 JαNEW and Vβ1.1 Jβ2.5. These TCR sequences were cloned into TCR cassette expression vectors (pTα and pTβ), and the resulting constructs (~15.0 kbp Tα ENV28 6C2 transgene and ~17.4 kbp Tβ ENV28 6C2 transgene) were co-microinjected into fertilized (BALB/cByJ x C57BL/6J)F2 oocytes. Founder mice were bred with BALB/cJ for germline transmission, and mice from line 16 (lineage 6C2.16) were selected. The Env28 TCR transgenic line 6C2.16 colony was backcrossed more than 12 generations with BALB/cJ inbred mice (H-2^d). In 2015, albino males hemizygous for the transgene were sent to The Jackson Laboratory Repository. Upon arrival, males were used to cryopreserve sperm. To establish the living mouse colony, an aliquot of the frozen sperm is used to fertilize oocytes from BALB/cJ inbred females (Stock No. 000651).

Of note, the donating investigator reports that, at least once during backcrossing, a hemizygous female was bred to a BALB/cJ inbred male (thus the Y chromosome of the congenic strain is of BALB/cJ origin).

Allele

Symbol: Tg(Tcra6C2,Tcrb6C2)16Chi
Name: transgene insertion 16, Francis Chisari
MGI accession ID: MGI:5648206
Generation method: Transgenic
Attributes: Inserted expressed sequence
Marker symbol: Tg(Tcra6C2,Tcrb6C2)16Chi
Marker name: transgene insertion 16, Francis Chisari
Chromosome: UN
Strain of origin: BALB/cByJ or C57BL/6J
Expressed genes: Tcra,T cell receptor alpha chain,mouse, laboratory; Tcrb,T cell receptor beta chain,mouse, laboratory
Molecular note: Rearranged Valpha4.1 JalphaNEW and Vbeta1.1 Jbeta2.5 sequences are obtained from L-restricted CD3<+> CD4<-> CD8<+> cytotoxic T lymphocytes (clone 6C2), which recognizes an epitope located between residues 28-39 of HBsAg. These TCR sequences were cloned into TCR cassette expression vectors (pTalpha and pTbeta), and co-injected. The primary investigator indicates that cells from founder lines 16 and 36 exhibit no differences in epitope recognition.
General note: TCR expression cassettes into which the TCRalpha and TCRbeta cDNA sequences had been cloned were co-injected into (BALB/cByJ x C57BL/6J)F2 fertilized oocytes.