Figure 4.
Figure 4. Evaluation of infiltrating donor-derived T cells in allogeneic target tissues. (A) Allogeneic CD4+ T cells precede the increase of donor CD8+ T cells in lymphoid organs during GVHD initiation. Shown are absolute numbers of donor-derived T cells in high-power fields (HPFs) by IFM. The shift from donor CD4+ (▪) to CD8+ T cells (▵) occurred first in the spleen on day 4 and later in other lymphoid organs. Donor CD4+ T cells appear in the small intestines after extensive proliferation in lymphoid organs not earlier than day 4 after HCT followed by infiltrating cytotoxic T cells (Standard deviation was calculated using cell counts from 3 to 4 HPFs of 2 experimental mice for each time point). (B) FACS analysis of donor (H-2Kq+) lymphocytes reveals changes in expression profiles of activation and homing markers. The up-regulation of CD69 in donor CD4+ and CD8+ T cells is followed by CD44 expression. Up-regulation of α4β7 differs between donor CD4+ and CD8+ T cells in the spleen. Shown is a representative experiment (of 2-3 independent experiments) in which cells were pooled from the spleens of allogeneic transplant recipients (n = 3-5/time point). The numbers in each quadrant represent percentages of cells. (C) Donor CD4+ T cells with gut-homing potential appear in PPs and mLNs until day 3 after HCT before donor CD4+ T cells start to appear in mucosal sites of the intestines. On day 3 after HCT the frequency of α4β7hi donor CD4+ T cells in PPs and mLNs is significantly higher in cells primed in vivo by PPs and mLNs versus spleen and cLNs. α4β7hi donor CD8+ T cells can be found in the spleen in addition to PPs and mLNs, but only a few are found in cLNs. Error bars indicate the standard deviation of 3 independent experiments. (D) CFSE proliferation analysis demonstrated that it takes more than 5 cell divisions before up-regulation of α4β7 integrin occurred in mLNs (> 98% of all α4β7hi). Peripheral LNs such as cLNs and iLNs do not contain gut-homing T cells. (PPs are not shown because of too-low cell yields for CFSE analysis.)

Evaluation of infiltrating donor-derived T cells in allogeneic target tissues. (A) Allogeneic CD4+ T cells precede the increase of donor CD8+ T cells in lymphoid organs during GVHD initiation. Shown are absolute numbers of donor-derived T cells in high-power fields (HPFs) by IFM. The shift from donor CD4+ (▪) to CD8+ T cells (▵) occurred first in the spleen on day 4 and later in other lymphoid organs. Donor CD4+ T cells appear in the small intestines after extensive proliferation in lymphoid organs not earlier than day 4 after HCT followed by infiltrating cytotoxic T cells (Standard deviation was calculated using cell counts from 3 to 4 HPFs of 2 experimental mice for each time point). (B) FACS analysis of donor (H-2Kq+) lymphocytes reveals changes in expression profiles of activation and homing markers. The up-regulation of CD69 in donor CD4+ and CD8+ T cells is followed by CD44 expression. Up-regulation of α4β7 differs between donor CD4+ and CD8+ T cells in the spleen. Shown is a representative experiment (of 2-3 independent experiments) in which cells were pooled from the spleens of allogeneic transplant recipients (n = 3-5/time point). The numbers in each quadrant represent percentages of cells. (C) Donor CD4+ T cells with gut-homing potential appear in PPs and mLNs until day 3 after HCT before donor CD4+ T cells start to appear in mucosal sites of the intestines. On day 3 after HCT the frequency of α4β7hi donor CD4+ T cells in PPs and mLNs is significantly higher in cells primed in vivo by PPs and mLNs versus spleen and cLNs. α4β7hi donor CD8+ T cells can be found in the spleen in addition to PPs and mLNs, but only a few are found in cLNs. Error bars indicate the standard deviation of 3 independent experiments. (D) CFSE proliferation analysis demonstrated that it takes more than 5 cell divisions before up-regulation of α4β7 integrin occurred in mLNs (> 98% of all α4β7hi). Peripheral LNs such as cLNs and iLNs do not contain gut-homing T cells. (PPs are not shown because of too-low cell yields for CFSE analysis.)

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal