Dynamics of Chimerism In Regulatory T Lymphocytes (Treg; CD4+/CD25+) After Allogeneic Stem Cell Transplantation

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 1328-1328
Author(s):  
Víctor Noriega ◽  
Carolina Martinez-Laperche ◽  
David Serrano ◽  
Gabriela Rodriguez-Macias ◽  
Mi Kwon ◽  
...  

Abstract Abstract 1328 Introduction: CD4+CD25+ regulatory T-cells (Treg) play an important role in inducing and maintaining allogeneic tolerance and can inhibit graft-versus-host disease (GVHD) after allogeneic stem cell transplantation (SCT). However, the dynamics of donor and recipient Treg cell populations after Allo-SCT has not been studied yet. Objective: To analyze the dynamics of chimerism in Tregs and compare it to that of T lymphocytes (TL; CD3+) and whole blood leukocytes (WBL). Material and Methods: The study includes 53 patients subjected to Allo-SCT (myeloablative and non-myeloablative). PB samples were obtained weekly during the first month and every 14 days after day +30 until complete chimerism (CC) was achieved, and at fixed time-points (+30, +60, +90, +180, +365) thereafter. TL and Tregs were purified from PB until CC was achieved using inmunomagnetic technology (Miltenyi Biotec; CD3+ Microbeads and CD4+/CD25+ Treg Isolation Kit, respectively). Chimerism analysis was performed by microsatellite PCR (STR-PCR; AmpFlSTR SGM Plus; Applied Biosystems) on genomic DNA obtained from WBL as well as on cell lysates obtained from purified TL and Tregs. Complete chimerism was considered in samples with percentage of recipient cells <1% (sensitivity of the STR-PCR) for WBL samples and <5% (minimum purity of 95% as estimated by flow cytometry) for purified cell lineages. Results: Median follow up for the whole cohort of patients was 338 days. CC was spontaneously achieved in WBL, TL and Tregs in 45/53 patients (85%) in a median time of 35.41, 38.8 and 42.5 days respectively. 5/3 patients (9%) suffered from graft failure/rejection showing mixed chimerism (MC) with increasing percentages of recipient cells both in WBL and leukocyte lineages. 3/53 patients (6%) maintained mixed chimerism (MC) in WBL and leukocyte lineages at day +180, with no signs of graft rejection or disease relapse. Analysis of the chimerism dynamics of those patients who spontaneously achieve CC revealed two different groups: Group 1 included 25/45 patients who achieved CC at the same time in WBL, TL and Tregs. Group 2 included 20/45 patients who achieved CC in WBL while maintaining MC in TL and Tregs. Interestingly enough, 9/20 patients from Group 2 maintained MC in Tregs 7–75 days after achieving CC in both WBL and TL (Figure 1). In a preliminary analysis, the small sample size precluded from obtaining statistically significant associations between the dynamics of chimerism in Tregs and the development of complications such as relapse or GVHD after SCT. Conclusions: To our knowledge, this is the first study dealing with Treg chimerism after SCT. We have shown it is feasible and can be performed on a routine basis together with standard lineage specific chimerism follow up. Although there is an association between chimerism dynamics in Tregs and TL, this is not absolute and a percentage of patients maintain residual Tregs of recipient origin after WTL and TL have become of complete donor origin. In this small cohort, Treg chimerism did not influence the development of post-SCT complications. Analysis of a larger and more homogeneous cohort would allow establishing the usefulness of Treg chimerism testing for the management of transplanted patients. Disclosures: No relevant conflicts of interest to declare.

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 4460-4460
Author(s):  
Paolo Anderlini ◽  
Sandra Acholonu ◽  
Grace-Julia Okoroji ◽  
Sergio A Giralt ◽  
Roy Jones ◽  
...  

Abstract Abstract 4460 Background: The role of DLIs in the management of recurrent HL following allo-SCT is not well established. Response rates in the 30–50% range have been reported (as high as 79% in a recent report), but their durability and impact on patient survival are not always clear (Peggs et al, BJH 2008; 143:468 and JCO 2011; 29: 971). We wish to update our initial report on this issue (Anderlini et al, BMT 2004; 34: 511). For more details on response criteria definitions, please refer to our original report. Patients and Methods: Between 1999 and 2010, 27 consecutive patients with relapsed/refractory HL following unmanipulated allo-SCT received a total of 55 DLIs as immunotherapy for treatment of progressive disease (PD). Their median age was 30 years (19–60; M/F 19/8), and 21/27 (78%) had a history of prior autologous SCT. Seventeen patients received more than one DLI (range 2–5). Seventeen had a matched sibling/parent donor and ten a matched unrelated donor. In all but two cases the conditioning regimen included fludarabine plus cyclophosphamide or melphalan plus/minus antithymocyte globulin. The median time to PD after allo-SCT was 5 months (range 1–21). In ten patients (37%) prior salvage chemotherapy was administered prior to at least one of their DLIs. The pre-DLI chimerism status was 2/27 mixed and 25/27 full donor. Results: Ten of 27 (37%) patients had a complete/partial response (CR/PR) following at least one of their DLIs. Six of 27 patients (22%) achieved CR/CRU (complete response, unknown), and four of 27(15%) achieved a PR. The median response duration was 7.5 months (range 0.5–20). Of all these ten responders, 100% developed graft-vs-host disease (GVHD) and half of them (50%) had received concomitant chemotherapy. Of the ten patients who received only one DLI, two (20%) had chemotherapy prior to their DLIs. One (10%) had a CRU and the other had stable disease (SD) (10%). Of the remaining eight that did not have chemo prior to DLI, 2 (20%) had CRU, 3 (30%) had PR, 2 (20%) had SD and 1 (10%) had PD. The median CD3+ cell dose administered was 49.8 × 10E6/ kg (range 0.05–285). GVHD developed (or flared) following the DLI in 45/55 cases (82%). After the DLI mixed chimerism (n=2) was converted to complete (or near-complete) donor chimerism. At the latest follow-up (March 2011), five patients (18%) are alive (two in CR). For these five survivors, the follow-up after the 1st DLI is 42 months (range 4–63). Four of them (80%) did not receive chemotherapy with their DLI(s), while one did. Twenty-two patients expired. Their median survival after the 1st DLI was 14 months (range 4–64). Causes of death included PD (n=15; 68%) and non-relapse mortality (n=7; 32%). Conclusions: Despite the limitations related to the small sample size, patient heterogeneity and concomitant chemotherapy administration, these data suggest that (a) DLIs for immunotherapy of recurrent HL following allo-SCT have significant (albeit not always durable) activity, and a subset of patients become long-term survivors (b) administration of multiple DLIs is feasible in selected patients (c) responses are associated with the development of GVHD (d) the role (if any) of concomitant chemotherapy is unclear and, lastly (e) PD remains the main cause of mortality following DLIs. The role of DLIs should be reassessed in the contest of new and effective agents now available in the salvage setting (SGN-35, panobinostat, etc). Disclosures: Off Label Use: Fludarabine, cyclophosphamide and melphalan as part of conditioning regimen for allogeneic stem cell transplantation.


Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 4515-4515
Author(s):  
Patrycja Zielinska ◽  
Malgorzata Krawczyk-Kulis ◽  
Miroslaw Markiewicz ◽  
Monika Dzierzak-Mietla ◽  
Anna Koclega ◽  
...  

Abstract Abstract 4515 Chronic lymphocytic leukemia (CLL) is an incurable disease when treated with standard chemotherapy. The only possibility to provide cure is allogeneic stem cell transplantation (allo-SCT). CLL patients aged less than 55 account for about 15% of patients and these cases allo-SCT should be taken into consideration. The indications for allo-SCT are as follows: del17p, resistance to chemoimmunotherapy, Richter’s syndrome or recurrent disease. A retrospective analysis of allo-SCT in 18 patients (10 males, 8 females) with CLL transplanted in years 2000–2010 was performed. The aim of the study was to assess of long term follow-up outcome of allo-SCT in CLL patients. The median age at diagnosis was 41ys (range: 35–51). The sibling donor was available in 16 cases (2 pts were mismatched), unrelated donors were in 2 cases (1 mismatched). Most of the pts (16 out of 18) were MRD positive when allotransplanted. Median lymphocytosis preceeding allo-SCT was 5.9G/l. Peripheral blood was the source of stem cells in 9 cases (50%), and bone marrow in the remaining 9 cases, 2 pts were transplanted with stem cells from bone marrow and peripheral blood. 4 pts (22%) underwent the allograft procedure twice or more. Reduced intensity conditioning with alemtuzumab was performed in 9 pts (50%), myeloablative regimen in 4 cases and RIC with rituximab in one case.The median number of CD34+cellsx10^6/kg was 4.1 (range: 0.86–9.64). All but one patient engrafted (this pt was transplanted again successfully in one year time). Acute graft-versus host disease (GvHD) was noted in 46% of pts (only in 2 pts grade IV). Extensive GvHD was observed only in 2 pts. Donor lymphocyte infusion (DLI) was performed in 8 pts (44%). With a median follow-up of 73 months (range: 9–89) for surviving patients, the five-year Kaplan-Meier of overall survival (OS) and progression free survival (PFS) was 55,5% and 34%, respectively. At five years, the cumulative probability of non-relapse mortality was 15%. Allogeneic stem cell transplantation remains the effective treatment in CLL for selected group of patients. Disclosures: No relevant conflicts of interest to declare.


Leukemia ◽  
2002 ◽  
Vol 16 (1) ◽  
pp. 13-21 ◽  
Author(s):  
N Schaap ◽  
A Schattenberg ◽  
E Mensink ◽  
F Preijers ◽  
M Hillegers ◽  
...  

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 5866-5866
Author(s):  
Yannick Le Bris ◽  
Berger Florian ◽  
Audrey Menard ◽  
Thierry Guillaume ◽  
Pierre Peterlin ◽  
...  

Abstract Introduction: After allogeneic stem cell transplantation (allo-SCT), engraftment can be assessed by quantitative polymerase chain reaction (qPCR) using differing donor/recipient markers (Alizadeh et al. Blood, 2002), identified in peripheral blood DNA cells before transplant. We report here on the concomitant examination of the proportions of donor and recipient DNA in peripheral whole blood (WB) and sorted CD3+ T-cells on days +60 and +90, looking at their impact on survival. Patients, material and methods: This monocentric study evaluated the impact on outcomes of early WB and sorted CD3+ T cells chimerism independently and of the four possible combinations of chimerism between WB and sorted CD3+ T-cells. All follow-up chimerism samples from allo-SCT patients performed in adults at Nantes University Hospital between October 2009 and October 2016 were reviewed, focusing on those where both PB and/or CD3+ T-cells were evaluated on days +60 (45-75) and/or +90 (75-120) after allo-SCT. A global cohort of 229 patients (239 grafts) was retrieved, which includes 52 patients evaluable on day +60 only, 67 evaluable on day +90 only and 120 evaluable on both days +60 and +90. A threshold of >95% donor DNA was considered for complete chimerism. Disease free survival (DFS) was calculated from the date of graft until relapse, death or last follow-up. Overall survival (OS) was calculated from the date of graft until death or last follow-up. Chi square tests were used to compare incidences. Log rang test and Kaplan Meier were used to evaluate DFS and OS. Results: The whole cohort comprised 62% males and had a median age of 58 years old (20-74) at the time of allo-SCT. Patients were treated for myeloid-lineage disease in 59% of the cases. Reduced-intensity conditioning was used in 89% (n=212), donors were familial in 45% (n=107), registry in 48% (n=114). Unrelated cord blood units were used in 8% of the cases (n=18). Post-transplant cyclophosphamide (PTCY) was performed in 48 procedures including 33 and 15 with haplo (HG) and matched donors respectively. Considering the 239 allograft procedures, the median follow-up was 5.8 years (95% CI: 3.1-5.8), the rate of relapse 27% and the rate of death 31%. Complete WB chimerism was observed for 80% and 71% of the cases on day +60 and day +90 respectively. Complete CD3+ chimerism was present for 53% and 51% of the grafts at days +30 and +90 respectively. Thus, cases displaying both complete WB and CD3+ chimerism on days +60 and +90 were 53% and 51% respectively, while 27% and 20% were documented with full WB and mixed CD3+ chimerism on days +60 and +90. Mixed chimerism was observed in both WB and CD3+ cells in 14% of the cases on day +60 and 22% on day +90. Finally, a small proportion of patients (6% and 7% at days +60 and +90) displayed an intriguing complete chimerism in CD3+ cells yet mixed WB chimerism. None of these features appeared associated to disease lineage (lymphoid or myeloid) nor cord blood allo-SCT. Interestingly, of the 27 grafts with myeloablative conditioning, only 14 had full WB/CD3+ engraftment on day +60 or +90, and thus all 27 were retained for the study. None of the four WB/CD3 chimerism combinations at the two times considered had an impact on DFS in this cohort. Surprisingly, although full or mixed WB chimerism had no impact on DFS and OS at days +60 and +90, the presence of a mixed CD3+ chimerism (vs full) at day+90 was associated with a significantly better OS (median: 5.8 months years [95%CI: -not reached] versus 3.1 years [95%CI: 2.2- 3.1]; p=0.025). CD3+ chimerism at day+60 has no impact on OS. All HG resulted in full CD3+ chimerism at both time points compared to non HG (100% vs 52%, p<0.0001). The same was almost true when considering PTCY procedures: 90% at day+60 and 92% at day +90. Of note, there was no influence on DFS nor OS of WB or CD3+ chimerism status when considering only HG or PTCY grafts vs others in this series. Discussion: In this large series, early WB chimerism status did not predict outcome. Surprisingly, mixed CD3+ chimerism at day+90 appears to be significantly associated with a longer OS, suggesting that remaining recipient memory lymphocytes could be beneficial. This result has to be confirmed prospectively. It remains also to define the place of donor lymphocyte infusions (DLI) to prevent relapse in patients with full or mixed CD3+ chimerism post-transplant (analyses of DLI received in our patients are on-going). Disclosures Moreau: Bristol-Myers Squibb: Honoraria; Takeda: Honoraria; Novartis: Honoraria; Amgen: Honoraria; Janssen: Honoraria, Speakers Bureau; Celgene: Honoraria.


Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4388-4388
Author(s):  
Nicolas Novitzky ◽  
Cecile E. Du Toit ◽  
Andrew B. McDonald

Abstract Introduction: The optimal schedule for the treatment of AML remains to be defined. No systematic analysis of the dose intensity of anthracycline agents during induction chemotherapy has been published. We have compared the outcome of 2 consecutive protocols where daunorubicin was given at two different dose intensities. Methods and patients: Consecutive patients with AML younger than 60 years and who had adequate organ function and were HIV non reactive were entered into a prospective study. Induction therapy included etoposide 100 mg/m2 (for 7 days), cytarabine 100 mg/m2 (as infusion for 7 days) and daunorubicin 75 mg/m2 (for 3 days) followed by one similar consolidation course where daunorubicin was reduced to 60 mg/ (for 3 days) (Group 1). Patients remaining in CR were offered autologous or allogeneic stem cell transplantation (if HLA donor available). They were compared to a previous cohort (Group 2; 1993–1998) who had received similar combination therapy but Daunorubicin was given at 45 mg/m2 and was followed by 2 consolidation courses. Group 2 patients achieving CR were also offered high dose therapy and stem cell grafts. Results: Seventy one individuals (Group 1) with AML with median age of 36 years (14–60) were studied. Fourteen had favourable cytogenetics but had other adverse prognostic factors; 22 had intermediate risk while 16 had adverse karyotypes and no results were available in 20 cases. Three patients had biphenotypic leukaemia. Circulating blast count greater than 50x109/L was present in 17 individuals. Remission was achieved in 80% (57 patients; 8 after 2 courses). Four patients did not receive consolidation because they had persistent fungal infection (n=2), cardiotoxicity (n=1) or refusal. Reduction in cardiac ejection fraction was seen in 3 individuals. Thirty nine patients underwent autologous (n=17) or allogeneic stem cell transplantation (n=22). At median follow up of 338 days, 39% survive disease free. Only two individuals of 19 who achieved CR but did not receive high dose therapy is alive (p < 0.05). Unfavourable cytogenetics but not older age (above median) was associated with adverse outcome. Kaplan and Meier analysis showed superior survival was associated with Allogeneic SCT (73%) than autologous (58%; p= 0.08). Patients in the historical control fraction, (Group 2) had no significant pre-treatment differences with Group 1. Remission in Group 2 was achieved in 59% (p= 0.03). CR occurred with a single course in 86% vs. 64% (p= 0.02) respectively. There were no differences in the toxicity profile between these two combinations. Disease recurred in 28% and 50% (p= 0.07) patients. For the 142 individuals, median follow up is 254 (range 19–4451) and 453 (12–1702; p= 0.01) days. Survival is 23% and 39% respectively, favouring patients treated in Group 1 (Log Rank; p= 0.03). Conclusions: We conclude that increasing daunorubicin during induction chemotherapy was not associated with excessive toxicity, while remission rates were of 80% and 58%. However, a particularly favourable outcome was seen in patients receiving allogeneic stem cell transplantation.


Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 5288-5288
Author(s):  
Ismael Buño ◽  
Pascual Balsalobre ◽  
Ainhoa Simón ◽  
Gustavo Iglesias ◽  
Cintia Manzano ◽  
...  

Abstract Background: The dynamics of chimerism evolution determines the success of allogeneic stem cell transplantation (SCT). Several factors such as the intensity of the conditioning regimen, the T-cell content of the graft or the GVHD prophylaxis, influence the degree of chimerism after SCT. Objective: To evaluate the dynamics of chimerism after different SCT settings (ablative, reduced intensity conditioning -RIC- and T-cell depleted -TCD-) and its influence in the success of the procedure. Patients and Methods: The study includes 68 SCT: 32 ablative (including 7 from HLA matched unrelated donors -UD-), 19 RIC and 17 TCD (including 8 from haploidentical donors and 2 from UD with 1 HLA disparity). Chimerism analysis was performed by FISH for the sex chromosomes or STR-PCR (sensitivity 1 Samples obtained on days +30, +100, +180, +365 and once a year thereafter, included bone marrow (BM) and peripheral blood (PB). Moreover, chimerism was analyzed in PB and leukocyte lineages (T lymphocytes CD3+, B lymphocytes CD19+ and myeloid cells CD15+ isolated (purity &gt;95%) by immunomagnetic means, AutoMACS, Miltenyi Biotec), every 2 weeks, starting on day +15 (except in ablative), and until complete chimerism (CC) was achieved. Results of chimerism follow-up were censored once the diagnosis of relapse or rejection was established. Results: The incidence (% patients) of mixed chimerism (MC) on day +30 (ablative: BM 27%, PB 15%; RIC: BM 40%, PB 41%, CD3 40%; TCD: BM 31%, PB 33%, CD3 54%), as well as its dynamics (MC on day +100: ablative BM 8%, PB 4%; RIC BM 8%, PB 15%, CD3 22%; TCD BM 20%, PB 40%, CD3 37%) were different in the three SCT settings. All except 2 CD15 samples analyzed showed CC. Moreover, the percentage of recipient cells (%R) was significantly higher after RIC and TCD than after ablative SCT, as well as in T lymphocytes than in BM or PB (7/8 cases with simultaneous studies showed MC in T lymphocytes and CC in PB). All RIC SCT evolved to CC by day +180 while TCD SCT showed persistent MC (2 patients with stable MC after one year). The incidence of rejection was greater after RIC (2/19) and TCD (4/17) than after ablative SCT (2/32). All these patients showed MC, mainly in T lymphocytes, which allowed early diagnosis and successful treatment with immunosuppression withdrawal and donor leukocyte infusion. Patients with CC in PB/T lymphocytes on day +30 had a higher incidence of GVHD&gt;I than those with MC. In the present series, however, a relationship between chimerism and relapse, disease free survival or overall survival, was not observed. Conclusions: SCT with greater incidence of MC (RIC and TCD) favor immune tolerance between donor and recipient which reduces the risk/severity of GVHD at the expense of a higher incidence of graft rejection.


Cancers ◽  
2021 ◽  
Vol 13 (22) ◽  
pp. 5640
Author(s):  
Michael Oertel ◽  
Jonas Martel ◽  
Jan-Henrik Mikesch ◽  
Sergiu Scobioala ◽  
Christian Reicherts ◽  
...  

Total body irradiation is an effective conditioning modality before autologous or allogeneic stem cell transplantation. With the whole body being the radiation target volume, a diverse spectrum of toxicities has been reported. This fact prompted us to investigate the long-term sequelae of this treatment concept in a large patient cohort. Overall, 322 patients with acute leukemia or myelodysplastic syndrome with a minimum follow-up of one year were included (the median follow-up in this study was 68 months). Pulmonary, cardiac, ocular, neurological and renal toxicities were observed in 23.9%, 14.0%, 23.6%, 23.9% and 20.2% of all patients, respectively. The majority of these side effects were grades 1 and 2 (64.9–89.2% of all toxicities in the respective categories). The use of 12 Gray total body irradiation resulted in a significant increase in ocular toxicities (p = 0.013) and severe mucositis (p < 0.001). Renal toxicities were influenced by the age at transplantation (relative risk: 1.06, p < 0.001) and disease entity. In summary, total body irradiation triggers a multifaceted, but manageable, toxicity profile. Except for ocular toxicities and mucositis, a 12 Gray regimen did not lead to an increase in long-term side effects.


2021 ◽  
Vol 27 ◽  
Author(s):  
A. Kopińska ◽  
A. Koclęga ◽  
A. Wieczorkiewicz-Kabut ◽  
K. Woźniczka ◽  
D. Kata ◽  
...  

Introduction: Refractory and relapsed Hodgkin lymphoma (R/R HL) is associated with poor prognosis, and allogeneic stem cell transplantation (allo-SCT) remains the only potentially curative approach.Aim: The aim of the study was to evaluate the feasibility of allotransplantation in R/R HL setting.Material: Overall, 24 patients (17 men and 7 women) at a median age of 27 years (range 18–44) underwent allo-SCT between 2002 and 2020.Results: Nineteen patients received prior autologous stem cell transplantation (ASCT1) whereas eight patients received second ASCT (ASCT2) after failure of ASCT1. Six patients received only brentuximab vedotin (BV; n = 4) or BV followed by checkpoint inhibitors (CPI; n = 2) before entering allo-SCT. Median time from ASCT1 to allo-SCT was 17.1 months. Fifteen patients received grafts from unrelated donors. Peripheral blood was a source of stem cells for 16 patients. Reduced-intensity conditioning was used for all patients. Disease status at transplant entry was as follows: complete remission (CR; n = 4), partial response (PR; n = 10), and stable disease (SD; n = 10). Acute and chronic graft-versus-host disease (GVHD) developed in 13 (54%) and 4 (16%) patients, respectively. Median follow-up for the entire cohort was 13.3 months. At the last follow-up, 17 (71%) patients died. The main causes of death were disease progression (n = 10), infectious complications (n = 6), and steroid-resistant GVHD (n = 1). Non-relapse mortality at 12 months was 25%. At the last follow-up, seven patients were alive; six patients were in CR, and one had PR. The 2-year overall survival (OS) was 40%.Conclusion: Chemosensitive disease at transplant was associated with better outcome. Allo-SCT allows for long-term survival in refractory and relapsed HL.


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