Gary D. Anderson

Identification of T-cell receptor Vβ deletion mutant mouse strain AU/ssJ (H-2q) which is resistant to collagen-induced arthritis

Our laboratory is involved in investigating the role of T-cell receptor (Tcr) in collagen-induced arthritis (CIA). During these studies we found AU/ssJ (H-2q) mice to be resistant to CIA like SWR (H-2q), as compared with other H-2q strains with wild-type Tcr like DBA/1 and B 10. Q. Upon screening with monoclonal antibodies F23.1 and KJ23a, AU/ssJ was found to be F23.1 negative (Vβ8 Tcr negative) and KJ23a positive (Vβ17a Tcr positive). Southern blot analysis on liver DNA using specific Tcr-Vβ probes confirmed the deletion of Vβ8 gene family and also showed that AU/ssJ mice have deletions of Vβ9, Vβ13, Vβ12, and Vβ11 genes of Tcr. Further, these mice show a restriction fragment length polymorphism pattern with Vβ10, Vβ6, and Vβ17 probes similar to SWR mice as compared with 1310 mice. Since SWR and AU/ssJ are from different backgrounds, these studies indicate that specific variable region β chain genes of Tcr are crucial for susceptibility to CIA in mice. Furthermore, these studies identify an additional inbred strain which has also deleted 50% of its Tcr-Vβ genes.

T-Cell Receptors and Collagen Induced Arthritis in H-2R Mice

Mouse strains B10, B10.RIII, RIIIS/J and the F1 and backcross progeny arising from them were tested for susceptibility to porcine type II collagen-induced arthritis (PII-CIA). The clinically severe arthritis of rapid onset that is characteristic of PII-immunized B10.RIII mice developed predominantly in hybrid offspring that had inherited at least one copy of wild type T cell receptor (TCR) genes (V beta b genotype) from the B10 or B10.RIII parent. The results indicate that, in the development of PII-CIA, mice expressing the H-2r/r haplotype preferentially utilize TCR V beta genes that are normally encoded within the TCR V beta genomic deletion region of RIIIS mice (V beta c). After aggressive immunization with PII, the use of alternative TCR V beta genes, encoded outside of the RIIIS deletion region, produced a high IgG antibody response that was cross-reactive with mouse type II collagen (MII) and equivalent to that of B10.RIII mice, but only a very mild, late onset arthritis of 56% (27/48) incidence in RIIIS male mice and 28% (10/35) incidence in RIIIS female mice. In comparison, B10.RIII mice routinely developed early onset of PII-CIA of significantly higher incidence (100%; p < 0.005) and four-fold greater severity, even after milder immunization protocols. The data are compatible with the proposal that the clinically weak CIA response of RIIIs mice may be primarily antibody driven while the severe CIA of B10.RIII mice reflects the added inflammatory effects of collagen-reactive effector-T cells in the joint.(ABSTRACT TRUNCATED AT 250 WORDS)

Human HLA-DQβ chain presents minor lymphocyte stimulating locus gene products and clonally deletes TCR Vβ6+, Vβ8.1+ T cells in single transgenic mice

Abstract Minor lymphocyte stimulating locus (Mls) gene products in association with mouse major histocompatibility complex (MHC) class II molecules are known to determine the repertoire of T-cell receptor (TCR) in mature T cells. In order to test whether human class II molecules can present mouse Mls, HLA-DQ β transgenic mice were generated. The expression and function of the DQ β transgene were studied in the progeny of one selected founder which was H-2 f and H-2E negative. In these mice, DQ β molecules pairing with mouse A α chain and invariant chain are expressed on the cell surface in a tissue-specific manner. When the DQ β gene was bred into the Mls-1 a strain DBA/1 (H-2 q ), T cells bearing V β 6 and V β 8.1 TCR were clonally deleted in the thymus of DQ β + transgenics but not in DQ β -negative full sibs. Thus, the data presented here clearly demonstrate that the human MHC DQ β chain can present Mls in the clonal deletion of T cells. Our results also suggest the requirement for an interaction between CD4 and class II molecules (α chain) for clonal deletion of T cells to occur.

Expression and function of mutant Ia antigen in transgenic mice.

Cell surface expression of Ia antigens requires the assembly of alpha and beta heterodimers. We have produced a double transgenic mouse with a wild form Ak alpha gene and a mutant Ak beta (Ak beta MB) gene with d-allele substitution at positions 63 and 65-67. Initial studies indicated that the Ak alpha and Ak beta MB transgenes are not expressed on the surface of lymphoid cells of the transgenic mice. However, when spleen cells were stimulated with LPS prior to FACS analyses, Ak/Ak MB assembly and subsequent surface expression was induced. The tail skins from transgenic founder mice were rejected by the parental mice indicating a role for the mutant antigen on the allograft. In addition, the Ak transgenic mice on H-2q/q background can partially delete V beta 6+ T cells, suggesting the presence of the transgene product in the thymus.