HomeScienceDendritic cell-derived IL-27 p28 regulates T cell program in pathogenicity and alleviates...

Dendritic cell-derived IL-27 p28 regulates T cell program in pathogenicity and alleviates acute graft-versus-host disease

[ad_1]

IL-27 p28 deficiency aggravates aGVHD in mice

To address the physiological role of IL-27 p28 in aGVHD pathogenesis, we first assessed the cellular source of IL-27 p28 during aGVHD. Results showed that DCs, rather than T cells, neutrophils, or monocytes, were the predominant source of IL-27 p28 in both murine aGVHD models and patients at the onset of aGVHD (Supplementary Fig. 1a, b). We therefore generated IL-27 p28 conditional knockout (p28 cKO) mice in DCs by crossing p28f/f mice with CD11c Cre mice.35 IL-27 p28 expression in the serum was markedly reduced in CD11c-p28f/f mice compared with WT mice at a steady-state or on day 7 after allogenic-bone marrow transplantation (BMT) (Supplementary Fig. 1c, d). Moreover, the levels of IL-27 heterodimer were also reduced in CD11c-p28f/f mice compared with WT mice (Supplementary Fig. 1e, f). In vitro mixed lymphocyte reaction (MLR) revealed that T cells from p28 cKO mice displayed aggravated alloreactivity than those from WT mice (Fig. 1a), as evidenced by enhanced cell proliferation (Fig. 1b, c). We then established an MHC-mismatched murine aGVHD model using CD11c-p28f/f or WT mice as donors. Compared with WT recipients, mice that received CD11c-p28f/f splenocytes showed accelerated aGVHD mortality (Fig. 1d). Histological analysis also revealed that recipients of CD11c-p28f/f splenocytes showed much more severe tissue damage (Fig. 1e, f). However, the mortality showed no significant difference when CD11c-p28f/f mice were used as recipients (Fig. 1g). Collectively, these results demonstrate that donor-derived IL-27 p28 protects from aGVHD development in mice.

Fig. 1
figure 1

IL-27 p28 deficiency aggravates aGVHD in mice. ac BALB/c DCs were cocultured with CFSE-labeled T cells (ratio 1:10) from CD11c-p28f/f mice or control littermates, respectively. Proliferations were assessed by a 3H-TdR or b, c flow cytometry 5 days post coculture. df BALB/c recipients were transplanted with 1 × 107 WT BMs and 5 × 106 splenocytes from either WT or CD11c-p28f/f mice (n = 10–14 per group). Overall survival curve is depicted (d). Representative H&E stained sections and histological scores of aGVHD tissues from recipients 14 days post-transplantation are shown (e, f). g C57BL/6 or CD11c-p28f/f recipients were lethal irradiation and received 1 × 107 BMs and 7.5 × 107 splenocytes from BALB/c mice (n = 15 per group). The overall survival curve is depicted. Data are representative of three independent experiments and presented as mean ± SD. *P < 0.05; **P < 0.01; ***P < 0.001

Loss of IL-27 p28 enhances T cell responses after allo-HSCT

To investigate the mechanisms by which IL-27 p28 alleviates aGVHD, we profiled the immune cell responses in aGVHD target tissues post transplantation. Donor T cells were substantially elevated in aGVHD target tissues from recipients that received CD11c-p28f/f grafts compared with those received WT grafts (Supplementary Fig. 2a). Notably, T cells from recipients transplanted with CD11c-p28f/f splenocytes showed more activated phenotypes, indicated by increased CD69 expression compared with those from WT recipients (Fig. 2a, b and Supplementary Fig. 2b). Furthermore, the frequencies of IFN-γ- and TNF-α-production in donor T cells were significantly upregulated in recipients of CD11c-p28f/f grafts compared with WT controls (Fig. 2c–f and Supplementary Fig. 2c). However, the IL-17A and IL-4 expression had no changes in aGVHD target tissues (Supplementary Fig. 2d). In addition, productions of IL-6, IFN-γ, and TNF-α were significantly upregulated, whereas IL-10 was downregulated in recipients of CD11c-p28f/f grafts compared with those that received WT grafts (Fig. 2g). Furthermore, recipients of CD11c-p28f/f grafts exhibited enhanced donor T cell alloreactivity in vivo (Fig. 2h). These findings demonstrate that loss of IL-27 p28 leads to enhanced alloreactive T cell responses which accelerates the aGVHD-related mortality.

Fig. 2
figure 2

Loss of IL-27 p28 enhances T cell responses after allo-HSCT. a, b BALB/c recipients were transplanted with 1 × 107 WT BMs together with 5 × 106 splenocytes from either C57BL/6 or CD11c-p28f/f mice. Immune cell subsets were examined 14 days post-transplantation. Representative flow cytometric plots and quantification of activated T cells in spleens (among H2-Kb+H2-Kd cells) from recipients (n = 6 per group) are depicted. cf Representative flow cytometric plots and quantification of IFN-γ-producing T cells (c, d) and TNF-α-producing T cells (e, f) in spleens (among H2-Kb+H2-Kd cells) from recipients (n = 6 per group) are depicted. g Serum from BALB/c recipients was collected 14 days post-transplantation and cytokines production was examined using LEGENDplex (n = 5–6 per group). h Splenocytes from aGVHD recipients were cocultured with irradiated BALB/c splenocytes. Proliferation rate was detected by 3H-TdR incorporation assay 3 days after coculture. i, j Treg cells were detected 14 days post-transplantation via FACS. Quantitative data (i) and representative plots (j) of donor Tregs are shown (n = 6 per group). k, l CFSE-labeled effector T cells (CD4+CD25 T cells) were cocultured with Treg cells sorted from the spleens of C57BL/6 or CD11c-p28f/f mice at the indicated ratios for 5 days. Representative figures (k) and frequency of cell proliferation are depicted (l) (n = 4 per group). m The frequency of IL-10+ Treg cells is shown (n = 4 per group). Data are representative of three independent experiments and presented as mean ± SD. *P < 0.05; **P < 0.01; ***P < 0.001

Tregs suppress conventional T cell activation and control GVHD.36 In our models, we observed that Tregs were substantially decreased in aGVHD target tissues of recipients that received CD11c-p28f/f grafts compared with those received WT grafts (Fig. 2i, j). However, CD11c-p28f/f Treg cells exhibited a similar apoptosis rate to that of WT recipients (Supplementary Fig. 2e), indicating that IL-27 p28 deficiency did not affect Treg cell survival in vivo. In contrast, Treg cells isolated from recipients of CD11c-p28f/f grafts displayed impaired suppressive effects toward CD4+ Teff cells compared to those from WT recipients (Fig. 2k, l). IL-27 has been identified as a differentiation factor that contributes to Tr1 cell generation.16,19,37 We found that IL-10 levels were significantly downregulated in the serum of recipients of CD11c-p28f/f grafts (Fig. 2g). However, no significant changes of Tr1 populations were observed in aGVHD target tissues between these two groups (Supplementary Fig. 2f), suggesting that the reduced IL-10 levels may be due to the decreased Treg cells. Indeed, IL-10 producing-Treg cells were significantly reduced in recipients of CD11c-p28f/f grafts (Fig. 2m). Together, these results demonstrate that the exacerbation of aGVHD mediated by IL-27 p28 deficiency associates with defective Treg cell function, which fails to suppress the alloreactive T cell responses.

Donor DC-derived IL-27 p28 deficiency and intrinsic functional defects of T cells are both responsible for exacerbated aGVHD

Based on our results, three hypotheses may explain the enhanced aGVHD by IL-27 p28 deficiency in CD11c positive cells. First, donor CD11c+ DC-derived IL-27 p28 directly affects allogenic-T cell activity during aGVHD. Second, splenic T cells from p28 cKO mice may have unleashed effector functions before transferring to aGVHD receipt. Third, a recent study reported that a certain subset of T cells also express CD11c.23 This minor population of T cells acquired IL-27 p28 deficiency may affect aGVHD development. To clarify whether donor T cells or DCs contribute to the effect of IL-27 p28, we transplanted recipients with T cell-depleted (TCD)-BMs and purified T cells from either WT or CD11c-p28f/f mice. As shown in Fig. 3a, recipients received TCD-BM from WT mice together with T cells from CD11c-p28f/f mice (hereinafter referred to as WK) had significantly shortened survival compared with those received TCD-BM and T cells both from WT mice (hereinafter referred to as WW). These results were consistent with our model in which BM and splenocytes were used as grafts, suggesting that T cells from p28 cKO mice are more pathogenic than those from WT mice, which contribute to the exacerbated aGVHD. Recipients of TCD-BM from CD11c-p28f/f mice and T cells from WT mice (hereinafter referred to as KW) showed a trend of shortened survival compared with the WW group (P = 0.0514), suggesting that donor DC-derived IL-27 p28 deficiency also contributes to the exacerbated aGVHD, albeit to a lesser extent (Fig. 3a). To mechanistically interrogate the role of DC-derived IL-27 p28 in aGVHD, we co-transferred bone marrow-derived DCs (BMDCs) from either WT or CD11c-p28f/f mice into WW recipients. We observed that co-transfer of BMDCs from CD11c-p28f/f mice exacerbated aGVHD, compared with those from WT mice, although the difference was not statistically significant (P = 0.082, Fig. 3b). These results indicate that donor DC-derived IL-27 p28 indeed contributes to the aGVHD development. To further clarify whether the aggravated aGVHD are due to pre-activation and distribution of T cell cargo before transfer or an altered differentiation upon transplantation, we compared the effects of donor-derived naive T cells in aGVHD induction. Interestingly, naive T cells from CD11c-p28f/f mice significantly enhanced aGVHD development than those from WT mice (Fig. 3c), indicating that the intrinsic functional defects occur before T cells are activated. We also observed an upregulation of co-stimulatory molecule CD80 expression on DCs (Fig. 3d). In addition, recipients transplanted with TCD-BM and T cells both from CD11c-p28f/f mice had the shortest survival compared with WW (P = 0.004) and KW groups (P = 0.04), and showed a trend of shorter survival than WK group (P = 0.1698). These results indicate that DC-derived IL-27 p28 and T cells of p28 cKO donors are both responsible for aGVHD pathogenesis.

Fig. 3
figure 3

Donor DC-derived IL-27 p28 deficiency and intrinsic functional defects of T cells are both responsible for exacerbated aGVHD. a BALB/c recipients received either WT or CD11c-p28f/f allografts of 5 × 106 TCD-BMs and 1 × 106 T cells as indicated (n = 10 per group). b WW recipients were injected with either 1 × 106 WT DCs or CD11c-p28f/f DCs at day 0, day1, and day2 post-BMT (n = 10 per group). The overall survival curve is depicted. c BALB/c recipients received 5 × 106 TCD-BMs from WT mice together with 1 × 106 naive T cells from WT or CD11c-p28f/f mice, respectively (n = 10 per group). di Splenocytes from normal donors were detected by flow cytometry. d CD80, CD86, and MHC-II expression on DCs. e Percentages of lymphocytes, CD3+ T, Tregs, and DCs. f The ratio of T cells in the spleen. g Percentages of T cell suesets. h CD69 expression on T cells. i IFN-γ, IL-17A, and IL-10 expression in T cells (n = 4 per group). Data are representative of three independent experiments and presented as mean ± SD. *P < 0.05; **P < 0.01; ***P < 0.001

We found that T cells at a steady-state did not express CD11c (<0.5%) but had very low levels in vitro (about 2% among total T cells) with allo-stimulation or in vivo (about 1% among total T cells) at 7 days after allo-BMT in both WT and CD11c-p28f/f mice (Supplementary Fig. 3a-c). Both CD11c+ and CD11c T cells did not secrete any detectable IL-27 p28 (data not shown). In addition, the IFN-γ production by CD11c+ and CD11c T cells were comparable between WT and CD11c-p28f/f mice under allo-stimulation (Supplementary Fig. 3d, e). Thus, CD11c+ T cell-deficient of IL-27 p28 unlikely contributes to aGVHD development. Of note, although WT and p28 cKO mice exhibited the equivalent levels of T cells, Treg cells, DCs, and CD4/CD8 ratio in the spleen at a steady-state (Fig. 3e, f), T cells from p28 cKO donors showed a higher activation phenotype, as evidenced by increased effector memory T cell populations (Fig. 3g). However, CD69 expression showed no difference among these T cells (Fig. 3h). Furthermore, increased proportions of IL-17A and IFN-γ producing T cells were observed in p28 cKO mice (Fig. 3i), indicating that T cells from p28 cKO mice are intrinsically different from those T cells from WT mice. DC-derived IL-27 p28 may mediate aGVHD development by affecting T cell pathogenicity during allo-HSCT.

IL-27 p28 deficiency impairs Treg cell generation and promotes pathogenic IL-1R2+TIGIT+ CD4+T cells in the thymus

Abnormalities in thymic development at least partially limites T cell maturation and functions.38 Our results are in agreement with prior studies in which DC-derived IL-27 p28 regulated T cell development in the thymus.35,39 However, the mechanism by which IL-27 p28 regulates thymic T cell differentiation remains undetermined. To address this, we applied single-cell RNA sequencing (scRNA-seq) to thymocytes collected from WT and CD11c-p28f/f mice, which allowed highly detailed profiling of immune cell subsets during development at the single-cell level. After quality control and filtering out poor-quality cells, a whole-transcriptome database of 17922 cells from the two groups was analyzed. We identified 9 clusters by t-Distributed Stochastic Neighbor Embedding (t-SNE), including T cells, B cells, DCs, macrophages, and fibroblasts (Supplementary Fig. 4a-c). T cells with specific expression of CD3d, CD4, or CD8a were distinguished from other clusters and were reclustered for further analysis (Fig. 4a). Violin plots revealed that cluster 5 represented double-negative (DN) T cells; clusters 0, 1, and 2 represented double-positive (DP) T cells; and clusters 3 and 4 represented CD8 and CD4 single-positive (SP) T cells, respectively (Fig. 4b). We observed a reduction in DP cells and elevation in CD4 and CD8 SP cells in IL-27 p28-deficient mice compared to WT mice (Fig. 4c). Flow cytometry confirmed that CD11c-p28f/f mice had reduced DP and increased CD4 and CD8 SP T cells compared with WT mice (Fig. 4d). We further divided CD4 SP T cells into three subclusters (Fig. 4e). Principal component analysis (PCA) showed a difference among these three clusters (Supplementary Fig. 5a). Further analysis showed that cluster 0 and cluster 1 represented conventional T cells, which was indicated by low expressions of Foxp3. Cluster 2 represented Treg cells, which was designated by high levels of Foxp3 and IL2ra expression (Supplementary Fig. 5b). CD4_cluster 0 and 2 were decreased dramatically in IL-27 p28-deficient mice (Fig. 4f), indicating that IL-27 p28 deficiency impairs Treg cell generation during thymic development. The expression levels of genes involved in Treg cell development, including Sell, Ccr7, Tnfrsf13b, Bcl11b, Stab1, Id3, Lef1, and Bach2 were pronouncedly reduced in Tregs from CD11c-p28f/f mice compared to those from WT mice (Fig. 4g).40,41,42,43,44,45,46,47 Subsequently, we found that the percentage of CD4_cluster 1 was 3% in WT mice while markedly increased to 40% in CD11c-p28f/f mice. Surprisingly, this T cell cluster was hyperactivated, with highly expressed activation marker genes, such as Tnfrsf9, Tight, Lag3, Pdcd1, Batf, Maf, Bhlhe40, Tnfrsf18, and Tnfrsf4, and downregulation of the naive gene Il7r.48 In addition, Il1r2 and Tigit were highly and specifically expressed in Cluster 1 (Fig. 4h, i). We therefore named cluster 1 cells as IL-1R2+TIGIT+ activated CD4+T cells. Flow cytometry data confirmed that IL-1R2+TIGIT+CD4+T subset was markedly increased in both thymus and spleen in CD11c-p28f/f donors than WT mice (Supplementary Fig. 6a, b). Enhanced Bhlhe40 expression in CD4+ T cells has been demonstrated to promote aGVHD pathogenesis.49 Other T cell effector genes, including Nr4a1 and Btla were increased, whereas Ccr7, Ccr9, and Klf2 were decreased (Supplementary Fig. 7a), suggesting that IL-27 p28 deficiency promotes pro-inflammatory and pathogenic fates of CD4+ T cells in the thymus. Pathway analysis performed by gene-set enrichment analysis (GSEA) comparing CD4 SP Cluster 1 with Cluster 0 revealed that the expression patterns of genes known to involve in cytokine production and cytokine-related responses were increased with IL-27 p28 deficiency (Fig. 4j). Signals include response to cytokine, cytokine binding, and cytokine receptor activity were enriched in IL-27 p28-deficient CD4 SP cells compared to WT CD4 SP cells (Supplementary Fig. 8). We further assessed the proportions of IL-1R2+TIGIT+CD4+ T cells in murine aGVHD models and clinical patients with or without aGVHD. Consistent with scRNA-seq, donor-derived IL-1R2+TIGIT+CD4+T were significantly increased in recipients of CD11c-p28f/f grafts compared with those of WT controls (Fig. 4k, l). Moreover, this pathogenic CD4+T subset was also increased in grade II-IV severe aGVHD patients compared with non-aGVHD patients after allo-HSCT (Fig. 4m, n). Our results therefore suggest that T cells are indeed defective in their steady-state in CD11c-p28f/f mice, as demonstrated by impaired Treg cell generation and increased IL-1R2+TIGIT+ pathogenic CD4+ T cells in the thymus, which are the main driving force of aGVHD development.

Fig. 4
figure 4

IL-27 p28 deficiency impairs Treg cell generation and promotes pathogenic IL-1R2+TIGIT+ CD4+T cells in the thymus. a Reclustering of T cell subpopulations in WT and CD11c-p28f/f mice. b Violin plots of the relative expressions of CD3D, CD4, and CD8. c Percent of different thymic T cell subsets determined by scRNA-seq analysis. d Populations and numbers of DP T cells in the thymus from WT and CD11c-p28f/f mice by flow cytometry. e t-SNE visualization of CD4 SP T cells. f Components of subclusters in CD4 SP cells. g Expression profile of genes involved in Treg development. h Volcano plot showing differential gene expression between Cluster 1 and Cluster 0 cells from CD11c-p28f/f and WT mice. i Violin plots showing the expression profile of T cell effector genes. Expression is measured as the log2-fold change. j GSEA of the upregulated gene set in Cluster 1 versus Cluster 0 in CD4 SP cells from CD11c-p28f/f relative to WT mice. k, l Populations of donor-derived IL-1R2+TIGIT+CD4+T cells were detected by FACS 7 days post-transplantation (n = 9–10 per group). m, n Percentages of IL-1R2+TIGIT+CD4+T cells in PBMCs were detected by FACS 30 days post-allo-HSCT. *P < 0.05; **P < 0.01; ***P < 0.001

IL-27 p28 deficiency restrains Treg cell differentiation via the IFN-γ/STAT1 signaling pathway

To further investigate the mechanism by which IL-27 p28 deficiency regulates Treg differentiation, we performed an in vitro Treg cell differentiation assay. We found that Treg cell differentiation was significantly impaired in CD11c-p28f/f mice (Fig. 5a). However, blocking p28 signaling using a p28 neutralizing antibody did not show similar effect (Fig. 5b). To further investigate whether IL-27 p28 regulates Treg cell differentiation in a cell-intrinsic manner, recipients were injected with TCD-BM cells from CD45.1 mice, along with an equal number of T cells from CD11c-p28f/f or CD45.1 congenic mice. Treg cell reconstitution was evaluated in vivo 14 days post transfer. The results showed that Treg cell reconstitution from IL-27 p28-deficient mice was significantly reduced compared to their CD45.1 counterparts (Fig. 5c), indicating a cell-intrinsic impairment of Treg cell development due to IL-27 p28 deficiency. To further evaluate the functions of Treg cells from WT or CD11c-p28f/f mice in controlling of aGVHD, we performed a Treg cell rescue experiment by co-transferring an equal number of Treg cells from WT or CD11c-p28f/f mice, respectively. The survival curve showed that co-transfer of Treg cells from WT mice significantly mitigated aGVHD, while co-transfer of Treg cells from CD11c-p28f/f mice had no protective effect against aGVHD (Fig. 5d). Moreover, Treg cells from CD11c-p28f/f mice also failed to control aGVHD induced by WT T cells, whilst WT Treg cells significantly prolonged the survival of the recipients (Fig. 5e). Therefore, these results suggest that IL-27 p28 deficiency dampened Treg cell functions during aGVHD development.

Fig. 5
figure 5

IL-27 p28 deficiency restrains Treg cell differentiation via the IFN-γ/STAT1 signaling pathway. a, b Splenocytes from WT or CD11c-p28f/f mice were induced for Treg polarization in the presence or absence of IL-27 p28 antibody (10 μg/ml). Representative figures and summary data of the frequency of Tregs are depicted. c BALB/c recipients were injected with 1 × 107 TCD-BM cells from CD45.1 mice, along with an equal number of T cells (2 × 106) from CD11c-p28f/f mice and CD45.1 congenic mice. Treg cell reconstitution was evaluated in vivo 14 days post transfer. d BALB/c recipients were transplanted with 5 × 106 TCD-BM cells plus 1 × 106 conventional T cells (Tconv) either from WT or CD11c-p28f/f mice and transferred with 7.5 × 105 Treg cells from WT or CD11c-p28f/f mice respectively. The overall survival curve is depicted (n = 10 per group). e BALB/c recipients were transplanted with 5 × 106 TCD-BM cells plus 1 × 106 Tconv cells from WT donors and transferred with or without 7.5 × 105 Tregs from WT or CD11c-p28f/f mice (n = 10 per group). fh Anti-IFN-γ, anti-IL-6R, or anti-IL-12 were added during Treg polarization at a concentration of 10 μg/ml. Percentage of Treg cells was detected by FACS (n = 3 per group). i IFN-γ production in the supernatant of polarized Tregs was detected by ELISA (n = 4 per group). j The expression of phosphorylated STAT1 among Treg populations is shown (n = 5 per group). k STAT1 inhibitor (10 μM) was added during Treg polarization. Percentages of Treg cells were detected by FACS (n = 3 per group). l, m BALB/c recipients were transplanted with 1 × 107 WT BMs together with 5 × 106 splenocytes from either WT or CD11c-p28f/f mice. Recipients were injected with αIFN-γ (250 μg per mouse) after BMT. Percentages of Tregs were detected 2 weeks after BMT (n = 4 per group). The overall survival curve is depicted (n = 10 per group). Data are representative of three independent experiments and presented as mean ± SD. *P < 0.05; **P < 0.01; ***P < 0.001

IFN-γ, IL-6, and IL-12 are cytokines that can inhibit Treg cell differentiation. We demonstrated that blocking IFN-γ, but not IL-6 or IL-12 could restore the inhibition of Treg cell differentiation caused by IL-27 p28 deficiency (Fig. 5f–h). We also observed a substantially elevated IFN-γ production during Treg cell differentiation after IL-27 p28 deficiency (Fig. 5i). However, IL-27 p28 expression was undetectable in the supernatants during Treg cell differentiation in both WT and CD11c-p28f/f mice (Data not shown). These results suggest that T cell-intrinsic IFN-γ signal is responsible for the impaired Treg cell differentiation. STAT1 is a key mediator of IFN-γ signaling that regulates the differentiation of Treg cells.50 We observed that phosphorylation of STAT1 was largely increased in Treg cells differentiated from CD11c-p28f/f mice compared with those from WT mice (Fig. 5j). In addition, STAT1 inhibitor added in the culture medium significantly reversed the IL-27 p28-mediated inhibition of Treg cell differentiation (Fig. 5k). Furthermore, blocking IFN-γ signaling in vivo increased Treg cell numbers in the spleen and liver and ameliorated aGVHD in mice receiving CD11c-p28f/f donor cells (Fig. 5l, m), suggesting that IL-27 p28 deficiency inhibits Treg differentiation and function in an IFN-γ-dependent manner.

IL-27 p28 is a valuable marker for predicting aGVHD after allo-HSCT in humans

Although the effect of IL-27 p28 in murine aGVHD has been reported, the clinical significance of IL-27 p28 in aGVHD patients remains undetermined. Hence, we detected IL-27 p28 expression in the serum of 67 patients who underwent allo-HSCT and found that patients with severe aGVHD (Grade II-IV, n = 27) displayed significantly decreased IL-27 p28 levels compared with patients with no/low-grade aGVHD (Grade 0-I, n = 40, P = 0.0015, Fig. 6a). When using 32.82 ng/ml as the cutoff value, IL-27 p28 levels could predict the occurrence of severe aGVHD based on ROC analysis (Fig. 6b). The sensitivity and specificity were 77.5% and 63%, respectively. Patients with high levels of IL-27 p28 had a significantly lower incidence of severe aGVHD but showed a similar OS rate compared with patients with low levels of IL-27 p28 (Fig. 6c, d). Consistently, univariate analyses showed that high levels of IL-27 p28 were significantly associated with a low incidence of severe aGVHD but not OS (Fig. 6e, f). High IL-27 p28 and disease status were independent factors for predicting severe aGVHD (HR = 5.88, 95% CI 1.606-21.525, P = 0.007, and HR = 4.363, 95% CI 1.059-17.984, P = 0.041, Fig. 6g) but not OS (Fig. 6h). IL-27 p28 showed similar levels among different primary diseases or cGVHD development (Supplementary Fig. 9a, b). Similar results were observed by evaluating IL-27 heterodimer in predicting aGVHD (Supplementary Fig. 10). Thus, IL-27 p28 or IL-27 may serve as useful predictors of severe aGVHD. In addition, we found that IL-27 p28 was positively correlated with IL-10 (r = 0.271, P = 0.026), whereas negatively correlated with IFN-γ (r = −0.283, P = 0.02; Fig. 6i, j), indicating that serum levels of IL-27 p28 are associated with Treg/Teff cell balance in humans.

Fig. 6
figure 6

IL-27 p28 is a valuable marker for predicting aGVHD after allo-HSCT in humans. a IL-27 p28 production in patients after allo-HSCT were examined by ELISA. b ROC curve was constructed to predict severe aGVHD occurrence. c, d The cumulative incidence of severe aGVHD (c) and overall survival (d) between patients with high and low IL-27 p28 levels are shown. e, f Univariate analyses of factors associated with severe aGVHD occurrence (e) or overall survival (f) after allo-HSCT. g, h Multivariate analyses of factors associated with severe aGVHD occurrence (g) or overall survival (h) after allo-HSCT. i, j The associations between the expression of IL-27 p28 and IL-10 (i), and IFN-γ (j) were analyzed. Data are presented as mean ± SD. **P < 0.01


[ad_2]

Source link

RELATED ARTICLES

LEAVE A REPLY

Please enter your comment!
Please enter your name here

Most Popular

Recent Comments

%d bloggers like this: