Terms Of Graft-versus-host Disease Research Articles

No Impact of Donor's Age-Related Clonal Hematopoiesis (ARCH) Observed on Graft-Versus-Host Disease Following Allogeneic Hematopoietic Stem Cell Transplantation: Result from Bar-Coded Error Corrected Sequencing in 33 Gene Mutations on 372 Pairs of Donor and Recipient

Introduction: Recent advances in next-generation sequencing (NGS) enables us to reveal the clinical relevance of age-related clonal hematopoiesis (ARCH) in healthy individuals without having hematologic disorders. It is plausible that transfer of ARCH from a donor to the recipient following allogeneic hematopoietic stem cell transplantation (HCT) results in delayed engraftment, thus increasing the risk of non-relapse mortality (NRM). A recent study reported that the presence of ARCH in the donor is associated with increasing risk of chronic graft-versus-host disease (GVHD) and reduced risk of relapse after allogeneic HCT (JCO 2018). However, detailed analyses are lacking with respect to engraftment, the severity of GVHD, organ-specific GVHD. The present study aimed to assess the impact of donor ARCH on HCT outcomes regarding overall and each organ-specific acute/chronic GVHD as well as engraftment kinetics. Methods and patients: The present study includes 372 pairs of donors and/or recipients who received HCT from 2000 till 2007. Genomic DNA samples were archived from the samples taken prior to HCT. For barcoded error-corrected sequencing, a molecular inversion probe capture protocol was modified and applied (Gen. Research 2013). In short, we designed probes targeting 33 genes covering ARCH mutations along with other AML related mutations. Barcoded NGS library was generated and processed. Following sequencing (NovaSeq, Illumina) with 150 bp pair-end mode, the analysis was performed using a customized computational pipeline. Recipients and transplant characteristics are as following: male/female (n=223/149); median age, 48 yrs; conditioning regimen, reduced-intensity conditioning (RIC, n=105)/myeloablative conditioning (MAC; n=267). Median age of donors was 48 yrs. Transplant outcomes were analyzed according to the presence of ARCH in the donor in terms of overall survival (OS), NRM, relapse incidence (RI), overall acute and chronic GVHD as well as organ-specific incidences of GVHD. Engraftment parameters such as neutrophil and platelet recovery were also analyzed. Results: 30 mutations were detected in 25 donor samples (6.7%). Consistent with previous studies, the most frequently mutated gene was DNMT3A (n=10/25, 40%). The median variant allele frequency (VAF) for 30 mutations in donor was 1.18% (ranges, 0.57%-13.29%). With respect to the presence or absence of ARCH in donors, there were no significant differences in baseline and transplant characteristics such as age, sex, conditioning regimen (RIC vs MAC), GVHD prophylaxis. The presence of ARCH in donor also did not show any difference in OS (p=0.66, HR 0.86 [0.44-1.68]), RI (p=0.71, HR 0.83 [0.31-2.21]) or NRM (p=0.7, HR 1.16 [0.54-2.51]). With respect to the engraftment, the median time to neutrophil (defined as ≥0.5x109/L) and platelet engraftment (≥20x109/L with transfusion independence) was not associated with the presence of donor ARCH. Neutrophil engraftment rate was 88.0% in ARCH and 91.6% in the non-ARCH group at day 30 with a median time of 18 days and 19 days (p=0.45). The median time to platelet engraftment was 15 days both in ARCH and in the non-ARCH group (p=0.43). In terms of GVHD, donor ARCH status did not affect incidences of aGVHD/cGVHD. Incidence of grade II to IV acute GVHD at day 100 was 69.9% and 66.2% in the ARCH and non-ARCH group (p=0.94). There were no differences between the two groups in organ-specific aGVHD of the skin (p=0.23), liver (p=0.98) or gut involvement (p=0.48). The cumulative incidence of chronic GVHD (based on the NIH consensus criteria) was 58.3% in ARCH and 58.6% in the non-ARCH group (p=0.80). No differences were noted in the severity of cGVHD between the 2 groups (p=0.87). No differences in organ-specific cGVHD were also noted for the ocular, oral, skin, liver, GI, lung or musculoskeletal involvement. Multivariate analyses confirmed that the donor ARCH was not significantly associated with survival, a/cGVHD and, engraftment after allogeneic HCT. Conclusion: The presence of ARCH in the donor does not affect the mortality, relapse or engraftment following allogeneic HCT. Our data suggest that donor ARCH has no apparent association with HCT outcomes, although there was a concern on the presence of ARCH in the donor's HSCs and its transfer through allogeneic HCT. It also does not seem to delay engraftment or increase the risk of non-relapse mortality following allogeneic HCT. Figure Disclosures Michelis: CSL Behring: Other: Financial Support. Mattsson:Gilead: Honoraria; Therakos: Honoraria; Celgene: Honoraria.

Influence of CD34+ and CD3+ Graft Content on Gvhd Development after Haploidentical Allogeneic Transplantation with Post-Transplant Cyclophosphamide

Introduction:CD34+ and CD3+ cell dose in peripheral blood stem cell (PBSC) graft have been related with increased incidence of graft versus host disease (GVHD) in some transplant settings. Our aim in this study was to evaluate the impact of CD34+ and CD3+ cells graft composition on GVHD incidence in the setting of haploidentical transplant (haplo-HSCT) with post-transplant cyclophosphamide (PT-Cy), in a multicenter analysis.Materials (or patients) and methods:We retrospectively evaluated 175 patients with hematologic malignancies who were treated with a haplo-HSCT from 2011 to 2014 with PBSC in GETH centers. All patients received Busulfan-Fludarabine as conditioning regimen and GVHD prophylaxis was performed with PT-Cy (50 mg/kg on days +3 and +4) and a calcineurin inhibitor plus mycophenolate from day +5. We analyzed the impact of CD34+ and CD3+ cell doses (low vs. high with cut-off at the median value, and the 25-75 percentiles) on the development of acute and chronic graft versus receptor disease. Death before GVHD development was considered as a competitive event. The analysis was adjusted for conditioning intensity. STATA software was used for data analysisResults:We analyzed 175 patients. Patient characteristics are resumed in table 1. Median CD34+ and CD3+ cell dose was 5.31x10e6/kg (p 25-75: 4.31-6.1) and 2.07x10e8/kg (p25-75: 1.19-2.94), respectively. Cumulative incidences of grade II-IV acute GVHD and all grades chronic GVHD was 32.6% and 30% respectively. The median follow-up was 12.3 months (r:6-46). No difference in terms of GVHD (acute and chronic), was found between low and high CD34+ cell doses. However, if we focus on T cells, high doses of CD3+, for values above the median, have been has been associated with increased incidence of acute GVHD II-IV (day 150: 47% vs 26% p=0.01, figure 2), as well as cGVHD (30 month: 39% vs 16%, p= 0.01 figure 1). If we focus the analyses regarding the intensity of conditioning regimen, the influence of CD3+ doses remains for the group of reduced-intensity conditioning (RIC) (aGVHD, day 150: 54% vs 24%, p=0.003; cGVHD, 30 month: 47% vs 16%, p=0.006 figure 2), but lost for patients who receive myeloablative intensity (aGVHD, day 150: 33% vs 33% p=0-8; cGVHD, 30 month: 21% vs 16%, p=0.81)Conclusion:In our series, in the setting of haplo-HSCT with Pt-Cy, the highest dose of lymphocytes was associated with a higher incidence of aGHVD II-IV and cGV HD, especially for the RIC procedures. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

The role of gamma delta T cells in haematopoietic stem cell transplantation.

Although haematopoietic stem cell transplantation (HSCT) is a potential curative treatment for haematological malignancies, it is still a procedure associated with substantial morbidity and mortality due to toxicity, graft-versus-host disease (GVHD) and relapse. Recent attempts of developing safer transplantation modalities increasingly focuses on selective cell depletion and graft engineering with the aim of retaining beneficial immune donor cells for the graft-versus-leukaemia (GVL) effect. In this context, the adoptive and especially innate effector functions of γδ T cells together with clinical studies investigating the effect of γδ T cells in relation to HSCT are reviewed. In addition to phospho-antigen recognition by the γδ T cell receptor (TCR), γδ T cells express receptors of the natural killer (NK) and natural cytotoxicity (NCR) families enabling them to recognize and kill leukaemia cells. Antigen recognition independent from the major histocompatibility complex (MHC) allows for the theoretical possibility of mediating GVL without an allogeneic response in terms of GVHD. Early studies on the impact of γδ T cells in HSCT have reported conflicting results. Recent studies, however, do suggest an overall favourable effect of high γδ T cell immune reconstitution after HSCT; patients with elevated numbers of γδ T cells had a significantly higher overall survival rate and a decreased rate of acute GVHD compared to patients with low or normal γδ T cell counts. Further research in terms of effector mechanisms, subtypes and tissue distribution during the course of HSCT is needed to assess the potentially beneficial effects of γδ T cells in this setting.

The effect of the CCR5-delta32 deletion on global gene expression considering immune response and inflammation

BackgroundThe natural function of the C-C chemokine receptor type 5 (CCR5) is poorly understood. A 32 base pair deletion in the CCR5 gene (CCR5-delta32) located on chromosome 3 results in a non-functional protein. It is supposed that this deletion causes an alteration in T-cell response to inflammation. For example, the presence of the CCR5-delta32 allele in recipients of allografts constitutes as an independent and protective factor associated with a decreased risk of graft-versus-host disease (GVHD) and graft rejection. However, the mechanism of this beneficial effect of the deletion regarding GVHD is unknown. In this survey we searched for a CCR5-delta32 associated regulation of critical genes involved in the immune response and the development of GVHD.MethodsWe examined CD34+ hematopoietic progenitor cells derived from bone marrow samples from 19 healthy volunteers for the CCR5-delta32 deletion with a genomic PCR using primers flanking the site of the deletion.Results12 individuals were found to be homozygous for CCR5 WT and 7 carried the CCR5-delta32 deletion heterozygously. Global gene expression analysis led to the identification of 11 differentially regulated genes. Six of them are connected with mechanisms of immune response and control: LRG1, CXCR2, CCRL2, CD6, CD7, WD repeat domain, and CD30L.ConclusionsOur data indicate that the CCR5-delta32 mutation may be associated with differential gene expression. Some of these genes are critical for immune response, in the case of CD30L probably protective in terms of GVHD.

High Response Rate and Improved Graft-Versus-Host Disease (GVHD) Following Bortezomib as Salvage Therapy after Reduced Intensity Conditioning Allogeneic Stem Cell Transplantation (RIC allo-SCT) for Multiple Myeloma (MM).

Bortezomib has been shown to be an efficient treatment for MM. Similarly, a potent allogeneic graft-versus-myeloma effect can be induced against MM. However, despite progress in terms of transplant-related mortality, a significant proportion of patients may still relapse or progress after RIC-allo-SCT. This retrospective report describes the results of 37 MM patients who received bortezomib (1.3 or 1.0 mg/m2 intravenously, days 1, 4, 8, and 11) as a salvage therapy after progression following RIC-allo-SCT. Median age was 49 years. All patients (100%) received and failed at least one autologous stem cell transplantation prior to RIC allo-SCT. Median time between the initial diagnosis of MM and allo-SCT was 11 (range, 6–96) months. Patients were not treated in a combined autologous/allo-SCT strategy. Before bortezomib initiation, 14 patients (38%) received and failed DLI. Also, 20 patients (54%) received and failed thalidomide (because of disease progression or toxicity). Overall, 32 patients (86%) received bortezomib salvage in progressive disease, while 5 patients (14%) were in PR. Median time between allo-SCT and bortezomib initiation was 20 (range, 1–65) months. At time of bortezomib initiation, the majority of patients (n=26; 70%) did not have any symptoms of chronic GVHD, while 8 patients (22%) had some form of limited chronic GVHD, and 3 patients (8%) had extensive signs. The median number of bortezomib cycles administered was 6 (range, 1–15). Peripheral neuropathy was frequently observeded after bortezomib (n=13; 35%; 4 grade 2 and 9 grade 1). Mild thrombocytopenia not requiring platelets transfusions was observed in 9 cases (24%). Fatigue was also observed in 7 patients (19%). None of the patient had to discontinue the treatment because of a life-threatening adverse event, and no treatment-related toxic deaths were observed. In terms of GVHD, patients did not experience reactivation or worsening of GVHD symptoms. Interestingly, two patients among the three patients with extensive GVHD signs at the beginning of bortezomib experienced a significant improvement and were staged as limited chronic GVHD at last follow-up. In all, 27 patients (73%; 95%CI, 59%–87%) achieved an objective disease response after starting of bortezomib (7 CR, 7 VGPR, and 13 PR). Prior use of thalidomide and/or DLI did not influence disease response to bortezomib. There was also no difference in disease response when using bortezomib in combination or without dexamethasone. With a median follow-up of 9 (range, 3–42) months from bortezomib initiation, 25 patients (68%; 95%CI, 53%–83%) still had a sustained objective disease response (5 CR, 5 VGPR, and 15 PR). Ten patients (27%) died and 27 are still alive with a median overall follow-up after allo-SCT of 80 (range, 18–153) months. The majority of deaths were directly attributed to disease progression (n=8; 80% of all deaths). Most importantly, patients achieving an objective disease response (CR, VGPR or PR) after introduction of bortezomib enjoyed a significantly higher overall survival as compared to non-responding patients (P=0.002). Collectively, these data demonstrate that bortezomib is a safe and potentially efficient option for MM patients failing RIC allo-SCT.

DPB1 disparities contribute to severe GVHD and reduced patient survival after unrelated donor bone marrow transplantation.

In order to evaluate the impact of HLA-DBP1 incompatibilities on the occurrence of acute graft-versus-host disease (GVHD) in unrelated hematopoietic cell transplantation, we studied 57 donor/recipient pairs characterized by their allelic identity for HLA-A, B, C, DRB1 and DQB1 and also for DRB3, 4, 5 loci and aimed to correlate DPB1 mismatches to already described risk factors for GVHD using multivariate Cox regression analysis. DPB1 identity between donor and recipient was observed in 24% and DPB1 compatibility (GVHD vector) in 42%. Two factors were independently associated with severe acute GVHD: two DP incompatibilities (RR = 8.25, 95% confidence interval (CI): 1.67-40.10, P = 0.010) and disease risk (RR = 10.23, 95% CI: 1.12-93.13, P = 0.012). Two DPB1 incompatibilities appeared also to be a factor in poorer survival independent of its effect on acute GVHD (RR = 4.97, 95% Cl: 1.80-13.71, P = 0.002). A correlation between acute GVHD and matching for each individual DPB1 polymorphic region and for residue 69 of the DP beta molecule, which seems to be a key residue in the alloimmune response, was not observed. Our data indicate that the outcome of unrelated hematopoietic cell transplantation in terms of GVHD but also survival, could be improved through HLA-DPB1 matching or at least by avoiding two DPB1 mismatches.