ULTRASOUND EXAMINATION OF BOWEL WALL THICKNESS IN PATIENTS WITH ACUTE INTESTINAL GRAFT-VERSUS-HOST DISEASE

Introduction. Haematopoietic stem cell transplantation is the treatment of choice in many patients with malignant diseases of the blood system. In such patients, acute graft-versus-host disease (GvHD) associated with intestinal damage constitutes one of the most serious complications. However, the volume of stool per day, which is currently used as the main diagnostic criterion for such conditions, does not always permit a timely diagnosis.Aim. To study the possibility of using intestine ultrasound examination for the diagnosis of acute intestinal GvHD.Materials and methods. The study included 50 patients having undergone transplantation of allogeneic haematopoietic stem cells, 40 of whom showed clinical signs of intestinal GvHD (diarrhoea> 500 ml/day). The control group included 10 patients who had undergone transplantation of allogeneic haematopoietic stem cells and exhibited no signs of gastrointestinal events. All patients underwent ultrasound measurement of intestinal wall thickness.Results. Patients were divided into three groups: those with acute GvHD, those with diarrhoea of viral or infectious origin, those with diarrhoea caused by the toxic effects of chemotherapy drugs. It is shown that the walls of all intestinal sections were signifi cantly thicker in patients with acute GvHD as compared to the control group and patients with diarrhoea caused by other reasons.Conclusion. The thickening of the caecum wall (more than 3.25 mm) as detected using the ultrasound method can be used as a diagnostic sign of intestinal GvHD.Conflict of interest: the authors declare no conflict of interest.Financial disclosure: the study had no sponsorship.

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Cotransplantation of Mesenchymal Stem Cells Prompted Platelets Recovery in HLA-Haploidentical/Mismatched Blood and Marrow Transplantation: A Randomized Phase II Study.

Blood ◽

10.1182/blood.v112.11.3284.3284 ◽

2008 ◽

Vol 112 (11) ◽

pp. 3284-3284 ◽

Cited By ~ 1

Author(s):

Kai yan Liu ◽

Xiao Jun Huang ◽

Yu hong Chen ◽

Huan Chen ◽

Lan ping Xu ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Graft Versus Host Disease ◽

Cumulative Incidence ◽

Acute Gvhd ◽

Control Group ◽

Study Group ◽

Graft Versus Host ◽

Matched Sibling ◽

Grade Ii

Abstract Up to now, HSCT has remained the standard and effective therapy for many hematopoietic malignant diseases and some non-malignant hematopoietic diseases. The HLA-matched sibling is the best source of donors; however, such donors are available for only approximately 30% of patients, especially in P.R.China. Virtually almost everyone possesses at least one HLA-haploidentical family donor. However, the use of such donors has presented a major challenge due to graft-versus-host disease (GVHD) and graft rejection. Peking University established a new approach for HLA-mismatched/hapoidentical HSCT using G-CSF primed bone marrow plus G-CSF-mobilized peripheral blood stem cells without ex vivo T cell depletion after the patients were treated with a modified BU/CY2+ATG regimen. Although the result was comparable to that in HLA matched sibling donor transplantation, there are still have room to improve the hemapoeitic/immune reconstitution, reduce the incidence and grade of GVHD further. Mesenchymal stem cell (MSC) is a pluripotent stem cell, supports hematopoeitc progenitors in vitro and posses potent immunosuppressive properties. We hypothesized that cotransplantation of culture-expanded MSCs and hematopoietic stem cells (HSCs) could facilitate engraftment and lessen graft-versus-host disease (GVHD). A phase I study has assessed the safety of contransplantation of MSC, and then a randomized, open-label phase II trial had been conducted in our institution during 2007.6–2008.6. Total twenty-nine patients with leukemia in CR were enrolled in this trial and randomized into study (n=14) or control group (n=15). Three patients in the study group had been found ineligible after randomized: one had relapsed leukemia just before conditioning, another had HLA identical donor and the third one is too overweighed to be infused of enough MSC. The remaining received the G-CSF-mobilized BM and PB from the HLA-mismatched/hapoidentical donor as have been published. Patients in the study group were given culture-expanded MSCs intravenously (3.0–5.0 × 105/kg) from the same donor or health volunteer within 24 hours before infusion of G-BM and PB. Baseline demographic and disease characteristics were balanced between the two groups. MSC infusions were well tolerated, without any infusion-related adverse events. All patients but one in the control group had achieved sustained neutrophil recovery. The median times to neutrophil (absolute neutrophil count ≥0.5 × 109/L) engraftment were 11 days (range, 11–19 days) and 11.5 days (range, 10–23 days) in the study and control groups respectively (p=1.0). There were 2 patients in the control group dead before the platelets engraftment, the median times to platelet recovery (platelet count ≥20 × 109/L ) were 13 days(range 9–52 days) and 19 days (range 10–36 days) (P=0.18). The platelet engraftment was prompted in the study group as for the time to count ≥50 ×109/L (21 days vs 26.5 days, P=0.047). Acute GVHD was observed in 7/11 of patients in study group, 2 had grade I, 5 had grade II. The cumulative incidence of grade II to IV acute GVHD was 48.9%. One patient in the control group had engraftment failure and another was dead of infection on +22 days. Of the thirteen remaining patients, nine patients experienced acute GVHD, 2 had grade I, 6 had grade II and 1 had grade IV. The cumulative incidence of grade II to IV acute GVHD was 61.1%. Our study demonstrated that cotransplantation of culture-expanded MSCs from donor or health volunteer in HLA-haploidentical/mismatched HSCT is feasible and safe. It is seems that MSCs are capable of accelerating hematopoietic cell engraftment, especially for platelet. Further study should be done to reveal whether MSC could reduce the incidence of acute GVHD.

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Mesenchymal Stem Cells for Treatment of Severe Acute Graft-Versus-Host Disease.

Blood ◽

10.1182/blood.v108.11.5304.5304 ◽

2006 ◽

Vol 108 (11) ◽

pp. 5304-5304 ◽

Cited By ~ 6

Author(s):

Katarina Le Blanc ◽

Francesco Frassoni ◽

Lynne Ball ◽

Edoardo Lanino ◽

Berit Sundberg ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Graft Versus Host Disease ◽

Acute Gvhd ◽

Chronic Gvhd ◽

Third Party ◽

Host Disease ◽

Graft Versus Host ◽

Acute Graft

Abstract Mesenchymal stem cells (MSC) from adult bone marrow have the capacity to differentiate into several mesenchymal tissues and inhibit T-cell alloreactivity in vitro. Within the EBMT MSC expansion consortium we have used MSC to treat grades III–IV acute graft-versus-host disease (GvHD) in 40 patients. The MSC dose was median 1.0 (range 0.4–9) 10^6 cells/kg body weight of the recipient. No side-effects were seen after MSC infusions. Nineteen patients received one dose, 19 patients received two doses, two patients received three and five doses respectively. MSC donors were in five cases HLA-identical sibling donors, 19 haploidentical donors and 41 third-party HLA-mismatched donors. Among the 40 patients treated for severe acute GvHD, 19 had complete responses, nine showed improvement, seven patients did not respond, four had stable disease and one patient was not evaluated due to short follow-up. Twenty-one patients are alive between six weeks up to 3.5 years after transplantation. Nine of these patients have extensive chronic GvHD. One patient with ALL has recurrent leukaemia and one patient has denovo AML of recipient origin. We conclude that MSC have immunomodulatory and tissue repairing effects and should be further explored as treatment of severe acute GvHD in prospective randomized trials.

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Mesenchymal Stem Cells for Treatment of Refractory Chronic Graft-Versus-Host Disease.

Blood ◽

10.1182/blood.v110.11.4968.4968 ◽

2007 ◽

Vol 110 (11) ◽

pp. 4968-4968

Author(s):

Weng Jianyu ◽

Xin Du ◽

Xiang Peng ◽

Zhang Xiumin ◽

Suijin Wu ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Graft Versus Host Disease ◽

Acute Gvhd ◽

Chronic Gvhd ◽

Immunosuppressive Agents ◽

Steroid Resistant ◽

Host Disease ◽

Graft Versus Host

Abstract Refractory extensive chronic graft-versus-host disease (GVHD) after allogeneic stem-cell transplantation (SCT) is associated with high mortality [Margolis J., SeminOncol 2000].However, conventional therapies including steroids are often unsuccessful in those patients with multiorgan involvement and are associated with significant therapy-related complications and poorly life quality. Mesenchymal stem cells (MSCs) have immunomodulatory effects [Tse WT et al., Transplantation 2003; Spees JI et al.,Proc Natl Acad Sci USA 2003]. Recently MSCs have been given intravenously to treat seven steroid resistant acute GVHD patients and one patient with chronic GVHD. MSCs effects in chronic GVHD is rarely known, although this successfully experience suggests that MSCs have been well tolerated and had a powerful immunosuppressive effects on acute GVHD. [Katarina Le Blanc et al., Lancet 2004; Olle Ringden., Transplantation 2006 ]. Here, we present our experience of using MSCs for treatment of Thirteen patients with refractory chronic GVHD. Between May 2005 and March 2007, thirteen patients (8 male, 5female) with hematological malignancies with a median age of 26(range:15 to 40) years who had received peripheral stem cells from sibling donors. All patients developed steroid resistant or extensive chronic GVHD, with progressive involvement of the skin(13), liver(10), oral mucosa(12),ocular glands(12), and thrombocytopenia (1) when the immunosuppressive agents were taped after five to twenty-four months. The MSC dose was median 1.0 ×106 cells/kg body weight of the recipient. In all, thirteen patients had at least received one dose, seven patients received more than two doses. MSC donors were in seven cases HLA-identical siblings, six unrelated mismatched donors. No side-effects were seen after MSCs infusions. All patients have responded after follow-up of the median time 15 months. One patient with moderate cGVHD had a complete responses, and discontinued all of the immunosuppressive agents without relapse more than 18.4 months after MSC infusion. Three moderate and two patients with severe chronic GVHD improved to mild degree, and six severe turned to moderate degree. Complete resolution was seen in gut(2/3), liver(5/10), skin(5/13), oral(6/12) and eye(2/12). One patient responded in skin, liver, oral mucose and eye, but developed in lung (bronchiolitis obliterans, BO) score of 2 which are considered severe chronic GVHD. Mean follow-up periods was 27m (rang: 14 to48m), Leukemia free survival(LFS)rate were 85%(11/13), and the overall survival (OS)rate were 92.3%(12/13). Our experience suggests that MSC infusion is a safe and effective adjunct therapy for refractory extensive chronic GVHD with resistance to conventional therapy. But more prospective, controlled studies with MSCs for treatment of GVHD should be performanced to evaluate this new treatment exactly.

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Extracellular Vesicles as Potential Biomarker for Acute Graft-Versus-Host-Disease

Blood ◽

10.1182/blood.v128.22.2239.2239 ◽

2016 ◽

Vol 128 (22) ◽

pp. 2239-2239

Author(s):

Giuseppe Lia ◽

Lucia Brunello ◽

Paola Omedè ◽

Monica Astolfi ◽

Daniela Drandi ◽

...

Keyword(s):

Stem Cells ◽

Extracellular Vesicles ◽

Graft Versus Host Disease ◽

Research Funding ◽

Acute Gvhd ◽

Serum Samples ◽

Hematopoietic Stem ◽

Host Disease ◽

Graft Versus Host ◽

Proportional Change

Abstract INTRODUCTION: Graft-versus-Host-Disease (GVHD) is the main cause of non-relapse mortality after Hematopoietic Stem Cells Transplantation (HSCT). Identyfing potential biomarkers of GVHD could be crucial to define patients at high risk of GVHD development and to better assess GVHD grading. One potential attractive biomarker may be represented by extracellular vesicles (EVs). EVs are membrane-enclosed structures secreted by many cell types and may be represented by exosomes, shedding vesicles (microvesicles; MVs) and apoptotic bodies. EVs are detectable in body fluids, in particular peripheral blood (PB) and urine; EVs play a key role in the regulation of physiological and pathological processes. The aim of this study was to investigate EVs in PB of allotransplanted patients (EVs count, size and phenotype) and analyze a potential correlation with acute and chronic GVHD (aGVHD, cGVHD). METHODS: At our center, between 2000 and 2008, 41 multiple myeloma patients underwent an allograft. Median age of the patients was 53 years (range 34-65). Donors were HLA-identical siblings in 83% of the transplants. Conditioning regimen was mostly non-myeloablative (32/41,78%) and PB stem cells were used as source in all patients. GVHD Prophylaxis consisted of cyclosporine and mycophenolic acid in 34/41 patients (83%). Disease status was partial or less than partial remission in 95% of the patients. For standard policy, serum samples were collected before and monthly after transplant up to 6 months or disease relapse. EVs extraction was feasible due to high EVs stability in frozen serum samples. EVs were extracted using a protamine-based precipitation method and analyzed by flow-cytometry (Guava EasyCyte Flow Cytometer) without using latex-activated beads. We investigated a panel of fluorescence antibodies against specific membrane proteins (CD44, CD138, CD146, KRT18, CD120a, CD8, CD30, CD106, CD25, CD31, CD144, CD86, and CD140a). At each time-point, we determined for each sample: total EVs concentration, fluorescence distribution and percentage of positive EVs for a given marker. Data were computed by logistic regression analysis; Odds Ratio (OR) was calculated as proportional change as compared with pre-transplant baseline level of each marker. RESULTS: In our cohort, the cumulative incidence of aGVHD and cGVHD was 56% (95% 40.7-71.8%) and 71% (95% CI 56.3-85.2%) at day 100 and 24 months, respectively. Four biomarkers (CD146, CD25, CD106, CD31) showed a potential correlation with acute GVHD development. Two biomarkers (CD146 and CD25) were associated with an increased risk of developing aGVHD (CD146 fluorescence, OR 2.94, p=0.040 and CD25 fluorescence, OR 1.61, p<0.001). Furthermore, CD106 positive concentration (OR 0.23, p=0.077) and CD31 fluorescence and positive concentration (OR 0.24, p=0.052; OR 0.50, p=0.067, respectively) were associated with a decreased risk of aGVHD. All the biomarkers associated with aGVHD showed a proportional change from baseline in signal level before GVHD onset (an increase in case of CD146 and CD25, a reduction with CD106 and CD31, respectively). No statistically significant association was observed between our panel of biomarkers and cGVHD. CONCLUSIONS: In this study, we observed a potential association between 4 biomarkers expressed on EVs' surface and aGVHD. CD146, CD31 and CD106 belong to Cell Adhesion Molecule family (MCAM-1, PECAM-1 and VCAM-1, respectively), which are crucial for endothelium and immune cells interaction. CD25 is IL2 Receptor and is a marker of immune activation and inflammation. All these proteins mentioned above, could have a role in acute GVHD pathogenesis. A perspective study is currently ongoing at our Center to validate these data. Disclosures Boccadoro: CELGENE: Honoraria, Research Funding; BMS: Honoraria, Research Funding; Novartis: Honoraria, Research Funding; Abbivie: Honoraria; Mundipharma: Research Funding; SANOFI: Honoraria, Research Funding; Amgen: Honoraria, Research Funding; Janssen: Honoraria, Research Funding.

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Butyrogenic bacteria after acute graft-versus-host disease (GVHD) are associated with the development of steroid-refractory GVHD

Blood Advances ◽

10.1182/bloodadvances.2019000362 ◽

2019 ◽

Vol 3 (19) ◽

pp. 2866-2869 ◽

Cited By ~ 10

Author(s):

Jonathan L. Golob ◽

Martha M. DeMeules ◽

Tillie Loeffelholz ◽

Z. Z. Quinn ◽

Michael K. Dame ◽

...

Keyword(s):

Stem Cells ◽

Graft Versus Host Disease ◽

Acute Gvhd ◽

Chronic Gvhd ◽

Epithelial Monolayer ◽

Graft Versus Host ◽

Key Points ◽

Colonic Stem Cells ◽

Steroid Refractory ◽

Acute Graft

Key Points The presence of butyrogenic bacteria after the onset of acute GVHD associates with subsequent steroid-refractory GVHD or chronic GVHD. Butyrate inhibits human colonic stem cells from forming an intact epithelial monolayer.

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Improved Immune Reconstitution with Low Incidence of Severe Graft-Versus-Host Disease after Transplantation of CD34+ or CD133+ Enriched Stem Cells with Add-Back of Ten Million T-Cells Per Kg.

Blood ◽

10.1182/blood.v104.11.1239.1239 ◽

2004 ◽

Vol 104 (11) ◽

pp. 1239-1239

Author(s):

Johann Greil ◽

Peter Lang ◽

Peter Bader ◽

Matthias Eyrich ◽

Paul G. Schlegel ◽

...

Keyword(s):

Stem Cells ◽

T Cells ◽

Stem Cell ◽

T Cell ◽

Acute Gvhd ◽

Median Number ◽

Control Group ◽

Graft Versus Host ◽

Short Course ◽

Unrelated Donors

Abstract Unrelated donors are commonly used for hematopoietic stem cell transplants, but graft-versus-host disease (GVHD) is a major problem. We investigated whether transplantation of CD34+ or CD133+ enriched stem cells with add-back of ten million T-cells per kg from unrelated donors would prevent acute GVHD in pediatric patients in combination with pharmacologic immunosuppression. Eighteen patients (1 CML in second chronic phase, 2 MDS, 4 ALL in CR1, 4 ALL in CR2, 1 JMML, 1 AML in CR1, 3 AML in CR2, 1 Wiskott-Aldrich syndrome and 1 NHL in CR2) were transplanted with G-CSF mobilized peripheral blood stem cells (PBSC) from HLA-matched unrelated donors (n = 18). Median of age was 8.9 years (0.5 to 18 years). Conditioning regimens were performed according to national therapy protocol guidelines. On the day of transplant patients received a median of 13.5 (4.5 to 30.0 x 106) CD34+ or CD133+ enriched stem cells and an aliquot of unmanipulated PBSC containing 10 x 106 T-cells per kg. GVHD prophylaxis consisted of cyclosporine A (CSA) and short course methotrexate (MTX) on day +1, +3 and +6. Engraftment was rapid with a median of 19.6 days in sixteen patients. Two patients failed to engraft at first. However, full donor chimerism and stable engraftment was achieved in both patients after cessation of CSA treatment and an additional stem cell boost without any reconditioning. One patient with ALL developed acute GVHD grade III (skin and gut) after cessation of CSA treatment, but responded well to treatment with CSA and steroids. None of the other 17 patients developed acute GVHD > grade I. Thirteen are alive and well with a median follow-up of 578 days (101 to 1095 days). Three patients died of severe infectious complications and two due to relapse (JMML, AML). Compared to a historical control group of patients transplanted with highly purified CD34+ selected cells, the group with add-back of 10 x 106 T-cells per kg showed significantly higher T-cell counts (p=0.008, Wilcoxon Rank sum test) on day 90 after transplantation with a median number of six T-cells/μl in the control group and a median number of 294 T-cells/μl in the study group. We conclude that add-back of 10 x 106 T-cells per kg in combination with CSA and short course MTX improves T-cell recovery and appears to be a safe T-cell dosage regarding acute GVHD in the setting of allogeneic peripheral blood stem cell transplantation from unrelated donors.

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Mesenchymal Stem Cells for Treatment of Severe Acute Graft-Versus-Host Disease.

Blood ◽

10.1182/blood.v108.11.2918.2918 ◽

2006 ◽

Vol 108 (11) ◽

pp. 2918-2918 ◽

Cited By ~ 2

Author(s):

Katarina Le Blanc ◽

Francesco Frassoni ◽

Lynne Ball ◽

Edoardo Lanino ◽

Berit Sundberg ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Graft Versus Host Disease ◽

Acute Gvhd ◽

Chronic Gvhd ◽

Third Party ◽

Host Disease ◽

Graft Versus Host ◽

Acute Graft

Abstract Mesenchymal stem cells (MSC) from adult bone marrow have the capacity to differentiate into several mesenchymal tissues and inhibit T-cell alloreactivity in vitro. Within the EBMT MSC expansion consortium we have used MSC to treat grades III-IV acute graft-versus-host disease (GvHD) in 40 patients. The MSC dose was median 1.0 (range 0.4–9) 10^6 cells/kg body weight of the recipient. No side-effects were seen after MSC infusions. Nineteen patients received one dose, 19 patients received two doses, two patients received three and five doses respectively. MSC donors were in five cases HLA-identical sibling donors, 19 haploidentical donors and 41 third-party HLA-mismatched donors. Among the 40 patients treated for severe acute GvHD, 21 had complete responses, eight showed improvement, eight patients did not respond, two had stable disease and one patient was not evaluated due to short follow-up. Twenty patients are alive between six weeks up to 3.5 years after transplantation. Nine of these patients have extensive chronic GvHD. One patient with ALL has recurrent leukaemia and one patient has denovo AML of recipient origin. We conclude that MSC have immunomodulatory and tissue repairing effects and should be further explored as treatment of severe acute GvHD in prospective randomized trials.

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Eosinophilia As a Biomarker Predicting the Occurrence of Chronic Graft Versus Host Disease,

Blood ◽

10.1182/blood.v118.21.4084.4084 ◽

2011 ◽

Vol 118 (21) ◽

pp. 4084-4084

Author(s):

Batia Ronit Avni ◽

Joycelynne Palmer ◽

Tracey Stiller ◽

Stephen J. Forman

Keyword(s):

Graft Versus Host Disease ◽

Acute Gvhd ◽

Immunosuppressive Drugs ◽

Control Group ◽

Host Disease ◽

Graft Versus Host ◽

Gvhd Prophylaxis ◽

Significant Difference ◽

Donor Type ◽

Female Donor

Abstract Abstract 4084 Introduction: Chronic graft-versus-host disease (cGVHD) is a major complication after allogeneic hematopoietic stem cell transplantation (HSCT) known to impair quality of life and functional status and adversely affects long-term survival. Given its impact on outcomes post HSCT, biological markers predicting cGVHD appearance could be useful. Eosinophilia has been reported to occur in the setting of both acute and cGVHD. Even though many physicians refer to eosinophilia as a predictor of GVHD, it is unclear whether eosinophilia can serve as a biomarker predicting occurrence of cGVHD in the general HSCT population. Objectives: In order to evaluate whether eosinophilia can predict the development of cGVHD, we have chosen to study a cohort of HSCT patients who developed eosinophilia while being tapered off their immunosuppressive drugs. Methods: From 01/2000 to 12/2009, a matched series of 112 patients who underwent allogeneic HSCT at City of Hope was identified. Of those, 56 patients presented with eosinophilia (>0.5×10^9 /L on two consecutive visits within 4 weeks, not related to relapse or documented allergy, drug reaction or parasitic infection) after day 120, while being tapered from their immunosuppressive drugs (case group). A control group (with no eosinophilia present on two consecutive visits within 4 weeks after day 120) was matched for the following criteria: age (±10 years), transplant year (± 2 years), HSC source (peripheral blood versus bone marrow), donor type (related vs. unrelated), female donor to male recipient, occurrence of acute GVHD and type of GVHD prophylaxis(tacrolimus/sirolimus vs. other). Results: A total of 112 patients were included in the analysis (Table no. 1). The most prevalent indication for HSCT was acute leukemia. Fifty seven percent of patients underwent myeloablative HSCT. There was no statistically significant difference between the case and control group for the matching criteria: age (p=0.88), HSC source, donor type, female donor to male recipient, occurrence of acute GVHD and type of GVHD prophylaxis (p=1). There was also no statistically significant difference between the groups for the different disease risk category (i.e. low risk, intermediate risk, high risk and non malignant) (p=0.4581). In the group of patients presenting with eosinophilia, 89% (50 out of 56) developed cGVHD, as opposed to 59% (33 out of 56) in the control group (p=0.0002, Chi square test). In this setting, eosinophilia was found to have a positive predictive value of 89%. In a multivariate logistic regression model, looking at the mentioned above known cGVHD risk factors, patients with eosinophilia were nearly 7 times more likely to develop cGVHD compared to those without eosinophila (p=0.0003, 95% CI 2.41–20.13). The median time from the development of eosinophilia to first GVHD sign was 20 days. Eighty four percent of patients both in the eosinophilia and the control groups presented with extensive GVHD grading, with skin, mouth and liver being the most prevalent sites for cGVHD occurrence. With a median follow up of 51 month for the entire cohort (range 5.5–132m) 80% of patients were alive in the eosinophilia group vs. 87% in the control group. Conclusions: The development of eosinophilia in the setting of tapering or discontinuation of immunosuppression may serve as a good predictor of cGVHD. Eosinophilia in this setting may be an indicator for early intervention. Disclosures: No relevant conflicts of interest to declare.

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Enterocytes Of Patients With Uncontrolled Acute Graft Versus Host Disease Of The Gut Undergo Massive Telomere Shortening Compared To Unaffected Controls

Blood ◽

10.1182/blood.v122.21.4467.4467 ◽

2013 ◽

Vol 122 (21) ◽

pp. 4467-4467

Author(s):

Sebastian Hummel ◽

Daniel Heudobler ◽

Elisabeth Huber ◽

Monica S Ventura Ferreira ◽

Dirk Fahrenkamp ◽

...

Keyword(s):

Stem Cell ◽

T Cell ◽

Immunosuppressive Therapy ◽

Graft Versus Host Disease ◽

Acute Gvhd ◽

Control Group ◽

Telomere Shortening ◽

Graft Versus Host ◽

History Of ◽

Acute Graft

Introduction Acute graft versus host disease of the gut (agGVHD) is caused by a T-cell mediated attack against the gut epithelia and remains one of the complications with the highest mortality after allogenic stem cell transplantation (allo-SCT). In particular, patients not responding to the initial immunosuppressive therapy during the first weeks have a poor outcome and frequently do not show complete recovery of their symptoms. The underlying mechanisms of refractory agGVHD are not fully understood. Telomere length (TL) reflects the replicative history of a cell. Reaching a critically short TL, cells stop dividing and enter cellular senescence. The aim of the study was to elucidate the role of telomere-mediated replicative exhaustion of enterocytes in affected patients with agGVHD compared to unaffected control patients after allo-SCT. Methods The study was carried out in a single-blinded fashion including 13 patients with refractory agGVHD and 7 patients undergoing allo-SCT, but without clinical history of GVHD. Proliferation score of gut enterocytes was assessed in a semi quantitative manner using MIB-1 immunohistochemistry staining. The TL of the enterocytes obtained from paraffin-embedded diagnostic colonoscopy/sigmoidoskopy biopsies were analyzed using confocal quantitative telomere FISH (cqFISH). Patients were matched according to age, clinical characteristics and onset of GVHD. Results Patients with agGVHD showed a significantly increased proliferation score (median: 2.0, range: 0.5-3.0, n= y, p=0.04) compared to the control group (median: 1.0, range: 0.5-2.0, n=y). TL of initial samples obtained during the first 100 days after transplantation did not differ significantly between patients with (mean±SE: 8.1±0.8 kb, n=13) and without GVHD (7.4±1.4 kb, n=7, p=0.65). Of 10 patients, longitudinal analysis was carried out. Five out of six patients with acute GVHD showed massive telomere shortening during the first weeks after development of GVHD (-2.9 kb, p=0.03), whereas no changes were observed in unaffected controls (-0.1 kb, n=3, p=0.75). This translates into a loss of 0.209 kb per week in patients with acute GVHD, compared to 0.004 kb in the control group. Conclusion In this preliminary study, we found that enterocytes of patients with refractory acute gut GVHD show increased proliferation and dramatically shortened TL in affected gut endothelium during the course of the disease. Increased cellular turnover due to T-cell mediated attack on the enteral stem cell pool and subsequent replicative exhaustion might provide a mechanism explaining eventual refractoriness of gut GVHD to immunosuppressive therapy. Based on our findings, TL measurement should be further explored prospectively as a prognostic biomarker in patients with agGVHD. Disclosures: No relevant conflicts of interest to declare.

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IL10-Transduced Mesenchymal Stem Cells Attenuate Experimental Acute Graft-Versus-Host Disease.

Blood ◽

10.1182/blood.v106.11.5247.5247 ◽

2005 ◽

Vol 106 (11) ◽

pp. 5247-5247

Author(s):

Chang-Ki Min ◽

Bo-Kyung Kim ◽

Gyeongsin Park ◽

Bin Cho ◽

Il-Hoan Oh

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Graft Versus Host Disease ◽

Murine Model ◽

Acute Gvhd ◽

Serum Levels ◽

Host Disease ◽

Graft Versus Host ◽

Adult Mesenchymal Stem Cells ◽

Immunosuppressive Microenvironment

Abstract Recent data suggest that, due to their immunosuppressive nature, adult mesenchymal stem cells (MSCs) may be of interest to enhance allogeneic hematopoietic engraftment and prevent graft-versus-host disease (GVHD). Using a murine model of acute GVHD, this study investigated whether the immunosuppressive properties of MSCs could reduce the severity of experimental GVHD. In a MHC-mismatched C57BL/6→B6D2F1 model, recipient animals were transplanted with bone marrow (BM) cells (10x106) plus 20x106 splenocytes after 1100 cGy on day 0. Various doses (1~3x106) of donor BM-derived MSCs were given intravenously from day 1 to day 5 and subsequently evaluated the clinical and immunologic parameters. MSCs did not attenuate the severity of acute GVHD. Using GFP expressing MSCs, we were unable to detect labeled cells in liver, spleen, lymph nodes and lung day 7 post-BMT. Because MSCs fail to display any immunosuppression in this experimental GVHD model and induce an immunosuppressive microenvironment through the production of IL10, we investigated whether the use of genetically modified MSCs expressing IL10 (IL10-transduced MSCs, IL10 MSCs) could improve the GVHD protection. Lethally irradiated recipients were transplanted and injected with 2x106 IL10 MSCs, MSC expressing MIG as a vector (MIG MSCs), or diluent on day 1. Compared with MIG MSCs or controls, recipients of IL10 MSCs demonstrated significantly reduced mortality at day 50 after BMT (percent survival, 0% or 10% vs 70%, P<0.001). The reduction in mortality was confirmed by the semi-quantitative GVHD score (P<0.01). It was associated with decreased serum levels of pro-inflammatory cytokines, IFNγ on day 7 (IL10 MSCs vs MIG MSCs; 297±116 pg/ml vs 681±87 pg/ml, P=0.015). Serum levels of TNFα or splenic donor CD3+ cell numbers were not different between the groups. Thus, benefical effects on GVHD could be observed when MSCs were engineered to express an anti-inflammatory cytokine, IL10. The mechanisms for the GVHD protection effect of IL10 MSCs in this murine model are currently under investigation.

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