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Bone marrow chimeras were generated using the Ly5.1/5.2 reconstitution system, as previously described [6].
Bone marrow chimeras were analyzed at 4 8 weeks post-bone marrow transfer (BMT).
Bone marrow chimeras were generated to test the possibility that lamin A/C expression is required in developing lymphocytes for normal T and B cell maturation.
Lmna-/ CD4+ and CD8+ T cell populations in mixed bone marrow chimeras were able to generate self-MHC class II and class I restricted T cell responses to LCMV infection.
Longitudinal sections of the optic nerve of 2, 6 and 12 months old wildtype, PD-1-/-, PLPtg and PLPtg/PD-1-/ mice and 10 months old bone marrow chimeras were analysed.
The bone marrow chimeras were transplanted with PD-1-/ or wildtype bone marrow at the age of 6 8 weeks and investigated at the age of 10 months (n = 8 9).
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The B cell reconstitution of the mixed bone marrow chimeras was determined 6 8 weeks after bone marrow injection by FACS analysis of blood samples using anti-IgMa, anti-IgMb, and anti-CD1d antibodies.
To determine if the expression of CD38 on p110δD910A Treg is governed by signaling within the T cells themselves or by an extrinsic factor, competitive bone-marrow chimeras were generated.
Groups of WT (CCR7+/+), CCR7-/ mice (n = 5-10 per timepoint) and bone-marrow chimeras were monitored for survival.
WT (CCR7+/+), CCR7-/ mice, or bone-marrow chimeras were given 0.05 U of sterile BLM sulfate (Blenoxane, Bristol-Meyers PharmaceutINals, Evansville, IN, USA) dissolved in phosphate-buffered saline (approximately 1.7 U/kg body weight) via an intratracheal injection.
Reconstitution of bone marrow chimera was verified by flow cytometry analysis: the intrahepatic CD11b+CD45+ cells isolated from Balb/c→B6 chimeras express high levels (>95%) of H2-Kd, and only a small fraction (∼4%) express H2-Kb at levels that are slightly above background.
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