Increased Th17/Treg Ratio In Liver Chronic Graft-Versus-Host-Disease

Abstract Background Chronic GVHD is the main cause of late non-relapse mortality and mortality after allogeneic stem cell transplantation (allo-SCT). In contrast to acute GVHD, pathophysiology of chronic GVHD remains poorly understood. Chronic GVHD presents clinical features that mimic autoimmune diseases. The identification of proinflammatory Th17 cells which contribute to autoimmune diseases pathophysiology, raised the issue of the role of Th17 cells in human chronic GVHD. Indeed, the contribution of Th17 cells in chronic GVHD was assessed in GVHD mouse models. Furthermore, theses studies indicates that the expansion of Th1 and Th17 cells is favored by the progressive loss CD4+CD25+Foxp3+ regulatory T cells (Treg) leading to chronic GVHD. This report investigated the role of Th17 cells and Treg in liver biopsies taken from patients with or without chronic GVHD. Patients and Methods Studies described in this report were performed in a single centre series of 17 patients who underwent allo-SCT for different hematological malignancies (cGVHD group). The median age of patients was 54 years (range, 27-71). The stem cell source was PBSCs in 9 cases (53%) and BM in 8 cases (47%). 13 patients received transplant from a matched-related donor, and 4 patients from an unrelated donor. A reduced-intensity conditioning regimen was used 8 patients (47%) and a myeloablative conditioning in 9 patients (53%). The control group included 8 patients without hematological malignancies who underwent liver biopsy for sleeve gastrectomy for morbid obesity (n=2) or adjacent tumor surgery (n=6). Immunohistochemistry was performed on deparaffinized tissues sections using an indirect immunoperoxydase method. A quantitative evaluation of antigens expression was performed by counting the number of positive cells in the whole biopsy at 200 magnifications for each sample. Results In the cGVHD group, based on standard pathology criteria, all 17 patients had a histologically proven liver chronic GVHD. Biopsies were taken at time of first hepatic symptoms declaration or during their reappearance and prior to corticosteroid treatment initiation or resumption. In the control group, all patients present histologically normal liver biopsy. In order to identify the Th17 cell population, biopsies were tested for expression of the CD161 and CCR6 markers, and ROR-gamma-t, the key transcription factor that orchestrates the differentiation of Th17 cells. Significantly higher numbers of ROR-gamma-t+, CD161+ and CCR6+ cells were counted in the liver of patients with chronic GVHD compared with liver of patients in the control group, mainly found in the portal space (p=0.0001, p=0.03 and p=0.03 for CD161, CCR6 and ROR-gamma-t, expression respectively). We also assessed the presence of T cells expressing Tbet (the transcription factor characterizing Th1 cells) and Foxp3 (the master regulator gene of Treg cells) in the liver biopsies. There was no difference in the number of Th1 and Treg cells between the 2 groups (p=0.88 and p=0.12 respectively). Finally we look at the Th17/Th1 and Th17/Treg ratios, considering Th17 cells as ROR- gamma-t positive cells as ROR- gamma-t is the more specific hallmark of Th17 cells. Both Th17/Th1 and Th17/Treg ratios were significantly increased in the liver of patients with liver cGVHD (p=0.005 and p=0.002 respectively). Conclusion The current study shed some light on the role of Th17 cells in the context of liver chronic GVHD. Using well-established specific markers, we show that Th17 cells infiltrate liver biopsies from patients with acute GVHD. In addition, Th17/Treg ratio was significantly increased in the liver of patients with liver chronic GVHD, suggesting a regulatory defect in liver chronic GVHD. These data raise the prospect of future innovative approaches to optimize immunosuppression regimens for the treatment or prophylaxis of chronic GVHD by targeting the Th17 response. Disclosures: Moreau: CELGENE: Honoraria, Speakers Bureau; JANSSEN: Honoraria, Speakers Bureau.

Download Full-text

Effect of Rituximab on the Incidence of GVHD in Lymphoma Patients who Received the BEAM-Conditioning and an Allogeneic Stem Cell Transplantation.

Blood ◽

10.1182/blood.v104.11.4980.4980 ◽

2004 ◽

Vol 104 (11) ◽

pp. 4980-4980

Author(s):

Issa F. Khouri ◽

Rima M. Saliba ◽

Daniel R. Couriel ◽

Grace-Julia Okoroji ◽

Sandra Acholonu ◽

...

Keyword(s):

Stem Cell ◽

B Cells ◽

Stem Cell Transplantation ◽

Cell Transplantation ◽

Allogeneic Stem Cell Transplantation ◽

Acute Gvhd ◽

Chronic Gvhd ◽

Conditioning Regimen ◽

Control Group ◽

Study Group

Abstract It has been postulated that B cells functioning as antigen-presenting cells may have an important role in the pathogenesis of GVHD. Depletion of donor cells from B-cells resulted in a low incidence of GVHD in mouse model (Schultz et al. BMT1995:16:289–289). More recently, we observed a lower incidence of chronic (and to a lesser extent acute GVHD) in patients with CLL who received an allogeneic stem cell transplantation after a non-myeloablative conditioning regimen containing rituximab (Exp Hematol32:28–35, 2004). The purpose of this study is to investigate the effect of rituximab on GVHD in the setting of a more intense chemotherapy with BEAM, in patients who received an allogeneic peripheral blood stem cell from HLA-identical siblings. To test this hypothesis, we retrospectively studied 11 consecutive patients with non-Hodgkin’s lymphoma who received BEAM/Rituximab at the M. D. Anderson Cancer Center. We attempted to match these patients by age, donor-recipient gender, and donor-recipient CMV reactivity to a historical control of 44 patients with lymphoma, who received BEAM alone as a conditioning regimen, without the Rituximab. Tacrolimus and methotrexate were used for GVHD prophylaxis in both groups. A total of 10 patients in the study group, could be matched with 19 patients in the control group and were included in the final analysis. The outcome of the 2 groups is shown below: Rituximab-Study Group Control Group -value P No. of patients 10 19 Median age 41 44 0.4 (range) (19–55) (19–60) Patient-Donor sex-matched 9(82%) 18(95%) 0.6 Median # CD34 + cells infused (106/kg) 5.1 4.73 0.1 Patient or Donor CMV+ 9(82%) 18(95%) 0.6 Patient and Donor CMV − 1(10%) 1(5%) Median # prior chemoregimens 3 3 0.9 range (1–8) (1–9) Median follow-up 17 38 range (8–48) (27–77) Acute GVHD 2–4 (n,%) 5(50%) 7(37%) 0.5 Acute GVHD 3–4 (n,%) 3(30%) 5(26%) 0.6 Chronic GVHD (n, % cumulative incidence) 8 (90% + 15) 10 (53% + 12 0.01 Our data suggest that the described protective effect of Rituximab against GVHD in mouse models or in the setting of non-myeloablative allogeneic transplantation, may be overcome by the BEAM. This more intense conditioning regimen may induce more GVHD by enhancing T-cell cytokines release and by causing more gastrointestinal toxicity, thus allowing for a greater antigen presentation.

Download Full-text

Plasmacytoid Dendritic Cells (PDC) and Th17 Immune Response Contribution in Gut Acute Graft-Versus-Host-Disease (GVHD)

Blood ◽

10.1182/blood.v118.21.2968.2968 ◽

2011 ◽

Vol 118 (21) ◽

pp. 2968-2968

Author(s):

Florent Malard ◽

Céline Bossard ◽

Jessy Arbez ◽

Patrice Chevallier ◽

Thierry Guillaume ◽

...

Keyword(s):

Immune Response ◽

Stem Cell ◽

Autoimmune Diseases ◽

Intestinal Mucosa ◽

Th17 Cells ◽

Acute Gvhd ◽

Type I ◽

Type I Ifn ◽

Th17 Response

Abstract Abstract 2968 Background. Acute GVHD after allogeneic stem cell transplantation (allo-SCT) is an exaggerated immune response against alloantigens involving dysregulation of inflammatory cytokine cascades. Previous studies established an important role of Th1 cells in acute GVHD pathophysiology. However, the identification of proinflammatory Th17 cells which contribute to autoimmune diseases pathophysiology, raised the issue of the role of Th17 cells in human acute GVHD. Indeed, the contribution of Th17 cells in acute GVHD was assessed in GVHD mouse models with conflicting results. In addition, the role of the PDC subset (the professional type I IFN-secreting cells), which play an important role in triggering Th17-related cytokines and autoimmune diseases, is not yet established in the acute GVHD setting. This report investigated the role of Th17 cells and their interaction with PDC in gastrointestinal (GI) biopsies taken from patients with or without acute GVHD. Patients and Methods. Studies described in this report were performed in a single centre series of 21 patients who underwent allo-SCT for different hematological malignancies (n=19) and severe aplastic anemia (n=2). The median age of patients was 53 years (range, 16–69). The stem cell source was PBSCs in 19 cases (85%), CB in 2 cases and BM in one case. Ten patients received transplant from a matched-related donor, and 11 patients from a matched-unrelated donor. A reduced-intensity conditioning regimen was used in the majority of cases (n=19; 90%) Immunohistochemistry was performed on deparaffinized tissues sections using an indirect immunoperoxydase method. A quantitative evaluation of antigens expression was performed by counting the number of positive cells in the whole biopsy at 200 magnifications for each sample. Results. In this cohort, based on standard pathology criteria, 16 patients had a histologically proven gastrointestinal acute GVHD. In all cases, biopsies were taken before initiation of systemic corticosteroid therapy. The remaining 5 patients did not have histological signs of acute GVHD (and did not develop clinical signs of acute GVHD) and thus, were used as controls. In order to identify the Th17 cell population, biopsies were tested for expression of the CD161 and CCR6 markers, and ROR-gamma-t, the key transcription factor that orchestrates the differentiation of Th17 cells. Significantly higher numbers of ROR-gamma-t+ and CD161+ cells were counted in the intestinal mucosa of patients with acute GVHD compared with intestinal mucosa of patients without acute GVHD, mainly found in the lamina propria but also in the epithelium of altered glands (p=0.016 and p=0.009 for ROR-gamma-t and CD161 expression respectively). Given the role of PDCs in triggering Th17-related cytokines, we sought next to determine the proportion of PDCs in intestinal biopsies from these same patients. This analysis showed a significant increase of CD123+ PDCs in the intestinal mucosa of patients with acute GVHD compared with mucosa of patients without acute GVHD (p=0.017). Moreover, we observed a significant correlation between the number of CD123+ PDCs and ROR-gamma-t or CD161 expressing cells in the intestinal mucosa of acute GVHD patients, highlighting the link between PDC and Th17 cells. Conclusion. The current study shed some light on the role of Th17 cells in the context of gastro-intestinal acute GVHD. Using well-established specific markers, we show that Th17 cells infiltrate intestinal biopsies from patients with acute GVHD. In addition, Th17 infiltration was paralleled by the infiltration of PDCs, suggesting a potential new pathophysiological link between PDCs and Th17 response in the context of gastro-intestinal acute GVHD. This is consistent with studies showing that PDCs can drive the differentiation of Th17 cells. Functional analyses are currently ongoing. Although the exact mechanism that links type I IFN production to PDC-mediated Th17 responses is still unclear in acute GVHD, these data raise the prospect of future innovative approaches to optimize immunosuppression regimens for the treatment or prophylaxis of acute GVHD by targeting PDCs and the Th17 response. Disclosures: No relevant conflicts of interest to declare.

Download Full-text

Outpatient Haploidentical Peripheral Blood Stem-Cell Transplantation with Post-Transplant Cyclophosphamide in Children and Adolescents

Blood ◽

10.1182/blood.v126.23.4389.4389 ◽

2015 ◽

Vol 126 (23) ◽

pp. 4389-4389

Author(s):

Oscar Gonzalez-Llano ◽

Elias Eugenio Gonzalez-Lopez ◽

Ana Carolina Ramirez-Cazares ◽

Edson Rene Marcos-Ramirez ◽

Guillermo J. Ruiz-Arguelles ◽

...

Keyword(s):

Stem Cell ◽

T Cell ◽

High Risk ◽

Children And Adolescents ◽

Hematological Malignancies ◽

Acute Gvhd ◽

Chronic Gvhd ◽

High Dose ◽

Post Transplant

Abstract Patients with high-risk hematological malignancies have a poor prognosis without a hematopoietic stem cell transplant. An HLA- haploidentical donor is available in 95% of the cases, and post-transplant cyclophosphamide permits the use of T-cell replete grafts in settings were ex-vivo manipulation is not available. The experience with HLA-haploidentical HSCT with PBSC and post-tranplant Cy in the pediatric and adolescent population is limited; we report the following experience. We retrospectively collected data on 25 patients (0 to 21 years old) with hematological malignancies, who underwent ambulatory haploidentical HSCT with post-transplant Cy from November 2011 to November 2014. The different conditioning regimens are described in Table 1. All patients received high-dose Cy(50mg/kg) on days +3 and +4. Cyclosporine A (CYA; 6mg/kg/d per os) and mycophenolate mofetil (MMF; 15mg/kg two times daily per os) were started on day +5. MMF was discontinued on day +35 and tapering of cyclosporin started day +90 in the absence of GVHD. All patients received anti-microbial prophylaxis for bacteria, fungal, herpes infection and Pneumocystis jiroveci according to institutional practices. First chimerism was performed at day +30, and second chimerism at day +100. Primary graft failure was defined when neutrophil counts did not exceed 0.5 x 109/L by day +30. Acute and chronic GVHD were graded according to NIH criteria. Patient, donor and stem-cell harvest characteristics are described in Table 1. All patients had high risk hematological malignancies. There were 5 patients who underwent their first transplant on 1st CR, 4 with ALL with high risk cytogenetics and 1 with AML. All other patients were defined as high risk because they were refractory/relapsed. Twenty-three patients (92%) had neutrophil engraftment after a median 17 (7-24) days. Platelet engraftment was observed in 20 (80%) patients after a median of 14.5 (11-23) days, 3 (12%) patients did not have platelet counts below 20,000/mcL. One patient was catalogued as a primary failure for not achieving neutrophil and platelet engraftment by day +30. One patient died before engraftment at day +10 of septic shock. Four patients (16%) died before day +30. The only patient that did not have a complete chimerism, had a diagnosis of AML and 30% of donor cells by day +30, by day +45 relapse of disease was documented. After a median follow-up of 157 days, 13 patients (52%) remain alive, with an estimated 1-year OS of 52% (95%CI: 30.4 - 65.6%).Nine patients (36%) died of complications (mainly infectious) not related to relapse at a median time of 66 days (10-579 days) from stem cell infusion. Nine patients (36%) relapsed in a median time of 105 days (45-288 days); three of those patients died at days +150, +113 and + 370 from transplant. Estimated 1 year event-free survival is 40.2% (95%CI: 41.3 - 75.8%) (Figures 1 and 2). Patients transplanted on 1st CR had a median follow-up of 664 days with an OS and EFS of 80% (4 patients), which was statistically different from the rest of the population (p=0.03) (Figure 3). Among those who engrafted (n=21), 9 cases (42.9%) had grade 2-4 acute GVHD and 4 cases (19%) of grade 3-4 acute GVHD. Three patients (14.3%) developed chronic GVHD, two had mild skin or liver cGVHD. One patient had severe (NIH stage 3) skin cGVHD, she was alive and with a grade 2 cGVHD until last follow up at day +893. Outpatient procedure, HLA-haploidentical HSCT including PBSC as a stem cell source, and post-transplant T-cell in vivo depletion using high-dose cyclophosphamide is feasible in children and adolescents, with acceptable rates of response and GVHD. Table 1. Patient, donor and harvest characteristics Variable N=25 Age, median(range in years) 10 (1-21) Gender, n(%) Male Female 17 (68%) 8 (32%) Diagnosis, n(%) ALL-B ALL-T AML CML 16 (64%) 2 (8%) 5 (20%) 2 (8%) Time from diagnosis to transplant (months) 17.2 (1.9-153.5) Conditioning regimen, n(%) Cy 1500mg/m2 + Flu 75mg/m2 + Bu 9.6mg/kg (IV) Cy 1050mg/m2 + Flu 75mg/m2 + Bu 12 mg/kg (oral) Cy/VP-16/RT Cy/Flu/Mel 16 (64%) 6 (24%) 2 (8%) 1 (4%) Donor, n(%) Mother Father Sister 20 (80%) 3 (12%) 2 (8%) Donor age, median(range in years) 38 (17-49) Infused CD34+ x 106/kg, median(range) 11 (3.2-20) Disclosures No relevant conflicts of interest to declare.

Download Full-text

Ursodeoxycholic acid for the prevention of hepatic complications in allogeneic stem cell transplantation

Blood ◽

10.1182/blood-2001-12-0159 ◽

2002 ◽

Vol 100 (6) ◽

pp. 1977-1983 ◽

Cited By ~ 159

Author(s):

Tapani Ruutu ◽

Britta Eriksson ◽

Kari Remes ◽

Eeva Juvonen ◽

Liisa Volin ◽

...

Keyword(s):

Stem Cell ◽

Stem Cell Transplantation ◽

Ursodeoxycholic Acid ◽

Cell Transplantation ◽

Allogeneic Stem Cell Transplantation ◽

Acute Gvhd ◽

Chronic Gvhd ◽

Treated Group ◽

Control Group ◽

To Receive

Abstract The role of ursodeoxycholic acid (UDCA) in the prevention of hepatic complications after allogeneic stem cell transplantation was studied in a prospective randomized open-label multicenter trial. A total of 242 patients were allocated to receive (n = 123) or not to receive (n = 119) UDCA in the dose of 12 mg/kg/d orally from the day preceding the conditioning until day 90 after transplantation. In the UDCA-treated group a significantly smaller proportion of patients developed a serum bilirubin level exceeding 50 μM (18 of 123 versus 31 of 119, P = .04), and similarly a smaller proportion of patients exceeded the alanine aminotransferase level of 100 U/L. There was no difference in the incidence of veno-occlusive disease of the liver. Compared to the control group, in the UDCA-treated group there was a nonsignificant trend toward a lower overall incidence of acute graft-versus-host disease (GVHD) and a significantly lower incidence of grade III to IV acute GVHD (5 of 123 versus 17 of 119,P = .01), stage II to IV liver and intestinal GVHD, and stage III to IV skin GVHD. There was no difference in the incidence of chronic GVHD or in the relapse rate. Among the patients given UDCA, the survival at 1 year was significantly better, 71% versus 55% (P = .02), and the nonrelapse mortality rate was lower, 19% versus 34% (P = .01), than in the control group. There were significantly more deaths in GVHD in the control group. In conclusion, UDCA administration reduced hepatic problems and severe acute GVHD and improved survival. These results suggest a role for UDCA in the prevention of transplant-related complications in allogeneic transplantation.

Download Full-text

Fludarabine and Treosulfan Conditioning for Allogeneic Stem-Cell Transplantation; a Dose- Intense Regimen with Limited Toxicity.

Blood ◽

10.1182/blood.v116.21.3473.3473 ◽

2010 ◽

Vol 116 (21) ◽

pp. 3473-3473 ◽

Cited By ~ 2

Author(s):

Avichai Shimoni ◽

Avital Rand ◽

Noga Shem-Tov ◽

Izhar Hardan ◽

Yulia Volchek ◽

...

Keyword(s):

Stem Cell ◽

High Risk ◽

Stem Cell Transplantation ◽

Allogeneic Stem Cell Transplantation ◽

Cumulative Incidence ◽

Graft Failure ◽

Hematological Malignancies ◽

Acute Gvhd ◽

Karnofsky Score

Abstract Abstract 3473 Allogeneic stem cell transplantation (SCT) is a potentially curative approach for patients (pts) with various hematological malignancies. However, standard myeloablative conditioning (MAC) is limited to younger, medically fit pts. Reduced-intensity conditioning (RIC) is feasible in pts not eligible for MAC, allowing engraftment and limiting organ toxicity, however, relapse rates are increased, especially in pts not in remission at the time of SCT. Novel approaches are in need to deliver dose intense conditioning, yet with RIC type toxicity, to these pts. Treosulfan is a bifunctional alkylating agent with cytotoxic and immunosuppressive effects. Prior studies demonstrated the feasibility of a fludarabine-treosulfan (FT) conditioning regimen, suggesting it is effective with limited toxicity. To better define the role of FT, we analyzed outcomes of a relatively large cohort of pts, treated in a single institution. Pts were eligible for FT only if considered not eligible for MAC, due to advanced age, comorbidities or extensive prior therapy (including a prior SCT). FT included fludarabine 30 mg/m2, on days -6 to -2, and treosulfan 10–12 gr/m2 on days -6 to -4. Pts with an unrelated donor or no prior chemotherapy were also given ATG (Fresenius, total dose 15 mg/kg). The study included 105 pts, median age 57 years (range, 20–76). Diagnoses included AML (n=56), MDS (n=33), CML (n=3), lymphoid malignancies (n=12) and myelofibrosis (n=1). The donor was an HLA-matched sibling (n=50) or matched unrelated [MUD, n=55; 10/10 match (n=43), 9/10 (n=7), 8/10 (n=5)]. Disease status was defined as early (n=17), intermediate (n=24) or advanced (n=64) by standard criteria. This was a relatively high-risk pt group as only 24 pts (23%) were in CR (first or later), 30 pts had a comorbidity score ≥3 and 22 pts had a Karnofsky score ≤80 at the time of SCT. Twenty-seven pts had a prior SCT; 13 autologous and 14 allogeneic (12 from a different donor). Ninety-five pts engrafted; 7 died prior to engraftment, 3 had early primary or secondary graft failure (MUD-1, mismatched MUD- 2 pts). The median time to neutrophil and platelet engraftment was 12 (range, 9–23) and 16 (range, 10–66) days, respectively. With a median follow-up of 16 months (range, 1–73), 57 pts are alive and 48 have died. The cumulative incidence of acute GVHD and grade III-IV acute GVHD was 26% and 11%, respectively. The cumulative incidence of chronic GVHD was 53%. In all, 8 pts died of complications of GVHD and 16 from other non-relapse causes, including multi-organ failure (n=4), CNS bleeding (n=2), infections (n=7), graft failure (n=2) and suicide (n=1). The cumulative incidence of non-relapse mortality 3-years after SCT was 25% (95%CI, 18–35). Twenty-four pts died of disease recurrence, cumulative incidence 30% (95%CI, 21–44) at 3 years. Considering that only 26% of the pts were in CR at the time of SCT, this rate seems promising. The median 3-year overall (OS) and progression-free survival were 45% (95%CI, 33–57) and 37% (95%CI, 26–49), respectively. The 2 most important factors predicting for OS in multivariate analysis were advanced disease and a low Karnofsky score at the time of SCT. Hazard ratios were 3.7 (p=0.03) and 4.2 (p=0.001), respectively. The 3-year OS was 71% (95%CI, 45–96), 33% (95%CI, 6–69) and 26% (95%CI, 6–46) for low, intermediate and advanced stage, respectively (p=0.03). Best results were obtained in pts with previously untreated MDS (n=33, including 28 pts with > 10% marrow blasts at SCT) and pts with high-risk AML in first CR (n=14), with 3-year OS rates of 61% (95%CI, 42–80) and 65% (95%CI, 38–93), respectively. Interestingly, a second SCT was not associated with a worse outcome. OS was 43% (42/78 alive), 46% (8/13 alive) and 40% (7/14 alive) in first SCT, a second SCT after a prior autologous SCT or a prior allogeneic SCT, respectively. In conclusion, treosulfan-based conditioning is a promising approach in pts with advanced hematological malignancies, not eligible for standard MAC. Favorable results were observed not only in pts with early stage disease, but also in previously untreated advanced MDS. Second SCT was also feasible with FT with a relatively favorable outcome. The low relapse risk suggests that this regimen can be considered a dose-intensive regimen, with a toxicity profile that resembles RIC regimens. The role of FT should be further defined in larger prospective studies. Disclosures: No relevant conflicts of interest to declare.

Download Full-text

Mature Circulating Endothelial Cells and Progenitors in Patients with Chronic Gvhd

Blood ◽

10.1182/blood.v118.21.4700.4700 ◽

2011 ◽

Vol 118 (21) ◽

pp. 4700-4700

Author(s):

Rocco Pastano ◽

Giovanna Andreola ◽

Patrizia Mancuso ◽

Federica Gigli ◽

Angelo Gardellini ◽

...

Keyword(s):

Endothelial Cells ◽

Stem Cell ◽

Stem Cell Transplantation ◽

Immunosuppressive Therapy ◽

Median Time ◽

Allogeneic Stem Cell Transplantation ◽

Acute Gvhd ◽

Chronic Gvhd ◽

Circulating Endothelial Cells

Abstract Abstract 4700 Acute and chronic graft-versus-host disease are a common complication of allogeneic stem cell transplantation. In animal models acute GVHD (aGVHD) is associated with increased neovascularization and number of circulating endothelial cells (CECs), while patients with sclerodermatous chronic GVHD (cGVHD) show a significant decrease in the number of circulating endothelial progenitor cells (EPCs) in peripheral blood as compared to patients with non sclerodermatous cGVHD or controls. In an attempt to evaluate the role of CECs and EPCs in patients with cGVHD, we analysed a total of 15 patients affected by hematological malignancies (3 Non-Hodgkin Lymphoma, 4 Hodgkin Disease, 4 Multiple Myeloma, 2 Myelodisplastic Syndrome, 2 Chronic Lymphocitic Leukemia) having undergone allogeneic stem cell transplantation following reduced intensity conditioning. Donors were HLA identical in 14 patients (12 sibling and 2 matched unrelated donors) and HLA aploidentical in 1. Acute GVHD and cGVHD were defined on the basis of time of manifestation, ≤100 days for aGVHD and >100 days for cGVHD. At the time of the blood sample collection, 8 patients, median age 42 years (28-51), with a median time after transplant of 177 days (21-1373), had no evidence of GVHD; of those 5/8 were evaluable for aGVHD and cGVHD, 2 only for aGVHD; 4/8 were on post-transplant calcineurin inhibitors immunosuppressive therapy; 7 other patients, median age 51 years (38-64), with a median time after transplant of 844 days (314-1779), were all evaluable for acute and cGVHD and had evidence of cGVHD as follows: sclerodermatous in 3 patients requiring systemic immunosuppressive therapy, oral mucosa lichen in 1 patient taking oral corticosteroid and cutaneous erythematous and dischromic cGVHD in the other 3 patients, with only 1 patient on systemic immunosuppressive therapy. Viable and apoptotic CECs and EPCs were evaluated by six color flow cytometry (Mancuso et al, Clin Cancer Res, 2009). Briefly, CECs were defined as DNA+CD45-CD31+ CD146+, EPCs as CD45- CD34+. The combination of Syto16 and 7-AAD was used to discriminate between viable (syto16bright/7-AAD-) and apoptotic (syto16weakly pos/7-AAD+) endothelial cells, and to exclude from analysis, platelets and endothelial macroparticles. The results, expressed as median of cells/mL, are summarized in the following table:Total CECsViable CECsApoptotic CECsEPCsHealthy Subject103 (33–322)21 (3–67)77* (28–303)31 (0–56)Patients with cGVHD46 (29–94)41 (25–68)5* (3–26)30 (0–213)Patients without GVHD138 (30–179)39 (10–153)68* (5–136)49 (0–355)*p<0.017 These preliminary data indicate a significant reduction in apoptotic circulating mature endothelial cells, likely reflecting a poor vascularization of multiple organs and tissues targeted by cGVHD and a trend towards a decreased number of EPCs in patients with cGVHD. A multicentric study is now planned to confirm these hypotheses and investigate a possible predictive/prognostic role of CEC and EPC counts in cGVHD. Disclosures: No relevant conflicts of interest to declare.

Download Full-text

Plasmacytoid Dendritic Cells (PDC) and Th17 Immune Response Contribution in Skin Acute Graft-Versus-Host-Disease (GVHD)

Blood ◽

10.1182/blood.v120.21.4103.4103 ◽

2012 ◽

Vol 120 (21) ◽

pp. 4103-4103

Author(s):

Florent Malard ◽

Céline Bossard ◽

Patrice Chevallier ◽

Thierry Guillaume ◽

Jacques Delaunay ◽

...

Keyword(s):

Immune Response ◽

Stem Cell ◽

Autoimmune Diseases ◽

Th17 Cells ◽

Acute Gvhd ◽

Unrelated Donor ◽

Type I ◽

Type I Ifn ◽

Th17 Response

Abstract Abstract 4103 Background. Acute GVHD after allogeneic stem cell transplantation (allo-SCT) is an exaggerated immune response against alloantigens involving dysregulation of inflammatory cytokine cascades. Previous studies established an important role of Th1 cells in acute GVHD pathophysiology. However, the identification of proinflammatory Th17 cells which contribute to autoimmune diseases pathophysiology, raised the issue of the role of Th17 cells in human acute GVHD. Indeed, the contribution of Th17 cells in acute GVHD was assessed in GVHD mouse models with conflicting results. In addition, the role of the PDC subset (the professional type I IFN-secreting cells), which play an important role in triggering Th17-related cytokines and autoimmune diseases, is not yet established in the acute GVHD setting. This report investigated the role of Th17 cells and their interaction with PDC in cutaneous biopsies taken from patients with or without acute GVHD. Patients and Methods. Studies described in this report were performed in a single centre series of 38 patients who underwent allo-SCT for different hematological malignancies (n=37) and severe aplastic anemia (n=1). The median age of patients was 52 years (range, 17–70). The stem cell source was PBSCs in 27 cases (71%), CB in 6 cases and BM in 5 cases. 11 patients received transplant from a matched-related donor, and 27 patients from an unrelated donor. A reduced-intensity conditioning regimen was used in the majority of cases (n=29; 76%) Immunohistochemistry was performed on deparaffinized tissues sections using an indirect immunoperoxydase method. A quantitative evaluation of antigens expression was performed by counting the number of positive cells in the whole biopsy at 200 magnifications for each sample. Results. In this cohort, based on standard pathology criteria, 29 patients had a histologically proven skin acute GVHD. In all cases, biopsies were taken before initiation of systemic corticosteroid therapy. The remaining 9 patients did not have histological signs of acute GVHD (and did not develop clinical signs of acute GVHD) and thus, were used as controls. In order to identify the Th17 cell population, biopsies were tested for expression of the CD161 and CCR6 markers, and ROR-gamma-t, the key transcription factor that orchestrates the differentiation of Th17 cells. Significantly higher numbers of ROR-gamma-t+, CD161+ and CCR6+ cells were counted in the skin of patients with acute GVHD compared with intestinal mucosa of patients without acute GVHD, mainly found in the lamina propria but also in the epithelium of altered glands (p=0.001, p<0.0001 and p=0.01 for ROR-gamma-t, CD161 and CCR6 expression respectively).Given the role of PDCs in triggering Th17-related cytokines, we sought next to determine the proportion of PDCs in cutaneous biopsies from these same patients. This analysis showed a significant increase of BDCA2+ PDCs in the skin of patients with acute GVHD compared with skin of patients without acute GVHD (p=0.03). Moreover, we observed a strong expression of the type I IFN-inducible protein Mx1 in the skin of patients with acute GVHD compared with skin of patients without acute GVHD, reflecting the high production of type I IFN by the BDCA2+ PDCs. Conclusion. The current study shed some light on the role of Th17 cells in the context of cutaneous acute GVHD. Using well-established specific markers, we show that Th17 cells infiltrate skin biopsies from patients with acute GVHD. In addition, Th17 infiltration was paralleled by the infiltration of PDCs, suggesting a potential new pathophysiological link between PDCs and Th17 response in the context of cutaneous acute GVHD. This is consistent with studies showing that PDCs can drive the differentiation of Th17 cells. Functional analyses are currently ongoing. These data raise the prospect of future innovative approaches to optimize immunosuppression regimens for the treatment or prophylaxis of acute GVHD by targeting PDCs and the Th17 response. Disclosures: No relevant conflicts of interest to declare.

Download Full-text

Regulation of Autoreactive CD4 T Cells by FoxO1 Signaling in CNS Autoimmunity

10.21203/rs.3.rs-322045/v1 ◽

2021 ◽

Author(s):

Emma E. Kraus ◽

Laura Kakuk-Atkins ◽

Marissa F. Farinas ◽

Mathew Jeffers ◽

Amy E. Lovett-Racke ◽

...

Keyword(s):

T Cells ◽

Transcription Factor ◽

Cd4 T Cells ◽

Th17 Cells ◽

Foxp3 Expression ◽

Treg Cells ◽

Th1 Cells ◽

T Effector Cells ◽

Cns Autoimmunity

Abstract BackgroundMyelin-specific CD4 T effector cells (Teffs), Th1 and Th17 cells, are encephalitogenic in experimental autoimmune encephalomyelitis (EAE), a well-defined murine model of multiple sclerosis (MS) and implicated in MS pathogenesis. Forkhead box O 1 (FoxO1) is a conserved effector molecule in PI3K/Akt signaling and critical in the differentiation of CD4 T cells into T helper subsets. However, it is still unclear whether FoxO1 may be a target for redirecting CD4 T cell differentiation and benefit CNS autoimmunity. MethodsUsing a selective FoxO1 inhibitor AS1842856, we determined the effects of FoxO1 inhibition in regulating myelin-specific Th1 and Th17 cells, and the transcriptional balance of T-bet and Foxp3 in myelin-specific CD4 T cells from EAE mice. The effects of AS1842856 in regulating the encephalitogenicity of myelin-specific T cells and the expansion of human Th1 cells from MS patients were also characterized. Furthermore, we characterized the potential role of FoxO1 in mediating PD-1 signaling in CD4 T cells, critical for regulating Teff and Treg cells. ResultsInhibition of FoxO1 suppressed the differentiation and expansion of Th1 cells. Moreover, the transdifferentiation of Th17 cells into encephalitogenic Th1-like cells was suppressed by FoxO1 inhibition upon reactivation of myelin-specific CD4 T cells from mice with EAE. When FoxO1 was inhibited in myelin-specific CD4 T cells, the transcriptional balance skewed from the Th1 transcription factor T-bet toward the Treg transcription factor Foxp3. Myelin-specific CD4 T cells treated with the FoxO1 inhibitor were less encephalitogenic in adoptive transfer EAE studies compared to control-treated cells. Inhibition of FoxO1 in T cells from MS patients significantly suppressed the expansion of Th1 cells. Furthermore, the immune checkpoint programmed cell death protein-1 (PD-1)-induced Foxp3 expression in CD4 T cells was impaired by FoxO1 inhibition, consistent with a bias toward Treg induction. ConclusionsThese data illustrate an important role of FoxO1 signaling in CNS autoimmunity via regulating autoreactive Teff and Treg balance.

Download Full-text

Role of Il-17 Varies at Different Periods After Hematopoietic Stem Cell Transplantation: Protection From Acute Graft-Versus-Host Disease and Exacerbation of Chronic Graft-Versus-Host Disease.

Blood ◽

10.1182/blood.v116.21.3741.3741 ◽

2010 ◽

Vol 116 (21) ◽

pp. 3741-3741 ◽

Cited By ~ 1

Author(s):

Rie Kuroda ◽

Ryosei Nishimura ◽

Katsuaki Sato ◽

Hideaki Maeba ◽

Kazuhito Naka ◽

...

Keyword(s):

T Cells ◽

Stem Cell ◽

Hematopoietic Stem Cell Transplantation ◽

Stem Cell Transplantation ◽

Immune Cells ◽

Acute Gvhd ◽

Chronic Gvhd ◽

Hematopoietic Stem ◽

Graft Versus Host

Abstract Abstract 3741 Th17 is a newly identified T cell lineage that secretes the proinflammatory cytokine IL-17. Th17 cells have been shown to play a crucial role in mediating autoimmune diseases such as experimental autoimmune encephalomyelitis (EAE), arthritis, and colitis. Anti-IL-17 therapy for some autoimmune diseases in clinical settings has been started and promising results have been reported. However the role of IL-17 on developing acute and chronic GVHD in hematopoietic stem cell transplantation (HSCT) is not yet fully understood. Interaction between IL-17 and IL-17 receptor is complicated because IL-17 is produced in various kinds of immune cells other than CD4+ T-cells, and IL-17 receptors express on not only immune cells but also various epithelial cells, including lung and intestine, both of which are target organs of GVHD. To explore the role of host derived or donor derived IL-17 separately in acute GVHD, lethally irradiated wild type (WT) or IL-17 knockout (KO) Balb/c (H-2d) were given WT or IL-17 KO C57BL/6 (H-2b) bone marrow (BM) cells with WT splenocytes to induce acute GVHD. Infused cell number of WT splenocytes in this study induced acute GVHD, but not lethal in IL-17 WT host mice. In contrast, IL-17 KO host mice receiving WT BM plus WT splenocytes developed severe acute gut GVHD and finally half of them died (p<0.05). To exclude the possibility that alloreactivity of host IL-17 KO derived dentritic cells (DCs) could be much more than that of WT DCs, mixed leukocyte reaction (MLR) was performed using stimulators from WT or IL-17 KO DCs and responders from WT CD4+ T-cells. No significant differences were observed between WT DCs and IL-17 KO DCs in thymidine uptake and percentage of responder cells producing IFN-g or TNF-a. Taken together, host-derived IL-17 has a protective effect against acute GVHD. Moreover similar results were observed when IL-17 KO Balb/c mice were given BM cells from another strain B10.D2 plus splenocytes shown in the figure below (p<0.05). Next, we compared the development of chronic GVHD between the lethally irradiated WT Balb/c mice given IL-17 KO C57BL/6 BM cells or WT BM cells with low dose WT splenocytes to induce sublethal acute GVHD and chronic GVHD subsequently. After day 60 the mice receiving WT BM cells plus WT splenocytes experienced weight loss accompanied by skin histological changes (p<0.05, shown in the figure below), while mice receiving IL-17 KO BM plus WT splenocytes showed minimal signs of GVHD as well as mice receiving IL-17 KO BM or WT BM alone. Increased number of donor-BM derived IL-17 producing cells was observed in the mice showing chronic GVHD compared to BM control (p<0.05). Moreover, a significant increase of T-cell proliferation was observed by adding rIL-17 into MLR culture (p<0.05). These results suggest that donor BM derived IL-17 producing cells involved in the pathogenesis of chronic GVHD by exacerbating the alloimmune response in part. In conclusion IL-17, especially from host-derived, has a protective effect against acute GVHD. On the contrary, donor BM derived IL-17 exacerbates chronic GVHD. Neutralizing IL-17 would be a potent strategy only for preventing chronic GVHD, not for acute GVHD. Disclosures: No relevant conflicts of interest to declare.

Download Full-text

Single Cord Blood Transplantation Versus HLA-Haploidentical Related Donor Transplantation Using Post-Transplant Cyclophosphamide in Patients with Hematological Malignancies

Blood ◽

10.1182/blood-2021-149403 ◽

2021 ◽

Vol 138 (Supplement 1) ◽

pp. 2927-2927

Author(s):

Fumiya Wada ◽

Junya Kanda ◽

Satoshi Yoshioka ◽

Takayuki Ishikawa ◽

Takashi Akasaka ◽

...

Keyword(s):

Stem Cell ◽

Board Of Directors ◽

Research Funding ◽

Hematological Malignancies ◽

Acute Gvhd ◽

Chronic Gvhd ◽

Advisory Committees ◽

Otsuka Pharmaceutical ◽

Related Donor ◽

Haplo Group

Abstract Introduction: Unrelated cord blood (UCB) and haploidentical related donor transplantation using post-transplant cyclophosphamide (PTCy-haplo) have become alternative options to treat patients with hematological malignancies without a human leukocyte antigen (HLA)-matched donor. Although a previous phase III study showed better outcomes after bone marrow transplantation (BMT) using PTCy-haplo than those after double unit UCB, a donor selection algorithm including peripheral blood stem cell transplant (PBSCT) using PTCy-haplo or high-risk patients remains to be clarified. With the aim to assess the relative efficacies of UCB and PTCy-haplo transplant, we performed a multi-center retrospective analysis to compare the clinical outcomes in patients with hematologic malignancies. Methods: We included 517 patients aged 16 years to 70 years with hematological malignancies who received a first stem cell transplantation using a single UCB unit or PTCy-haplo between 2013 and 2019 in Kyoto Stem Cell Transplantation Group (KSCTG), which is a multi-center group of 18 transplantation centers in Japan. The primary endpoint was to compare overall survival (OS) between the UCB and PTCy-haplo groups. Secondary endpoints were relapse-free survival (RFS), GVHD- and relapse-free survival (GRFS), relapse, non-relapse mortality (NRM), neutrophil engraftment, platelet engraftment, grade II-IV acute graft-versus-host disease (GVHD), grade III-IV acute GVHD, chronic GVHD, and extensive chronic GVHD. Results: Among 517 patients, 460 received single UCB transplant and 57 received PTCy-haplo related donor transplant (53 PBSCT and 4 BMT). The median age of the UCB and PTCy-haplo groups was 55 and 52 years, respectively (P= 0.770). Myeloablative conditioning (MAC) was used more often in the UCB group than in the PTCy-haplo group (P< 0.001). In the PTCy-haplo cohort, the total dose of Cy was 80mg/m 2 (43.9%) or 100mg/m 2 (56.1%), and the median duration of tacrolimus and mycophenolate mofetil use was 173 and 34 days, respectively. The median follow-up periods of survivors were 2.2 and 2.4 years, respectively. OS in the UCB group was comparable to that in the PTCy-haplo group (2-year OS; 53.7%, 53.6%, P= 0.71 and adjusted hazard ratio (aHR) 1.00, P= 0.99), with similar risks of relapse (aHR 0.91, P= 0.99) and NRM (aHR 0.93, P= 0.69). This result was consistent regardless of disease risk. Neutrophil and platelet engraftment were significantly lower (92.2% vs 94.7%, P= 0.042 and 79.8% vs 82.5%, P= 0.003) and the incidence of acute GVHD in the UCB group tended to be higher (gradeⅡ-Ⅳ; aHR 1.64, p= 0.055, grade III-IV; aHR 3.60, P= 0.073) in the UCB group than those in the PTCy-haplo group. However, the incidences of chronic or extensive chronic GVHD after UCB transplant were comparable (chronic; aHR 0.89, p=0.68, extensive chronic; aHR 0.69, P= 0.35) to those after PTCy-haplo transplant. In the subgroup analysis of disease-specific comparison, acute leukemia patients have a significantly lower risk of relapse with UCB transplant (UBC vs PTCy-haplo, HR 0.55, P= 0.030) and lymphoma patients tended to have good results after PTCy-haplo transplant in terms of higher OS (UCB vs PTCy-haplo, aHR 2.13, P= 0.162) and lower relapse (aHR 3.43, P= 0.100). In terms of Cy toxicity in the PTCy-haplo group, transplant with a reduced dose of Cy (80 mg/m 2) tended to show higher OS (2-year OS, 64.8% vs 45.0%) and significantly lower NRM (2-year NRM, 8.0% vs 31.2%) without an increase in gradeⅡ-IV acute GVHD compared to those after standard-dose Cy (100 mg/m 2). Conclusion: Our findings suggest that UCB transplant gives outcomes comparable to PTCy-haplo transplant for patients without an HLA-matched sibling or unrelated donor. Disclosures Kanda: Amgen Astellas BioPharma: Honoraria; Astellas Pharma Inc.: Consultancy, Honoraria; Bristol-Myers Squibb Co: Honoraria; CHUGAI PHARMACEUTICAL Co., Ltd.: Honoraria; DAIICHI SANKYO Co., Ltd.: Honoraria, Membership on an entity's Board of Directors or advisory committees; Eisai: Research Funding; Janssen Pharmaceutical K.K.: Honoraria, Membership on an entity's Board of Directors or advisory committees; Kyowa Kirin Co., Ltd.: Honoraria; Megakaryon Co: Honoraria, Membership on an entity's Board of Directors or advisory committees; NextGeM Inc: Patents & Royalties; Novartis Pharma K.K.: Honoraria; Ono Pharma Inc.: Honoraria; Otsuka Pharmaceutical Co., Ltd.: Honoraria; Sanofi K.K.: Honoraria; Sumitomo Dainippon Pharma Co., Ltd.: Honoraria; SymBio Pharmaceuticals, Ltd.: Membership on an entity's Board of Directors or advisory committees; Takeda Pharmaceutical Company Limited: Honoraria, Membership on an entity's Board of Directors or advisory committees; TEIJIN PHARMA LIMITED.: Honoraria. Imada: Celgene Co., Ltd.: Honoraria; Bristol-Myers Squibb K.K.: Honoraria; Astellas Pharma Inc.: Honoraria; Chugai Pharmaceutical Co., Ltd.: Honoraria; Sumitomo Dainippon Pharma Co., Ltd.: Honoraria; Otsuka Pharmaceutical Co. Ltd.: Honoraria; Takeda Pharmaceutical Co. Ltd.: Honoraria; Novartis Pharma K.K.: Honoraria. Kondo: Asahi Kasei Pharmaceutical: Research Funding; Otsuka Pharmaceutical Co., Ltd.: Honoraria; Chugai Pharma: Honoraria; MSD Pharmaceutical: Honoraria; Sumitomo Dainippon Pharma: Honoraria. Takaori-Kondo: Bristol-Myers K.K.: Honoraria; ONO PHARMACEUTICAL CO., LTD.: Research Funding; Celgene: Research Funding.

Download Full-text