Characterization of the ovalbumin‐specific TCR transgenic line OT‐I: MHC elements for positive and negative selection

Abstract

Proliferative responses were performed as previously described. 10 Briefly, lymph nodes from OT-I mice were removed and single cell Immunology and Cell Biology (2000) Summary The present report provides the first extensive characterization of the OT-I TCR transgenic line, which produces MHC class I-restricted, ovalbumin-specific, CD8 + T cells (OT-I cells). These cells are shown to be positively selected in vivo in H-2 b C57BL/6 mice and in bm5 mice, which express the K bm5 mutant molecule. In contrast, OT-I cells were not selected by mutant K b molecules in bm1, bm3, bm8, bm10, bm11 or bm23 mice. Interestingly, however, when positive selection was examined in vitro in foetal thymic organ culture (FTOC), bm1 and bm8 were still poorly selective, but the bm3 haplotype now selected as efficiently as B6. The ability to select in vitro correlated with the capacity to present the ovalbumin (OVA) peptide to OT-I cells, as measured by induction of an OVA-specific proliferative response. These results suggest that a lower affinity TCR:MHC interaction may be necessary for positive selection in FTOC compared with selection in situ.

Development of a Novel CD4+ TCR Transgenic Line That Reveals a Dominant Role for CD8+ Dendritic Cells and CD40 Signaling in the Generation of Helper and CTL Responses to Blood-Stage Malaria

“…1B, 1D). Similar to previously described TCR transgenic mice, total cellularity was reduced in the thymus, indicative of efficient positive selection (16,59). …”

Type I Interferons Mediate the Innate Cytokine Response to Recombinant Fowlpox Virus but Not the Induction of Plasmacytoid Dendritic Cell-Dependent Adaptive Immunity

“…Control mice were produced in-house as F1 progeny from successful matings of normal C57BL/6J (B6) and SV129 mice purchased from Laboratory Animal Services ([LAS] University of Adelaide, S. A., Australia). OT-I T-cell receptor (TCR) transgenic (Tg) mice (8), which contain Tg CD8 ϩ T cells that recognize the dominant chicken ovalbumin peptide consisting of residues 257 to 264 (OVA [257][258][259][260][261][262][263][264] ) in the context of H2-K b , and B6 bm1 mice, which harbor a single natural mutation in the binding groove of H-2K b and are therefore unable to present the H-2K b -restricted OVA 257-264 peptide to CD8 ϩ T cells (39), were both kindly provided by W. R. Heath (Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia). Congenic CD45.1 ϩ B6.SJL mice were purchased from the Animal Resource Centre (Canning Vale, W. A., Australia).…”

Dendritic Cell Immunization Route Determines Integrin Expression and Lymphoid and Nonlymphoid Tissue Distribution of CD8 T Cells

“…Because BMDC were unable to up-regulate ␣ 4 ␤ 7 , this raised the question of whether the endogenous cells in the LN that imprinted expression of this integrin in vivo also were responsible for presenting Ag. To address this, we used irradiated C57BL/6 mice reconstituted with bone marrow from bm1 mice, which have a mutated H-2K b molecule that is unable to present OVA 257 to OT-1 cells (26). After i.p.…”