Abrogation of donor T-cell IL-21 signaling leads to tissue-specific modulation of immunity and separation of GVHD from GVL - PubMed (original) (raw)

. 2011 Jul 14;118(2):446-55.

doi: 10.1182/blood-2010-07-294785. Epub 2011 May 19.

Lucy W Kappel, Nury L Yim, Rebecca A Nejat, Gabrielle L Goldberg, Odette M Smith, Uttam K Rao, Lindsay Dykstra, Il-Kang Na, Amanda M Holland, Jarrod A Dudakov, Chen Liu, George F Murphy, Warren J Leonard, Glenn Heller, Marcel R M van den Brink

Affiliations

Abrogation of donor T-cell IL-21 signaling leads to tissue-specific modulation of immunity and separation of GVHD from GVL

Alan M Hanash et al. Blood. 2011.

Abstract

IL-21 is a proinflammatory cytokine produced by Th17 cells. Abrogation of IL-21 signaling has recently been shown to reduce GVHD while retaining graft-versus-leukemia/lymphoma (GVL) responses. However, the mechanisms by which IL-21 may lead to a separation of GVHD and GVL remain incompletely understood. In a murine MHC-mismatched BM transplantation model, we observed that IL-21 receptor knockout (IL-21R KO) donor T cells mediate decreased systemic and gastrointestinal GVHD in recipients of a transplant. This reduction in GVHD was associated with expansion of transplanted donor regulatory T cells and with tissue-specific modulation of Th-cell function. IL-21R KO and wild-type donor T cells showed equivalent alloactivation, but IL-21R KO T cells showed decreased infiltration and inflammatory cytokine production within the mesenteric lymph nodes. However, Th-cell cytokine production was maintained peripherally, and IL-21R KO T cells mediated equivalent immunity against A20 and P815 hematopoietic tumors. In summary, abrogation of IL-21 signaling in donor T cells leads to tissue-specific modulation of immunity, such that gastrointestinal GVHD is reduced, but peripheral T-cell function and GVL capacity are retained. IL-21 is thus an exciting target for therapeutic intervention and improvement of clinical transplantation outcomes.

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Figures

Figure 1

Figure 1

IL-21R is up-regulated on donor T cells early after BMT but is not required for efficient alloactivation. (A) WT B6 T cells were cultured in vitro for 24 hours in the presence or absence of PMA and ionomycin, then stained for FACS analysis of IL-21R levels. (B) WT B6 BM-TCD and T cells (1 × 106) were transplanted into lethally irradiated BALB/c recipients, which were killed on day 4 or day 7 after BMT. Splenocytes were harvested and stained for FACS analysis of donor T-cell IL-21R levels. Data are combined from 2 independent transplantations per time point, with 8-10 total recipients per group. **P < .01 and ***P < .001.

Figure 2

Figure 2

IL-21R KO T cells cause decreased GVHD and lead to increased regulatory T cells. (A) Lethally irradiated BALB/c mice were reconstituted with either BM-TCD only (n = 9), 1 × 106 IL-21R KO T cells + BM-TCD (n = 15), or 1 × 106 WT T cells + BM-TCD (n = 15). Curves represent combined data from 2 independent experiments. (B) Lethally irradiated BALB/c mice received either BM-TCD only (n = 9), 5 × 105 IL-21R KO CD4 T cells + BM-TCD (n = 15), or 5 × 105 WT CD4 T cells + BM-TCD (n = 15). Curves represent combined data from 2 independent experiments. (C,D) Transplant recipients were scored on a weekly basis for 5 clinical GVHD parameters: weight loss, activity, kyphosis, fur ruffling, and skin flaking. Scores range from 0 to 2 for each parameter, and animals are killed once a total score of 5 is attained. Curves represent the average score of each group at each time point. Shown are combined data from 2 independent experiments. (E) Lethally irradiated BALB/c mice were reconstituted with B6 BM-TCD and 1 × 106 B6 WT or 1 × 106 B6 IL-21R KO T cells. Spleens were harvested on days 7, 14, and 21 after BMT and stained for FACS analysis of donor CD4+CD25+Foxp3+ T-regs or CD4+Foxp3− nonregulatory T cells. Data represent 2 combined independent experiments on day 7 (n > 19), day 14 (n = 18), and day 21 (n = 15). (F) Lethally irradiated BALB/c mice were reconstituted with CD45.1 B6 BM-TCD and 5 × 105 CD45.2 B6 MACS-purified T cells. Transplanted T-cell populations were either unfractionated or depleted of T-regs by sorting for CD25− cells. Spleens were harvested on day 21 after BMT and analyzed by FACS for Foxp3 expression among donor CD4 T cells. Data represent 2 combined independent experiments with 8-11 total recipients per group. *P < .05, **P < .01, and ***P < .001 for WT versus KO T cells.

Figure 3

Figure 3

IL-21 signaling is not required for GVL against hematopoietic malignancy. (A) Lethally irradiated BALB/c mice were transplanted with either B6 BM-TCD alone or with A20 tumor cells (2.5-5 × 105) with or without WT versus IL-21R KO T cells (0.5-1 × 106). Recipients were followed for mortality, and curves represent combined data from 4 independent experiments (n = 19-20 for non–T-cell groups, n = 39 for T-cell groups). (B) Lethally irradiated B6 × DBA/2 F1 mice were transplanted with either B6 BM-TCD alone or with P815 tumor cells (1 × 103) with or without WT versus IL-21R KO T cells (1 × 106). Recipients were followed for mortality, and curves represent combined data from 2 independent experiments (n = 10 for non–T-cell groups, n = 20 for T-cell groups). ***P < .001 for WT versus KO T cells.

Figure 4

Figure 4

IL-21R KO T cells have decreased intestinal infiltration and LPAM expression after BMT. (A) Lethally irradiated BALB/c mice were transplanted with B6 BM-TCD and WT or IL-21R KO T cells (1 × 106). Skin, liver, small intestine (S.I.) and large intestine (L.I.) were harvested 3 weeks after BMT. H&E-stained slides were scored for histopathologic damage and lymphocyte infiltration. Shown is the mean of each group from 2 combined independent experiments (n > 12). (B) Lethally irradiated BALB/c mice were transplanted with B6 BM-TCD and WT B6 T cells (1 × 106). Lamina propria lymphocytes from small and large intestines were isolated 1 week after BMT and stained for IL-21R expression on donor T cells. Shown are combined data from 2 independent experiments with 7 total recipients per group. (C) Lethally irradiated LP mice were transplanted with B6 BM-TCD and WT or IL-21R KO B6 T cells (1 × 106). Liver, small intestine, and large intestine were harvested 3 weeks after BMT, and H&E-stained slides were scored for histopathologic damage. Shown are combined data from 2 independent experiments with 17 total recipients per group. (D) Lethally irradiated BALB/c mice were transplanted with B6 BM-TCD and WT or IL-21R KO T cells (1 × 106). Splenocytes were harvested 1 week after BMT and stained for FACS analysis of donor T-cell CD103, CCR9, and LPAM. Shown are combined data from 4 to 5 independent experiments with 20-25 total recipients per group. *P < .05 and ***P < .001 for WT versus KO T cells (A,C,D) or WT T cells in small versus large intestine (B).

Figure 5

Figure 5

IL-21R KO T cells are decreased in MLNs after BMT. (A) Lethally irradiated BALB/c mice were transplanted with B6 BM-TCD and WT or IL-21R KO T cells (1 × 106). Spleens and MLNs were harvested 1 week after BMT and stained for FACS analysis of donor CD4 and CD8 T cells. Shown are combined data from 4 independent experiments with 20 total recipients per group. (B) CD45.1 B6 and IL-21R KO (CD45.2) T cells were cotransferred into lethally irradiated BALB/c recipients. Spleens and MLNs were harvested 4 hours after transfer and stained for FACS analysis of donor T cells. The ratio of spleen to MLN donor T cells was calculated, and connecting lines indicate WT (CD45.1) and KO (CD45.2) donor T cells isolated from individual recipients. Shown is 1 of 3 independent experiments. (C) Freshly isolated MLNs were harvested from untransplanted WT or CD45.1 B6 and IL-21R KO B6 mice and stained for FACS analysis of T-cell CCR7. Shown are combined results of 4 independent experiments with a total of 14 mice per group. *P < .05 and ***P < .001 for WT versus KO T cells.

Figure 6

Figure 6

IL-21R KO CD4 T-cell IFN-γ and TNF-α are maintained in the spleen but reduced in MLNs. Lethally irradiated BALB/c mice were transplanted with B6 BM-TCD and WT or IL-21R KO T cells (1 × 106). (A) MLNs and (B) spleens were harvested 1 week after BMT and stained for intracellular cytokine FACS analysis of donor CD4 T cells. Shown are combined data from 4 independent experiments with 20 total recipients per group. **P < .01 and ***P < .001 for WT versus KO T cells.

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References

    1. Shlomchik WD. Graft-versus-host disease. Nat Rev Immunol. 2007;7(5):340–352. - PubMed
    1. van den Brink MR, Burakoff SJ. Cytolytic pathways in haematopoietic stem-cell transplantation. Nat Rev Immunol. 2002;2(4):273–281. - PubMed
    1. Hill GR, Ferrara JL. The primacy of the gastrointestinal tract as a target organ of acute graft-versus-host disease: rationale for the use of cytokine shields in allogeneic bone marrow transplantation. Blood. 2000;95(9):2754–2759. - PubMed
    1. Petrovic A, Alpdogan O, Willis LM, et al. LPAM (alpha 4 beta 7 integrin) is an important homing integrin on alloreactive T cells in the development of intestinal graft-versus-host disease. Blood. 2004;103(4):1542–1547. - PubMed
    1. Reddy P. Pathophysiology of acute graft-versus-host disease. Hematol Oncol. 2003;21(4):149–161. - PubMed

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