Cornelia S. Link

Abundant cytomegalovirus (CMV) reactive clonotypes in the CD8(+) T cell receptor alpha repertoire following allogeneic transplantation.

Allogeneic stem cell transplantation is potentially curative, but associated with post‐transplantation complications, including cytomegalovirus (CMV) infections. An effective immune response requires T cells recognizing CMV epitopes via their T cell receptors (TCRs). Little is known about the TCR repertoire, in particular the TCR‐α repertoire and its clinical relevance in patients following stem cell transplantation. Using next‐generation sequencing we examined the TCR‐α repertoire of CD8+ T cells and CMV‐specific CD8+ T cells in four patients. Additionally, we performed single‐cell TCR‐αβ sequencing of CMV‐specific CD8+ T cells. The TCR‐α composition of human leucocyte antigen (HLA)‐A*0201 CMVpp65– and CMVIE‐specific T cells was oligoclonal and defined by few dominant clonotypes. Frequencies of single clonotypes reached up to 11% of all CD8+ T cells and half of the total CD8+ T cell repertoire was dominated by few CMV‐reactive clonotypes. Some TCR‐α clonotypes were shared between patients. Gene expression of the circulating CMV‐specific CD8+ T cells was consistent with chronically activated effector memory T cells. The CD8+ T cell response to CMV reactivation resulted in an expansion of a few TCR‐α clonotypes to dominate the CD8+ repertoires. These results warrant further larger studies to define the ability of oligoclonally expanded T cell clones to achieve an effective anti‐viral T cell response in this setting.

Abundant CMV reactive clonotypes in the CD8+ T cell receptor alpha repertoire following allogeneic transplantation

Allogeneic stem cell transplantation is potentially curative, but associated with post‐transplantation complications, including cytomegalovirus (CMV) infections. An effective immune response requires T cells recognizing CMV epitopes via their T cell receptors (TCRs). Little is known about the TCR repertoire, in particular the TCR‐α repertoire and its clinical relevance in patients following stem cell transplantation. Using next‐generation sequencing we examined the TCR‐α repertoire of CD8+ T cells and CMV‐specific CD8+ T cells in four patients. Additionally, we performed single‐cell TCR‐αβ sequencing of CMV‐specific CD8+ T cells. The TCR‐α composition of human leucocyte antigen (HLA)‐A*0201 CMVpp65– and CMVIE‐specific T cells was oligoclonal and defined by few dominant clonotypes. Frequencies of single clonotypes reached up to 11% of all CD8+ T cells and half of the total CD8+ T cell repertoire was dominated by few CMV‐reactive clonotypes. Some TCR‐α clonotypes were shared between patients. Gene expression of the circulating CMV‐specific CD8+ T cells was consistent with chronically activated effector memory T cells. The CD8+ T cell response to CMV reactivation resulted in an expansion of a few TCR‐α clonotypes to dominate the CD8+ repertoires. These results warrant further larger studies to define the ability of oligoclonally expanded T cell clones to achieve an effective anti‐viral T cell response in this setting.

Generation of high-avidity, WT1-reactive CD8+ cytotoxic T cell clones with anti-leukemic activity by streptamer technology

Antje Tunger, Rebekka Wehner, Malte von Bonin, Denise K€ uhn, Falk Heidenreich, Sarah Matko, Magdalena Nauerth, Elke R€ ucker-Braun, Sevina Dietz, Cornelia S. Link, Anne Eugster, Marcus Odendahl, Dirk H. Busch, Torsten Tonn, Ezio Bonifacio, Lothar Germeroth, Johannes Schetelig, Michael P. Bachmann, Martin Bornh€auser and Marc Schmitz Institute of Immunology, Medical Faculty, TU Dresden, Dresden, Germany; National Center for Tumor Diseases, University Hospital Carl Gustav Carus, TU Dresden, Germany; Department of Medicine I, University Hospital of Dresden, Dresden, Germany; German Cancer Consortium (DKTK), Dresden, Germany; German Cancer Research Center (DKFZ), Heidelberg, Germany; Center for Regenerative Therapies Dresden (CRTD), Medical Faculty, TU Dresden, Dresden, Germany; Institute of Transfusion Medicine, German Red Cross Blood Donation Service North-East, Dresden, Germany; Institute for Medical Microbiology, Immunology and Hygiene, TU Munich, Munich, Germany; Juno Therapeutics GmbH, G€ottingen, Germany; Department of Radioimmunology, Institute of Radiopharmaceutical Cancer Research, Helmholtz Center Dresden-Rossendorf, Dresden, Germany

Association of Oesophageal Varices and Splanchnic Vein Thromboses in Patients with JAK2-Positive Myeloproliferative Neoplasms: Presentation of Two Cases and Data from a Retrospective Analysis

Background: Oesophageal varices and gastrointestinal bleeding are common complications of liver cirrhosis. More rarely, oesophageal varices occur in patients with non-cirrhotic portal hypertension that results from thromboses of portal or splanchnic veins. Case Report: We describe 2 young men who initially presented with varices as a result of portal vein thromboses. In the clinical follow-up, both were tested positive for a JAK2 mutation and consequently diagnosed with myeloproliferative neoplasms (MPNs). In an attempt to characterise the frequency of gastrointestinal complications in patients with JAK2-positive MPNs, we retrospectively analysed all known affected patients from our clinic for the diagnosis of portal vein thromboses and oesophageal varices. Strikingly, 48% of those who had received an oesophagogastroduodenoscopy had detectable oesophageal or gastric varices, and 82% of those suffered from portal or splanchnic vein thromboses. Conclusion: While the association between JAK2, myeloproliferative disease and thrombotic events is well established, patients with idiopathic oesophageal varices are not regularly tested for JAK2 mutations. However, the occurrence of oesophageal varices may be the first presenting symptom of a MPN with a JAK2 mutation, and affected patients may profit from a close haematological monitoring to assure the early detection of developing MPN.

Long-term follow-up of patients with relapsed or refractory non-Hodgkin’s lymphoma receiving allogeneic stem cell transplantation

Long-term follow-up of patients with relapsed or refractory non-Hodgkin’s lymphoma receiving allogeneic stem cell transplantation

Assessment of the T cell receptor repertoire in long-term platelet donors by next generation sequencing.

Temporary suppression of circulating antiplatelet alloantibodies by the massive infusion of fresh, stored, or lyophilized platelets. Transfusion, 18, 429–435. Narvios, A., Reddy, V., Martinez, F. & Lichtiger, B. (2005) Slow infusion of platelets: a possible alternative in the management of refractory thrombocytopenic patients. American Journal of Hematology, 79, 80. Stanworth, S.J., Navarrete, C., Estcourt, L. & Marsh, J. (2015) Platelet refractoriness–practical approaches and ongoing dilemmas in patient management. British Journal of Haematology, 171, 297–305. Vassallo, R. (2013) Management of the platelettransfusion-refractory patient. In: Platelet Transfusion Therapy (eds. by Sweeney, J.D. & Lozano, M.), pp. 321–358. AABB Press, Bethesda (MD).

Anti-CD20 immunotherapy as a bridge to tolerance, after allogeneic stem cell transplantation for patients with chronic lymphocytic leukaemia: results of the CLLX4 trial

Bashford-Rogers, R.J., Nicolaou, K.A., Bartram, J., Goulden, N.J., Loizou, L., Koumas, L., Chi, J., Hubank, M., Kellam, P., Costeas, P.A. & Vassiliou, G.S. (2016) Eye on the B-ALL: B-cell receptor repertoires reveal persistence of numerous B-lymphoblastic leukemia subclones from diagnosis to relapse. Leukemia, 30, 2312– 2321. Bolotin, D.A., Poslavsky, S., Mitrophanov, I., Shugay, M., Mamedov, I.Z., Putintseva, E.V. & Chudakov, D.M. (2015) MiXCR: software for comprehensive adaptive immunity profiling. Nature Methods, 12, 380–381. Ding, L.W., Sun, Q.Y., Tan, K.T., Chien, W., Thippeswamy, A.M., Eng Juh Yeoh, A., Kawamata, N., Nagata, Y., Xiao, J.F., Loh, X.Y., Lin, D.C., Garg, M., Jiang, Y.Y., Xu, L., Lim, S.L., Liu, L.Z., Madan, V., Sanada, M., Fernandez, L.T., Preethi, H., Lill, M., Kantarjian, H.M., Kornblau, S.M., Miyano, S., Liang, D.C., Ogawa, S., Shih, L.Y., Yang, H. & Koeffler, H.P. (2017) Mutational landscape of pediatric acute lymphoblastic leukemia. Cancer Research, 77, 390–400. van Dongen, J.J., Langerak, A.W., Bruggemann, M., Evans, P.A., Hummel, M., Lavender, F.L., Delabesse, E., Davi, F., Schuuring, E., GarciaSanz, R., van Krieken, J.H., Droese, J., Gonzalez, D., Bastard, C., White, H.E., Spaargaren, M., Gonzalez, M., Parreira, A., Smith, J.L., Morgan, G.J., Kneba, M. & Macintyre, E.A. (2003) Design and standardization of PCR primers and protocols for detection of clonal immunoglobulin and T-cell receptor gene recombinations in suspect lymphoproliferations: report of the BIOMED-2 Concerted Action BMH4-CT98-3936. Leukemia, 17, 2257–2317. Gawad, C., Pepin, F., Carlton, V.E., Klinger, M., Logan, A.C., Miklos, D.B., Faham, M., Dahl, G. & Lacayo, N. (2012) Massive evolution of the immunoglobulin heavy chain locus in children with B precursor acute lymphoblastic leukemia. Blood, 120, 4407–4417. Kim, E., Hurtz, C., Koehrer, S., Wang, Z., Balasubramanian, S., Chang, B.Y., Muschen, M., Davis, R.E. & Burger, J.A. (2017) Ibrutinib inhibits pre-BCR+ B-cell acute lymphoblastic leukemia progression by targeting BTK and BLK. Blood, 129, 1155–1165. Langerak, A.W. & van Dongen, J.J.M. (2012) Multiple clonal Ig/TCR products: implications for interpretation of clonality findings. Journal of Hematopathology, 5, 35–43. Muschen, M. (2015) Rationale for targeting the pre-B-cell receptor signaling pathway in acute lymphoblastic leukemia. Blood, 125, 3688–3693. Qian, M., Zhang, H., Kham, S.K., Liu, S., Jiang, C., Zhao, X., Lu, Y., Goodings, C., Lin, T.N., Zhang, R., Moriyama, T., Yin, Z., Li, Z., Quah, T.C., Ariffin, H., Tan, A.M., Shen, S., Bhojwani, D., Hu, S., Chen, S., Zheng, H., Pui, C.H., Yeoh, A.E. & Yang, J.J. (2017) Whole-transcriptome sequencing identifies a distinct subtype of acute lymphoblastic leukemia with predominant genomic abnormalities of EP300 and CREBBP. Genome Research, 27, 185–195. van der Velden, V.H., Szczepanski, T., Wijkhuijs, J.M., Hart, P.G., Hoogeveen, P.G., Hop, W.C., van Wering, E.R. & van Dongen, J.J. (2003) Age-related patterns of immunoglobulin and Tcell receptor gene rearrangements in precursorB-ALL: implications for detection of minimal residual disease. Leukemia, 17, 1834–1844.

Validation of the Revised Pretransplant Assessment of Mortality Score in Patients with Acute Myelogenous Leukemia Undergoing Allogeneic Hematopoietic Stem Cell Transplantation

Despite recent advances, allogeneic hematopoietic stem cell transplantation (allo-HSCT) continues to be accompanied by a high rate of morbidity and mortality. Several scores have been developed to predict outcome after allo-HSCT. The recently revised Pretransplant Assessment of Mortality (PAM) score is based on patient age, donor type, disease risk, cytomegalovirus (CMV) serostatus of patient and donor, and forced expiratory volume in 1 second (FEV1). The aim of this study was to analyze the predictive power of the PAM score in an independent large cohort of patients with acute myelogenous leukemia (AML). We selected adult patients with AML who underwent a first allo-HSCT at the University Hospital of Dresden, a tertiary care hospital with a large transplantation program. All adult patients treated between January 1, 2003, and July 1, 2015, were included. The PAM score was calculated as described previously. Overall survival (OS), cumulative incidence of relapse (CIR), and nonrelapse mortality (NRM) after allo-HSCT were analyzed. Age, AML type, sex match, CMV match, donor type, European Leukemia Net risk classification, type of conditioning, disease stage, and PAM score as a continuous variable were selected a priori for multivariate Cox regression analyses. A total of 544 patients met the inclusion criteria. The median patient age was 57 years. With a median follow-up of 47 months (range, 1 to 161 months), the estimated OS for the whole cohort at 4 years was 43%, with a CIR of 30% and an NRM of 31%. The probability of OS at 4 years was 65% for patients with a PAM score of 0, 52% in those with a PAM score of 1, 33% in those with a PAM score of 2, and 22% in those with a PAM score of 3 (P < .001, log-rank test). Both the CIR and NRM increased with higher PAM scores (P = .005 and P < .001, respectively, Gray test). In multivariate analysis, age (hazard ratio [HR], 1.02 per year; P = .004), disease stage (primary induction failure versus first complete remission (CR1); HR, 1.5; P = .03), and the PAM score (HR 1.04; P = .03) had a significant impact on OS. This is the first independent validation of the revised PAM score allowing for simple and valid estimation of transplantation outcomes. It can serve as an important tool in counseling patients with AML, as well as in designing future trials.

Mass spectrometry-based identification of a naturally presented receptor tyrosine kinase-like orphan receptor 1-derived epitope recognized by CD8+ cytotoxic T cells

Despite recent treatment improvements with the approval of cell signaling pathway inhibitors, additional treatment options are needed for patients with relapsed/refractory high-risk chronic lymphocytic leukemia (CLL). An emerging immunotherapeutic strategy for CLL is based on the adoptive transfer