Journal: The Journal of Experimental Medicine

Article Title: The Effect of Graft-versus-Host Disease on T Cell Production and Homeostasis

doi:

Figure Lengend Snippet: The influence of GVHD on T cell development in allogeneic chimeras. Irradiated euthymic (A and C) or thymectomized (B and D) A.BY (Thy1.2, Ly5.2) recipients received a graft containing 10 7 T cell–depleted B6.PL (Thy1.1, Ly5.2) bone marrow cells (as a source of hematopoietic progenitors) with or without mature T cells (0.4 or 1.6 × 10 6 ) harvested from the LNs of B6.SJL donors (Thy1.2, Ly5.1). Chimeras' spleen cells were analyzed by three-color staining on day 100 ± 5 posttransplant. The origin of recipient T cells was determined according to their Thy1/Ly5 phenotype. Results are presented as the mean ± SD (three to four mice per group). Note that the ordinate scale is different in A and B versus C and D. nd, not determined.

Article Snippet: Bone marrow cells were obtained from the tibias and femurs of donor mice and T cell depleted with a specific anti-Thy1.2 mAb (5a-8; mouse IgG) or with a rabbit anti–mouse T cells (Thy1) antiserum, both obtained from Cedarlane Labs., and rabbit serum (Low-Tox-M rabbit complement; Cedarlane Labs.) as a source of complement.

Techniques: Irradiation, Staining

Journal: The Journal of Experimental Medicine

Article Title: The Effect of Graft-versus-Host Disease on T Cell Production and Homeostasis

doi:

Figure Lengend Snippet: Skewing of Vβ usage by CD4 + and CD8 + T cells in athymic GVHD + chimeras. Three-color staining was performed using Abs against CD4 or CD8, Thy1.1 or Thy1.2, and specific Vβ elements. Chimeras were studied on day 100 ± 5. Results are presented as the mean ± SD (three to four mice per group).

Article Snippet: Bone marrow cells were obtained from the tibias and femurs of donor mice and T cell depleted with a specific anti-Thy1.2 mAb (5a-8; mouse IgG) or with a rabbit anti–mouse T cells (Thy1) antiserum, both obtained from Cedarlane Labs., and rabbit serum (Low-Tox-M rabbit complement; Cedarlane Labs.) as a source of complement.

Techniques: Staining

Journal: The Journal of Experimental Medicine

Article Title: The Effect of Graft-versus-Host Disease on T Cell Production and Homeostasis

doi:

Figure Lengend Snippet: Expansion of T cells from GVHD + mice after adoptive transfer into normal hosts. Three-color staining was performed on spleen cell suspensions using Abs specific for CD4 or CD8, Ly5.1 or Ly5.2, and Thy1.1 or Thy1.2. (A) Numbers of CD4 + and CD8 + T cells of B6.PL origin (Thy1.1, Ly5.2) found in the spleen of day 100 nonthymectomized GVHD + recipients (negative controls). (B) Numbers of CD4 + and CD8 + T cells of B6.PL origin (Thy1.1, Ly5.2) found in the spleen of irradiated/thymectomized B6.SJL or A.BY secondary hosts, on day 30 after injection of 10 7 T cell–depleted C57BL/6 bone marrow cells + 2 × 10 6 T cells harvested from the spleen of day 70 GVHD + mice (test group). (C) Numbers of Thy1.2 + Ly5.2 + T cells found in the spleen of irradiated/thymectomized B6.SJL or A.BY hosts, on day 30 after injection of 10 7 T cell–depleted B6.PL bone marrow cells + 2 × 10 6 T cells harvested from the spleen of normal syngeneic donors (A.BY for A.BY secondary recipients, C57BL/6 for B6.SJL secondary recipients) (positive controls). GVHD + mice were prepared as in Fig. C (irradiation followed by transplantation of 10 7 B6.PL bone marrow cells + 0.4 × 10 6 B6.SJL LN T cells). Results are presented as the mean ± SD (three to four mice per group). * P < 0.05, ** P < 0.005 relative with numbers of CD4 + or CD8 + T cells in GVHD + mice (Student's t test). No significant difference was found between T cell numbers presented in B and C.

Article Snippet: Bone marrow cells were obtained from the tibias and femurs of donor mice and T cell depleted with a specific anti-Thy1.2 mAb (5a-8; mouse IgG) or with a rabbit anti–mouse T cells (Thy1) antiserum, both obtained from Cedarlane Labs., and rabbit serum (Low-Tox-M rabbit complement; Cedarlane Labs.) as a source of complement.

Techniques: Adoptive Transfer Assay, Staining, Irradiation, Injection, Transplantation Assay

Journal: The Journal of Experimental Medicine

Article Title: The Effect of Graft-versus-Host Disease on T Cell Production and Homeostasis

doi:

Figure Lengend Snippet: Adoptive transfer of normal host (A.BY)-tolerant T cells into nonthymectomized GVHD + mice and thymectomized GVHD − recipients. (A) Number of T cells in the spleen of day 100 GVHD + mice. GVHD + mice were prepared as in Fig. C (irradiation followed by transplantation of 10 7 B6.PL bone marrow cells + 0.4 × 10 6 B6.SJL T cells). (B) Number of T cells in the spleen of day 100 GVHD + mice previously injected, on day 60, with 5 × 10 6 host-tolerant T cells of C57BL/6 (Thy1.2,Ly5.2) origin. (C) Number of T cells in the spleen of thymectomized A.BY mice 40 d after irradiation and injection of 10 7 T cell–depleted B6.SJL bone marrow cells + 5 × 10 6 host-tolerant B6.PL T cells. Host (A.BY)-tolerant T cells used in B and C experiments were obtained from the spleen of nonthymectomized A.BY hosts, 60 d after irradiation and injection of T cell–depleted C57BL/6 or B6.PL bone marrow cells. Results are presented as the mean ± SD (three to four mice per group).

Article Snippet: Bone marrow cells were obtained from the tibias and femurs of donor mice and T cell depleted with a specific anti-Thy1.2 mAb (5a-8; mouse IgG) or with a rabbit anti–mouse T cells (Thy1) antiserum, both obtained from Cedarlane Labs., and rabbit serum (Low-Tox-M rabbit complement; Cedarlane Labs.) as a source of complement.

Techniques: Adoptive Transfer Assay, Irradiation, Transplantation Assay, Injection