Figure 1
Figure 1. Donor T cells with a distinct phenotype infiltrate the CNS during acute GVHD. Flow cytometric analysis and histopathological CNS findings in syngeneic and allogeneic transplanted animals. Lethal GVHD (0.5 × 106 T cells) assessed at days 7, 14, and 21. (A-G) Mononuclear cells were isolated from brains of mice given syngeneic (5 × 106 Thy1.1 BALB/c T cell–depleted bone marrow (TCD BM) cells + 5 × 105 CD5+ Thy1.1 T cells transplanted into Thy1.2 BALB/c recipients) or allogeneic (5 × 106 Ly5.1 B6 TCD BM cells + 5 × 105 CD5+ B6 T cells transplanted into Thy1.2 BALB/c recipients) HSCT after complete perfusion on days 7 (n = 10-13), 14 (n = 13-14), and 21 (n = 48). (A) Representative flow cytometry plots detailing expression of CD3. (B) Absolute number of CNS infiltrating donor CD3+ T cells. (C) Origin of allogeneic CD3+ T cells (donor = CD45.2+H-2b+). (D) Absolute number of CD4+ or CD8+ allogeneic CD3+ T cells. (E) Absolute number of CD4+ and CD8+ T cells in syngeneic and allogeneic HSCT recipients in spleen (n = 8-10). (F) Representative plots and absolute numbers of naive (N, CD62L+CD44-), effector memory (EM, CD62L+CD44+), and central memory (CM, CD62L-CD44+) T cells in allo-HSCT recipients at day 21 (n = 31-37). (G) Expression of PSGL-1, α4-Integrin, LFA-1, LPAM, CXCR3, and CCR4 on CD4+ and CD8+ T cells in CNS and spleen at day 21 (n = 10-15). (H-M) Coronal sections were taken from paraffin-embedded brains 21 days after syn- (n = 10) or allo-HSCT (n = 10). (H) CD3 or CD4 staining. (I) Hematoxylin and eosin–stained section from an untreated mice demonstrating observed areas of the brain examined; cortex (Co), hippocampus (Hc), midbrain (Mb: thalamus, basal ganglia, hypothalamus), cerebellum (Ce), medulla oblongata (Mo), and choroid plexus (CP). Quantification of total CD3+ T cells (J) and CD4+ T cell (K) numbers in indicated areas. (L) Relative and (M) absolute distribution of CD3+ T cells in meninges/ependyma, parenchyma, and parenchymal vessels. Bar graphs represent mean ± standard error of the mean. All data are representative of at least 2 independent experiments. *P < .05; **P < .01; ***P < .001.

Donor T cells with a distinct phenotype infiltrate the CNS during acute GVHD. Flow cytometric analysis and histopathological CNS findings in syngeneic and allogeneic transplanted animals. Lethal GVHD (0.5 × 106 T cells) assessed at days 7, 14, and 21. (A-G) Mononuclear cells were isolated from brains of mice given syngeneic (5 × 106 Thy1.1 BALB/c T cell–depleted bone marrow (TCD BM) cells + 5 × 105 CD5+ Thy1.1 T cells transplanted into Thy1.2 BALB/c recipients) or allogeneic (5 × 106 Ly5.1 B6 TCD BM cells + 5 × 105 CD5+ B6 T cells transplanted into Thy1.2 BALB/c recipients) HSCT after complete perfusion on days 7 (n = 10-13), 14 (n = 13-14), and 21 (n = 48). (A) Representative flow cytometry plots detailing expression of CD3. (B) Absolute number of CNS infiltrating donor CD3+ T cells. (C) Origin of allogeneic CD3+ T cells (donor = CD45.2+H-2b+). (D) Absolute number of CD4+ or CD8+ allogeneic CD3+ T cells. (E) Absolute number of CD4+ and CD8+ T cells in syngeneic and allogeneic HSCT recipients in spleen (n = 8-10). (F) Representative plots and absolute numbers of naive (N, CD62L+CD44-), effector memory (EM, CD62L+CD44+), and central memory (CM, CD62L-CD44+) T cells in allo-HSCT recipients at day 21 (n = 31-37). (G) Expression of PSGL-1, α4-Integrin, LFA-1, LPAM, CXCR3, and CCR4 on CD4+ and CD8+ T cells in CNS and spleen at day 21 (n = 10-15). (H-M) Coronal sections were taken from paraffin-embedded brains 21 days after syn- (n = 10) or allo-HSCT (n = 10). (H) CD3 or CD4 staining. (I) Hematoxylin and eosin–stained section from an untreated mice demonstrating observed areas of the brain examined; cortex (Co), hippocampus (Hc), midbrain (Mb: thalamus, basal ganglia, hypothalamus), cerebellum (Ce), medulla oblongata (Mo), and choroid plexus (CP). Quantification of total CD3+ T cells (J) and CD4+ T cell (K) numbers in indicated areas. (L) Relative and (M) absolute distribution of CD3+ T cells in meninges/ependyma, parenchyma, and parenchymal vessels. Bar graphs represent mean ± standard error of the mean. All data are representative of at least 2 independent experiments. *P < .05; **P < .01; ***P < .001.

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