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      Higher-order connections between stereotyped subsets: implications for improved patient classification in CLL

      1 , 1 , 2 , 1 , 3 , 4 , 1 , 5 , 6 , 7 , 8 , 9 , 10 , 11 , 12 , 2 , 13 , 14 , 15 , 16 , 17 , 18 , 9 , 19 , 20 , 21 , 16 , 22 , 23 , 24 , 11 , 25 , 9 , 5 , 6 , 26 , 27 , 28 , 29 , 27 , 28 , 30 , 25 , 31 , 18 , 23 , 32 , 5 , 1 , 22 , 33 , 2 , 34 , 3 , 35 , 19 , 36 , 37 , 38 , 39 , 40 , 32 , 41 , 42 , 43 , 44 , 45 , 22 , 27 , 28 , 46 , 30 , 31 , 11 , 20 , 21 , 47 , 45 , 26 , 48 , 49 , 7 , 17 , 9 , 50 , 8 , 5 , 6 , 4 , 4 , 15 , 51 , 15 , 2 , 52 , 14 , 1 , 2
      Blood
      American Society of Hematology

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          Abstract

          Chronic lymphocytic leukemia (CLL) is characterized by the existence of subsets of patients with (quasi)identical, stereotyped B-cell receptor (BcR) immunoglobulins. Patients in certain major stereotyped subsets often display remarkably consistent clinicobiological profiles, suggesting that the study of BcR immunoglobulin stereotypy in CLL has important implications for understanding disease pathophysiology and refining clinical decision-making. Nevertheless, several issues remain open, especially pertaining to the actual frequency of BcR immunoglobulin stereotypy and major subsets, as well as the existence of higher-order connections between individual subsets. To address these issues, we investigated clonotypic IGHV-IGHD-IGHJ gene rearrangements in a series of 29 856 patients with CLL, by far the largest series worldwide. We report that the stereotyped fraction of CLL peaks at 41% of the entire cohort and that all 19 previously identified major subsets retained their relative size and ranking, while 10 new ones emerged; overall, major stereotyped subsets had a cumulative frequency of 13.5%. Higher-level relationships were evident between subsets, particularly for major stereotyped subsets with unmutated IGHV genes (U-CLL), for which close relations with other subsets, termed “satellites,” were identified. Satellite subsets accounted for 3% of the entire cohort. These results confirm our previous notion that major subsets can be robustly identified and are consistent in relative size, hence representing distinct disease variants amenable to compartmentalized research with the potential of overcoming the pronounced heterogeneity of CLL. Furthermore, the existence of satellite subsets reveals a novel aspect of repertoire restriction with implications for refined molecular classification of CLL.

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          Most cited references80

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          Potent neutralizing antibodies against SARS-CoV-2 identified by high-throughput single-cell sequencing of convalescent patients’ B cells

          Summary The COVID-19 pandemic urgently needs therapeutic and prophylactic interventions. Here we report the rapid identification of SARS-CoV-2 neutralizing antibodies by high-throughput single-cell RNA and VDJ sequencing of antigen-enriched B cells from 60 convalescent patients. From 8,558 antigen-binding IgG1+ clonotypes, 14 potent neutralizing antibodies were identified with the most potent one, BD-368-2, exhibiting an IC50 of 1.2 ng/mL and 15 ng/mL against pseudotyped and authentic SARS-CoV-2, respectively. BD-368-2 also displayed strong therapeutic and prophylactic efficacy in SARS-CoV-2-infected hACE2-transgenic mice. Additionally, the 3.8Å Cryo-EM structure of a neutralizing antibody in complex with the spike-ectodomain trimer revealed the antibody’s epitope overlaps with the ACE2 binding site. Moreover, we demonstrated that SARS-CoV-2 neutralizing antibodies could be directly selected based on similarities of their predicted CDR3H structures to those of SARS-CoV neutralizing antibodies. Altogether, we showed that human neutralizing antibodies could be efficiently discovered by high-throughput single B-cell sequencing in response to pandemic infectious diseases.
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            Guidelines for diagnosis, indications for treatment, response assessment and supportive management of chronic lymphocytic leukemia

            The previous edition of the consensus guidelines of the International Workshop on Chronic Lymphocytic Leukemia (iwCLL), published in 2008, has found broad acceptance by physicians and investigators caring for patients with CLL. Recent advances including the discovery of the genomic landscape of the disease, the development of genetic tests with prognostic relevance, and the detection of minimal residual disease (MRD), coupled with the increased availability of novel targeted agents with impressive efficacy, prompted an international panel to provide updated evidence- and expert opinion-based recommendations. These recommendations include a revised version of the iwCLL response criteria, an update on the use of MRD status for clinical evaluation, and recommendations regarding the assessment and prophylaxis of viral diseases during management of CLL.
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              Diversity in the CDR3 region of V(H) is sufficient for most antibody specificities.

              J. Xu, M Davis (2000)
              All rearranging antigen receptor genes have one or two highly diverse complementarity determining regions (CDRs) among the six that typically form the ligand binding surface. We report here that, in the case of antibodies, diversity at one of these regions, CDR3 of the V(H) domain, is sufficient to permit otherwise identical IgM molecules to distinguish between a variety of hapten and protein antigens. Furthermore, we find that somatic mutation can allow such antibodies to achieve surprisingly high affinities. These results are consistent with a model in which the highly diverse CDR3 loops are the key determinant of specificity in antigen recognition in both T cell receptors (TCR) and antibodies, whereas the germline-encoded CDR1 and CDR2 sequences are much more cross-reactive.
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                Journal
                Blood
                American Society of Hematology
                0006-4971
                1528-0020
                March 11 2021
                March 11 2021
                : 137
                : 10
                : 1365-1376
                Affiliations
                [1 ]Institute of Applied Biosciences, Centre for Research and Technology Hellas, Thessaloniki, Greece;
                [2 ]Department of Molecular Medicine and Surgery, Karolinska Institute, Stockholm, Sweden;
                [3 ]Department of Molecular Biology and Genetics, Democritus University of Thrace, Alexandroupolis, Greece;
                [4 ]International ImMunoGeneTics Information System (IMGT), Laboratoire d’ImmunoGénétique Moléculaire (LIGM), Institut de Génétique Humaine (IGH), Unité Mixte de Recherche (UMR), Centre National de la Recherche Scientifique (CNRS), Université de Montpellier, Montpellier, France;
                [5 ]Department of Internal Medicine, Hematology and Oncology, Faculty of Medicine, Masaryk University–University Hospital Brno, Brno, Czech Republic;
                [6 ]Center of Molecular Medicine, Central European Institute of Technology, Masaryk University, Brno, Czech Republic;
                [7 ]Department of Haematology, Royal Bournemouth Hospital, Bournemouth, United Kingdom;
                [8 ]The Feinstein Institute for Medical Research, Northwell Health, Manhasset, NY;
                [9 ]MLL Munich Leukemia Laboratory, Munich, Germany;
                [10 ]University Hospital Medical Center, Ulm, Germany;
                [11 ]Deparment of Hematology, Rigshospitalet, Copenhagen University Hospital, Copenhagen, Denmark;
                [12 ]Department of Immunology, Mayo Clinic, Rochester, MN;
                [13 ]Science for Life Laboratory, Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden;
                [14 ]Strategic Research Program on CLL, B-Cell Neoplasia Unit, Division of Experimental Oncology, Università Vita-Salute San Raffaele/Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS) Ospedale San Raffaele, Milan, Italy;
                [15 ]Laboratory Medical Immunology, Department of Immunology, Erasmus MC, University Medical Center (UMC), Rotterdam, The Netherlands;
                [16 ]Assistance Publique–Hôpitaux de Paris (AP-HP), Hôpital Pitié-Salpêtrière, Department of Biological Hematology, Sorbonne Université, UMR_S 1138, Centre de Recherche des Cordeliers, Paris, France;
                [17 ]Department of Internal Medicine III, Ulm University, Ulm, Germany;
                [18 ]National Research Center for Hematology, Moscow, Russia;
                [19 ]Department of Experimental Medicine, University of Genoa, Genoa, Italy;
                [20 ]Centro de Investigacion Biomedica en Red en Oncologia (CIBERONC), Madrid, Spain;
                [21 ]Institut d’Investigacions Biomediques August Pi I Sunyer, Barcelona, Spain;
                [22 ]Molecular Pathology Unit, Haematology Department, Niguarda Cancer Center, Niguarda Hospital, Milan, Italy;
                [23 ]Department of Laboratory Medicine, Medical University of Vienna, Vienna, Austria;
                [24 ]Faculty of Pharmacy, Ss Cyril and Methodius University of Skopje, Skopje, Republic of Northern Macedonia;
                [25 ]Clinical Haematology, Belfast City Hospital, Belfast Health and Social Care Trust, Belfast, United Kingdom;
                [26 ]Second Medical Department, University Hospital Schleswig-Holstein, Campus Kiel, Kiel, Germany;
                [27 ]Hematology and Clinical Immunology Unit, Department of Medicine (DIMED), University of Padua, Padua, Italy;
                [28 ]Veneto Institute of Molecular Medicine, Padua, Italy;
                [29 ]Division of Immunology, Transplantation and Infectious Diseases, IRCCS San Raffaele Scientific Institute, Milan, Italy;
                [30 ]Division of Hematology, Department of Translational Medicine, University of Eastern Piedmont–Ospedale Maggiore della Carità, Novara, Italy;
                [31 ]General Anatomopathology and Molecular Oncogenetics, Azienda Ospedaliero Universitaria (AOU), City of Health and Science of Turin, Turin, Italy;
                [32 ]First Department of Propaedeutic Medicine, University of Athens, Athens, Greece;
                [33 ]Experimental Hematooncology Department, Medical University of Lublin, Lublin, Poland;
                [34 ]Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Belgrade, Serbia;
                [35 ]MTA-SE Momentum Molecular Oncohematology Research Group, First Department of Pathology and Experimental Cancer Research, Semmelweis University, Budapest, Hungary;
                [36 ]UO Molecular Pathology, IRCCS Ospedale Policlinico San Martino, Genoa, Italy;
                [37 ]Department of Hematology, Amsterdam Infection and Immunity Institute, Cancer Center Amsterdam, Amsterdam UMC–University of Amsterdam, Amsterdam, The Netherlands;
                [38 ]Department of Hematology, Faculty of Medicine, Ss Cyril and Methodius University of Skopje, Skopje, Republic of Northern Macedonia;
                [39 ]Division of Hematology, Oncology Institute of Southern Switzerland, Bellinzona, Switzerland;
                [40 ]Department of Medicine, Faculty of Medicine, Kuwait University, Safat, Kuwait;
                [41 ]The Center for the Study of Haematological Malignancies, Nicosia, Cyprus;
                [42 ]Karaiskakio Foundation, Nicosia, Cyprus;
                [43 ]Laboratorio de Citogenètica Molecular, Laboratori de Citologia Hematològica, Servei de Patologia i Servei de Hematologia, Hospital del Mar, Barcelona, Spain;
                [44 ]Clinic for Hematology, Clinical Center of Serbia, Belgrade, Serbia;
                [45 ]Hammersmith Hospital, London, United Kingdom;
                [46 ]Hematocrit (HCT) Unit, Hematology Department, G. Papanicolaou Hospital, Thessaloniki, Greece;
                [47 ]Hospital Clinic of Barcelona, University of Barcelona, Barcelona, Spain;
                [48 ]University Hospital Cologne, Cologne, Germany;
                [49 ]Department I of Internal Medicine, University of Cologne, Cologne, Germany;
                [50 ]Department of Immunology, Mayo Clinic, Scottsdale, AZ;
                [51 ]Hematology Department, Nikea General Hospital, Pireaus, Greece; and
                [52 ]Clinical Genetics, Karolinska University Laboratory, Karolinska University Hospital, Stockholm, Sweden
                Article
                10.1182/blood.2020007039
                7976441
                32992344
                cb0feaae-ae87-4a15-86cd-d4735a9b62c5
                © 2021
                History

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