NOD.129(B6)-Ctla4^tm1All/DoiJ

Stock number: 005144
Product name: NOD.Ctla4^null
Research areas: Cancer Research, Diabetes and Obesity Research, Immunology, Inflammation and Autoimmunity Research

Description

These Ctla4 knock-out mice develop develop a fatal lymphoproliferative disorder, and are suitable for use in applications related to the regulation of self tolerance and susceptibility to autoimmune diseases.

Detailed description

Heterozygous Ctla4^tm1All mice are viable, fertile, and normal in size and exhibit diabetes incidence comparable to NOD.

Ctla4^tm1All homozygous mice on various backgrounds, including C57BL/6, BALB/c, 129 and NOD, die at 3-4 weeks of age due to massive polyclonal expansion of T-cells and massive lymphocyte infiltration into non-lymphoid organs, such as heart, liver, lung and pancreas. Treatment with murine Ctla4Ig or depletion of CD4⁺ T-cells blocks this lymphoproliferative disease. Splenic and lymph node T-cells display an activated phenotype characterized as CD69⁺, CD25⁺, CD44^hi, Mel14^lo, CD45RB^lo and Sca1^hi and have down regulated the TCR. Lymph node and spleen cells cultured from Ctla4^tm1All homozygous mice spontaneously proliferate while wild-type controls do not. Cells from whole lymph nodes or purified T-cells are unresponsive to anti CD28mAb stimulation, while rIl2 increases the proliferative response. Homozygous Ctla4^tm1All NOD mice also transgenically expressing the BDC2.5 alpha and beta TCR chains do not exhibit the lymphoproliferative phenotype. Other TCR transgenes exert a similar effect on differing backgrounds. However, all of the mice develop aggressive autoimmunity and rapidly develop diabetes by7 weeks of age.In contrast, mice heterozygous for Ctla4^tm1All and transgenically expressing the BDC2.5 TCR have late and relatively rare diabetes.

NOD.129(B6)-Ctla4^tm1All/DoiJ may be useful for studying the role of Ctla4/Cd28:Cd80/Cd86 in the regulation of self tolerance and susceptibility to autoimmune diseases.

Development

A construct containing a neomycin expression cassette inserted into exon 3 of Ctla4 (cloned from 129/Sv mouse) was transfected into R1 ((129X1/SvJ x 129S1/Sv)F1-Kitl +) embryonic stem cells (ES cells). These ES cells were injected into C57BL/6 blastocysts. To establish a colony and germ line transmission chimeric mice were backcrossed to C57BL/6 (Chambers et, al, 1997). Luhder et al. 2000, subsequently backcrossed this mutation to NOD/LtJ. In 2004, the Type 1 Diabetes Resource received NOD.129(B6)- Ctla4^tm1All /DoiJ heterozygotes at generation N18.

Allele

Symbol: Ctla4^tm1All
Name: targeted mutation 1, James P Allison
MGI accession ID: MGI:2180670
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Ctla4
Marker name: cytotoxic T-lymphocyte-associated protein 4
Chromosome: 1
Strain of origin: (129X1/SvJ x 129S1/Sv)F1-Kitl⁺
Molecular note: The insertion of neomycin selection cassette under the control of a PGK promoter disrupted exon 3.