CD3/CD28-costimulated T1 and T2 subsets: differential in vivo allosensitization generates distinct GVT and GVHD effects

Abstract

Adoptive T-cell therapy using CD3/CD28 costimulation likely requires in vivo generation of antigen specificity. Because CD28 promotes T H 1/T C 1 (T1) or T H 2/T C 2 (T2) differentiation, costimulation may generate donor T1 or T2 cells capable of differentially mediating allogeneic graft-versus-tumor (GVT) effects and graft-versus-host disease (GVHD). Costimulation under T1 or T2 conditions indeed generated murine T H 1/T C 1 cells secreting interleukin-2/interferon-␥ (IL-2/ IFN-␥) or T H 2/T C 2 cells secreting IL-4/IL-5/IL-10. In vivo, allogeneic T1 cells expanded, maintained T1 secretion, and acquired allospecificity involving IFN-␥ and IL-5. In contrast, allogeneic T2 cells expanded less and maintained T2 secretion but did not develop significant allospecificity. Allogeneic, but not syngeneic, T1 cells mediated a GVT effect against host-type breast cancer cells, as median survival time (MST) increased from 25.6 ؎ 2.6 (tumor controls) to 69.2 ؎ 5.9 days (P < 1.2 ؋ 10 ؊9 ). This T1-associated GVT effect operated independently of fasL because T1 cells from gld mice mediated tumor-free survival. In contrast, allogeneic T2 cells mediated a modest, noncurative GVT effect (MST, 29 ؎ 1.3 days; P < .0019). T1 recipients had moderate GVHD (histologic score, 4 of 12) that contributed to lethality after bone marrow transplantation; in contrast, T2 recipients had minimal GVHD (histologic score, 1 of 12). CD3/CD28 costimulation, therefore, generates T1 or T2 populations with differential in vivo capacity for expansion to alloantigen, resulting in differential GVT effects and GVHD.

Activation of Th1 and Tc1 cell adenosine A2A receptors directly inhibits IL-2 secretion in vitro and IL-2-driven expansion in vivo

“…Costimulated Tc1 and Tc2 subsets each possessed cytolytic activity (Figure 7). Similar to prior results, 17 costimulated Tc2 cells were more potent than Tc1 cells with respect to exocytosis-based killing, whereas costimulated Tc1 cells were modestly enriched for fas-based killing. Strikingly, adenosine A 2A receptor activation at doses of CGS (10 Ϫ8 and 10 Ϫ6 M) that potently regulated cytokine secretion did not modulate Tc1 or Tc2 cytolytic function in either exocytosisor fas-based killing assays.…”

“…This result predicts that adenosine receptor modulation might be used to treat IL-2-driven diseases involving primarily Th1 and Tc1 cellular pathways, such as graft-versus-host disease (GVHD). 17,32 Conversely, it is also interesting to speculate that the mechanism accounting for antitumor effects after IL-2 administration 33 may operate in part through abrogation of adenosinemediated T-cell inhibition.…”

Pharmacologic Inhibition of JAK1/JAK2 Signaling Reduces Experimental Murine Acute GVHD While Preserving GVT Effects

“…Building upon this prior study, we now show that the preserved GVT activity by ruxolitinib is associated with maintained in vivo polarization of donor T cells toward the Th1 and Th17 phenotypes that are generally recognized as efficient mediators of antitumor effects (36,37). Moreover, our observations are in agreement with a previous report from Choi and colleagues (19) that described JAK1/JAK2 inhibition by ruxolitinib as an approach to block the IFNgR-CXCR3 axis and prevent T-cell migration into GVHD organs.…”

Ex Vivo Rapamycin Generates Apoptosis-Resistant Donor Th2 Cells That Persist In Vivo and Prevent Hemopoietic Stem Cell Graft Rejection

“…Anti-CD3-and anti-CD28-coated beads (CD3/CD28 beads) were produced as previously detailed (26); T cells were stimulated with CD3/CD28 beads (T cell:bead ratio, 1:3) in complete medium (CM) containing RPMI 1640 (Mediatech), 10% FCS (Gemini Bio-Products), penicillin-streptomycin-glutamine, nonessential amino acids (Invitrogen Life Technologies), 2-ME (5 ϫ 10 Ϫ5 M; Invitrogen Life Technologies), and N-acetyl-L-cysteine (3.3 mM; BristolMyers Squibb). For Th2/Tc2 cultures, CM was supplemented with rhIL-2 (1000 IU/ml for Th2/Tc2 or purified CD8 ϩ Tc2 cultures; 20 IU/ml for purified CD4 ϩ Th2 cultures), rhIL-7 (20 ng/ml), and rmIL-4 (1000 IU/ml); for Th1 cultures, CD4 ϩ T cells were expanded in CM supplemented with rhIL-2 (20 IU/ml), rhIL-7 (20 ng/ml), 11B11 (10 g/ml) and rmIL-12 (10 ng/ml; cytokines obtained from PeproTech).…”

“…Because the number of donor T cells capable of IFN-␥ secretion in response to alloantigen is a suitable measure of graft-vs-host reactivity (GVHR) (25,26), we reasoned that quantification of host T cells capable of IFN-␥ secretion in response to donor alloantigen might represent a marker of HVGR. To quantify HVGR, posttransplant T cells were harvested, stimulated with either syngeneic host APC or the relevant allogeneic APC, and then subjected to surface flow cytometry for detection of T cell lineage markers (CD4 ϩ or CD8 ϩ subsets) and the host genetic MHC marker (H-2 d ), with concomitant capture detection of IFN-␥ using anti-CD45, anti-IFN-␥ bispecific Ab.…”