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Gamma delta (γδ) T cells form an unconventional subset of T lymphocytes that express a T cell receptor (TCR) consisting of γ and δ chains. Unlike conventional αβ T cells, γδ T cells share the immune signature of both the innate and the adaptive immunity. These features allow γδ T cells to act in front-line defense against infections and tumors, rendering them an attractive target for immunotherapy. The role of γδ T cells in the immune response to cytomegalovirus (CMV) has been the focus of intense research for several years, particularly in the context of transplantation, as CMV reactivation remains a major cause of transplant-related morbidity and mortality. Therefore, a better understanding of the mechanisms that underlie CMV immune responses could enable the design of novel γδ T cell-based therapeutic approaches. In this regard, the advent of next-generation sequencing (NGS) and single-cell TCR sequencing have allowed in-depth characterization of CMV-induced TCR repertoire changes. In this review, we try to shed light on recent findings addressing the adaptive role of γδ T cells in CMV immunosurveillance and revisit CMV-induced TCR reshaping in the era of NGS. Finally, we will demonstrate the favorable and unfavorable effects of CMV reactive γδ T cells post-transplantation.
Ahmed Gaballa; Faisal Alagrafi; Michael Uhlin; Arwen Stikvoort. Revisiting the Role of γδ T Cells in Anti-CMV Immune Response after Transplantation. Viruses 2021, 13, 1031 .
AMA StyleAhmed Gaballa, Faisal Alagrafi, Michael Uhlin, Arwen Stikvoort. Revisiting the Role of γδ T Cells in Anti-CMV Immune Response after Transplantation. Viruses. 2021; 13 (6):1031.
Chicago/Turabian StyleAhmed Gaballa; Faisal Alagrafi; Michael Uhlin; Arwen Stikvoort. 2021. "Revisiting the Role of γδ T Cells in Anti-CMV Immune Response after Transplantation." Viruses 13, no. 6: 1031.
The role of γδ T cells in antitumor immunity has been under investigation for the past two decades, but little is known about their contribution to clinical outcomes in patients. Here, we set out to define the clonotypic, phenotypic, and functional features of γδ T cells in peripheral blood, ascites, and metastatic tumor tissue from patients with advanced epithelial ovarian cancer. T cell receptor (TCR) sequencing of the γ chain revealed that tumor-infiltrating γδ T cells have a unique and skewed repertoire with high TCR diversity and low clonality. In contrast, ascites-derived γδ T cells presented a lower TCR diversity and higher clonality, suggesting a TCR-dependent clonal focusing at this site. Further investigation showed that tumor samples had abundant γδ T cells with a tissue-resident, activation-associated phenotype, less usage of Vγ9 and an impaired response to adaptive-associated stimuli, implying an innate-like activation pathway, rather than an adaptive TCR-engaging pathway, at these tumor sites. Furthermore, high γδ T cell cytokine responsiveness upon stimulation was associated with a favorable outcome for patients in terms of both overall survival and reduced residual tumor burden after primary surgery. Last, the functionality of γδ T cells and patient survival were negatively affected by the proportions of CD39-expressing T cells, highlighting the potential of CD39 as a target to improve γδ T cell responses and unleash their antitumor capabilities.
Emelie Foord; Lucas C. M. Arruda; Ahmed Gaballa; Charlotte Klynning; Michael Uhlin. Characterization of ascites- and tumor-infiltrating γδ T cells reveals distinct repertoires and a beneficial role in ovarian cancer. Science Translational Medicine 2021, 13, eabb0192 .
AMA StyleEmelie Foord, Lucas C. M. Arruda, Ahmed Gaballa, Charlotte Klynning, Michael Uhlin. Characterization of ascites- and tumor-infiltrating γδ T cells reveals distinct repertoires and a beneficial role in ovarian cancer. Science Translational Medicine. 2021; 13 (577):eabb0192.
Chicago/Turabian StyleEmelie Foord; Lucas C. M. Arruda; Ahmed Gaballa; Charlotte Klynning; Michael Uhlin. 2021. "Characterization of ascites- and tumor-infiltrating γδ T cells reveals distinct repertoires and a beneficial role in ovarian cancer." Science Translational Medicine 13, no. 577: eabb0192.
Background and objectives Commercial blood bags are predominantly made of polyvinyl chloride (PVC) plasticized with di(2‐ethylhexyl) phthalate (DEHP). DEHP is favourable for storage of red blood cells (RBC). Historically, removal of DEHP from blood bags has been linked to unacceptable haemolysis levels. Oncoming regulatory restrictions for DEHP due to toxicity concerns increase the urgency to replace DEHP without compromising RBC quality. Di(2‐ethylhexyl) terephthalate (DEHT) is one suggested substitute. The aim of this study was to compare PVC‐DEHT to PVC‐DEHP blood bags using additive solutions saline–adenine–glucose–mannitol (SAGM) and phosphate–adenine–glucose–guanosine–saline–mannitol (PAGGSM), to determine whether DEHT can maintain acceptable component quality. Materials and methods RBC concentrates (N = 64), platelet concentrates (N = 16) and fresh frozen plasma (N = 32) were produced from whole blood collected into either DEHT or DEHP plasticized systems. Using a pool‐and‐split study design, pairs of identical RBC content were created within each plasticizer arm and assigned either SAGM or PAGGSM. Storage effects were assessed weekly for 49 days (RBC), 7 days (platelets) and before/after freezing (plasma). Results Though haemolysis was slightly higher in DEHT, all study arms remained below half of the European limit 0·8%. K+ was lower in DEHT than in DEHP independent of additive solution. The metabolic parameters were not influenced by choice of plasticizer. Platelet activation/metabolism and plasma content were similarly preserved. Conclusion Our study demonstrates that the plasticizer DEHT provides adequate blood component quality. We propose DEHT as a strong future candidate for replacement of DEHP in blood bags.
Linda Larsson; Per Sandgren; Sara Ohlsson; Julia Derving; Tove Friis‐Christensen; Felicia Daggert; Naïma Frizi; Stefan Reichenberg; Sonia Chatellier; Beatrice Diedrich; Jovan Antovic; Stella Larsson; Michael Uhlin. Non‐phthalate plasticizer DEHT preserves adequate blood component quality during storage in PVC blood bags. Vox Sanguinis 2020, 116, 60 -70.
AMA StyleLinda Larsson, Per Sandgren, Sara Ohlsson, Julia Derving, Tove Friis‐Christensen, Felicia Daggert, Naïma Frizi, Stefan Reichenberg, Sonia Chatellier, Beatrice Diedrich, Jovan Antovic, Stella Larsson, Michael Uhlin. Non‐phthalate plasticizer DEHT preserves adequate blood component quality during storage in PVC blood bags. Vox Sanguinis. 2020; 116 (1):60-70.
Chicago/Turabian StyleLinda Larsson; Per Sandgren; Sara Ohlsson; Julia Derving; Tove Friis‐Christensen; Felicia Daggert; Naïma Frizi; Stefan Reichenberg; Sonia Chatellier; Beatrice Diedrich; Jovan Antovic; Stella Larsson; Michael Uhlin. 2020. "Non‐phthalate plasticizer DEHT preserves adequate blood component quality during storage in PVC blood bags." Vox Sanguinis 116, no. 1: 60-70.
PD-1/PD-L1 blockade has revolutionized the field of immunooncology. Despite the relative success, the response rate to anti-PD-1 therapy requires further improvements. Our aim was to explore the enhancement of T-cell function by using novel PD-1-blocking proteins and compare with clinically approved monoclonal antibodies (mAbs). We isolated T-cells from the ascites and tumor of 17 patients with advanced epithelial ovarian cancer (EOC) and analyzed the effects using the mAbs nivolumab and pembrolizumab and two novel engineered ankyrin repeat proteins (DARPin® proteins). PD-1 blockade with either mAb or DARPin® molecule significantly increased the release of IFN-γ, granzyme B, IL-2, and TNF-α, demonstrating successful reinvigoration. The monovalent DARPin® protein was less effective compared to its bivalent equivalent, demonstrating that bivalency brings an additional benefit to PD-1 blockade. Overall, we found a higher fold increase of lymphokine secretion in response to the PD-1 blockade by tumor-derived T-cells; however, the absolute amounts were significantly lower compared to the release from ascites-derived T-cells. Our results demonstrate that PD-1 blockade can only partially reinvigorate functionally suppressed T-cells from EOC patients. This warrants further investigation preferably in combination with other therapeutics. The study provides an early pilot proof-of-concept for the potential use of DARPin® proteins as eligible alternative scaffold proteins to block PD-1.
Emelie Foord; Charlotte Klynning; Esther Schoutrop; Judith M. Förster; Jennifer Krieg; Anette Mörtberg; Mischa R. Müller; Christel Herzog; Dieter Schiegg; Denis Villemagne; Ulrike Fiedler; Dan Snell; Benjamin Kebble; Jonas Mattsson; Victor Levitsky; Michael Uhlin. Profound Functional Suppression of Tumor-Infiltrating T-Cells in Ovarian Cancer Patients Can Be Reversed Using PD-1-Blocking Antibodies or DARPin® Proteins. Journal of Immunology Research 2020, 2020, 1 -12.
AMA StyleEmelie Foord, Charlotte Klynning, Esther Schoutrop, Judith M. Förster, Jennifer Krieg, Anette Mörtberg, Mischa R. Müller, Christel Herzog, Dieter Schiegg, Denis Villemagne, Ulrike Fiedler, Dan Snell, Benjamin Kebble, Jonas Mattsson, Victor Levitsky, Michael Uhlin. Profound Functional Suppression of Tumor-Infiltrating T-Cells in Ovarian Cancer Patients Can Be Reversed Using PD-1-Blocking Antibodies or DARPin® Proteins. Journal of Immunology Research. 2020; 2020 ():1-12.
Chicago/Turabian StyleEmelie Foord; Charlotte Klynning; Esther Schoutrop; Judith M. Förster; Jennifer Krieg; Anette Mörtberg; Mischa R. Müller; Christel Herzog; Dieter Schiegg; Denis Villemagne; Ulrike Fiedler; Dan Snell; Benjamin Kebble; Jonas Mattsson; Victor Levitsky; Michael Uhlin. 2020. "Profound Functional Suppression of Tumor-Infiltrating T-Cells in Ovarian Cancer Patients Can Be Reversed Using PD-1-Blocking Antibodies or DARPin® Proteins." Journal of Immunology Research 2020, no. : 1-12.
CD8+ T cell exhaustion is a hallmark of many cancers and chronic infections. In mice, T cell factor 1 (TCF-1) maintains exhausted CD8+ T cell responses, whereas thymocyte selection-associated HMG box (TOX) is required for the epigenetic remodeling and survival of exhausted CD8+ T cells. However, it has remained unclear to what extent these transcription factors play analogous roles in humans. In this study, we mapped the expression of TOX and TCF-1 as a function of differentiation and specificity in the human CD8+ T cell landscape. Here, we demonstrate that circulating TOX+ CD8+ T cells exist in most humans, but that TOX is not exclusively associated with exhaustion. Effector memory CD8+ T cells generally expressed TOX, whereas naive and early-differentiated memory CD8+ T cells generally expressed TCF-1. Cytolytic gene and protein expression signatures were also defined by the expression of TOX. In the context of a relentless immune challenge, exhausted HIV-specific CD8+ T cells commonly expressed TOX, often in clusters with various activation markers and inhibitory receptors, and expressed less TCF-1. However, polyfunctional memory CD8+ T cells specific for cytomegalovirus (CMV) or Epstein-Barr virus (EBV) also expressed TOX, either with or without TCF-1. A similar phenotype was observed among HIV-specific CD8+ T cells from individuals who maintained exceptional immune control of viral replication. Collectively, these data demonstrate that TOX is expressed by most circulating effector memory CD8+ T cell subsets and not exclusively linked to exhaustion.
Takuya Sekine; André Perez-Potti; Son Nguyen; Jean-Baptiste Gorin; Vincent H. Wu; Emma Gostick; Sian Llewellyn-Lacey; Quirin Hammer; Sara Falck-Jones; Sindhu Vangeti; Meng Yu; Anna Smed-Sörensen; Ahmed Gaballa; Michael Uhlin; Johan K. Sandberg; Christian Brander; Piotr Nowak; Paul A. Goepfert; David A. Price; Michael R. Betts; Marcus Buggert. TOX is expressed by exhausted and polyfunctional human effector memory CD8+ T cells. Science Immunology 2020, 5, eaba7918 .
AMA StyleTakuya Sekine, André Perez-Potti, Son Nguyen, Jean-Baptiste Gorin, Vincent H. Wu, Emma Gostick, Sian Llewellyn-Lacey, Quirin Hammer, Sara Falck-Jones, Sindhu Vangeti, Meng Yu, Anna Smed-Sörensen, Ahmed Gaballa, Michael Uhlin, Johan K. Sandberg, Christian Brander, Piotr Nowak, Paul A. Goepfert, David A. Price, Michael R. Betts, Marcus Buggert. TOX is expressed by exhausted and polyfunctional human effector memory CD8+ T cells. Science Immunology. 2020; 5 (49):eaba7918.
Chicago/Turabian StyleTakuya Sekine; André Perez-Potti; Son Nguyen; Jean-Baptiste Gorin; Vincent H. Wu; Emma Gostick; Sian Llewellyn-Lacey; Quirin Hammer; Sara Falck-Jones; Sindhu Vangeti; Meng Yu; Anna Smed-Sörensen; Ahmed Gaballa; Michael Uhlin; Johan K. Sandberg; Christian Brander; Piotr Nowak; Paul A. Goepfert; David A. Price; Michael R. Betts; Marcus Buggert. 2020. "TOX is expressed by exhausted and polyfunctional human effector memory CD8+ T cells." Science Immunology 5, no. 49: eaba7918.
The role of gamma delta (γδ) T cells in human cytomegalovirus (HCMV) immune surveillance has been the focus of research interest for years. Recent reports have shown a substantial clonal proliferation of γδ T cells in response to HCMV, shedding light on the adaptive immune response of γδ T cells. Nevertheless, most efforts have focused on Vδ2negγδ T cell subset while less attention has been given to investigate other less common γδ T cell subsets. In this regard, a distinct subpopulation of γδ T cells that expresses the CD8 coreceptor (CD8+γδ T cells) has not been thoroughly explored. Whether it is implicated in HCMV response and its ability to generate adaptive response has not been thoroughly investigated. In this study, we combined flow cytometry and immune sequencing of the TCR γ-chain (TRG) to analyze in-depth bone marrow (BM) graft γδ T cells from CMV seropositive (CMV+) and CMV seronegative (CMV-) donors. We showed that the frequency of CD8+γδ T cells was significantly higher in CMV+ grafts compared to CMV- grafts (P<0.001). Further characterization revealed that CD8+γδ T cells from CMV+ grafts express Vγ9- and preferentially differentiated from a naive to terminal effector memory phenotype (CD27low/-CD45RO-). In line with these findings, TRG immune sequencing revealed clonal focusing and reduced usage of the Vγ9/JP gene segment in a CMV+ graft. Furthermore, CD8+γδ T cells showed an enhanced response to TCR/CD3 and cytokine stimulation in contrast to CD8-γδ T cells. We conclude that γδ T cells in BM grafts are reshaped by donor CMV serostatus and highlight the potential adaptive role of CD8+γδ T cells in HCMV immune response.
Ahmed Gaballa; Lucas C. M. Arruda; Emelie Foord (Former Surname Rådestad); Michael Uhlin. CD8+γδ T Cells Are More Frequent in CMV Seropositive Bone Marrow Grafts and Display Phenotype of an Adaptive Immune Response. Stem Cells International 2019, 2019, 1 -13.
AMA StyleAhmed Gaballa, Lucas C. M. Arruda, Emelie Foord (Former Surname Rådestad), Michael Uhlin. CD8+γδ T Cells Are More Frequent in CMV Seropositive Bone Marrow Grafts and Display Phenotype of an Adaptive Immune Response. Stem Cells International. 2019; 2019 ():1-13.
Chicago/Turabian StyleAhmed Gaballa; Lucas C. M. Arruda; Emelie Foord (Former Surname Rådestad); Michael Uhlin. 2019. "CD8+γδ T Cells Are More Frequent in CMV Seropositive Bone Marrow Grafts and Display Phenotype of an Adaptive Immune Response." Stem Cells International 2019, no. : 1-13.
Allogeneic hematopoietic stem cell transplantation (HSCT) using αβ T-/B-cell-depleted grafts recently emerged as a transplant strategy and highlighted the potential role of γδ T cells on HSCT outcomes. Our aim was to scrutinize available evidence of γδ T-cell impact on relapse, infections, survival, and acute graft-versus-host disease (aGVHD). We performed a systematic review and meta-analysis of studies assessing γδ T cells in HSCT. We searched PubMed, Web of Science, Scopus, and conference abstracts from inception to March 2019 for relevant studies. We included all studies that assessed γδ T cells associated with HSCT. Data were extracted independently by 2 investigators based on strict selection criteria. A random-effects model was used to pool outcomes across studies. Primary outcome was disease relapse. We also assessed infections, survival, and aGVHD incidence. The review was registered with PROSPERO (CRD42019133344). Our search returned 2412 studies, of which 11 (919 patients) were eligible for meta-analysis. Median follow-up was 30 months (interquartile range, 22-32). High γδ T-cell values after HSCT were associated with less disease relapse (risk ratio [RR], 0.58; 95% confidence interval [95% CI], 0.40-0.84; P = .004; I2 = 0%), fewer viral infections (RR, 0.59; 95% CI, 0.43-0.82; P = .002; I2 = 0%) and higher overall (HR, 0.28; 95% CI, 0.18-0.44; P < .00001; I2 = 0%) and disease-free survivals (HR 0.29; 95% CI, 0.18-0.48; P < .00001; I2 = 0%). We found no association between high γδ T-cell values and aGVHD incidence (RR, 0.72; 95% CI, 0.41-1.27; P = .26; I2 = 0%). In conclusion, high γδ T cells after HSCT is associated with a favorable clinical outcome but not with aGVHD development, suggesting that γδ T cells have a significant effect on the success of HSCT. This study was registered with PROSPERO as #CRD42019133344.
Lucas C. M. Arruda; Ahmed Gaballa; Michael Uhlin. Impact of γδ T cells on clinical outcome of hematopoietic stem cell transplantation: systematic review and meta-analysis. Blood Advances 2019, 3, 3436 -3448.
AMA StyleLucas C. M. Arruda, Ahmed Gaballa, Michael Uhlin. Impact of γδ T cells on clinical outcome of hematopoietic stem cell transplantation: systematic review and meta-analysis. Blood Advances. 2019; 3 (21):3436-3448.
Chicago/Turabian StyleLucas C. M. Arruda; Ahmed Gaballa; Michael Uhlin. 2019. "Impact of γδ T cells on clinical outcome of hematopoietic stem cell transplantation: systematic review and meta-analysis." Blood Advances 3, no. 21: 3436-3448.
Mucositis is a common complication after allogeneic hematopoietic stem cell transplantation (HSCT), and is caused by a combination of conditioning-induced mucosal damage and severe neutropenia. The symptoms include oral and abdominal pain, inability to swallow food and fluids, and severe diarrhoea. Severe mucositis is associated with increased risk of Graft-versus-Host disease and infection. Granulocyte transfusions (GCX) could be a treatment option, and our objective was to study its feasibility and potential benefits. This retrospective, single-centre study included 30 patients receiving GCX because of severe oral mucositis after HSCT during 2005-2017. Clinical outcome, response to GCX, change in opiate administration and adverse events were studied. Twenty-seven patients received GCX from donors pre-treated with steroids and G-CSF, and three from donors pre-treated with steroids only. Overall response was 83% (24/29 evaluable patients). Fifteen patients reached a complete response. In 14 of 24 responders, a reduction of the administration of opiate pain relief was seen. In eight patients this reduction was ≥50% of the dose. Adverse events (AEs) were reported in 14 cases, and were mild to moderate, and well manageable with symptomatic treatment. No life-threatening or fatal AEs were recorded. These results indicate that GCX could be a safe and effective treatment for oral mucositis after HSCT with the potential to reduce the necessity of opiate analgesic treatment in this disorder. No severe AEs were seen in this study, but the risk for severe pulmonary AEs after GCX needs to be considered.
Sofia Berglund; Emma Watz; Mats Remberger; Karin Garming Legert; Ulla Axdorph‐Nygell; Mikael Sundin; Michael Uhlin; Jonas Mattsson. Granulocyte transfusions could benefit patients with severe oral mucositis after allogeneic hematopoietic stem cell transplantation. Vox Sanguinis 2019, 114, 769 -777.
AMA StyleSofia Berglund, Emma Watz, Mats Remberger, Karin Garming Legert, Ulla Axdorph‐Nygell, Mikael Sundin, Michael Uhlin, Jonas Mattsson. Granulocyte transfusions could benefit patients with severe oral mucositis after allogeneic hematopoietic stem cell transplantation. Vox Sanguinis. 2019; 114 (7):769-777.
Chicago/Turabian StyleSofia Berglund; Emma Watz; Mats Remberger; Karin Garming Legert; Ulla Axdorph‐Nygell; Mikael Sundin; Michael Uhlin; Jonas Mattsson. 2019. "Granulocyte transfusions could benefit patients with severe oral mucositis after allogeneic hematopoietic stem cell transplantation." Vox Sanguinis 114, no. 7: 769-777.
There is a growing concern for shortage in future blood supply, caused by a predicted decrease in eligible blood donors and simultaneous increase in recipients. Cryopreservation of split red blood cell units could increase stock supply by reducing waste of rare blood. This would be particularly useful in paediatric care where very small volumes often are transfused. The aim of this study was to develop a cryopreservation protocol for split units using the closed-system automated cell processor ACP215, as such protocols are currently missing. Using a pool-and-split design, red blood cell units (N = 8) were glycerolized and frozen, either as standard volume reference units, or further divided into three smaller split units each. After thawing/deglycerolization, the supernatant of the smaller splits was reduced by additional centrifugation, and new SAGM was added to 60% haematocrit. The units were analysed for storage lesion effects up to ten days post-thawing. Haemolysis and extracellular potassium ion levels were lower in the split units than in the whole units from day three onwards. The metabolic parameters pH, ATP, glucose and lactate were also lower, though likely caused by lower additive solution pH rather than storage. Split units of red blood cells can be successfully cryopreserved using the ACP215. In addition to favourably low haemolysis and potassium, they also have higher haematocrit than corresponding whole units and enable involvement of fewer donors at repeated transfusions. These characteristics are all desirable features in paediatric care.
Linda Larsson; Stella Larsson; Julia Derving; Emma Watz; Michael Uhlin. A novel protocol for cryopreservation of paediatric red blood cell units allows increased availability of rare blood types. Vox Sanguinis 2019, 114, 711 -720.
AMA StyleLinda Larsson, Stella Larsson, Julia Derving, Emma Watz, Michael Uhlin. A novel protocol for cryopreservation of paediatric red blood cell units allows increased availability of rare blood types. Vox Sanguinis. 2019; 114 (7):711-720.
Chicago/Turabian StyleLinda Larsson; Stella Larsson; Julia Derving; Emma Watz; Michael Uhlin. 2019. "A novel protocol for cryopreservation of paediatric red blood cell units allows increased availability of rare blood types." Vox Sanguinis 114, no. 7: 711-720.
Lymphocyte reconstitution is pivotal for successful long-term outcome after allogeneic hematopoietic stem cell transplantation (HSCT), and conditioning regimen and post-transplantation immunosuppression are risk factors for prolonged immunodeficiency. Nevertheless, the effects of different immunosuppressive protocols on lymphocyte output and replicative capacity have not been investigated. Here we assessed T cell receptor excision circles (TREC), kappa-deleting recombination excision circles (KREC), and T cell telomere length (TL) as proxy markers for immune reconstitution in patients in a prospective randomized trial comparing graft-versus-host disease (GVHD) prophylaxis after transplantation (cyclosporine/methotrexate versus tacrolimus/sirolimus; n = 200). Results showed that medians of TREC, KREC, and TL were not significantly different between the prophylaxis groups at any assessment time point during follow-up (24 months), but the kinetics of TREC, KREC, and TL were significantly influenced by other transplantation-related factors. Older recipient age, the use of antithymocyte globulin before graft infusion, and use of peripheral blood stem cell grafts were associated with lower TREC levels, whereas acute GVHD transiently affected KREC levels. Patients with lymphocyte excision circle levels above the median at ≤6 months post-transplantation had reduced transplantation-related mortality and superior 5-year overall survival (P < .05). We noticed significant T cell telomere shortening in the patient population as a whole during follow-up. Our results suggest that lymphocyte reconstitution after transplantation is not altered by different immunosuppressive protocols. This study has been registered at ClinicalTrials.gov (identifier: NCT00993343).
Johan Törlén; Ahmed Gaballa; Mats Remberger; Lisa-Mari Mörk; Berit Sundberg; Jonas Mattsson; Michael Uhlin. Effect of Graft-versus-Host Disease Prophylaxis Regimens on T and B Cell Reconstitution after Allogeneic Hematopoietic Stem Cell Transplantation. Biology of Blood and Marrow Transplantation 2019, 25, 1260 -1268.
AMA StyleJohan Törlén, Ahmed Gaballa, Mats Remberger, Lisa-Mari Mörk, Berit Sundberg, Jonas Mattsson, Michael Uhlin. Effect of Graft-versus-Host Disease Prophylaxis Regimens on T and B Cell Reconstitution after Allogeneic Hematopoietic Stem Cell Transplantation. Biology of Blood and Marrow Transplantation. 2019; 25 (6):1260-1268.
Chicago/Turabian StyleJohan Törlén; Ahmed Gaballa; Mats Remberger; Lisa-Mari Mörk; Berit Sundberg; Jonas Mattsson; Michael Uhlin. 2019. "Effect of Graft-versus-Host Disease Prophylaxis Regimens on T and B Cell Reconstitution after Allogeneic Hematopoietic Stem Cell Transplantation." Biology of Blood and Marrow Transplantation 25, no. 6: 1260-1268.
Allogeneic hematopoietic stem cell transplantation (HSCT) is associated with several potentially lethal complications. Higher levels of CD3+ T-cells in the graft have been associated with increased risk of graft-versus-host disease (GVHD), but also beneficial graft-versus-leukemia effect and reduced infections. To tackle post-transplant complications, donor lymphocyte infusions have been used but with an increased risk of GVHD. To reduce this risk, we performed depletion of αβ T-cells and treated 12 patients post-HSCT suffering from infections and/or poor immune reconstitution. The αβ T-cell depleted cell products were characterized by flow cytometry. The median log depletion of αβ T-cells was −4.3 and the median yield of γδ T-cells was 73.5%. The median CD34+ cell dose was 4.4 × 106/kg. All 12 patients were alive 3 months after infusion and after 1 year, two patients had died. No infusion-related side effects were reported and no severe acute GVHD (grade III-IV) developed in any patient post-infusion. Overall, 3 months after infusion 11 out of 12 patients had increased levels of platelets and/or granulocytes. In conclusion, we describe the use of αβ T-cell depleted products as stem cell boosters with encouraging results.
Emelie Foord (Former Surname Rådestad); Mikael Sundin; Johan Törlén; Sarah Thunberg; Björn Önfelt; Per Ljungman; Emma Watz; Jonas Mattsson; Michael Uhlin. Individualization of Hematopoietic Stem Cell Transplantation Using Alpha/Beta T-Cell Depletion. Frontiers in Immunology 2019, 10, 189 .
AMA StyleEmelie Foord (Former Surname Rådestad), Mikael Sundin, Johan Törlén, Sarah Thunberg, Björn Önfelt, Per Ljungman, Emma Watz, Jonas Mattsson, Michael Uhlin. Individualization of Hematopoietic Stem Cell Transplantation Using Alpha/Beta T-Cell Depletion. Frontiers in Immunology. 2019; 10 ():189.
Chicago/Turabian StyleEmelie Foord (Former Surname Rådestad); Mikael Sundin; Johan Törlén; Sarah Thunberg; Björn Önfelt; Per Ljungman; Emma Watz; Jonas Mattsson; Michael Uhlin. 2019. "Individualization of Hematopoietic Stem Cell Transplantation Using Alpha/Beta T-Cell Depletion." Frontiers in Immunology 10, no. : 189.
Although the impact of donor graft composition on clinical outcomes after hematopoietic stem cell transplantation (HSCT) has been studied, little is known about the role of intragraft γδ TCR repertoire on clinical outcomes following HSCT. Using a high-throughput sequencing platform, we sought to analyze the TCR γ-chain (TRG) repertoire of γδ T cells within donor stem cell grafts and address its potential impact on clinical response in the corresponding patients. A total of 20 peripheral blood stem cell grafts were analyzed, and donors were classified as CMV+/−. The respective acute myeloid leukemia recipients were followed for disease relapse and acute graft-versus-host disease (aGvHD) development post-HSCT. In all samples, TRG repertoire showed a reduced diversity and displayed overrepresented clones. This was more prominent in grafts from CMV+ donors, which presented a more private repertoire, lower diversity, skewed distribution, and reduced usage of the V9-JP pairing. Grafts given to nonrelapse patients presented a more public repertoire and increased presence of long sequence clonotypes. Variable-joining gene segment usage was not associated with aGvHD development, but a higher usage of V2-JP1 pairing and lower usage of V4-J2/V5-J2/V8-JP2 were observed in grafts given to nonrelapse patients. Our work identified five private overrepresented and one public CDR3 sequence (CATWDGPYYKKLF) associated with CMV infection, in addition to 12 highly frequent public sequences present exclusively in grafts given to nonrelapse patients. Our findings show that, despite CMV infection reshaping the TRG repertoire, TRG composition is not associated with aGvHD development, and several public sequences are associated with clinical remission.
Lucas C. M. Arruda; Ahmed Gaballa; Michael Uhlin. Graft γδ TCR Sequencing Identifies Public Clonotypes Associated with Hematopoietic Stem Cell Transplantation Efficacy in Acute Myeloid Leukemia Patients and Unravels Cytomegalovirus Impact on Repertoire Distribution. The Journal of Immunology 2019, 202, 1859 -1870.
AMA StyleLucas C. M. Arruda, Ahmed Gaballa, Michael Uhlin. Graft γδ TCR Sequencing Identifies Public Clonotypes Associated with Hematopoietic Stem Cell Transplantation Efficacy in Acute Myeloid Leukemia Patients and Unravels Cytomegalovirus Impact on Repertoire Distribution. The Journal of Immunology. 2019; 202 (6):1859-1870.
Chicago/Turabian StyleLucas C. M. Arruda; Ahmed Gaballa; Michael Uhlin. 2019. "Graft γδ TCR Sequencing Identifies Public Clonotypes Associated with Hematopoietic Stem Cell Transplantation Efficacy in Acute Myeloid Leukemia Patients and Unravels Cytomegalovirus Impact on Repertoire Distribution." The Journal of Immunology 202, no. 6: 1859-1870.
Background and objective Efficient pathogen inactivation (PI) offers the possibility of increasing the number of buffy coats per pool without the concurrent increased risk of pathogen transmission. Here, we describe the findings of in vitro analyses of platelets from pools of eight buffy coats treated with amotosalen and UVA light (INTERCEPT Blood System for Platelets) using INTERCEPT disposable processing sets with plastic materials sourced from alternate suppliers and split afterwards to obtain two therapeutic transfusion doses. Methods Double‐dose platelet concentrates were prepared from pools of eight buffy coats in additive solution (SSP+) using either previous 6‐lead or new 8‐lead pooling sets and PI processing sets in previous or alternate supplier sourced plastics (AS). Platelets were treated with the INTERCEPT Blood System then stored for up to 7 days and tested for in vitro quality. Results All tested units (n = 30) were in conformity with European guidelines. Using AS sets more effectively maintained glucose reserves (P < 0·01), reduced lactate production (P < 0·01), reduced CD62P expression (P < 0·01) and downregulated levels of surface CD42b (P < 0·01) overtime. AS set maintained JC‐positive cells (NS) between day 2 and day 7 and sustained platelet integrin activation (PAC‐1) between day 2 and day 7 (NS). Overall sCD40L and PGDF accumulated in an equivalent way (P < 0·01) within series. Summary/conclusions In summary, our data demonstrate that PI treatment using AS sets, in combination with the new pooling set for double‐dose platelet preparation, maintained the platelet in vitro quality over 7 days of storage.
S. Ohlsson; B. Diedrich; M. Uhlin; P. Sandgren. Optimized processing for pathogen inactivation of double‐dose buffy‐coat platelet concentrates: maintained in vitro quality over 7‐day storage. Vox Sanguinis 2018, 113, 611 -621.
AMA StyleS. Ohlsson, B. Diedrich, M. Uhlin, P. Sandgren. Optimized processing for pathogen inactivation of double‐dose buffy‐coat platelet concentrates: maintained in vitro quality over 7‐day storage. Vox Sanguinis. 2018; 113 (7):611-621.
Chicago/Turabian StyleS. Ohlsson; B. Diedrich; M. Uhlin; P. Sandgren. 2018. "Optimized processing for pathogen inactivation of double‐dose buffy‐coat platelet concentrates: maintained in vitro quality over 7‐day storage." Vox Sanguinis 113, no. 7: 611-621.
Michael Uhlin; Mohamed Abumaree; Essam M. Abdelalim. Therapeutic Use of Extraembryonic-Derived Tissues. Stem Cells International 2018, 2018, 1 -2.
AMA StyleMichael Uhlin, Mohamed Abumaree, Essam M. Abdelalim. Therapeutic Use of Extraembryonic-Derived Tissues. Stem Cells International. 2018; 2018 ():1-2.
Chicago/Turabian StyleMichael Uhlin; Mohamed Abumaree; Essam M. Abdelalim. 2018. "Therapeutic Use of Extraembryonic-Derived Tissues." Stem Cells International 2018, no. : 1-2.
The use of CD19 chimeric antigen receptor (CAR) T cells to treat B-cell malignancies has proven beneficial. Several groups use serum to produce CD19 CAR T cells. Today, ready-to-use serum-free media that require no addition of serum are commercially available. Therefore, it becomes important to evaluate the production of CD19 CAR T cells with and without the addition of serum. T cells from buffy coats were cultured in AIM-V and TexMACS (TM) supplemented with 5% human serum (A5% and TM5%, respectively), and in TM without serum. Cells were activated with OKT3 and expanded in interleukin (IL)-2. Viral transduction was performed in RetroNectin-coated plates using the spinoculation method. CD19 CAR T cells were tested for their viability, expansion, transduction efficacy, phenotype and cytotoxicity. CD19 CAR T cells expanded in A5% and TM5% showed significantly better viability and higher fold expansion than cells expanded in TM. TM promoted the expansion of CD8+ T cells and effector phenotype of CD19 CAR T cells. The transduction efficacy and the cytotoxic function were comparable between the different media. Higher CD107a+ cells were detected in TM and TM5%, whereas higher IL-2+ and IL-17+ cells were detected in A5%. CD19 CAR exhibited co-expression of inhibitory receptors such as TIM-3+LAG-3+ and/or TIM-3+PD-1+. Our results indicate that serum supplementation promotes better CD19 CAR T-cell expansion and viability in vitro. CD19 CAR T cells produced in TM medium showed lower CD4/CD8 ratio, which warrants further evaluation in clinical settings. Overall, the choice of culture medium impacts CD19 CAR T-cell end product.
Rehab Alnabhan; Ahmed Gaballa; Lisa-Mari Mörk; Jonas Mattsson; Michael Uhlin; Isabelle Magalhaes. Media evaluation for production and expansion of anti-CD19 chimeric antigen receptor T cells. Cytotherapy 2018, 20, 941 -951.
AMA StyleRehab Alnabhan, Ahmed Gaballa, Lisa-Mari Mörk, Jonas Mattsson, Michael Uhlin, Isabelle Magalhaes. Media evaluation for production and expansion of anti-CD19 chimeric antigen receptor T cells. Cytotherapy. 2018; 20 (7):941-951.
Chicago/Turabian StyleRehab Alnabhan; Ahmed Gaballa; Lisa-Mari Mörk; Jonas Mattsson; Michael Uhlin; Isabelle Magalhaes. 2018. "Media evaluation for production and expansion of anti-CD19 chimeric antigen receptor T cells." Cytotherapy 20, no. 7: 941-951.
The composition of the graft used for allogeneic hematopoietic stem cell transplantation (HSCT) is important for the treatment outcome. Different apheresis devices may yield significant differences in peripheral blood stem cell graft cellular composition. We compared stem cell grafts produced by Cobe Spectra (Cobe) and Spectra Optia (Optia) with use of the mononuclear cell (MNC) protocol, and evaluated clinical outcome parameters such as graft-versus-host disease (GvHD), transplant-related mortality (TRM), relapse, and overall survival. During 5 years, 31 Cobe Spectra and 40 Spectra Optia grafts were analyzed for CD34, CD3, CD4, CD8, CD19, and CD56 cell content. Clinical outcome parameters were correlated and compared between the two patient groups using different apheresis devices. Optia grafts contained fewer lymphocytes compared to Cobe (p < 0.001). Optia grafts had a significantly lower incidence of acute GvHD Grades II through IV (Cobe 45% vs. Optia 23%; p = 0.039) and TRM (16% vs. 2.5%; p < 0.05) but higher chronic GvHD (32% vs. 67%; p = 0.005) compared to Cobe grafts. Finally, the multivariate analysis showed a significant correlation among the different apheresis devices and both acute GvHD II through IV and severe chronic GvHD. The multivariate analysis also showed a significant correlation between the CD3+ cell dose and the incidence of severe acute GvHD. Optia-obtained grafts yielded a lower acute GvHD Grades II-IV and TRM risk, but had no impact on relapse or overall survival in this study. Understanding and further improvement of peripheral blood stem cell (PBSC) apheresis techniques may be used in the future to personalize HSCT by, for example, fine-tuning the GvHD incidence.
T. Wang; M. Remberger; U. Axdorph Nygell; M. Sundin; A. Björklund; J. Mattsson; M. Uhlin; E. Watz. Change of apheresis device decreased the incidence of severe acute graft-versus-host disease among patients after allogeneic stem cell transplantation with sibling donors. Transfusion 2018, 58, 1442 -1451.
AMA StyleT. Wang, M. Remberger, U. Axdorph Nygell, M. Sundin, A. Björklund, J. Mattsson, M. Uhlin, E. Watz. Change of apheresis device decreased the incidence of severe acute graft-versus-host disease among patients after allogeneic stem cell transplantation with sibling donors. Transfusion. 2018; 58 (6):1442-1451.
Chicago/Turabian StyleT. Wang; M. Remberger; U. Axdorph Nygell; M. Sundin; A. Björklund; J. Mattsson; M. Uhlin; E. Watz. 2018. "Change of apheresis device decreased the incidence of severe acute graft-versus-host disease among patients after allogeneic stem cell transplantation with sibling donors." Transfusion 58, no. 6: 1442-1451.
Gammadelta (γδ) T cells are found in both blood and tissues and have antiviral and antitumor properties. The frequency of γδ T cells in umbilical cord blood (UCB) is low, and the majority express δ1, in contrast to blood, whereas the main subset is δ2γ9 T cells. UCB γδ T cells are functionally immature, which together with their scarcity complicates the development of UCB γδ T cell therapies. We aimed to develop an effective expansion protocol for UCB γδ T cells based on zoledronate and IL-2. We found that culture with 5 μM zoledronate and 200 IU IL-2/ml medium for 14 days promoted extensive proliferation. The majority of the cultured cells were γ9δ2 T cells. The fold expansion of this, originally infrequent, subset was impressive (median and maximum fold change 253 and 1085, resp.). After culture, the cells had a polyclonal γδ T cell repertoire and the main memory subset was central memory (CD45RO+ CD27+). The cells produced cytokines such as IL-1B, IL-2, and IL-8 and displayed significant tumor-killing capacity. These results show that development of in vitro expanded UCB γδ T cell therapies is feasible. It could prove a valuable treatment modality for patients after umbilical cord blood transplantation.
Sofia Berglund; Ahmed Gaballa; Piamsiri Sawaisorn; Berit Sundberg; Michael Uhlin. Expansion of Gammadelta T Cells from Cord Blood: A Therapeutical Possibility. Stem Cells International 2018, 2018, 1 -15.
AMA StyleSofia Berglund, Ahmed Gaballa, Piamsiri Sawaisorn, Berit Sundberg, Michael Uhlin. Expansion of Gammadelta T Cells from Cord Blood: A Therapeutical Possibility. Stem Cells International. 2018; 2018 ():1-15.
Chicago/Turabian StyleSofia Berglund; Ahmed Gaballa; Piamsiri Sawaisorn; Berit Sundberg; Michael Uhlin. 2018. "Expansion of Gammadelta T Cells from Cord Blood: A Therapeutical Possibility." Stem Cells International 2018, no. : 1-15.
Acute graft-versus-host disease (aGVHD) is one of the main major complications of post hematopoietic stem cell transplantation (HSCT). Identifying patients at risk of severe aGVHD may lead to earlier intervention and treatment, resulting in increased survival and a better quality of life. We aimed to identify biomarkers in donor grafts and patient plasma around the time of transplantation which might be predictive of aGVHD development. We build upon our previously published methods by using multiplex assays and multi-colour flow cytometry. We identified five easily assessable cellular markers in donor grafts that combined could potentially be used to calculate risk for severe aGVHD development. Most noteworthy are the T cell subsets expressing IL-7 receptor-α (CD127) and PD-1. Additionally, we identified a potential role for elevated TNFα levels in both graft and patient before HSCT in development of aGVHD.
Arwen Stikvoort; Ahmed Gaballa; Martin Solders; Iris Nederlof; Björn Önfelt; Berit Sundberg; Mats Remberger; Mikael Sundin; Jonas Mattsson; Michael Uhlin. Risk Factors for Severe Acute Graft-versus-Host Disease in Donor Graft Composition. Biology of Blood and Marrow Transplantation 2018, 24, 467 -477.
AMA StyleArwen Stikvoort, Ahmed Gaballa, Martin Solders, Iris Nederlof, Björn Önfelt, Berit Sundberg, Mats Remberger, Mikael Sundin, Jonas Mattsson, Michael Uhlin. Risk Factors for Severe Acute Graft-versus-Host Disease in Donor Graft Composition. Biology of Blood and Marrow Transplantation. 2018; 24 (3):467-477.
Chicago/Turabian StyleArwen Stikvoort; Ahmed Gaballa; Martin Solders; Iris Nederlof; Björn Önfelt; Berit Sundberg; Mats Remberger; Mikael Sundin; Jonas Mattsson; Michael Uhlin. 2018. "Risk Factors for Severe Acute Graft-versus-Host Disease in Donor Graft Composition." Biology of Blood and Marrow Transplantation 24, no. 3: 467-477.
CD19 chimeric antigen receptor (CAR) T cell immunotherapy has demonstrated dramatic results for the treatment of B cell malignancies such as chronic lymphocytic leukemia (CLL). As T cell defects are common in patients with CLL, we compared the T cells from these patients with healthy donors (HDs), and subsequently the CD19 CAR T cells produced from patients and HDs. Despite initial differences when comparing the phenotype of circulating T cells in patients with CLL and HDs, the CD19 CAR T cells manufactured from patients’ or HDs’ cells showed a similar phenotype (effector memory or terminally differentiated), both were specifically activated by and killed CD19+ target cells, and secreted cytokines (ie, IL-2, TNF, and IFN-γ). The frequency of CD19 CAR T cells producing IFN-γ was significantly higher in cells produced from patients as compared with those produced from HDs. Furthermore, our data showed that the polyfunctional profile of CD19 CAR+ T cells was differently modulated by CD19+ K562 cells and autologous B cells. The increased IFN-γ production by CD19 CAR T cells produced from patients with CLL after in vitro stimulation, may if this is also the case in vivo, contribute to a higher risk of a cytokine release syndrome in patients. The different impact by CD19+ target cells on the polyfunctional profile of CD19 CAR T cells in vitro underlines the importance of the choice of CD19+ target cells when assessing CD19 CAR T cells functions.
Isabelle Magalhaes; Ingrid Kalland; James N. Kochenderfer; Anders Österborg; Michael Uhlin; Jonas Mattsson. CD19 Chimeric Antigen Receptor T Cells From Patients With Chronic Lymphocytic Leukemia Display an Elevated IFN-γ Production Profile. Journal of Immunotherapy 2018, 41, 73 -83.
AMA StyleIsabelle Magalhaes, Ingrid Kalland, James N. Kochenderfer, Anders Österborg, Michael Uhlin, Jonas Mattsson. CD19 Chimeric Antigen Receptor T Cells From Patients With Chronic Lymphocytic Leukemia Display an Elevated IFN-γ Production Profile. Journal of Immunotherapy. 2018; 41 (2):73-83.
Chicago/Turabian StyleIsabelle Magalhaes; Ingrid Kalland; James N. Kochenderfer; Anders Österborg; Michael Uhlin; Jonas Mattsson. 2018. "CD19 Chimeric Antigen Receptor T Cells From Patients With Chronic Lymphocytic Leukemia Display an Elevated IFN-γ Production Profile." Journal of Immunotherapy 41, no. 2: 73-83.
Immunodeficiency associated with mutations in the DNA cross-link repair 1C gene (DCLRE1C) can have variable clinical presentations including severe combined immunodeficiency (SCID), Omenn syndrome, atypical SCID or common variable immunodeficiency (CVID) (1-3). DCLRE1C encodes the protein Artemis, a nuclease with intrinsic 5'-3' exonuclease activity on single-stranded DNA that is involved in non-homologous end joining (NHEJ). Artemis is essential for V(D)J recombination of the immunoglobulin and T-cell receptor genes that occur during B- and T-cell development.
Mikael Sundin; Michael Uhlin; Ahmed Gaballa; Kim Ramme; Antonios Kolios; Per Marits; Jakob Nilsson. Late presenting atypical severe combined immunodeficiency (SCID) associated with a novel missense mutation in DCLRE1C. Pediatric Allergy and Immunology 2017, 29, 108 -111.
AMA StyleMikael Sundin, Michael Uhlin, Ahmed Gaballa, Kim Ramme, Antonios Kolios, Per Marits, Jakob Nilsson. Late presenting atypical severe combined immunodeficiency (SCID) associated with a novel missense mutation in DCLRE1C. Pediatric Allergy and Immunology. 2017; 29 (1):108-111.
Chicago/Turabian StyleMikael Sundin; Michael Uhlin; Ahmed Gaballa; Kim Ramme; Antonios Kolios; Per Marits; Jakob Nilsson. 2017. "Late presenting atypical severe combined immunodeficiency (SCID) associated with a novel missense mutation in DCLRE1C." Pediatric Allergy and Immunology 29, no. 1: 108-111.