| 1. |
- Andersson, Jennie, 1978, et al.
(författare)
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Eosinophils from hematopoietic stem cell recipients suppress allogeneic T cell proliferation.
- 2014
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Ingår i: Biology of blood and marrow transplantation : journal of the American Society for Blood and Marrow Transplantation. - : Elsevier BV. - 1523-6536. ; 20:12, s. 1891-8
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Tidskriftsartikel (refereegranskat)abstract
- Eosinophilia has been associated with less severe graft-versus-host disease (GVHD), but the underlying mechanism is unknown. We hypothesized that eosinophils diminish allogeneic T cell activation in patients with chronic GVHD. The capacity of eosinophils derived from healthy subjects and hematopoietic stem cell (HSC) transplant recipients, with or without chronic GVHD, to reduce allogeneic T cell proliferation was evaluated using a mixed leukocyte reaction. Eosinophil-mediated inhibition of proliferation was observed for the eosinophils of both healthy subjects and patients who underwent HSC transplantation. Eosinophils from patients with and without chronic GVHD were equally suppressive. Healthy eosinophils required cell-to-cell contact for their suppressive capacity, which was directed against CD4(+) T cells and CD8(+) T cells. Neither eosinophilic cationic protein, eosinophil-derived neurotoxin, indoleamine 2,3-dioxygenase, or increased numbers of regulatory T cells could account for the suppressive effect of healthy eosinophils. Real-time quantitative PCR analysis revealed significantly increased mRNA levels of the immunoregulatory protein galectin-10 in the eosinophils of both chronic GVHD patients and patients without GVHD, as compared with those from healthy subjects. The upregulation of galectin-10 expression in eosinophils from patients suggests a stimulatory effect of HSC transplantation in itself on eosinophilic galectin-10 expression, regardless of chronic GVHD status. To conclude, eosinophils from HSC transplant recipients and healthy subjects have a T cell suppressive capacity.
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| 2. |
- Lingblom, Christine, 1984, et al.
(författare)
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Eosinophils from eosinophilic oesophagitis patients have T cell suppressive capacity and express FOXP3.
- 2017
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Ingår i: Clinical and Experimental Immunology. - : Oxford University Press (OUP). - 1365-2249 .- 0009-9104. ; 187:3, s. 455-465
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Tidskriftsartikel (refereegranskat)abstract
- Eosinophilic esophagitis (EoE) is an antigen-driven T cell-mediated chronic inflammatory disease where food and environmental antigens are thought to have a role. Human eosinophils express the immunoregulatory protein galectin-10 and have T cell suppressive capacity similar to regulatory T cells (Tregs ). We hypothesized that one function of eosinophils in EoE might be to regulate the T cell-driven inflammation in the oesophagus. This was tested by evaluating the suppressive capacity of eosinophils isolated from the blood of adult EoE patients in a mixed lymphocyte reaction. In addition, eosinophilic expression of forkhead box protein 3 (FOXP3), the canonical transcription factor of Tregs , was determined by conventional and imaging flow cytometry, quantitative polymerase chain reaction (qPCR), confocal microscopy and immunoblotting. It was found that blood eosinophils from EoE patients had T cell suppressive capacity, and that a fraction of the eosinophils expressed FOXP3. A comparison of EoE eosinophils with healthy control eosinophils indicated that the patients' eosinophils had inferior suppressive capacity. Furthermore, a higher percentage of the EoE eosinophils expressed FOXP3 protein compared with the healthy eosinophils, and they also had higher FOXP3 protein and mRNA levels. FOXP3 was found in the cytosol and nucleus of the eosinophils from both the patients and healthy individuals, contrasting with the strict nuclear localization of FOXP3 in Tregs . To conclude, these findings suggest that the immunoregulatory function of eosinophils may be impaired in EoE.
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| 3. |
- Danielsson Norén, Kristina, et al.
(författare)
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15-Lipoxygenase-2 expression in human macrophages induces chemokine secretion and T cell migration.
- 2008
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Ingår i: Atherosclerosis. - : Elsevier BV. - 1879-1484 .- 0021-9150. ; 199:1, s. 34-40
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Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND: We determined previously that hypoxia results in increased 15-lipoxygenase type 2 (15-LOX-2) expression and CXCL8 secretion in macrophages. This study sought to determine whether 15-LOX-2 expression links directly with the secretion of inflammatory molecules in macrophages and also investigated its subsequent effects on T cell migration. METHODS: Adenovirus-mediated gene delivery caused overexpression of 15-LOX-2 in human macrophages. We used cytometric bead array to measure chemokine secretion, and assessed T cell migration by counting cells in chemotaxis chambers. Expression of chemokine receptors was determined by FACS analysis. Using siRNA, we reduced 15-LOX-2 expression in human macrophages. We used scrambled siRNA as control. RESULTS: Macrophages that overexpress 15-LOX-2 showed increased secretion of chemokine CXCL10 after 24h incubation. In addition, preconditioned medium from 15-LOX-2-overexpressing cells increased T cell migration and surface expression of CXCR3, the CXCL10 receptor. Knockdown of 15-LOX-2 expression decreased CXCL10 secretion from hypoxic macrophages and also reduced T cell migration. CONCLUSION: In macrophages, overexpression of 15-LOX-2 results in increased secretion of CXCL10 and CCL2. Products released in response to increased 15-LOX-2 activation lead to increased expression of CD69, the T cell activation marker as well as increased T cell migration. Therefore, increased expression of 15-LOX-2 induced by hypoxia may participate in T cell recruitment in diseases such as atherosclerosis.
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| 4. |
- Ebefors, Kerstin, 1977, et al.
(författare)
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Role of glomerular proteoglycans in IgA nephropathy
- 2011
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Ingår i: PLoS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 6:4
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Tidskriftsartikel (refereegranskat)abstract
- Mesangial matrix expansion is a prominent feature of the most common form of glomerulonephritis, IgA nephropathy (IgAN). To find molecular markers and improve the understanding of the disease, the gene and protein expression of proteoglycans were investigated in biopsies from IgAN patients and correlated to clinical and morphological data. We collected and microdissected renal biopsies from IgAN patients (n = 19) and from healthy kidney donors (n = 14). Patients were followed for an average time of 4 years and blood pressure was according to target guidelines. Distinct patterns of gene expression were seen in glomerular and tubulo-interstitial cells. Three of the proteoglycans investigated were found to be of special interest and upregulated in glomeruli: perlecan, decorin and biglycan. Perlecan gene expression negatively correlated to albumin excretion and progress of the disease. Abundant decorin protein expression was found in sclerotic glomeruli, but not in unaffected glomeruli from IgAN patients or in controls. Transforming growth factor beta (TGF-beta), known to interact with perlecan, decorin and biglycan, were upregulated both on gene and protein level in the glomeruli. This study provides further insight into the molecular mechanisms involved in mesangial matrix expansion in IgAN. We conclude that perlecan is a possible prognostic marker for patients with IgAN. In addition, the up-regulation of biglycan and decorin, as well as TGF-beta itself, indicate that regulation of TGF-beta, and other profibrotic markers plays a role in IgAN pathology.
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| 5. |
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| 6. |
- Ingelsten, Madeleine, 1978, et al.
(författare)
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Is Indoleamine 2,3-Dioxygenase Important for Graft Acceptance in Highly Sensitized Patients After Combined Auxiliary Liver-Kidney Transplantation?
- 2009
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Ingår i: Transplantation. - 0041-1337. ; 88:7, s. 911-919
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Tidskriftsartikel (refereegranskat)abstract
- Background. In the clinical setting, transplanted liver seems to protect other grafts from the same donor from rejection. Our previous findings suggest that an auxiliary liver transplantation a few hours before a renal transplantation not only inhibits hyperacute antibody-mediated rejection but also improves long-term kidney graft survival in sensitized recipients. Here, we investigated indoleamine 2,3-dioxygenase (IDO) activity, as one potential mechanism for liver-induced long-term acceptance of kidney grafts. Methods. Tryptophan degradation was measured to estimate IDO activity in patient sera and cell culture supernatants with high performance liquid chromatography. Gene expression in the grafted organs and cell lysates was studied using real time polymerase chain reaction analysis. Results. Tryptophan degradation increased in peripheral blood from patients undergoing combined auxiliary liver-kidney transplantation, whereas it decreased in patients after regular renal transplantation. A 100-fold increase in IDO mRNA, preceded by upregulation of the IDO-inducing cytokines tumor necrosis factor-[alpha], interleukin-1[beta], and interferon-[gamma], was observed in the transplanted organs after graft reperfusion in patients undergoing combined graft transplantation. Subsequent studies in vitro revealed that immature dendritic cells, but not hepatocytes, strongly activated IDO on maturation with tumor necrosis factor-[alpha], interleukin-1[beta], and interferon-[gamma]. Finally, serum from liver-transplanted patients elicited an even stronger IDO-activity in such cytokine-stimulated dendritic cells. Conclusions. Taken together these findings suggest that the liver-induced long-term acceptance seen in human combined auxiliary liver and kidney transplantation is at least partly mediated by IDO activity. (C) 2009 Lippincott Williams & Wilkins, Inc.
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| 7. |
- Ingelsten, Madeleine, 1978, et al.
(författare)
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Liver-derived IL-10 as a potential inhibitor of a TH1-deviating crosstalk between donor DC and recipient NK cells during combined liver-kidney transplantation
- 2009
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Ingår i: The Transplantation society XI Basic science symposium, European society for organ transplantation I Basic science meeting, Brussels 2009..
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Konferensbidrag (refereegranskat)abstract
- During transplantation DCs within the allograft become activated and migrate to secondary lymphoid organs (SLO) where they activate NK cells and alloreactive T cell. Previous findings suggest that a liver graft is less prone to induce rejection compared to a renal graft but the mechanisms are still unclear. We compared changes in graft gene expression of inflammatory mediators known to participate in transplantation-induced DC activation in the liver and kidney in patients undergoing combined liver-kidney transplantation (n=8). Real-time PCR analysis revealed a similar up-regulation of mRNA for the DC-activation-associated inflammatory mediators TNF-α, IL-1β and IFN-gamma after reperfusion in the liver and kidney. While no reperfusion-dependent mRNA increase for IL-10 was found in transplated grafts, serial measurement of cytokine levels in peripheral blood revealed that IL-10 levels increased 60-fold in serum shortly (within 1 h) after liver, but before kidney, reperfusion and returned to pre-transplant levels at day 2 post-transplantation (n=3). No significant changes were seen in IL-12 levels. DCs matured with a cocktail consisting of IFN-γ, TNF-α and IL-1β demonstrated a strong and sustained production of CXCR3-ligands which are indispensable for DC-mediated recruitment of blood NK cells into SLO and their subsequent boosting of TH1-deviated alloresponses. Addition of IL-10 during DC maturation significantly decreased the production of all three CXCR3-ligands (CXCL9, CXCL10 and CXCL11). These findings indicate that IL-10 is selectively and rapidly released during liver transplantation, probably due to accumulation during the ischehmia period. This release may potentially inhibit a TH1-deviating crosstalk between donor DC and recipient NK cells.
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| 8. |
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| 9. |
- Ingelsten, Madeleine, 1978
(författare)
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Molecular mechanisms behind the liver-induced acceptance of renal grafts in highly sensitized patients
- 2009
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Preformed antibodies directed at donor HLA are considered an absolute contraindication for kidney transplantation, because of the high risk of rejection. Thus, patients with high levels of HLA antibodies have little chance of receiving a kidney transplant. Recently it was demonstrated that an auxiliary liver graft from the same donor may protect a subsequently grafted kidney from these harmful antibodies. The aim of this thesis was to elucidate the mechanisms behind the kidney protection afforded by the auxiliary liver graft in highly sensitized patients. We focused on the activation of dendritic cells, because these cells, which reside in all peripheral tissues, play a key role in the initiation of an immune response. This thesis demonstrates that gene expression in the liver graft correlates with clinical outcome: In patients without an acute rejection episode, 14 out of 45 investigated immunological genes were significantly higher expressed in the liver graft 4h after reperfusion, compared with patients that experienced an acute rejection episode within the first month. This indicates that high- and low-risk patients can be identified within hours after transplantation. One gene of particular interest was indoleamine 2,3-dioxygenase (IDO), which is a tolerance-inducing enzyme previously found to play a key role in maintenance of semi-allogeneic pregnancy in mice. In our study, mRNA levels of IDO were strongly upregulated in patients after combined auxiliary liver-kidney transplantation and IDO expression in the liver graft correlated with clinical outcome. Furthermore, IDO activity was higher in patients after combined auxiliary liver-kidney transplantation and liver transplantation compared with patients undergoing kidney transplantation. Strongly increased serum levels of the anti-inflammatory cytokine interleukin (IL) 10 were also found after liver but not kidney transplantation. IL-10 has several immune inhibitory effects on dendritic cells. We found that IL-10 inhibited the production of chemokines MIG, IP-10 and I-TAC in monocyte-derived dendritic cells in vitro. When comparing different cytokine cocktails for dendritic cell maturation, we showed that MIG, IP-10 and I-TAC were essential for dendritic cell-mediated recruitment of natural killer (NK) cells. This is considered important for the initiation of a type 1 T helper cell response. We also showed that IL-10 treated dendritic cells, which expressed less of these chemokines, had reduced potential to activate NK cells. Thus, the liver provides IDO and IL-10, both of which have the ability to reduce the immunostimulatory ability of dendritic cells, giving them a tolerance-promoting profile. We therefore suggest that the protective effect of an auxiliary liver in presensitized patients may, at least in part, be mediated by the liver-specific expression of IDO and IL-10.
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| 10. |
- Ingelsten, Madeleine, 1978, et al.
(författare)
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Postischemic Inflammatory Response in an Auxiliary Liver Graft Predicts Renal Graft Outcome in Sensitized Patients
- 2011
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Ingår i: Transplantation. - 0041-1337 .- 1534-6080. ; 91:8, s. 888-894
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Tidskriftsartikel (refereegranskat)abstract
- Background. The liver is considered a tolerogenic organ that favors the induction of peripheral tolerance and protects other organs from the same donor from rejection. This has been exploited in combined auxiliary liver-kidney transplantation, where a renal graft is transplanted against a positive crossmatch under the protection of a liver transplanted from the same donor. Methods. To elucidate mechanisms behind the liver protective effect, we studied early transcriptional changes of inflammatory mediators in the grafts during combined auxiliary liver-kidney transplantation using microarrays and real-time polymerase chain reaction. The results were correlated to clinical data. Results. Liver and kidney grafts both exhibited an upregulation of the leukocyte-recruiting chemokines CCL2, CCL3, and CCL4. Notably, liver grafts strongly upregulated CCL20, a dendritic cell, and T-cell recruiting chemokine. By comparing the gene expression in liver grafts with the clinical outcome, we found that 14 of 45 investigated inflammatory genes were expressed significantly higher in patients without early rejection when compared with those with early rejections. This included the above-mentioned chemokines and the T-cell-recruiting CX3CL1, NFKB1, and the tolerance-inducing gene indoleamine 2,3-dioxygenase. Conclusions. In this study, the protective role of the liver was associated with a proinflammatory reaction within this organ after ischemia-reperfusion. In particular, we found an increased expression of leukocyte-recruiting chemokines in patients without rejection, indicating a protective role of host inflammatory cells infiltrating the auxiliary liver graft in presensitized patients. Second, gene expression profiling of transplant biopsies shortly after reperfusion predicted the risk of early rejection in these patients.
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| 11. |
- Ingelsten, Madeleine, 1978, et al.
(författare)
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Rapid Increase of Interleukin-10 Plasma Levels After Combined Auxiliary Liver-Kidney Transplantation in Presensitized Patients
- 2014
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Ingår i: Transplantation. - : Ovid Technologies (Wolters Kluwer Health). - 0041-1337. ; 98:2, s. 208-215
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Tidskriftsartikel (refereegranskat)abstract
- Background. After transplantation, donor dendritic cells (DCs) in the grafted organ are activated by an ischemia/reperfusion-induced inflammatory process that induces their migration to the recipient's secondary lymphoid tissues. The subsequent interaction between migrated and mature donor DCs, recipient T cells, and natural killer (NK) cells is proposed to be crucial in directing host immune reactions toward allograft rejection. A liver transplant is less prone to induce rejection compared with most other solid organ transplants, and simultaneous transplantation of liver and kidney is known to improve the clinical outcome of kidney transplantation. Methods and Results. Here we show that liver as well as combined auxiliary liver-kidney transplantation in patients induces a rapid increase in plasma interleukin-10 (IL-10) to levels that are significantly higher than those seen after standard kidney transplantation. Addition of IL-10 during in vitro maturation of human monocyte-derived DCs with ischemia/reperfusion-associated factors was found to affect phenotypic DC maturation significantly. Addition of IL-10 inhibited DC production of the NK cell- and T cell-recruiting chemokines CXCL9, CXCL10 and CXCL11. Conclusion. Our findings indicate that liver transplantation induces a substantial systemic release of IL-10, which may inhibit T cell- and NK cell- mediated rejection processes toward the transplanted liver and concurrently transplanted kidney.
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| 12. |
- Ingelsten, Madeleine, 1978, et al.
(författare)
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The role of IL-10 in the liver tolerance effect
- 2009
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Ingår i: World congress of Nephrology, Milan 2009.
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Konferensbidrag (refereegranskat)abstract
- Introduction and aim: A liver graft is less prone to induce rejection compared to a renal graft and previous findings suggest that an auxiliary liver transplantation a few hours before a renal transplantation improves kidney graft survival in highly sensitized patients. Dendritic cells (DC) are important initiators of rejection. During transplantation DC within the allograft become activated and migrate to secondary lymphoid organs where they activate NK cells and alloreactive T cell. The aim of this study was to investigate the role of DC maturation in the liver mediated acceptance of a renal graft. Methods: Real-time PCR was used to compare changes in graft gene expression of IFN-γ, TNF-α, IL-1β and IL-10 in the liver and kidney in patients undergoing combined auxiliary liver-kidney transplantation (n=8). These are inflammatory mediators known to participate in transplantation-induced DC activation. Presence of interleukin (IL) 10 and 12 was measured in peripheral blood in these patients. Monocyte-derived DCs was matured with a cytokine-cocktail consisting of IFN-γ, TNF-α and IL-1β +/- IL-10. The effect of these cytokines on DC secretion of chemokines MIG, IP-10 and I-TAC was investigated with ELISA. Results: The DC-activation-associated inflammatory mediators TNF-α, IL-1β and IFN-γ were similarly upregulated in both liver and kidney grafts after reperfusion. While no reperfusion-dependent mRNA increase for IL-10 was found in transplated grafts, serial measurement of cytokine levels in peripheral blood revealed that IL-10 levels increased 60-fold in serum shortly (within 1 h) after liver, but before kidney, reperfusion and returned to pre-transplant levels at day 2 post-transplantation (n=3). No significant changes were seen in IL-12 levels. DCs matured with a cocktail consisting of IFN-γ, TNF-α and IL-1β demonstrated a strong production of CXCR3-ligands (MIG 280 ng/ml, IP-10 19 ng/ml, I-TAC 619 pg/ml) which was sustained even after removal of the cytokine-cocktail. These chemokines are indispensable for DC-mediated recruitment of blood NK cells into secondary lymphoid organs and their subsequent boosting of TH1-deviated alloresponses. Addition of IL-10 during DC maturation significantly decreased the production of all three CXCR3-ligands (MIG -33%, IP-10 -25%, I-TAC -67%). Conclusions: These findings indicate that IL-10 is selectively and rapidly released during liver transplantation, probably due to accumulation during the ischehmia period. This release may potentially inhibit a TH1-deviating crosstalk between donor DC and recipient NK cells, suggesting a role for IL-10 in the liver-mediated acceptance of a renal graft in highly sensitized patients.
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| 13. |
- Slavica, Lucija, et al.
(författare)
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Indoleamine 2,3-dioxygenase Expression and Functional Activity in Dendritic Cells Exposed to Cholera Toxin
- 2012
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Ingår i: Scandinavian Journal of Immunology. - : Wiley. - 0300-9475. ; 76:2, s. 113-122
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Tidskriftsartikel (refereegranskat)abstract
- Indoleamine 2,3-dioxygenase (IDO), a tryptophan-metabolizing enzyme expressed by dendritic cells (DC), has the potential to inhibit T cell responses and to promote tolerance. In contrast, cholera toxin (CT), the enterotoxin produced by Vibrio cholerae, promotes T cell responses, partly through its ability to induce DC maturation and promote antigen presentation. We hypothesized that the adjuvant activity of CT is associated with a lack of induction of IDO in DC. To test this hypothesis, monocyte-derived DC were pulsed with CT, and the IDO mRNA expression, IDO functional activity and cytokine production were measured as well as the ability of DC to induce T cell responses in vitro. Cholera toxin exposure induced enhanced levels of IDO mRNA in DC but no functional IDO protein activity. Cholera toxin pulsing however primed DC for CD40L-induced IDO protein activity. CD40L stimulation of CT-pulsed DC induced a modest IL-12p40 production, but not IL-12p70 or IL-23 secretion. Furthermore, CT-pulsed DC induced strong allogeneic and autologous T cell responses in vitro, which were not affected by the IDO-specific inhibitor 1-methyl tryptophan. Our results show that CT per se does not induce the expression of functional IDO protein, although it primes DC for CD40L-mediated IDO production and IL-12p40 secretion. Furthermore, CT-treated DC were equally powerful in their T cell stimulatory capacity as cytokine-matured DC.
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