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1.
Figure 3

Figure 3. Role of the TCR Tg in endogenous TCR expression.. From: Endogenous TCR Recombination in TCR Tg Single RAG-Deficient Mice Uncovered by Robust In Vivo T Cell Activation and Selection.

(a) Endogenous TCR rearrangements in donor T cells from OT-1 TCR Tg RAG-1- and aHY TCR Tg RAG-2-deficient mice after transfer into allogeneic hosts. One million LN T-cells from OT-1 RAG-1-deficient or aHY RAG-2-deficient mice were transferred into allogeneic RAG-20/0 γc0/0 H2a hosts. At 9 (OT-1) and 30 (aHY) weeks after transfer, mice were sacrificed. Total RNA was extracted from spleen cells of each hosts and Immunoscope analyses were performed for Vα chains of aHY donor T cell TCRs (left panel) and Vβ chains of OT-1 donor T cell TCRs (right panel). (b) Total mRNA was extracted from the spleen of a RAG-20/0 mouse, retro-transcribed, and amplified using several Vα (left panel) and Vβ (right panel) specific primers. CDR3 length profiles are shown. (c) Shows co-expression of Vα11 and Vβ3 TCR Tg chains with the non-Tg Vα2 or Vβ5.1.2, 6 and 8.1.2.3 TCR chains analyzed by flow cytometry on CD3+CD4+ or CD3+CD8+ LN T-cells from 5CC7 TCR Tg RAG-20/0 mice.

Caroline Montaudouin, et al. PLoS One. 2010;5(4):e10238.
2.
Figure 2

Figure 2. Pre-existence of endogenous TCR expression in the thymus and peripheral lymphoid tissues of 5CC7 RAG-20/0 mice.. From: Endogenous TCR Recombination in TCR Tg Single RAG-Deficient Mice Uncovered by Robust In Vivo T Cell Activation and Selection.

(a–b) Thymus and peripheral cells from 5CC7 RAG-20/0 mice were analyzed for the presence of non-Tg TCR expression both by Flow Cytometry and Immunoscope analyses. (a) Top dot plots show the expression of the Vα11 and Vβ3 TCR Tg chains by CD3lo/hiDN (CD4CD8), DP (CD4+CD8+), SPCD4+ and SPCD8+ thymocytes. Lower panels show CDR3 length profiles for the Vα11 Tg and other non-Tg Vα transcripts present in retro-transcribed and amplified mRNA from total thymocytes using several Vα specific primers. (b) Top dot plots show the expression of the Vα11 and Vβ3 TCR Tg chains by CD3+CD4+ or CD3+CD8+ spleen T-cells. Bottom panels show CDR3 length profiles for the Vα11 Tg and other non-Tg Vα transcripts present in retro-transcribed and amplified mRNA from total spleen cells using several Vα specific primers. (c) T-cells from 5CC7 RAG-deficient donors were transferred into allogeneic or syngeneic hosts. Panels show the numbers of endogenous (filled line) and Tg (dotted line) TCR expressing CD4+ (circles) or CD8+ (squares) T-cells present in the inoculum and 5 and 28 weeks after injection in allogeneic (upper graph) and syngeneic (lower graph) hosts.

Caroline Montaudouin, et al. PLoS One. 2010;5(4):e10238.
3.
Figure 1

Figure 1. Emergence of non Tg TCR expressing T-cells after transfer into allogeneic hosts.. From: Endogenous TCR Recombination in TCR Tg Single RAG-Deficient Mice Uncovered by Robust In Vivo T Cell Activation and Selection.

One million CFSE-labeled LN T-cells from 5CC7 RAG-2-deficient mice were injected into syngeneic H-2a or allogeneic H-2b RAG-20/0γc0/0 hosts. Five (a,b) and 28 (b,c and d) weeks later, mice were sacrificed and spleen cells were analyzed by Flow Cytometry or Immunoscope. (a) Dot plots show CFSE dilution and Vα11 TCR Tg chain expression by gated CD3+CD4+ or CD3+CD8+ 5 weeks after transfer. (b) Dot plots show expression of Vα11 and Vβ3 TCR Tg chains by gated CD3+CD4+ or CD3+CD8+ at 5 and 28 weeks after transfer; (c) Dot plots show that 28 weeks after transfer into allogeneic hosts both CD3+CD4+ and CD3+CD8+ Vβ3 T-cells were panβ+, i.e. they expressed a αβ TCR. Each dot plot is representative of 3 to 6 different mice (d) Twenty-eight weeks after transfer, mRNA was extracted from CD4 or CD8 donor T-cells recovered from pooled allogeneic hosts. mRNA were retro-transcribed and amplified using several Vα (top panels) and Vβ (bottom panels) specific primers. CDR3 length profiles obtained by Immunoscope are shown, with CDR3 size on x-axis (peaks are separated by 1 amino acid), and arbitrary scale of fluorescence on y-axis (proportional to the RT-PCR product amount). The bold framed profiles correspond to the Vα11 or Vβ3 Tg transcripts.

Caroline Montaudouin, et al. PLoS One. 2010;5(4):e10238.

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