On a few occasions NIAID has detected the suppression of or the loss of DO11.10 expression in lines containing this transgene. They are investigating this further and increasing the frequency of their quality control for these mice. However, it would be advisable to re-test the lines upon receipt. Any mice found not expressing the Tg will be replaced at no cost to the investigator. The preferred method of testing is flow cytometry of blood using the clonotypic monoclonal antibody KJ1-26 from Pharmingen. The Rag2 Knockout/Transgenic DO11.10 T Cell Receptor line is homozygous for a transgene that encodes T cell receptor that is specific for the chicken ovalbumin (OVA) peptide, amino acids 323-339, presented by the MHC class II molecule I-Ad. It is also deficient in the Rag2 gene (recombinase activating gene 2) and therefore does not develop endongenous mature T or B cells. Virtually all peripheral CD4+ T cells in these mice express the transgenic TCR. A few CD4- cells also express the transgene. These mice are useful for in vitro
studies of CD4+ T cell differentiation into TH1 and TH2 populations, and as a source of donor cells for adoptive transfer into BALB/c mice for in vivo
studies of tolerance and immune responses. Applications include studies of inflammation, T cell development, and tolerance.
Origin: The Transgenic DO11.10 T Cell Receptor mouse was generated by Dr. Ken Murphy in the Laboratory of Dr. Dennis Loh at Washington University, St. Louis in 1989. The line was made with the rearranged Vb 8.1 and Va 13 T cell receptor genes of the Kappler and Marrack OVA-specific T cell hybridoma DO11.10 using (C57BL/6 x C3H/HeJ)F1 x BALB/c donor embryos. The founder line has two copies of the Vα gene and four copies of the Vβ gene cointegrated onto the distal part of chromosome 18. It was initially backcrossed to BALB/c 12 times. The line was then transferred to Dr. Ethan Shevach at NIH and subsequently embryo transferred into the NIAID Repository at Taconic. The N12 mice were crossed twice to a homozygous Rag2 knockout mouse on a BALB/cAnTac background and Rag2-/-, DO11.10 positive offspring were selected. These mice were then intracrossed to produce the line homozygous for the TCR transgene. TCR homozygotes were selected by test mating the mice to wild type partners and identifying the animals that only gave rise to transgene positive offspring. This strain is designated as N14.
Availability: This model is no longer available.
Murphy KM, Heimberger, and Loh DY: Induction by Antigen of Intrathymic Apoptosis of CD4+ CD8+ TCRlo Thymocytes in vivo
. Science 1990; 250:1720-3
Iwabuchi K, Nakayama KI, McCoy RL, Wang F, Nishimura T, Habu S, Murphy KM and Loh DL: Cellular and peptide requirements for in vitro
clonal deletion of immature cytes. Proc. Nat. Acad. Sci. USA, 1992 Oct; 89: 9000-4
Hsieh CS, Macatonia SE, Tripp CS, Wolf SF, O'Garra A, Murphy KM: Development of TH1 CD4+ T cells through IL-12 produced by Listeria-induced macrophages. Science, 1993 Apr 23;260(5107):547-9