VSports在线直播 - Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The . gov means it’s official. Federal government websites often end in . gov or . mil VSports app下载. Before sharing sensitive information, make sure you’re on a federal government site. .

Https

The site is secure. The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely. V体育官网.

. 2021 Aug 26;138(8):722-737.
doi: 10.1182/blood.2020006375.

IFN-λ therapy prevents severe gastrointestinal graft-versus-host disease

Andrea S Henden  1   2   3 , Motoko Koyama  4 , Renee J Robb  1 , Adriana Forero  5 , Rachel D Kuns  1 , Karshing Chang  1 , Kathleen S Ensbey  4 , Antiopi Varelias  1 , Stephen H Kazakoff  6 , Nicole Waddell  6 , Andrew D Clouston  7 , Rabina Giri  8 , Jakob Begun  8 , Bruce R Blazar  9 , Mariapia A Degli-Esposti  10   11 , Sergei V Kotenko  12   13 , Steven W Lane  14 , Kate L Bowerman  15 , Ram Savan  5 , Philip Hugenholtz  15 , Kate H Gartlan  1   3 , Geoffrey R Hill  1   4   16
Affiliations

"VSports" IFN-λ therapy prevents severe gastrointestinal graft-versus-host disease

Andrea S Henden et al. Blood. .

Abstract

Immunopathology and intestinal stem cell (ISC) loss in the gastrointestinal (GI) tract is the prima facie manifestation of graft-versus-host disease (GVHD) and is responsible for significant mortality after allogeneic bone marrow transplantation (BMT) VSports手机版. Approaches to prevent GVHD to date focus on immune suppression. Here, we identify interferon-λ (IFN-λ; interleukin-28 [IL-28]/IL-29) as a key protector of GI GVHD immunopathology, notably within the ISC compartment. Ifnlr1-/- mice displayed exaggerated GI GVHD and mortality independent of Paneth cells and alterations to the microbiome. Ifnlr1-/- intestinal organoid growth was significantly impaired, and targeted Ifnlr1 deficiency exhibited effects intrinsic to recipient Lgr5+ ISCs and natural killer cells. PEGylated recombinant IL-29 (PEG-rIL-29) treatment of naive mice enhanced Lgr5+ ISC numbers and organoid growth independent of both IL-22 and type I IFN and modulated proliferative and apoptosis gene sets in Lgr5+ ISCs. PEG-rIL-29 treatment improved survival, reduced GVHD severity, and enhanced epithelial proliferation and ISC-derived organoid growth after BMT. The preservation of ISC numbers in response to PEG-rIL-29 after BMT occurred both in the presence and absence of IFN-λ-signaling in recipient natural killer cells. IFN-λ is therefore an attractive and rapidly testable approach to prevent ISC loss and immunopathology during GVHD. .

PubMed Disclaimer

"VSports手机版" Figures

None
Graphical abstract
Figure 1.
Figure 1.
IFN-λ signaling in recipient tissue determines GVHD severity. (A) Survival by Kaplan-Meier estimates for B6.WT (n = 31) and B6.Ifnlr1–/– (Ifnlr1–/–, n = 31) recipient mice lethally irradiated (1000 cGy) and transplanted with BALB/c-derived BM and T cells. A non-GVHD control group received T cell–depleted BM only (TCD; n = 10). Data combined from 5 experiments. (B) Clinical GVHD scores + SEM. (C) Representative images of colon and SI at day 7 after BMT. (D) Semiquantitative GVHD histopathology scores at day 7 after BMT (WT and Ifnlr1–/–, n = 9; TCD, n = 6, combined from 2 experiments). (E) Serum fluorescein isothiocyanate (FITC) dextran at day 7 post-BMT (WT, n = 10; Ifnlr1–/–, n = 9; combined from 2 experiments). (F) Serum IFN-γ, IL-6, tumor necrosis factor (TNF), and IL-17A at day 4 post-BMT (WT & Ifnlr1–/–, n = 23; TCD, n = 10; combined from 3 experiments). (G) IL-28A/B measured in sera, SI, and colon from naive and 24 hours postirradiation (1000 cGy) WT mice (n = 9, combined from 2 experiments). SI and colon mucosal homogenates were prepared, and the IL28-A/B amounts in mucosal supernatants corrected for each gram of tissue. (H) IL-28A/B concentration in serum and SI mucosal homogenates as for panel G at days 1, 3, and 7 after lethal irradiation (1000 cGy) and transplantation with BALB/c BM and T cells or TCD only (n = 9, combined from 2 experiments). (I-J) B6D2F1 recipients were transplanted with BM and T cells from WT or Ifnlr1–/– donors. Survival (I) and GVHD clinical scores (J) (GVHD groups, n = 12; TCD, n = 8; combined from 2 experiments). Data are presented as mean ± SEM. P values were calculated by using the 2-tailed Mann-Whitney t test. Kaplan-Meier survival was compared by using the log-rank Mantel-Cox test. *P <.05, **P < .01, ***P <.001, ****P <.0001.
Figure 2.
Figure 2.
Enhanced GVHD in Ifnlr1−/− recipients is dependent on signaling in hematopoietic and nonhematopoietic cells. (A) Transplant schema for creation of BM chimeras and secondary transplants. (B) Representative images of colon at day 7 post-BMT. (C) Semiquantitative colon GVHD histopathology scores at day 7 post-BMT (n = 11 for WT → WT and WT → Ifnlr1–/–, n = 15 for Ifnlr1–/– → WT and n = 14 for Ifnlr1–/– → Ifnlr1–/–, combined from 2 experiments). (D) Serum IFN-γ and IL-6 at day 4 post-BMT. (E) Quantitative polymerase chain reaction enumeration of Ifnlr1 transcripts from naive WT homogenized tissue normalized to expression in liver (n = 3). Data are presented as mean ± SEM. P values were calculated by using analysis of variance and Tukey’s multiple comparison. *P <.05, **P < .01, ***P < .001, ****P < .0001.
Figure 3.
Figure 3.
Ifnlr1-signaling in recipient NK cells is responsible for the protection from GVHD mediated by hematopoietic cells. (A-B) Donor BALB/cLuc T-cell expansion in WT and Ifnlr1–/– recipients determined by bioluminescence 7 days post-BMT. Representative images (A) and quantification (n = 9 per group, combined from 3 experiments) (B). (C) Quantification of donor T cells in spleen at day 4 post-BMT (n = 8 per group, combined from 2 experiments). (D) Quantification of donor T cells in colon at day 4 post-BMT (n = 6, combined from 2 experiments). (E) Proportion of T cells at day 4 post-BMT that are donor vs host (n = 8, combined from 2 experiments). (F-G) Functional capacity of splenic DCs from naive B6 WT or Ifnlr1–/– mice to stimulate BALB/c. CD4+ (F) or CD8+ (G) T cells in a mixed lymphocyte reaction (n = 3; data are from 1 of 2 representative experiments). BALB/c recipients were transplanted with WT or Ifnlr1–/– BM + T cells at day 0 and B6.TeaLuc T cells (reactive to BALB/c I-Ed derived TEa peptide expressed in donor B6 I-Ab) were transferred at day 12 to assess donor-derived APC function in the GI tract, as determined by bioluminescence of antigen-specific TEa T cells. (H-I) Representative bioluminescence (H) and quantification (n = 10, combined from 2 experiments) (I). (J) Proportions of donor T cells producing IFN-γ in spleen at day 4 post-BMT (n = 8, combined from 2 experiments). (K) Dividing capacity of splenic BALB/c T cells transplanted into WT or Ifnlr1–/– recipients calculated at day 4 by carboxyfluorescein diacetate succinimidyl ester dilution (n = 14, combined from 3 experiments). (L) Proportion of AnnexinV+7AAD– apoptotic donor T cells at day 4 as in panel K) (n = 7, combined from 2 experiments). (M) Number of NKp46+ cells in naive recipient mice (n = 8). (N) B6 WT or Ifnlr1–/– recipients received αNK1.1 or IgG isotype and were transplanted with BALB/c BM + T cells. IFN-γ was determined in sera at day 4 post-BMT (n = 9 per group, combined from 2 experiments). B6 CD45.2+ WT or Ifnlr1–/– recipients received αNK1.1 or IgG Isotype and were transplanted with CD45.1+ allogeneic BALB/c BM and CD45.1+CD45.2+ syngeneic B6 BM. (O) Representative fluorescence-activated cell sorting plots from NK-depleted recipients showing syngeneic vs allogeneic cells in spleen 48 hours post-BMT. (P) Index of cytotoxicity as described in "Methods" (n = 8, combined from 2 experiments). (Q) Index of cytotoxicity in spleen of WT and Ifnlr1–/– recipient mice in addition to NKp46Cre.Ifnlr1fl.fl-negative and -positive recipients (n = 12, combined from 2 experiments). (R) Number of NKp46+ NK cells in NKp46Cre.Ifnlr1fl.fl-negative and -positive mice 48 hours after 1000 cGy irradiation. Data are presented as mean ± SEM. P values were calculated by using the 2-tailed Mann-Whitney t test. *P < .05, **P < .01, ***P < .001, ****P < .0001.
Figure 4.
Figure 4.
Ifnlr1-signaling in Lgr5+ ISCs provides nonhematopoietic protection from GVHD. (A) Heat map displaying differentially abundant operational taxonomic units consistently increased or decreased in separately housed or cohoused B6 WT and/or Ifnlr1–/– mice. Cohousing was performed for 4 weeks before transplantation (n = 10 per strain and housing condition, combined from 2 experiments). (B) Principal component analysis of fecal microbial composition for mice as in panel A. (C) Survival of recipients in panel A transplanted with BALB/c BM + T cells. (D) Representative images. (E-F) Numbers (E) and size (F) of GI organoids grown from colonic crypt isolates and enumerated at day 5 (n = 6-7, combined from 3 experiments). (G-H) Representative images (G) and semiquantitative GVHD histopathology scores (H) at day 7 after BMT from tamoxifen-treated Cre-positive or Cre-negative Lgr5Cre.Ifnlr1fl.fl recipient mice (n = 10, combined from 2 experiments). (I) Numbers of GI organoids grown from colonic crypt isolates and enumerated at day 5 from mice as in panels D-E (n = 6, combined from 2 experiments). Data are presented as mean ± SEM. P values were calculated by using the 2-tailed Mann-Whitney t test. Survival calculated by using the log-rank Mantel-Cox test. *P <.05, **P < .01. PCI, first principal component; PC2, second principal component.
Figure 5.
Figure 5.
IFN-λ treatment produces a proliferative phenotype in GI stem cells. PEG-rIL-29 (5 μg) or PBS was given intraperitoneally for 3 days before gut harvest and evaluation of GI epithelial function. (A-C) Representative images (A), numbers (B), and size of GI organoids (C) grown from colonic crypt isolates (n = 9 per group, combined from 3 experiments). (D-E) Numbers of SI organoids (n = 5, combined from 3 experiments) (D) and number of stem cells (CD45.2–/EpCAM+/GFP+) (E) isolated from digested gut of Lgr5-EGFP-IREScreERT2 mice (n = 5, combined from 3 experiments). (F) Number of organoids grown from FACS sorted single stem cells (n = 4, combined from 3 experiments). (G) Number of colonic organoids from WT and IL-22–/–- mice treated with PBS or PEG-rIL-29 (n = 4, combined from 2 experiments). (H) Number of colon and SI organoids from Ifnar–/– mice treated with PBS or PEG-rIL-29 (n = 6, combined from 3 experiments). (I) RNA-seq from sort purified colonic epithelial cells and stem cells derived from either PEG-rIL-29–treated or PBS-treated mice. Heat map showing top 300 genes significantly differentially expressed in colonic epithelial cells (Lgr5–) and stem cells (Lgr5+) after in vivo PEG-rIL-29 vs PBS treatment (2420 genes total). Expression of the same genes from ISCs derived from PBS- or IL-29–treated mice included for comparison (n = 5 per group). (J) Normalized messenger RNA transcript counts for Ifnlr1, Il10rb, Ifnar1, and Ifnar2 in colonic epithelial cells (Lgr5–) and stem cells (Lgr5+). (K) Functional enrichment analysis of differentially expressed genes: canonical pathway enrichment analysis (log2 fold-change >0.58, and adjusted P value <.05) in sorted Lgr5+ and Lgr5– epithelial cells after in vivo PEG-rIL-29 treatment relative to genotype-matched PBS-treated samples using Ingenuity pathway analysis. Enrichment of canonical pathways associated with immune responses (left) and regulation of cellular proliferation (right). Bubbles represent significant pathway enrichment, as determined by Fisher’s exact test. Bubble diameter represents the log10 P value as determined by Fisher’s exact test. Crosses signify a lack of significant pathway enrichment, color indicates predicted pathway activation (red) or predicted inhibition (blue), and white bubbles represent significant functional enrichment of pathways with no available prediction patterns. (L) Venn diagram of overlap of differentially expressed genes in Lgr5+ and Lgr5– cells as for panel K. Data are presented as mean ± SEM. P values were calculated by using the 2-tailed Mann-Whitney t test. For RNA-seq, differentially regulated genes were determined after filtering for genes with >5 cpm and fold change differences >1.2 and corrected P values (false discovery rate) <.05. *P <.05, **P < .01, ***P < .001.
Figure 6.
Figure 6.
Predicted activation state of PEG-rIL-29–treated Lgr5+ and Lgr5– sorted GI epithelial cells. Cytokines (A), transcriptional regulators (B), and kinases (C) with significantly associated transcriptional changes in sorted Lgr5+ and Lgr5– intestinal epithelial cells after in vivo PEG-rIL-29-administration using Ingenuity pathway analysis. Bubble plot representation of significant enrichment scores (activation z score >2) in at least one treatment condition (ie, PEG-rIL-29). Color indicates predicted activation (red) or predicted inhibition (green), and bubble diameter represents the -log10 P value as determined by Fisher’s exact test. Crosses signify a lack of significant activation scores at specific time points.
Figure 7.
Figure 7.
IFN-λ treatment protects from GVHD within the GI tract. PEG-rIL-29 or PBS was given as previously described on days −2, −1, and 0 to BMT recipients. (A) Survival by Kaplan-Meier analysis of B6 recipients transplanted with BALB/c BM + T cells (n = 10, combined from 2 experiments). (B) Survival by Kaplan-Meier analysis of B6D2F1 recipients transplanted with B6 BM + T cells (n = 10, combined from 2 experiments). (C) B6 recipients were transplanted with Balb/c BM + T cells. Serum IFN-γ and IL-6 at day 4 after BMT as in panel A (n = 15, combined from 3 experiments). (D) Representative images of colon and SI at day 7 post-BMT. (E) Semiquantitative GVHD histopathology scores at day 7 post-BMT (n = 10, combined from 2 experiments). (F) Paneth cell numbers. (G) Representative images of proliferation in colon and SI using Ki-67 at day 7 after BMT. (H-I) Quantification of Ki-67 expression at day 7 post-BMT in colon (n = 10, combined from 2 experiments) (H) and in SI (as in panel H) (I). (J) B6 recipients were transplanted with BALB/c BM ± T cells and crypt isolates obtained 7 days later. Colon organoids were quantified (n = 4, combined from 2 replicate experiments). (K-L) Representative images (K) and enumeration (L) of Lgr5+ ISCs in tissue sections at day 7 post-BMT from ileum from PBS or PEG-rIL-29 recipients of BALB/c BM + T cells after NK-cell depletion with NK1.1 on day −1 (1 mg), day +3 (0.5 mg), and day +6 (0.5 mg), (n = 9, combined from 2 experiments). (M) B6D2F1 recipients were treated with PBS or PEG-rIL-29, then transplanted with BM ± T cells from B6.WT donors, together with recipient type BCR-ABL nup98hoxA9 leukemia expressing GFP. (M) The number of GFP+ leukemia cells was determined in peripheral blood thereafter (n = 10, combined from 2 experiments). (N) Death from leukemia in recipients transplanted as in panel M. Data are presented as mean ± SEM. P values were calculated by using the 2-tailed Mann-Whitney t test. Survival was calculated by using the log-rank Mantel-Cox test. *P < .05, **P < .01, ***P < .001, ****P < .0001. DAPI, 4′,6-diamidino-2-phenylindole; TCD, T cell–depleted.
See this image and copyright information in PMC

"VSports手机版" Comment in

References

    1. Jenq RR, Ubeda C, Taur Y, et al. . Regulation of intestinal inflammation by microbiota following allogeneic bone marrow transplantation. J Exp Med. 2012;209(5):903-911. - PMC (V体育平台登录) - PubMed
    1. Zeiser R, Socié G, Blazar BR.. Pathogenesis of acute graft-versus-host disease: from intestinal microbiota alterations to donor T cell activation. Br J Haematol. 2016;175(2):191-207. - "VSports app下载" PubMed
    1. Simms-Waldrip TR, Sunkersett G, Coughlin LA, et al. . Antibiotic-induced depletion of anti-inflammatory Clostridia is associated with the development of graft-versus-host disease in pediatric stem cell transplantation patients. Biol Blood Marrow Transplant. 2017;23(5):820-829. - PubMed
    1. Koyama M, Mukhopadhyay P, Schuster IS, et al. . MHC Class II antigen presentation by the intestinal epithelium initiates graft-versus-host disease and is influenced by the microbiota. Immunity. 2019;51(5):885-898.e7. - PMC - PubMed
    1. Burman AC, Banovic T, Kuns RD, et al. . IFNgamma differentially controls the development of idiopathic pneumonia syndrome and GVHD of the gastrointestinal tract. Blood. 2007;110(3):1064-1072. - PubMed

Publication types

VSports app下载 - MeSH terms