Detection of Circulating Antigen-Specific CD8+ Lymphocytes against Leukemia Associated Antigens WT1, PR1 and Survivin Using HLA 0201 Pentamers After Allogeneic Stem Cell Transplantation.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 5115-5115
Author(s):  
Juana Serrano-Lopez ◽  
Joaquin Sanchez Garcia ◽  
Josefina Serrano ◽  
Carmen Martin ◽  
Rafael Rojas ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Stem Cell ◽  
Stem Cell Transplantation ◽  
Peripheral Blood ◽  
Median Percentage ◽  
Blood Samples ◽  
Hematological Disorders ◽  
Multiparametric Flow Cytometry ◽  
Leukemia Antigen

Abstract Abstract 5115 INTRODUCTION Allogeneic stem cell transplantation (allo-SCT) is a potentially curative treatment option for patients with hematological disorders. Alloreactive donor-derived T lymphocytes exert a beneficial graft-versus-leukemia (GVL) effect through the recognition of leukemia-restricted (or preferentially expressed) antigens as Wilms tumor protein (WT1), survivin (SURV) or proteinase (PR1). Currently research in transplant immunology focuses in enhancing GVL while preventing the deleterious graft-versus-host disease (GVHD) that could be achieved by manipulating donor-derived antigen-specific T-populations. In this study we tested the presence of peripheral blood leukemia-associated antigen-specific CD8+ T-lymphocytes during post allo-SCT follow-up. PATIENTS AND METHODS Forty-three consecutive HLA*0201 patients (homo or heterozygotous) undergoing conventional myeloablative (n=24) or non-myeloablative (n=19) allo-SCT as treatment of hematological disorders were included. Allogeneic donor was an HLA-identical sibling in 26 cases (60.5%) and unrelated in 17 cases (39.5%). Hematopoietic stem cell source included mobilized peripheral blood (n=20), bone marrow (n=18) and umbilical cord blood (n=5). As GVHD prophylaxis regimens Cyclosporine plus Methotrexate (n=20) or Cyclosporine plus Mofetil micofenolate (n=23) were employed. In addition, 22 patients received rabbit antithymocyte globulin at 6-8mg/kg. At last follow-up four patients had relapsed 9-14 months after allo-SCT. We sought for leukemia-antigen specific CD8 lymphocytes in peripheral blood samples drawn within a median of 7 months (range 2-38) when lymphocyte recovery had occurred and complete donor chimerism was achieved. We used four color multiparametric flow cytometry in a FACSCanto II acquiring at least 5 ×105 viable (Propidium Iodide low) lymphoid gated events, stained with MnAbs: CD8-FITC and CD3PE/APC MnAb. To identify leukemia-antigen specific CD8 lymphocytes we used class I HLA pentamers 0201 APC or PE conjugated (Proimmune, London, UK) against the following nonapeptides: Proteinase 1: VLQELNVTV (169-177) WT1: RMFPNAPYL (126-134) and SURV: ELTLGEFLKL (95-104). As positive staining control we used CMV pp65: NLVPMVATV (495-503) and as negative controls we used irrelevant nonapeptide and peripheral blood samples from patients lacking HLA* 0201 genotype. RESULTS Detection of donor-derived CD8+ lymphocytes against CMV pp65 occurred in 61% of recruited patients with a median percentage of 0.1% (range 0.03-13 over CD3+CD8+ events). Likewise, it was possible to detect CD8+ lymphocytes specific for PR1, WT1 and SURV in 65.2%, 47.8% y 39.1% of recruited patients respectively. Median percentage of PR1 and WT1 leukemia-antigen specific lymphocytes was 0.1% (range 0.04-1% over CD3+CD8+ events) and for WT1 0.1% (range: 0.01-0.2%). Detection of leukemia-antigen specific CD8+ lymphocytes was not significantly associated with clinical variables such as conditioning regimen (conventional or non-myeloablative), Disease status at transplant, donor type (sibling or unrelated), ATG use or HLA-disparity degree. The presence of WT1 specific CD8+ lymphocytes was significantly more frequent in patients undergoing allo-SCT for lymphoid hematological malignancy (p=0.04). By contrast, the presence of circulating anti-PR1 specific CD8+ lymphocytes was not more frequently found in patients undergoing allo-SCT for myeloid malignancies. Of note, none of the four patients who eventually relapsed harbored circulating leukemia-specific CD8+ lymphocytes. CONCLUSIONS Multiparametric flow cytometry is a useful tool to detect and quantify rare donor-derived CD8+ lymphocytes specific for leukemia-associated antigens as PR1, WT1 or SURV. The presence of these populations in peripheral blood is not associated to conventional clinical variables and in our series anti-WT1 CD8+ lymphocytes were more frequently detected in patients receiving allo-SCT for lymphoid malingnacies. By contrast, larger series are needed to assess if the lack of these leukemia-associated antigen-specific CD8 lymphocytes in peripheral blood could identify patients in a higher risk of relapse. Financial support This study was supported by a grant of Conserjeria de Salud, Junta de Andalucia 2006/0355. J. Serrano López is a post-doc fellow from Fundación Española de Hematología y Hemoterapia Disclosures No relevant conflicts of interest to declare.

Alternative Donor Hematopoietic Stem Cell Transplantation for Children with Severe Aplastic Anemia.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 4348-4348
Author(s):  
Meerim Park ◽  
Kyung Nam Koh ◽  
Keun Wook Bae ◽  
Mee Jeong Lee ◽  
Ho Joon Im ◽  
...  
Keyword(s):  
Stem Cell ◽  
Hematopoietic Stem Cell Transplantation ◽  
Stem Cell Transplantation ◽  
Aplastic Anemia ◽  
Cell Transplantation ◽  
Hematopoietic Stem Cell ◽  
Peripheral Blood ◽  
Hematopoietic Stem ◽  
Matched Sibling

Abstract Abstract 4348 Background Hematopoietic stem cell transplantation (HSCT) from matched sibling donor is the standard first-line treatment for children with severe aplastic anemia (SAA). However, the management of SAA lacking a suitable donor remains a great challenge. For those children, HSCT using unrelated donor or mismatched related donor could be a therapeutic alternative. The purpose of this study is to evaluate the outcome in children with SAA who received HSCT from donors other than matched sibling. Patients and Method Between March 2003 and July 2009, 17 patients received HSCT from alternative donors (AD) at Asan Medical Center. We reviewed their medical records and analyzed their transplant-related parameters and outcome. Results Of a total of 17 patients, 11 were male and the median age at HSCT was 9.0 years, ranging from 3.0 to 16.7 years. Four patients had Fanconi anemia and 13 had acquired SAA including 2 who developed SAA after liver transplantation. Donors included unrelated bone marrow (U-BM) in 5, unrelated peripheral blood (U-PB) in 6, unrelated cord blood (U-CB) in 2 and related haploidentical peripheral blood (H-PB) in 4. Of 17 patients, 15 (88%) achieved sustained engraftment. Of 15 with engraftment, only 1 patient who received HSCT from U-CB died of severe GI GVHD and the other 14 patients remain on stable normal counts without transfusion support. All 2 patients (1 U-BM, 1 H-PB) who failed to engraft were dead despite DLI or 2nd HSCT. With a median follow-up of 31.9 months, the Kaplan-Meier estimated overall survival at 2 years was 76.6%. Conclusion In children with SAA, HSCT from AD including haploidentical family donor could be considered as a treatment option if the patients have no matched sibling donor. Given the limitation of this study such as small number of patients and short follow-up period, further trial will be necessary. Disclosures: No relevant conflicts of interest to declare.

Rapid Prediction of Peripheral Blood CD34 Counts Using a Multiple Regression Model Derived from Automated Hematology Analyzer Cell Population Data

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 3099-3099 ◽  
Author(s):  
Thomas Porturas ◽  
Mary Sell ◽  
Leah Irwin ◽  
Una O'Doherty ◽  
Carlos Hipolito Villa
Keyword(s):  
Flow Cytometry ◽  
Stem Cell ◽  
Multiple Regression ◽  
Peripheral Blood ◽  
Population Data ◽  
Blood Samples ◽  
Peripheral Blood Cd34 ◽  
Cell Counts ◽  
Flow Cytometric ◽  
Hematology Analyzer

Abstract Background: Although peripheral blood CD34+ stem cell counts by flow cytometry correlate well with yields, the time, complexity, and cost associated with flow cytometry limits its utility. Rapid, cost-effective, surrogate predictors (with <1hr turnaround) would allow for same-visit analyses and alteration of collection and mobilization strategies, particularly for the optimal use of time-sensitive and costly agents such as plerixafor. We previously demonstrated that morphologic parameters of neutrophil-like cells measured by hematology analyzers correlated with CD34 counts. We aimed to improve these models by using multiple regression analyses on data from a common hematology analyzer. Methods: Patients undergoing stem cell apheresis were evaluated over a 6 month period. The day prior to initiation of apheresis, and on the morning of initial collection, peripheral blood samples were drawn into EDTA collection tubes and flow cytometric CD34 measurement and/or CBCs were performed on the Beckman Coulter DxH 800 hematology analyzer per standard protocol. CD34 cells were counted by flow cytometric ISHAGE protocols. Data from the DxH (48 variables per specimen) were exported into a data matrix with the corresponding flow cytometric data. Multiple regression analysis was performed using a step-wise method with log(peripheral CD34) as the dependent variable (SPSS, IBM). Data were randomly selected into a training-set of 70% of cases and a test-set of 30% of cases for validation. The derived model was further tested against peripheral blood data from the morning of collection to predict harvest yields. Further analyses were performed using Prism (GraphPad). Results: Tandem peripheral blood CD34 counts and CBC cell-population data were obtained from 69 blood samples in 64 patients. The population included patients with multiple myeloma (45), non-Hodgkin lymphoma (12), Hodgkin lymphoma (5), and amyloidosis (2). 41% of patients were female. In the test data set examining collection yields, 37 patients were mobilized with GCSF (+/- chemotherapy) alone, while 17 had plerixafor added to the regimen. 33 of these patients had same-day CBC data available for model prediction. The median processed volume was 15 L (range 5.9 to 19.7). The model to predict peripheral CD34 counts incorporated 3 variables from the hematology analyzer data (SD-V-EGC, SD-C-EGC, and NE#). Interestingly, the model included two variables descriptive of the morphology of early granulocytic cells. The model demonstrated an R value of 0.829 (adjusted R2 = 0.670, figure 1a). In testing the morning-of-collection model-predicted peripheral CD34, we found the model performed similarly to flow cytometry in predicting 1st collection yields. Furthermore, the CD34 prediction using the model (Figure 1 b) resulted in similar correlation with first-collection yields in patients treated with plerixafor versus patients not treated with plerixafor, in contrast to day-prior CD34 counts by flow-cytometry (Figure 1c). Two outliers for CD34 cell yield based on model predicted peripheral CD34 were identified. In one patient, the processed volume was very low (6.8 L, <5% percentile), while the second had a low mononuclear cell collection efficiency (35%) compared to the mean in this population (58.7%±23.3%). Threshold values for the model accurately identified patients appropriate for collection initiation (or plerixafor administration). Conclusion: Using data from a common, automated CBC analyzer, we developed a rapid, less-costly, and simple model to predict CD34 cell counts and 1st harvest yields. Because the measurement results can be obtained within the same clinic visit, and can be repeated with each CBC, the model is particularly useful to guide optimal use of plerixafor. We also envision that the model is useful for quality assurance of collection by identifying patients in whom cell yields were sub-optimal with respect to predicted CD34 cell counts. Additional studies to test the model in a larger population are ongoing. We propose that this model (and similarly derived models) can be implemented in clinical planning algorithms to improve the efficiency and cost of stem cell collection by apheresis. Acknowledgments: We would like to acknowledge and the nurses and staff of the apheresis unit and the stem cell and flow cytometry laboratories at the Hospital of the University of Pennsylvania for their contributions. Figure 1. Figure 1. Disclosures No relevant conflicts of interest to declare.

scholarly journals Case Report: Graft Versus Tumor Effect After Non-Myeloablative Allogeneic Stem-Cell Transplantation in a Patient With Brentuximab-Vedotin Refractory Sezary Syndrome

2021 ◽  
Vol 11 ◽  
Author(s):  
Georg-Nikolaus Franke ◽  
Konstantin Dumann ◽  
Madlen Jentzsch ◽  
Astrid Monecke ◽  
Christine Doehring ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Stem Cell ◽  
Stem Cell Transplantation ◽  
Cell Transplantation ◽  
Peripheral Blood ◽  
Conditioning Regimen ◽  
Brentuximab Vedotin ◽  
Sézary Syndrome ◽  
Sezary Syndrome

Sezary Syndrome (SS) is a rare leukemic variant of primary cutaneous T-cell lymphoma. Relapsed or refractory disease is generally considered incurable by conventional therapeutic approaches, although durable responses can be achieved with novel monoclonal antibodies. Allogeneic hematopoietic stem cell transplantation (alloHSCT) may have potential value by inducing graft vs-lymphoma (GvL) effects, but there is currently no consensus regarding the timing of alloHSCT or type of conditioning regimen. Here we present the case of a male patient who achieved a complete remission (CR) of primary refractory SS after non-myeloablative alloHSCT. Patient: Two years prior to HSCT, the patient had been refractory to CHOEP-based chemotherapy, interferon, extracorporeal photopheresis (ECP), and bexarotene. Directly prior to alloHSCT brentuximab-vedotin (BV) was applied resulting in a partial remission of the skin compartment and overall in a stable disease. Prior to HSCT, flow cytometry of the bone marrow and peripheral blood showed an infiltration with T-cells positive for CD5, CD4, low CD3, low CD2 and negative for CD7, CD38, HLA-DR and CD8. The trephine biopsy showed a 7% infiltration of SS cells. The CD4:CD8 ratio in peripheral blood (pb) was massively increased at 76.67, with 63.5% of white blood cells expressing a SS immune phenotype. The conditioning regimen included 30 mg/m2 fludarabine on days -5, -4 and -3 and total body irradiation with 2 Gy on day -1. Immunosuppression consisted of cyclosporine A from day-1 and mycophenolate mofetil from day 0. The patient received 6.55x106 CD34+ cells and 1.11x108 CD3+ cells/kg body weight. Bone marrow evaluation on day 28 still showed persistent SS cells by flow cytometry. After tapering immunosuppression until day 169, the CD4:CD8 ratio in pb normalized. CR was documented on day 169 after alloHSCT and is now ongoing for almost 3 years after alloHSCT. Conclusions: We confirm that an alloHSCT can be a curative option for refractory patients with SS. The achievement of a CR after tapering the immunosuppressive therapy indicates a significant role of the GvL effect. In present treatment algorithms for patients with SS, the timing of an alloHSCT and the intensity of conditioning should be further explored.

Peripheral-blood stem cell transplantation in pediatric recipients in Peru: Experience of a reference center.

2019 ◽  
Vol 37 (15_suppl) ◽  
pp. e21530-e21530
Author(s):  
Essy Maradiegue ◽  
Oliver Guillermo Sulca-Huamani ◽  
Alvaro Jesus Quincho-Lopez ◽  
Kelly Jasmin Meza ◽  
Liliana Vasquez ◽  
...  
Keyword(s):  
Stem Cell ◽  
Stem Cell Transplantation ◽  
Cell Transplantation ◽  
Peripheral Blood ◽  
Peripheral Blood Stem Cell ◽  
Blood Stem Cell ◽  
Neoplastic Disease ◽  
Reference Center ◽  
Blood Stem Cell Transplantation

e21530 Background: Stem cell transplantation (SCT) is a curative treatment for children with refractory/relapsed malignancies. However, data on pediatric transplantation in developing countries like Peru is still needed. The aim of this study was to describe and assess the survival and complications of children with neoplastic disease who have received peripheral-blood stem cell transplantation (PBSCT) at a reference center in Peru. Methods: A retrospective longitudinal study was performed by reviewing medical histories of 20 children (≤14 years) with neoplastic disease who underwent related allo- or auto-PBSCT from October 2014 to December 2017 of INEN (Instituto Nacional de Enfermedades Neoplásicas). In the cases of ALL > 3 years, radiotherapy was not part of the conditioning regimen. The Kaplan-Meier method was used to determine the overall survival (OS) and event-free survival (EFS). Data were analyzed using Stata/MP 14.0. Results: We found 13 males and 7 females with a median age of 9.5 years (1-14), of whom 18 received allo-SCT and 2 auto-SCT. Diagnosis: 14 ALL(66.6%), 1 AML(4.7%), 1 NHL(4.7%), 2 biphenotypic leukemia(9.5%), 1 juvenile chronic myelomonocytic leukemia(4.7%) and 1 germ cell tumor(4.7%) who performed two auto-transplants. Complete remission (CR) was achieved before transplantation in 19 cases: 8(38%) 1CR; 8(38%) 2CR and 3(14.2%) 3CR. A median of 7.14x106CD34+/kg (3.97-13.05) (auto- vs allo-; 6.35 vs 7.27) were collected and median time to myeloid engrafment: 11 days. At a median follow-up of 12 months (1-39) the 2-year OS and EFS were 75.7% (95% confidence interval [CI], 38-92%) and 51.6% (95% CI, 22-74%) respectively. Considering only ALL > 3 years, OS and EFS were 51.9% and 45.1%, respectively. Acute graft-versus-host disease (GVHD), chronic GVHD, transplantation-related mortality (TRM) and relapse were 27.7%, 11.1%, 4.7%, and 33.3%, respectively. Conclusions: We are encouraged by this first experience in our country. OS and EFS are comparable with results at the international level. However, survival of children with ALL > 3 years who did not receive radiotherapy is less than international standards. Studies with a higher number of cases and longer follow-up time are needed to reach definitive conclusions.

The Outcome of Allogeneic Stem Cell Transplantation in Chronic Lymphocytic Leukaemia: A Single Centre 10-Year-Experience

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 4515-4515
Author(s):  
Patrycja Zielinska ◽  
Malgorzata Krawczyk-Kulis ◽  
Miroslaw Markiewicz ◽  
Monika Dzierzak-Mietla ◽  
Anna Koclega ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Stem Cell ◽  
Stem Cell Transplantation ◽  
Cell Transplantation ◽  
Allogeneic Stem Cell Transplantation ◽  
Peripheral Blood ◽  
Conflicts Of Interest ◽  
Cumulative Probability

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.

scholarly journals AUTOLOGOUS HEMATOPOIETIC STEM CELL TRANSPLANTATION FOR HIGH-RISK ACUTE LYMPHOBLASTIC LEUKEMIA: NON-RANDOMIZED STUDY WITH A MAXIMUM FOLLOW-UP OF MORE THAN 22 YEARS

2014 ◽  
Vol 6 (1) ◽  
pp. e2014047 ◽  
Author(s):  
Grzegorz Helbig ◽  
Malgorzata Krawczyk-Kulis ◽  
Malgorzata Kopera ◽  
Krystyna Jagoda ◽  
Patrycja Rzepka ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Stem Cell ◽  
Acute Lymphoblastic Leukemia ◽  
High Risk ◽  
Hematopoietic Stem Cell Transplantation ◽  
Stem Cell Transplantation ◽  
Peripheral Blood ◽  
Lymphoblastic Leukemia ◽  
Hematopoietic Stem

Objective. To evaluate the efficacy and toxicity of autologous hematopoietic stem cell transplantation (AHSCT) for high-risk acute lymphoblastic leukemia (ALL). Material and methods. Overall, 128 high-risk ALL patients at a median age of 26 years (range 18-56 years) at diagnosis received AHSCT between 1991-2008. Induction treatment was anthracycline-based in all patients. Conditioning regimen consisted of CAV (cyclophosphamide, cytarabine, etoposide) in 125 patients whereas 3 subjects received cyclophosphamide and TBI (total body irridation). Bone marrow was stored for 72 hours in 4oC and re-infused 24 hours after conditioning completion. Bone marrow was a source of stem cells in 119 patients, peripheral blood in 2 and 7 subjects received both bone marrow and peripheral blood. Results. With a median follow-up after AHSCT of 1.6 years (range 0.1-22.3 years), the probability of leukemia-free survival (LFS) for the whole group at 10 years was 27% and 23% at 20 years. Transplant-related mortality at 100 days after AHSCT was 3.2%.. There was a strong tendency for better LFS for MRD-negative patients if compared with patients who had positive or unknown MRD status at AHSCT (32% vs 23% and 25%, respectively; p=0.06). There was no difference in LFS between B- and T-lineage ALL as well as between patients transplanted in first complete remission (CR1) and CR2. LFS at 10 years for patients with detectable BCR-ABL at transplant was 20% and this was comparable with subjects with negative and missing BCR-ABL status (26% and 28%; p=0.97). Conclusions. The results of AHSCT for high-risk ALL remains unsatisfactory with low probability of long-term LFS.

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