2,369 results on '"Immunophenotyping methods"'
Search Results
102. Longitudinal characterization of phenotypic profile of T cells in chronic hepatitis B identifies immune markers associated with HBsAg loss.
- Author
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Xiong S, Zhu D, Liang B, Li M, Pan W, He J, Wang H, Sutter K, Dittmer U, Lu M, Liu D, Yang D, Liu J, and Zheng X
- Subjects
- Adult, Biomarkers blood, Female, Flow Cytometry methods, HLA-DR Antigens immunology, Hepatitis B, Chronic immunology, Humans, Immunophenotyping methods, Male, Hepatitis B Surface Antigens immunology, Hepatitis B, Chronic blood, T-Lymphocytes immunology
- Abstract
Background: The current desirable endpoint of treatment against chronic hepatitis B virus infection (cHBV) is to achieve a functional cure, which is defined as HBsAg loss (sAg-L) with or without anti-HBs seroconversion. However, the immunological features that are associated with functional cure have not been studied in detail., Methods: 172 cHBV patients (67 HBeAg+ and 105 HBeAg-), including 141 HBsAg retained (sAg-R) patients (115 chronic hepatitis and 26 asymptomatic carriers), 31 sAg-L patients, and 24 healthy individuals (vaccinated but not infected with HBV) were examined for their T cell phenotypic profile and HBV-specific T cell responses by flow cytometry. 18 cHBV patients with low serum HBsAg levels were also longitudinally followed for their T cell phenotypic profile and HBV-specific T cell responses up to 60 weeks., Findings: sAg-L patients showed distinct CD4
+ and CD8+ T cell phenotype fingerprints compared to those of sAg-R patients, as mainly indicated by the upregulation of HLA-DR on both CD4+ and CD8+ T cells, and a potent HBcAg-specific CD8+ T cell response. The changes in the T cell phenotype in cHBV patients were even more profound during rapid HBsAg decrease and was associated with interferon α treatment. The expression of HLA-DR (r = 0·3269, p = 0·0037), CD95 (r = 0·2796, p = 0·0151), CD40L (r = 0·2747, p = 0·0156), CTLA-4 (r = 0·2786, p = 0·0148), TIM-3 (r = 0·3082, p = 0·0068), CD107a (r = 0·3597, p = 0·0013) on CD4+ T cells, and HLA-DR (r = 0·3542, p = 0·0016), CD69 (r = 0·2507, p = 0·0279), CD107a (r = 0·2875, p = 0·0112) on CD8+ T cells were positively correlated with the rate of HBsAg decrease. The expression of HLA-DR (r = 0·2846, p = 0·0009) and CD95 (r = 0·2442, p = 0·0049) on CD8+ T cells were positively correlated with the magnitude of the HBcAg-specific T cell responses in cHBV patients. Importantly, CTLA-4, CD95 and CD107a expression on CD4+ T cells, as well as HLA-DR and TIM-3 expression on CD8+ T cells in combination with HBsAg quantification were identified as potential predictive factors for sAg-L within 48 weeks in cHBV patients., Interpretation: The onset of HBsAg decrease and subsequent loss in cHBV patients on treatment is associated with significant alterations of both CD4+ and CD8+ T cell phenotypes. Characterization of the T cell phenotype in cHBV patients may present predicative value for sAg-L., Funding: National Natural Science Foundation of China, National Scientific and Technological Major Project of China, Integrated Innovative Team for Major Human Diseases Program of Tongji Medical College, "Double-First Class" Project for the International Cooperation Center on Infection and Immunity, HUST., Competing Interests: Declaration of Competing Interest The authors have declared that no conflict of interest exists., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)- Published
- 2021
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103. Value of flow cytometry for MRD-based relapse prediction in B-cell precursor ALL in a multicenter setting.
- Author
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Modvig S, Hallböök H, Madsen HO, Siitonen S, Rosthøj S, Tierens A, Juvonen V, Osnes LTN, Vålerhaugen H, Hultdin M, Matuzeviciene R, Stoskus M, Marincevic M, Lilleorg A, Ehinger M, Norén-Nystrøm U, Toft N, Taskinen M, Jónsson OG, Pruunsild K, Vaitkeviciene G, Vettenranta K, Lund B, Abrahamsson J, Porwit A, Schmiegelow K, and Marquart HV
- Subjects
- Adolescent, Adult, Child, Child, Preschool, Female, Flow Cytometry methods, Humans, Immunophenotyping methods, Infant, Male, Middle Aged, Recurrence, Young Adult, Neoplasm, Residual drug therapy, Neoplasm, Residual pathology, Precursor Cell Lymphoblastic Leukemia-Lymphoma drug therapy, Precursor Cell Lymphoblastic Leukemia-Lymphoma pathology, Precursor Cells, B-Lymphoid drug effects, Precursor Cells, B-Lymphoid pathology
- Abstract
PCR of TCR/Ig gene rearrangements is considered the method of choice for minimal residual disease (MRD) quantification in BCP-ALL, but flow cytometry analysis of leukemia-associated immunophenotypes (FCM-MRD) is faster and biologically more informative. FCM-MRD performed in 18 laboratories across seven countries was used for risk stratification of 1487 patients with BCP-ALL enrolled in the NOPHO ALL2008 protocol. When no informative FCM-marker was available, risk stratification was based on real-time quantitative PCR. An informative FCM-marker was found in 96.2% and only two patients (0.14%) had non-informative FCM and non-informative PCR-markers. The overall 5-year event-free survival was 86.1% with a cumulative incidence of relapse (CIR
5y ) of 9.5%. FCM-MRD levels on days 15 (HzR 4.0, p < 0.0001), 29 (HzR 2.7, p < 0.0001), and 79 (HzR 3.5, p < 0.0001) associated with hazard of relapse adjusted for age, cytogenetics, and WBC. The early (day 15) response associated with CIR5y adjusted for day 29 FCM-MRD, with higher levels in adults (median 2.4 × 10-2 versus 5.2 × 10-3 , p < 0.0001). Undetectable FCM- and/or PCR-MRD on day 29 identified patients with a very good outcome (CIR5y = 3.2%). For patients who did not undergo transplantation, day 79 FCM-MRD > 10-4 associated with a CIR5y = 22.1%. In conclusion, FCM-MRD performed in a multicenter setting is a clinically useful method for MRD-based treatment stratification in BCP-ALL.- Published
- 2021
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104. Primary cutaneous follicle center lymphoma with extensive plasmacytic differentiation and t(14;18) in both the lymphoid and plasma cell components.
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Kelley JT, Brown NA, Hristov AC, and Bresler SC
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- Aged, Biopsy, Needle methods, Cell Differentiation, Diagnosis, Differential, Female, Humans, Immunoglobulin lambda-Chains metabolism, Immunohistochemistry methods, Immunophenotyping methods, In Situ Hybridization, Fluorescence methods, Lymphocytes metabolism, Lymphoma, B-Cell, Marginal Zone pathology, Lymphoma, Follicular metabolism, Neprilysin metabolism, Plasma Cells metabolism, Proto-Oncogene Proteins c-bcl-2 metabolism, Proto-Oncogene Proteins c-bcl-6 metabolism, Translocation, Genetic, Lymphocytes pathology, Lymphoma, B-Cell, Marginal Zone diagnosis, Lymphoma, Follicular diagnosis, Plasma Cells pathology, Skin Neoplasms pathology
- Abstract
Primary cutaneous follicle center lymphoma (PCFCL) is the most common cutaneous B-cell lymphoma. The typical immunophenotype includes expression of both CD20 and BCL6, with the majority of cases lacking expression of CD10, BCL2, and the characteristic t(14;18)/IGH-BCL2 rearrangement seen in systemic follicular lymphoma (FL). Plasmacytic differentiation (PD) is an uncommon finding in both systemic and cutaneous FLs and presents a diagnostic challenge when present, leading to the potential for misdiagnosis as marginal zone lymphoma (MZL). Limited reports have described light chain restriction in the plasma cell component of FLs with PD, and rare cases of PCFCL with PD have been described. While the IGH-BCL2 translocation has been identified in a subset of FLs with PD, the presence of the BCL2 translocation in monotypic plasma cells of PCFCL has not been previously described to our knowledge. Here, we report a case of PCFCL with extensive PD in a 77-year-old woman that was favored to represent primary cutaneous MZL on an initial punch biopsy. Excisional biopsy, however, revealed that the atypical lymphocytes expressed CD10, BCL6, and BCL2, while the plasma cell component demonstrated light-chain lambda restriction. FISH studies showed the presence of an IGH-BCL2 translocation in both the lymphocytic and plasmacytic components., (© 2021 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)
- Published
- 2021
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105. Histomorphological and immunophenotypical spectrum of cutaneous myoepitheliomas: A series of 35 cases.
- Author
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Plaza JA, Brenn T, Chung C, Salim S, Linos KD, Jour G, Duran Rincon J, Wick M, Sangueza M, and Gru AA
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- Actins metabolism, Adenoma, Pleomorphic pathology, Adult, Anion Exchange Protein 1, Erythrocyte metabolism, Awareness, Biomarkers metabolism, Biomarkers, Tumor metabolism, Calcium-Binding Proteins metabolism, Carcinoma pathology, Chloride-Bicarbonate Antiporters metabolism, Desmin metabolism, Diagnosis, Differential, Female, Glial Fibrillary Acidic Protein metabolism, Humans, Immunohistochemistry methods, Keratins metabolism, Male, Membrane Proteins metabolism, Microfilament Proteins metabolism, Mucin-1 metabolism, Myoepithelioma pathology, Myoepithelioma ultrastructure, S100 Proteins metabolism, Calponins, Immunophenotyping methods, Myoepithelioma diagnosis, Myoepithelioma metabolism, Skin Neoplasms pathology
- Abstract
Myoepithelial tumors comprise a group of mesenchymal lesions that show heterogeneous histomorphological features, including dual epithelial, neural, and myoid differentiation. Cutaneous myoepithelioma is a rare neoplasm that is composed primarily of myoepithelial cells and represents one end of a histopathological spectrum of cutaneous myoepithelial neoplasms including chondroid syringoma and myoepithelial carcinoma. These tumors display a wide histopathological spectrum and immunophenotypical profile often showing epithelial and myoepithelial differentiation. In this series, we studied 35 cases of cutaneous myoepitheliomas. Our cases highlighted the broad histopathological range where most cases showed a non-infiltrative and non-encapsulated tumor exclusively located in the dermis and with no subcutaneous involvement. The majority of our cases had a solid growth pattern (syncytial pattern) and the remainder of cases had a multinodular growth pattern. The tumor cells were epithelioid in 23 cases, spindled in eight cases and there was a mixture of epithelioid and spindled cells in four cases. Mitotic figures ranged from 0 to 5 per 10 HPF. By immunohistochemistry epithelial membrane antigen (EMA) was expressed in 59% of cases S100 was positive in 88% of cases, CAM 5.2 was positive in 16% of cases, AE1/AE3 was positive in 44% of cases, p63 was positive in 17% of cases, smooth muscle actin was positive in 38% of cases, desmin was positive in 6% of cases, calponin was positive in 22% of cases, and glial fibrillary acidic protein was positive in 36% of cases. In addition, there were five cases without EMA, keratin, or p63 expression that only showed S100 expression. We describe a large series of cutaneous myoepitheliomas delineating their histomorphological spectrum and immunophenotypical profile. Awareness of some of the unusual histopathological features and the heterogeneous immunohistochemical may pose difficulties for the diagnosis., (© 2020 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)
- Published
- 2021
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106. CD319 (SLAMF7) an alternative marker for detecting plasma cells in the presence of daratumumab or elotuzumab.
- Author
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Soh KT, Tario JD Jr, Hahn T, Hillengass J, McCarthy PL, and Wallace PK
- Subjects
- ADP-ribosyl Cyclase 1 antagonists & inhibitors, Adolescent, Adult, Antibodies, Monoclonal adverse effects, Antibodies, Monoclonal blood, Antibodies, Monoclonal, Humanized adverse effects, Antibodies, Monoclonal, Humanized blood, Female, Flow Cytometry methods, Gene Expression Regulation, Neoplastic genetics, Granulocytes metabolism, Granulocytes pathology, Humans, Immunophenotyping methods, Intercellular Adhesion Molecule-1 blood, Male, Middle Aged, Monocytes metabolism, Monocytes pathology, Multiple Myeloma blood, Multiple Myeloma pathology, Neoplasms, Plasma Cell pathology, Plasma Cells drug effects, Plasma Cells pathology, Young Adult, Biomarkers, Tumor blood, Multiple Myeloma drug therapy, Neoplasms, Plasma Cell blood, Signaling Lymphocytic Activation Molecule Family blood
- Abstract
Background: Daratumumab is an anti-CD38 immunotherapeutic drug that has increasingly been used to treat patients with heavily pre-treated and relapsed/refractory multiple myeloma. In so doing, the detection of CD38 antigen on plasma cells by flow cytometry is impeded. We hypothesized that alternative markers can be used in place or in addition to CD38 when detecting plasma cells post-treated with daratumumab., Methods: A total of 16 alternative markers were tested using 22 bone marrow aspirates from patients with plasma cell neoplasm. The ability of selected markers to discern plasma cells from other hematopoietic cells were evaluated. The stability of tested markers when stored at 4 or 25°C after T = 0, 24, 48, and 72 h was also established. Finally, selected markers were incorporated into a panel used for monitoring multiple myeloma measurable residual disease to test their utility to identify plasma cells in the presence of daratumumab and/or elotuzumab (anti-CD319) drugs., Results: Out of the 16 tested markers, CD319, CD54, CD229, CD317, and p63 were expressed by >90% of the plasma cells. Only CD319, CD54, and CD229 achieved 100% detection sensitivity. Further analysis showed that CD319 was better than CD229 and CD54 at resolving plasma cells from background hematopoietic cells, with CD54 being the worst (resolution metric, mean ± SD: CD319 [2.04 ± 0.86]; CD229 [1.47 ± 0.45]; and CD54 [1.22 ± 0.60]). CD229 was expressed by >90% of T lymphocytes, whereas CD319 was expressed preferentially by the CD8+ T cells and less frequently in CD4+ T cells. Additionally, CD229 was found on >60% of B and NK cells, as well as minor subsets of monocytes and granulocytes. CD319 was expressed on most NK cells and a minor subset of B cells, granulocytes, and monocytes. Even though CD229 and CD319 were expressed by different leukocyte subsets, their expression levels were highest on plasma cells. The expression of CD138 on plasma cells was significantly lower after storage at 4°C, while the expression levels of CD38, CD229, and CD319 remained stable at 4 or 25°C. Using limiting dilution experiments, the treatment of cells with daratumumab severely impeded the detection of CD38 antigen on plasma cells, whereas elotuzumab treatment did not block detection of CD319 on plasma cells., Conclusions: CD319 is a suitable alternative to CD38 for identifying plasma cells. Our results showed that a panel used for monitoring multiple myeloma measurable residual disease could be modified by using CD319 alone or in combination with CD38 to detect PCs in daratumumab or elotuzumab treated patients., (© 2020 International Clinical Cytometry Society.)
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- 2021
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107. Monitoring treatment with 5-Azacitidine by flow cytometry predicts duration of hematological response in patients with myelodysplastic syndrome.
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Subirá D, Alhan C, Oelschlaegel U, Porwit A, Psarra K, Westers TM, Golbano N, Nilsson L, van de Loosdrecht AA, and de Miguel D
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- Aged, Antimetabolites, Antineoplastic therapeutic use, Blood Cells drug effects, Blood Cells pathology, Bone Marrow drug effects, Bone Marrow pathology, Female, Humans, Immunophenotyping methods, Male, Middle Aged, Myelodysplastic Syndromes blood, Myelodysplastic Syndromes diagnosis, Prognosis, Treatment Outcome, Azacitidine therapeutic use, Drug Monitoring methods, Flow Cytometry methods, Myelodysplastic Syndromes drug therapy
- Abstract
5-Azacitidine (AZA) therapy is used in high-risk myelodysplastic syndrome (MDS) patients who often show abnormalities in their immunophenotype. We explored the potential impact of AZA on these immunophenotypic abnormalities in serial bone marrow studies performed in 81 patients from five centers. We compared the immunophenotypic features before and after therapy with AZA, established definitions consistent with flow cytometry immunophenotyping (FCI) improvement, and explored its clinical significance. After a median of 6 cycles of AZA, 41% of patients showed a FCI improvement and this finding associated with best possible clinical response (P < 0.001). FCI improvement also correlated with hematological improvement (HI) (53/78 patients; 68%), independently of their eligibility for stem cell transplantation. Among patients who achieved a HI after 6 cycles of AZA, the probability of maintaining this response at 12 cycles of AZA was twice as large (67%) for those patients who also achieved a FCI improvement after 6 cycles of AZA as compared to patients who did not (33%, P < 0.01). These findings support that monitoring of the immunophenotypic abnormalities during therapy with AZA may assist in redefining the quality of response in patients with MDS.
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- 2021
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108. Immunophenotypic switch in cutaneous T-cell lymphoma: A series of three cases and review of the literature.
- Author
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Bitar C, Hile G, Brown NA, Fullen DR, Lowe L, Tejasvi T, Wilcox RA, Harms PW, Chan MP, Bresler SC, and Hristov AC
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- Adolescent, Adult, Aged, Aged, 80 and over, Awareness, Biopsy methods, Cell Transformation, Neoplastic pathology, Child, Preschool, Diagnosis, Differential, Disease Progression, Fatal Outcome, Female, Gene Rearrangement genetics, Genes, T-Cell Receptor genetics, Genotype, Humans, Lymphoma, T-Cell, Cutaneous genetics, Lymphoma, T-Cell, Cutaneous immunology, Male, Middle Aged, Mycosis Fungoides drug therapy, Mycosis Fungoides metabolism, Mycosis Fungoides radiotherapy, Skin Neoplasms immunology, Treatment Outcome, Cell Transformation, Neoplastic immunology, Immunophenotyping methods, Lymphoma, T-Cell, Cutaneous diagnosis, Mycosis Fungoides pathology, Skin Neoplasms pathology
- Abstract
Primary cutaneous T-cell lymphoma (CTCL) comprises a heterogeneous group of neoplasms with variable clinical behavior. Immunophenotypic switch (IS) is a phenomenon that occurs during lymphoma progression and is defined by an alteration in the immunophenotypic expression of a tumor with retention of its genotypic signature. This has been well-recognized in hematopoietic neoplasms; however, it has been rarely reported in CTCL and its clinical implications are not well understood. We present the clinical, histopathologic, immunophenotypic, and genetic findings of three cases of CTCL that demonstrated IS post treatment with variable outcomes. We add our cases to the small number previously reported to increase awareness of this phenomenon and its diagnostic challenge., (© 2021 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)
- Published
- 2021
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109. Diagnostic performance of the ClearLLab 10C B cell tube.
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Espasa A, Torrents S, Morales-Indiano C, Rico LG, Bardina J, Ancochea A, Bistué-Rovira À, Linio R, Raya M, Vergara S, Juncà J, Grifols JR, Petriz J, Soria MG, and Sorigue M
- Subjects
- Antigens, CD blood, Antigens, CD19 blood, Antigens, CD20 blood, B-Lymphocytes pathology, Female, Flow Cytometry methods, Humans, Immunophenotyping methods, Leukemia, Lymphocytic, Chronic, B-Cell blood, Leukemia, Lymphocytic, Chronic, B-Cell diagnosis, Leukemia, Lymphocytic, Chronic, B-Cell pathology, Leukemia, Lymphoid blood, Leukemia, Lymphoid pathology, Lymphoma blood, Lymphoma pathology, Lymphoproliferative Disorders diagnosis, Lymphoproliferative Disorders immunology, Lymphoproliferative Disorders pathology, Male, Neprilysin blood, B-Lymphocytes immunology, Flow Cytometry instrumentation, Immunophenotyping instrumentation, Lymphoproliferative Disorders blood
- Abstract
Introduction: Pre-analytical and analytical errors can threaten the reliability of flow cytometry (FC) results. A potential solution to some of these is the use of dry, pre-mixed antibodies, such as the ClearLLab 10C system. The purpose of the present study was to compare the diagnostic performance of the ClearLLab 10C B cell tube with that of our standard laboratory practice., Methods: We compared the diagnoses made with the ClearLLab 10C B cell tube (experimental strategy) with those made with standard laboratory practice (standard strategy). Samples were selected aiming for representation of the full spectrum of B cell disorders, with an emphasis on mature B cell malignancies, as well as healthy controls., Results: We included 116 samples (34 normal controls, 4 acute lymphoblastic leukemias, 54 mature lymphoproliferative disorders in peripheral blood and bone marrow, 3 myelomas, 6 bone marrow samples with involvement by lymphoma and 1 with elevated hematogone count, 14 lymph node samples, 1 cerebrospinal fluid, and 1 pleural effusion). There were two diagnostic errors (1.7%). The agreement between the two strategies in the percentage of CD19 cells and fluorescence intensity of CD5, CD19, CD20, CD200, and CD10 was very good., Conclusions: In this study, the ClearLLab 10C B cell tube performed similarly to our standard laboratory practice to diagnose and classify mature B cell malignancies., (© 2020 International Clinical Cytometry Society.)
- Published
- 2021
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110. Cell surface markers for immunophenotyping human pluripotent stem cell-derived cardiomyocytes.
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Boheler KR and Poon EN
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- Animals, Cell Differentiation physiology, Humans, Immunophenotyping methods, Phenotype, Antigens, Surface metabolism, Biomarkers metabolism, Myocytes, Cardiac metabolism, Pluripotent Stem Cells metabolism
- Abstract
Human pluripotent stem cells (hPSC) self-renew and represent a potentially unlimited source for the production of cardiomyocytes (CMs) suitable for studies of human cardiac development, drug discovery, cardiotoxicity testing, and disease modelling and for cell-based therapies. However, most cardiac differentiation protocols yield mixed cultures of atrial-, ventricular-, and pacemaker-like cells at various stages of development, as well as non-CMs. The proportions and maturation states of these cell types result from disparities among differentiation protocols and time of cultivation, as well as hPSC reprogramming inconsistencies and genetic background variations. The reproducible use of hPSC-CMs for research and therapy is therefore limited by issues of cell population heterogeneity and functional states of maturation. A validated method that overcomes issues of cell heterogeneity is immunophenotyping coupled with live cell sorting, an approach that relies on accessible surface markers restricted to the desired cell type(s). Here we review current progress in unravelling heterogeneity in hPSC-cardiac cultures and in the identification of surface markers suitable for defining cardiac identity, subtype specificity, and maturation states.
- Published
- 2021
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111. Single-cell immunophenotyping of the skin lesion erythema migrans identifies IgM memory B cells.
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Jiang R, Meng H, Raddassi K, Fleming I, Hoehn KB, Dardick KR, Belperron AA, Montgomery RR, Shalek AK, Hafler DA, Kleinstein SH, and Bockenstedt LK
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- Adult, Aged, Female, Histocompatibility Antigens Class II genetics, Histocompatibility Antigens Class II immunology, Histocompatibility Antigens Class II metabolism, Humans, Immunoglobulin M genetics, Immunoglobulin M metabolism, Lyme Disease, Male, Middle Aged, RNA-Seq, Skin cytology, Transcriptome genetics, Transcriptome immunology, B-Lymphocytes cytology, B-Lymphocytes immunology, B-Lymphocytes metabolism, Erythema Chronicum Migrans immunology, Erythema Chronicum Migrans pathology, Immunophenotyping methods, Single-Cell Analysis methods
- Abstract
The skin lesion erythema migrans (EM) is an initial sign of the Ixodes tick-transmitted Borreliella spirochetal infection known as Lyme disease. T cells and innate immune cells have previously been shown to predominate the EM lesion and promote the reaction. Despite the established importance of B cells and antibodies in preventing infection, the role of B cells in the skin immune response to Borreliella is unknown. Here, we used single-cell RNA-Seq in conjunction with B cell receptor (BCR) sequencing to immunophenotype EM lesions and their associated B cells and BCR repertoires. We found that B cells were more abundant in EM in comparison with autologous uninvolved skin; many were clonally expanded and had circulating relatives. EM-associated B cells upregulated the expression of MHC class II genes and exhibited preferential IgM isotype usage. A subset also exhibited low levels of somatic hypermutation despite a gene expression profile consistent with memory B cells. Our study demonstrates that single-cell gene expression with paired BCR sequencing can be used to interrogate the sparse B cell populations in human skin and reveals that B cells in the skin infection site in early Lyme disease expressed a phenotype consistent with local antigen presentation and antibody production.
- Published
- 2021
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112. Macrophage Plasticity and Polarization Are Altered in the Experimental Model of Multiple Sclerosis.
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Leuti A, Talamonti E, Gentile A, Tiberi M, Matteocci A, Fresegna D, Centonze D, and Chiurchiù V
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- Animals, Encephalomyelitis, Autoimmune, Experimental pathology, Female, Immunophenotyping methods, Macrophages pathology, Mice, Mice, Inbred C57BL, Multiple Sclerosis pathology, Cell Polarity physiology, Encephalomyelitis, Autoimmune, Experimental immunology, Macrophages immunology, Multiple Sclerosis immunology, Neuronal Plasticity physiology
- Abstract
Multiple sclerosis (MS) is an immune-mediated demyelinating disease of the central nervous system. MS is characterized by infiltrations of leukocytes such as T and B lymphocytes and macrophages. Macrophages have been identified as major effectors of inflammation and demyelination in both MS and its animal model, experimental autoimmune encephalomyelitis (EAE). However, the activation and heterogeneity of macrophages in MS has been poorly investigated. Thus, in this study, we evaluated M1 and M2 macrophages immunophenotype from EAE and control mice by analyzing over 30 surface and intracellular markers through polychromatic flow cytometry, qRT-PCR, and ELISA assay. We showed that M1 macrophages possessed a higher proinflammatory profile in EAE compared to control mice, since they expressed higher levels of activation/co-stimulatory markers (iNOS, CD40, and CD80) and cytokines/chemokines (IL-6, IL-12, CCL2, and CXCL10), whereas M2 lost their M2-like phenotype by showing a decreased expression of their signature markers CD206 and CCL22, as well as a concomitant upregulation of several M1 makers. Furthermore, immunization of M1 and M2 macrophages with MOG35-55 led to a significant hyperactivation of M1 and a concomitant shift of anti-inflammatory M2 to pro-inflammatory M1 macrophages. Overall, we provide evidence for a phenotypic alteration of M1/M2 balance during MS, which can be of crucial importance not only for a better understanding of the immunopathology of this neurodegenerative disease but also to potentially develop new macrophage-centered therapeutic strategies.
- Published
- 2021
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113. Comprehensive genomic and immunophenotypic analysis of CD4 T cell infiltrating human triple-negative breast cancer.
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Zhang H, Qin G, Yu H, Han X, and Zhu S
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- Female, Follow-Up Studies, Humans, Lymphocyte Activation, Middle Aged, Prognosis, Triple Negative Breast Neoplasms genetics, Triple Negative Breast Neoplasms immunology, Biomarkers, Tumor genetics, CD4-Positive T-Lymphocytes immunology, Genomics methods, Immunophenotyping methods, Lymphocytes, Tumor-Infiltrating immunology, T-Lymphocytes, Regulatory immunology, Triple Negative Breast Neoplasms pathology
- Abstract
The aim of this study is to investigate the gene expression module of tumor-infiltrating CD4
+ T cells and its potential roles in modulating immune cell functions in triple-negative breast cancer. Differentially expressed genes were identified by comparison of the expression profile in CD4+ T cells isolated from tumor tissues and peripheral blood of TNBC patients respectively. The differential expression analysis was conducted using R, and then the functional and pathway enrichment of the DEGs were analyzed using GSEA, followed by integrated regulatory network construction and genetic analysis of tumor-infiltrating immune cells based on a scientific deconvolution algorithm. As a result, abundant Treg and exhausted lymphocytes were detected, accompanied by largely decreased of effector/memory and cytotoxic T cells. Immune-related gene correlation analysis showed that the extent of follicular helper T cells gene expression signatures were inversely associated with those of CD4+ naive T cells and CD4+ memory resting T cells, but positively correlated with that of CD4+ memory activated T cells. In addition, we found five core genes including IFNG, CTLA4, FAS, CXCR6, and JUN were significantly over expressed in CD4+ TILs which may contribute to exhaustion of lymphocytes and participate in biological processes associated with regulation of chemotaxis. Study provides a comprehensive understanding of the roles of DEGs associated with the chemotactic and exhausted immunophenotypes of CD4+ TILs that are a valuable resource from which future investigation may be carried out to better understand the mechanisms that promote TNBC progression.- Published
- 2021
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114. Deep immune profiling of whole blood to identify early immune signatures that correlate to patient outcome after major trauma.
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Mendoza AE, Raju Paul S, El Hechi M, Naar L, Nederpelt C, Mikdad S, van Erp I, Hess JM, Velmahos GC, Poznansky M, and Reeves P
- Subjects
- Acute Kidney Injury blood, Acute Kidney Injury immunology, Acute Kidney Injury prevention & control, Adult, Case-Control Studies, Clinical Decision-Making methods, Critical Illness, Female, Healthcare-Associated Pneumonia blood, Healthcare-Associated Pneumonia immunology, Healthcare-Associated Pneumonia prevention & control, Humans, Injury Severity Score, Intensive Care Units statistics & numerical data, Length of Stay statistics & numerical data, Male, Middle Aged, Prospective Studies, Risk Assessment methods, Wounds and Injuries complications, Wounds and Injuries diagnosis, Wounds and Injuries immunology, Acute Kidney Injury epidemiology, Healthcare-Associated Pneumonia epidemiology, Immunophenotyping methods, Wounds and Injuries blood
- Abstract
Background: Major injury results in an early cascade of immunologic responses that increase susceptibility to infection and multiorgan dysfunction. Detailed immune profiling by mass cytometry has the potential to identify immune signatures that correspond to patient outcomes. Our objective was to determine the prognostic value of immune signatures early after major trauma injury., Methods: Trauma patients (n = 17) were prospectively enrolled between September 2018 and December 2019. Serial whole blood samples were obtained from trauma patients (mean Injury Severity Score, 26.2; standard error of the mean, 3.7) at Days 1 and 3 after injury, and from age- and sex-matched uninjured controls using a standardized protocol for fixation, storage, and labeling. Computational analyses including K-nearest neighbor automated clustering of immune cells and Spearman's correlation analysis were used to identify correlations between cell populations, clinical measures, and patient outcomes., Results: Analysis revealed nine immune cell clusters that correlated with one or more clinical outcomes. On Days 1 and 3 postinjury, the abundance of immature neutrophil and classical monocytes exhibited a strong positive correlation with increased intensive care unit and hospital length of stay. Conversely, the abundance of CD4 T-cell subsets, namely Th17 cells, is associated with improved patient outcomes including decreased ventilator days (r = -0.76), hospital-acquired pneumonia (r = -0.69), and acute kidney injury (r = -0.73)., Conclusion: Here, we provide a comprehensive multitime point immunophenotyping analysis of whole blood from patients soon after traumatic injury to determine immune correlates of adverse outcomes. Our findings indicate that alterations in myeloid-origin cell types may contribute to immune dysfunction after injury. Conversely, the presence of effector T cell populations corresponds with decreased hospital length of stay and organ dysfunction. Overall, these data identify novel immune signatures following traumatic injury that support the view that monitoring of immune (sub)-populations may provide clinical decision-making support for at-risk patients early in their hospital course., Level of Evidence: Prognostic/Epidemiologic, Level IV., (Copyright © 2021 Wolters Kluwer Health, Inc. All rights reserved.)
- Published
- 2021
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115. Single-cell profiling of myasthenia gravis identifies a pathogenic T cell signature.
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Ingelfinger F, Krishnarajah S, Kramer M, Utz SG, Galli E, Lutz M, Zwicky P, Akarca AU, Jurado NP, Ulutekin C, Bamert D, Widmer CC, Piccoli L, Sallusto F, Núñez NG, Marafioti T, Schneiter D, Opitz I, Lanzavecchia A, Jung HH, De Feo D, Mundt S, Schreiner B, and Becher B
- Subjects
- Adult, Aged, Aged, 80 and over, Autoantibodies, Autoimmunity, B-Lymphocytes immunology, Biomarkers, Female, Humans, Machine Learning, Male, Middle Aged, Myasthenia Gravis blood, Receptors, Cholinergic immunology, T-Lymphocytes immunology, Thymectomy, Thymus Gland, Immunophenotyping methods, Myasthenia Gravis immunology, Myasthenia Gravis pathology, Single-Cell Analysis, T-Lymphocytes pathology
- Abstract
Myasthenia gravis (MG) is an autoimmune disease characterized by impaired neuromuscular signaling due to autoantibodies targeting the acetylcholine receptor. Although its auto-antigens and effector mechanisms are well defined, the cellular and molecular drivers underpinning MG remain elusive. Here, we employed high-dimensional single-cell mass and spectral cytometry of blood and thymus samples from MG patients in combination with supervised and unsupervised machine-learning tools to gain insight into the immune dysregulation underlying MG. By creating a comprehensive immune map, we identified two dysregulated subsets of inflammatory circulating memory T helper (Th) cells. These signature Th
CD103 and ThGM cells populated the diseased thymus, were reduced in the blood of MG patients, and were inversely correlated with disease severity. Both signature Th subsets rebounded in the blood of MG patients after surgical thymus removal, indicative of their role as cellular markers of disease activity. Together, this in-depth analysis of the immune landscape of MG provides valuable insight into disease pathogenesis, suggests novel biomarkers and identifies new potential therapeutic targets for treatment.- Published
- 2021
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116. From the archives of MD Anderson Cancer Center: A case of concurrent follicular lymphoma and Langerhans cell sarcoma with a review of the literature.
- Author
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Tashakori M, Thakral B, Tang G, Khoury JD, and Medeiros LJ
- Subjects
- Adult, Aged, B-Lymphocytes pathology, Cell Dedifferentiation genetics, Cell Transdifferentiation genetics, Female, Histiocytes pathology, Histiocytosis, Langerhans-Cell metabolism, Histiocytosis, Langerhans-Cell pathology, Humans, Immunohistochemistry methods, Immunophenotyping methods, Langerhans Cell Sarcoma metabolism, Langerhans Cell Sarcoma pathology, Langerhans Cells pathology, Lymphoma, Follicular metabolism, Lymphoma, Follicular pathology, Male, Middle Aged, Mitogen-Activated Protein Kinases genetics, Mitogen-Activated Protein Kinases metabolism, Neoplasm Staging methods, Histiocytosis, Langerhans-Cell diagnosis, Langerhans Cell Sarcoma diagnosis, Lymphoma, Follicular diagnosis, Neoplasms, Second Primary pathology
- Abstract
Transdifferentiation of follicular lymphoma to a Langerhans cell neoplasm is rarely reported and not well understood. Here we present a case, review the literature and discuss some of the biological underpinnings of lineage switch of B cells to histiocytes/Langerhans cells. A 31-year-old woman had follicular lymphoma (FL) and Langerhans cell sarcoma (LCS) co-localized above and below diaphragm. The FL was low-grade, had typical morphologic features, and was positive for CD10, BCL-2, and BCL-6. The LCS was cytologically atypical with necrosis and a high mitotic rate, and the immunophenotype supported Langerhans cell lineage positive for CD1a, CD207/langerin, and S-100 protein. Both tumors carried IGH-BCL2 and the LCS cells had immunophenotypic evidence of a residual B cell program, supporting the notion that these neoplasms are clonally related. The case reported is unusual because the patient was young and both diseases presented simultaneously, before any therapy. In addition, immunohistochemical analysis showed that the LCS was negative for BRAF V600E and phospho-ERK, suggesting that the LCS belongs to the known subset of Langerhans cell tumors lacking BRAF V600E and MAP2K1 mutations. Concurrent occurrence of FL and Langerhans cell neoplasm is an unusual phenomenon, with 10 cases reported previously: 4 Langerhans cell histiocytosis and 6 Langerhans cell sarcoma, including this case., (Copyright © 2021 Elsevier Inc. All rights reserved.)
- Published
- 2021
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117. The "Intermediate" CD14 + CD16 + monocyte subpopulation plays a role in IVIG responsiveness of children with Kawasaki disease.
- Author
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Kim YS, Yang HJ, Kee SJ, Choi I, Ha K, Ki KK, Jeong IS, and Cho HJ
- Subjects
- Biomarkers, Pharmacological analysis, Child, Preschool, Female, Fever blood, Fever immunology, Flow Cytometry methods, GPI-Linked Proteins immunology, Humans, Immunologic Factors administration & dosage, Immunologic Factors immunology, Immunophenotyping methods, Male, Patient Acuity, Treatment Outcome, Immunoglobulins, Intravenous administration & dosage, Immunoglobulins, Intravenous immunology, Lipopolysaccharide Receptors immunology, Monocytes immunology, Monocytes pathology, Mucocutaneous Lymph Node Syndrome blood, Mucocutaneous Lymph Node Syndrome diagnosis, Mucocutaneous Lymph Node Syndrome immunology, Mucocutaneous Lymph Node Syndrome therapy, Receptors, IgG immunology
- Abstract
Background: Kawasaki disease (KD) is an acute, self-limited febrile illness of unknown cause. Intravenous immunoglobulin (IVIG)-resistance are related to greater risk for permanent cardiac complications. We aimed to determine the correlation between monocytes and the phenotype of KD in relation to IVIG responsiveness in children., Materials and Methods: The study cohort included 62 patients who were diagnosed with KD, 20 non febrile healthy controls (NFC), and 15 other febrile controls (OFC). In all enrolled patients, blood was taken at least 4 times and laboratory tests were performed. In addition, subtypes of monocytes were characterized via flow cytometry., Results: The numbers of intermediate monocytes were significantly lower in IVIG-resistant group compared to IVIG-responsive group before IVIG infusion (p < 0.0001). After infusion, intermediate monocytes decreased in the responsive group, while a trend of increase was observed in the resistant group. Only intermediate monocytes were significant in logistic regression with adjusted OR of 0.001 and p value of 0.03., Conclusions: CD14 + CD16 + intermediate monocyte may play an important role in IVIG responsiveness among KD children. Low starting levels of intermediate monocytes, followed by a dramatic increase post-IVIG infusion during acute phase of KD are associated with IVIG-resistance. Functional studies on intermediate monocyte may help to reveal the pathophysiology.
- Published
- 2021
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118. Comparison of minimal residual disease detection in multiple myeloma between the DuraClone and EuroFlow methods.
- Author
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Yoroidaka T, Narita K, Takamatsu H, Fujisawa M, Nakao S, and Matsue K
- Subjects
- Antibodies immunology, Humans, Multiple Myeloma immunology, Neoplasm, Residual immunology, Plasma Cells immunology, Flow Cytometry methods, Immunophenotyping methods, Multiple Myeloma diagnosis, Neoplasm, Residual diagnosis
- Abstract
In this study, the minimal residual disease (MRD) levels in patients with multiple myeloma (MM) were assessed by comparing the new 8-color single-tube multiparameter flow cytometry method (DuraClone), which reduces the cost of antibodies and labor burden of laboratories, with the EuroFlow next-generation flow (NGF) method. A total of 96 samples derived from 69 patients with MM were assessed to determine the total cell acquisition number (tCAN), percentages of total and normal plasma cells (PCs), and MRD levels using two methods. We found that the tCAN was significantly higher with EuroFlow-NGF than with DuraClone (median 8.6 × 10
6 vs. 5.7 × 106 ; p < 0.0001). In addition, a significant correlation in the MRD levels between the two methods was noted (r = 0.92, p < 0.0001). However, in the qualitative analysis, 5.2% (5/96) of the samples showed discrepancies in the MRD levels. In conclusion, the DuraClone is a good option to evaluate MRD in multiple myeloma but it should be used with caution.- Published
- 2021
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119. Advanced Flow Cytometry Assays for Immune Monitoring of CAR-T Cell Applications.
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Blache U, Weiss R, Boldt A, Kapinsky M, Blaudszun AR, Quaiser A, Pohl A, Miloud T, Burgaud M, Vucinic V, Platzbecker U, Sack U, Fricke S, and Koehl U
- Subjects
- Antigens, Neoplasm immunology, Antigens, Neoplasm metabolism, Biomarkers, Cell Survival, Humans, Immunotherapy, Adoptive methods, Receptors, Antigen, T-Cell genetics, Receptors, Antigen, T-Cell metabolism, Receptors, Chimeric Antigen genetics, Receptors, Chimeric Antigen metabolism, Flow Cytometry methods, Immunophenotyping methods, T-Lymphocytes immunology, T-Lymphocytes metabolism
- Abstract
Adoptive immunotherapy using chimeric antigen receptor (CAR)-T cells has achieved successful remissions in refractory B-cell leukemia and B-cell lymphomas. In order to estimate both success and severe side effects of CAR-T cell therapies, longitudinal monitoring of the patient's immune system including CAR-T cells is desirable to accompany clinical staging. To conduct research on the fate and immunological impact of infused CAR-T cells, we established standardized 13-colour/15-parameter flow cytometry assays that are suitable to characterize immune cell subpopulations in the peripheral blood during CAR-T cell treatment. The respective staining technology is based on pre-formulated dry antibody panels in a uniform format. Additionally, further antibodies of choice can be added to address specific clinical or research questions. We designed panels for the anti-CD19 CAR-T therapy and, as a proof of concept, we assessed a healthy individual and three B-cell lymphoma patients treated with anti-CD19 CAR-T cells. We analyzed the presence of anti-CD19 CAR-T cells as well as residual CD19+ B cells, the activation status of the T-cell compartment, the expression of co-stimulatory signaling molecules and cytotoxic agents such as perforin and granzyme B. In summary, this work introduces standardized and modular flow cytometry assays for CAR-T cell clinical research, which could also be adapted in the future as quality controls during the CAR-T cell manufacturing process., Competing Interests: TM, MB, and MK are employees of Beckman Coulter Life Sciences. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Blache, Weiss, Boldt, Kapinsky, Blaudszun, Quaiser, Pohl, Miloud, Burgaud, Vucinic, Platzbecker, Sack, Fricke and Koehl.)
- Published
- 2021
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120. Attenuation of GARP expression on regulatory T cells by protein transport inhibitors.
- Author
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Anvari S, Schuster K, Grimbergen A, Davis CM, and Makedonas G
- Subjects
- Brefeldin A pharmacology, Cells, Cultured, Cytokines metabolism, Enterotoxins immunology, Healthy Volunteers, Humans, Lymphocyte Activation, Membrane Proteins antagonists & inhibitors, Membrane Proteins metabolism, Monensin pharmacology, Primary Cell Culture, T-Lymphocytes, Regulatory drug effects, T-Lymphocytes, Regulatory metabolism, Cytokines analysis, Immunophenotyping methods, Membrane Proteins analysis, T-Lymphocytes, Regulatory immunology
- Abstract
An integrated understanding of the functional capacities of cells in the context of their physical parameters and molecular markers is increasingly demanded in immunologic studies. Regulatory T cells (Tregs) are a subpopulation of T cells involved in immune response modulation and mediating tolerance to self-antigen with their absence leading to a loss of tolerance. Glycoprotein repetitions A predominant (GARP) is a key marker for activated Tregs, but its detection may also be useful in determining the functional capacities of the cell. This study aims to deduce the optimal stimulation period and the impact of protein transport inhibitors (PTIs), commonly used in the detection of intracellular cytokines, on GARP detection. Through flow cytometric analysis we analyzed different cell culture conditions for optimal GARP expression on activated Tregs. Healthy donor PBMCs were stimulated with either Staphylococcal Enterotoxin B (SEB) or PMA/Ionomycin (PMA/Iono), in the presence and absence of PTIs monensin and/or brefeldin A (BFA) and GARP expression was assessed on CD4+ CD25+ FOXP3+ Tregs. The optimal stimulation period for the detection of GARP was highest at 24-h. Furthermore, we determined that GARP expression on Tregs is significantly reduced when cells are treated with the PTIs monensin and/or BFA following PMA/Iono stimulation. This effect was not seen following SEB stimulation. Therefore, due to the effects of PTIs, alternative methods should be considered when performing simultaneous analysis for cytokine expression and GARP expression on Tregs., (Copyright © 2021 Elsevier B.V. All rights reserved.)
- Published
- 2021
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121. Mast cell differentiation of leukemic blasts in diverse myeloid neoplasms: A potential pre-myelomastocytic leukemia condition.
- Author
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Panda D, Chatterjee G, Khanka T, Ghogale S, Badrinath Y, Deshpande N, Sardana R, Chaturvedi A, Rajpal S, Shetty D, Patkar NV, Gujral S, Subramanian PG, and Tembhare PR
- Subjects
- Adolescent, Adult, Aged, Antigens, CD metabolism, Bone Marrow metabolism, Bone Marrow pathology, Child, Female, Hematologic Neoplasms metabolism, Hematologic Neoplasms pathology, Humans, Immunophenotyping methods, Leukemia, Myeloid, Acute metabolism, Leukemia, Myeloid, Acute pathology, Leukemia, Myelomonocytic, Chronic metabolism, Leukemia, Myelomonocytic, Chronic pathology, Male, Mast Cells metabolism, Mastocytosis, Systemic metabolism, Mastocytosis, Systemic pathology, Middle Aged, Myeloproliferative Disorders metabolism, Myeloproliferative Disorders pathology, Primary Myelofibrosis metabolism, Cell Differentiation physiology, Mast Cells pathology, Primary Myelofibrosis pathology
- Abstract
Introduction: Myeloid neoplasm with blasts showing mast cell (MC)-differentiation and MC-component less than 10% of all nucleated cells but not fulfilling the criteria for systemic mastocytosis with associated hematological neoplasm (SM-AHN) or myelomastocytic leukemia (MML) has not been described in the literature. Herein, we report a study of diverse myeloid malignancies with blasts showing MC-differentiation but not meeting the criteria for SM-AHN or MML. We also evaluated the utility of flow-cytometric immunophenotyping (FCI) in the characterization of immature-MCs (iMCs)., Methods: We identified nine patients of myeloid neoplasms and studied their morphological, FCI, immunohistochemistry, cytogenetic and molecular characteristics. We also compared the immunophenotypic features of MCs from patient samples with control samples., Results: The study included patients with newly-diagnosed acute myeloid leukemia (n = 4), chronic myelomonocytic leukemia (n = 1), and chronic myeloid leukemia on follow-up (n = 4) showing MC differentiation in leukemic-blasts. These patients had mildly increased MCs (range, 0.5%-3%) in bone-marrow morphology, including immature-forms and did not meet the criteria for either SM-AHN or MML. On FCI, iMCs were positive for bright-CD117, heterogeneous-CD34, dim-to-negative-HLADR, and moderate-CD203c expression. Expression-levels of CD123 and CD38 were higher (p < 0.001) but CD33 and CD45 were lower in iMCs compared to mature-MC from control samples (p = 0.019 and p = 0.0037)., Conclusion: We reported a rare finding of MC differentiation of leukemic blasts in diverse myeloid neoplasms and proposed it as a potential pre-myelomastocytic leukemia condition. We described the distinct immunophenotypic signature of immature-MCs using commonly used markers and highlighted the utility of FCI for the diagnosis of this entity., (© 2020 International Clinical Cytometry Society.)
- Published
- 2021
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122. Hodgkin and Reed-Sternberg-like cells within a recurrent primary cutaneous marginal zone lymphoma.
- Author
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Blaise B, Wilson ML, and Guitart J
- Subjects
- Aged, 80 and over, Biopsy, Fatal Outcome, Female, Hodgkin Disease metabolism, Hodgkin Disease pathology, Humans, Immunophenotyping methods, In Situ Hybridization methods, Lymphoma, B-Cell, Marginal Zone metabolism, Lymphoma, B-Cell, Marginal Zone pathology, Neoplasm Recurrence, Local radiotherapy, Lymphoma, B-Cell, Marginal Zone diagnosis, Neoplasm Recurrence, Local pathology, Reed-Sternberg Cells pathology, Skin Neoplasms pathology
- Published
- 2021
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123. Impact of laboratory work-up of lymphoma guidelines on cytopathology practices.
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Davey DD
- Subjects
- Biopsy, Fine-Needle methods, Biopsy, Large-Core Needle methods, Diagnosis, Differential, Flow Cytometry methods, Humans, Immunohistochemistry methods, Immunophenotyping methods, Lymphoma classification, Molecular Diagnostic Techniques methods, Laboratories, Lymph Nodes pathology, Lymphoma diagnosis, Lymphoma pathology, Practice Guidelines as Topic standards
- Abstract
A multi-society expert panel recently published evidence-based guidelines and recommendations for the primary diagnosis and classification of lymphoma, which included a public comment period. The guideline concludes: "primary diagnosis and classification of lymphoma can be achieved with a variety of specimens." The guideline recommends that fine-needle aspiration biopsy (FNAB) cytomorphology not be used without ancillary testing, and that either flow cytometry or immunohistochemical immunophenotyping be performed for any type of specimen. Either excisional or core biopsy should be obtained when there is a high suspicion of lymphoma, and excision is recommended when feasible for Hodgkin lymphoma primary diagnosis. The use of cerebrospinal fluid to diagnose central nervous system lymphoma is discussed. For any patient with initial negative biopsy or fluid specimens and a high suspicion of lymphoma, additional tissue samples should be obtained. Additional molecular testing and good practice statements are summarized in the guideline. Cytopathologists should continue to advocate for judicious use of lymph node FNAB as an excellent triage tool that may require additional tissue biopsy for definitive diagnosis., (Copyright © 2021 American Society of Cytopathology. Published by Elsevier Inc. All rights reserved.)
- Published
- 2021
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124. Detection of engineered T cells in FFPE tissue by multiplex in situ hybridization and immunohistochemistry.
- Author
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Wright JH, Huang LY, Weaver S, Archila LD, McAfee MS, Hirayama AV, Chapuis AG, Bleakley M, Rongvaux A, Turtle CJ, Chanthaphavong RS, Campbell JS, and Pierce RH
- Subjects
- Animals, Biopsy, Cell Engineering, Coculture Techniques, Genetic Vectors genetics, Hepatitis B Virus, Woodchuck genetics, Humans, In Situ Hybridization, Fluorescence, Lentivirus genetics, Lymph Nodes pathology, Male, Mice, Mice, Transgenic, Models, Animal, Paraffin Embedding, Receptors, Chimeric Antigen genetics, Receptors, Chimeric Antigen immunology, Skin cytology, Skin immunology, Skin pathology, T-Lymphocytes metabolism, T-Lymphocytes transplantation, Tissue Fixation, Transduction, Genetic, Transplantation Chimera, Immunohistochemistry methods, Immunophenotyping methods, Receptors, Chimeric Antigen analysis, T-Lymphocytes immunology
- Abstract
Identifying engineered T cells in situ is important to understand the location, persistence, and phenotype of these cells in patients after adoptive T cell therapy. While engineered cells are routinely characterized in fresh tissue or blood from patients by flow cytometry, it is difficult to distinguish them from endogenous cells in formalin-fixed, paraffin-embedded (FFPE) tissue biopsies. To overcome this limitation, we have developed a method for characterizing engineered T cells in fixed tissue using in situ hybridization (ISH) to the woodchuck hepatitis post-transcriptional regulatory element (WPRE) common in many lentiviral vectors used to transduce chimeric antigen receptor T (CAR-T) and T cell receptor T (TCR-T) cells, coupled with alternative permeabilization conditions that allows subsequent multiplex immunohistochemical (mIHC) staining within the same image. This new method provides the ability to mark the cells by ISH, and simultaneously stain for cell-associated proteins to immunophenotype CAR/TCR modified T cells within tumors, as well as assess potential roles of these cells in on-target/off-tumor toxicity in other tissue., (Copyright © 2020 Elsevier B.V. All rights reserved.)
- Published
- 2021
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125. Lymphocytic variant of hypereosinophilic syndrome: A report of seven cases from a single institution.
- Author
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Hu Z, Wang W, Thakral B, Chen Z, Estrov Z, Bueso-Ramos CE, Verstovsek S, Medeiros LJ, and Wang SA
- Subjects
- Adult, Aged, Cytokines metabolism, Female, Flow Cytometry methods, Humans, Hypereosinophilic Syndrome metabolism, Immunophenotyping methods, Male, Middle Aged, T-Lymphocytes metabolism, Hypereosinophilic Syndrome diagnosis, Hypereosinophilic Syndrome pathology, T-Lymphocytes pathology
- Abstract
Background: Lymphocytic variant of hypereosinophilic syndrome (L-HES) is a subtype of HES driven by cytokines produced by clonal T-cells. Due to the rarity of its occurrence and challenges in diagnosis, this subtype of HES is under recognized., Methods and Results: We report seven patients with L-HES, diagnosed from a group of 136 patients who were referred to our institution for the work-up of hypereosinophilia. The clinical presentation, symptoms and signs were heterogeneous and uncharacteristic; indistinguishable from idiopathic HES. Flow cytometry immunophenotypic analysis revealed aberrant T-cells in all patients, with a Th2 immunophenotype, CD2 + CD3-CD4 + CD5 + CD7dim+/-CD8- in six of seven (86%) cases. CD10 was partially expressed in one of seven (14%) cases, and clonal TCR gene rearrangement was detected by PCR in five of seven (71%) patients. All patients were treated with corticosteroids and two of seven (29%) patients received anti-IL5 antibody therapy. With a median follow-up time of 7.5 years (2.3-14.1 years), one (11%) patient developed peripheral T-cell lymphoma 6.1 years after the initial diagnosis of L-HES and responded well to chemotherapy. All patients were alive at the last follow-up., Conclusion: In conclusion, a combination of flow cytometry immunophenotyping and molecular analysis allows the identification of aberrant T-cells, facilitating a diagnosis of L-HES in patients with eosinophilia. A correct diagnosis is essential for the proper management of these patients., (© 2020 International Clinical Cytometry Society.)
- Published
- 2021
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126. Cutaneous follicle center lymphomas with plasmacytic differentiation.
- Author
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Skala SL, Harms PW, Fullen DR, Brown NA, Tejasvi T, Wilcox RA, Boyer DF, and Hristov AC
- Subjects
- Adult, Aged, Biopsy, Cell Differentiation, Female, Humans, Immunoglobulin lambda-Chains metabolism, Immunophenotyping methods, Lymphoma, B-Cell pathology, Lymphoma, Follicular surgery, Lymphoma, Follicular ultrastructure, Male, Middle Aged, Plasma Cells metabolism, Treatment Outcome, Immunoglobulin kappa-Chains metabolism, Lymphoma, B-Cell diagnosis, Lymphoma, Follicular diagnosis, Plasma Cells pathology, Skin Neoplasms pathology
- Abstract
Follicle center lymphomas, including primary cutaneous follicle center lymphoma (PCFCL), may rarely show plasmacytic differentiation. Such cases can pose a diagnostic challenge and can be mistaken for other lymphomas that more commonly include plasma cells. Here, we report four cases of PCFCL and one case of systemic follicular lymphoma involving the skin with associated monotypic plasma cells, including the clinical, morphologic and immunophenotypic features., (© 2020 John Wiley & Sons A/S . Published by John Wiley & Sons Ltd.)
- Published
- 2021
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127. Flow cytometric evaluation of TRBC1 expression in tissue specimens and body fluids is a novel and specific method for assessment of T-cell clonality and diagnosis of T-cell neoplasms.
- Author
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Berg H, Otteson GE, Corley H, Shi M, Horna P, Jevremovic D, and Olteanu H
- Subjects
- Adolescent, Adult, Aged, Aged, 80 and over, Antibodies, Monoclonal metabolism, Female, Flow Cytometry methods, Humans, Immunophenotyping methods, Male, Middle Aged, Receptors, Antigen, T-Cell, gamma-delta metabolism, Young Adult, Body Fluids metabolism, Lymphoma, T-Cell diagnosis, Lymphoma, T-Cell metabolism, Receptors, Antigen, T-Cell, alpha-beta metabolism, T-Lymphocytes metabolism
- Abstract
Background: Flow cytometric detection of T-cell clonality is challenging. The current available methodology for T-cell receptor (TCR) Vβ repertoire evaluation is a complex assay and has limited sensitivity especially for detecting low levels of disease. Therefore, there is an unmet need for a reliable, simple, and rapid assay to identify T-cell clonality. The rearrangement of the TCRB gene involves the random and mutually exclusive expression of one of two constant β chain genes (TRBC1 and TRBC2), analogous to the kappa and lambda gene utilization by B cells., Methods: Here, we used a single TRBC1 antibody, in conjunction with other T-cell associated markers, to detect T-cell clonality in tissue biopsies and body fluids. A total of 143 tissue/body fluid specimens from 46 patients with a definitive diagnosis of a T-cell neoplasm and 97 patients with no T-cell malignancy were analyzed with a cocktail of monoclonal antibodies including CD2/CD3/CD4/CD5/CD7/CD8/CD45/TCRγδ/TRBC1., Results: We examined TRBC1 expression on neoplastic T-cell populations identified based on their immunophenotypic aberrancies, and monotypic TRBC1 expression was identified in all 46 known T-cell lymphoma cases. We applied a similar gating strategy to the 97 cases without T-cell neoplasms, and arbitrarily dissected T-cell populations into immunophenotypically distinct subsets; in this group, we found that all cases revealed an expected polytypic TRBC1 expression in all subsets., Conclusions: Single TRBC1 antibody detection of T-cell clonality by flow cytometry is a simple, rapid, and robust assay that could be routinely utilized in flow cytometric laboratories., (© 2020 International Clinical Cytometry Society.)
- Published
- 2021
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128. Analysis of Tumor Microenvironment Characteristics in Bladder Cancer: Implications for Immune Checkpoint Inhibitor Therapy.
- Author
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Chen X, Chen H, He D, Cheng Y, Zhu Y, Xiao M, Lan H, Wang Z, and Cao K
- Subjects
- Algorithms, Humans, Immune Checkpoint Inhibitors therapeutic use, Immunophenotyping methods, Prognosis, Urinary Bladder Neoplasms drug therapy, Biomarkers, Tumor immunology, Machine Learning, Tumor Microenvironment immunology, Urinary Bladder Neoplasms immunology
- Abstract
The tumor microenvironment (TME) plays a crucial role in cancer progression and recent evidence has clarified its clinical significance in predicting outcomes and efficacy. However, there are no studies on the systematic analysis of TME characteristics in bladder cancer. In this study, we comprehensively evaluated the TME invasion pattern of bladder cancer in 1,889 patients, defined three different TME phenotypes, and found that different subtypes were associated with the clinical prognosis and pathological characteristics of bladder cancer. We further explored the signaling pathways, cancer-immunity cycle, copy number, and somatic mutation differences among the different subtypes and used the principal component analysis algorithm to calculate the immune cell (IC) score, a tool for comprehensive evaluation of TME. Univariate and multivariate Cox regression analyses showed that ICscore is a reliable and independent prognostic biomarker. In addition, the use of anti-programmed death-ligand (PD-L1) treatment cohort, receiver operating characteristic (ROC) curve, Tumor Immune Dysfunction and Exclusion (TIDE), Subnetwork Mappings in Alignment of Pathways (SubMAP), and other algorithms confirmed that ICscore is a reliable prognostic biomarker for immune checkpoint inhibitor response. Patients with higher ICscore showed a significant therapeutic advantage in immunotherapy. In conclusion, this study improves our understanding of the characteristics of TME infiltration in bladder cancer and provides guidance for more effective personalized immunotherapy strategies., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be constructed as a potential conflict of interest., (Copyright © 2021 Chen, Chen, He, Cheng, Zhu, Xiao, Lan, Wang and Cao.)
- Published
- 2021
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129. Utility of FLAER and CD157 in a five-color single-tube high sensitivity assay, for diagnosis of Paroxysmal Nocturnal Hemoglobinuria (PNH)-A standalone flow cytometry laboratory experience.
- Author
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Seth N, Mahajan V, Kedia S, Sutar A, and Sehgal K
- Subjects
- Adolescent, Adult, Aged, Aged, 80 and over, Blood Cell Count, Child, Child, Preschool, Female, Flow Cytometry standards, GPI-Linked Proteins metabolism, Hemoglobinuria, Paroxysmal etiology, Humans, Immunophenotyping methods, Immunophenotyping standards, Infant, Leukocytes metabolism, Male, Middle Aged, Prognosis, Sensitivity and Specificity, Young Adult, ADP-ribosyl Cyclase metabolism, Antigens, CD metabolism, Biomarkers, Flow Cytometry methods, Hemoglobinuria, Paroxysmal diagnosis, Hemoglobinuria, Paroxysmal metabolism
- Abstract
Background: FLAER-based flow cytometry assay is considered the gold standard for diagnosis of paroxysmal nocturnal hemoglobinuria (PNH). CD157 is a recently reported marker for GPI-anchored protein found both on neutrophils and monocytes. This study highlights the robustness of FLAER and CD157 combination to identify PNH clones in a high sensitivity assay. Though rare, the data shown highlight the presence of CD157 negativity in few cases re-emphasizing the importance of FLAER for PNH diagnosis., Methods: A single 5-color tube-FLAER Alexa488/ CD157PE/ CD15PECy5/ CD64PE-Cy7 & CD45APCH7-was used for a high sensitivity PNH assay., Results: Of 364 cases, 59(16.2%) cases had PNH clone in both granulocytes and monocytes. PNH clone sizes ranged from 0.02% to 96.6% in granulocytes and 0.07% to 96.3% in monocytes based on their FLAER-negative, CD157-negative phenotype. Twenty-two of the 59 PNH cases (37.3%) had WBC clone size of <1%. In addition, there were 10 cases which showed absence of CD 157 expression on both granulocytes and monocytes but on FLAER staining showed normal staining patterns. Three of these ten cases also showed a PNH clone based on absence of FLAER expression on both granulocytes and monocytes., Conclusion: FLAER and CD157 is a robust combination for diagnosis of clinical and subclinical PNH. Absence of CD157 expression in normal WBCs, though rare, should be kept in mind and re-emphasizes the importance of FLAER for the high sensitivity PNH assay., (© 2020 John Wiley & Sons Ltd.)
- Published
- 2021
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130. Secondary cutaneous involvement by direct extension in high-grade B-cell lymphomas.
- Author
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Subtil A and Gru AA
- Subjects
- Aged, Antineoplastic Combined Chemotherapy Protocols therapeutic use, Biopsy methods, Bone Marrow Transplantation methods, Cyclophosphamide therapeutic use, Cytogenetic Analysis methods, Doxorubicin therapeutic use, Humans, Immunohistochemistry methods, Immunophenotyping methods, Lymphadenopathy pathology, Lymphoma, Large B-Cell, Diffuse drug therapy, Lymphoma, Large B-Cell, Diffuse metabolism, Male, Neoplasm Grading methods, Neoplasm Recurrence, Local, Prednisone therapeutic use, Rituximab therapeutic use, Treatment Outcome, Treatment Refusal, Vincristine therapeutic use, Hodgkin Disease pathology, Lymphadenopathy diagnosis, Lymphoma, Large B-Cell, Diffuse pathology, Skin Neoplasms secondary
- Abstract
Cutaneous involvement by a systemic lymphoma via direct extension is a very rare phenomenon, historically described in patients who had advanced-stage disease with bulky lymphadenopathy. Since its original description, most cases of cutaneous lymphomatous spread via direct extension are attributable to Hodgkin disease. Here, we report the occurrence of direct cutaneous dissemination in the setting of nodal high-grade B-cell lymphomas, specifically diffuse large B-cell lymphoma (DLBCL) and B-cell lymphoma, unclassifiable, with features intermediate between DLBCL and classical Hodgkin lymphoma (also called gray-zone lymphoma). Both cases are adequately documented with clinical, histopathologic, and immunophenotypic data, as well as additional cytogenetic analysis., (© 2020 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)
- Published
- 2021
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131. Comparison between the histopathological and immunophenotypical features of hypopigmented and nonhypopigmented mycosis fungoides: A retrospective study.
- Author
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Jayasinghe DR, Dissanayake K, and de Silva MVC
- Subjects
- Adolescent, Adult, Biopsy, CD8-Positive T-Lymphocytes pathology, Child, Child, Preschool, Cross-Sectional Studies, Epidermis immunology, Epidermis microbiology, Female, Humans, Infant, Infant, Newborn, Lymphoma, T-Cell, Cutaneous pathology, Male, Middle Aged, Mycosis Fungoides pathology, Mycosis Fungoides ultrastructure, Pigmentation Disorders pathology, Retrospective Studies, Young Adult, Epidermis pathology, Immunophenotyping methods, Mycosis Fungoides diagnosis, Mycosis Fungoides genetics, Skin Neoplasms pathology
- Abstract
Background: Mycosis fungoides (MF), the commonest primary cutaneous T-cell lymphoma, has classic and variant types which include hypopigmented MF (HMF). Previous studies have identified distinct clinicopathological profiles in HMF. This study aims to objectively compare the clinicopathological features of HMF with non-HMF lesions in order to characterize salient features of HMF., Methods: This cross-sectional, retrospective study analyzed biopsy specimens of 87 patients with MF. HMF and non-HMF groups were compared using clinical data, immunophenotypic features and scores given for six histopathological features: dermal infiltrate, basilar and superficially extending epidermotropism, Pautrier microabscesses and dermal and epidermotropic lymphocytic atypia., Results: Seventy-six patients had HMF. Presentation in females (59.21%; p = .04) and patch stage (88.16%; p = .01) in HMF were significant, and HMF presented at a younger mean age when compared to non-HMF. Both groups had equal intensity of epidermotropism, with HMF showing milder dermal infiltrates and significantly less dermal atypia. Pautrier microabscesses were significantly commoner in non-HMF (LR 10.76; p < .01). 94.74% of HMF were CD4-/CD8+., Conclusion: HMF presents at a lower age and earlier stage with female predominance compared to non-HMF. Because of milder dermal infiltrates, less dermal atypia, and Pautrier microabscesses, the diagnosis of HMF requires correlation with clinical features and careful assessment of epidermotropic cells., (© 2020 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)
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- 2021
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132. Plasma cell dedifferentiation in refractory multiple myeloma.
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Arbab A, Saada V, Cotteret S, Marzac C, and Ghez D
- Subjects
- Aged, Antibodies, Monoclonal administration & dosage, Antibodies, Monoclonal adverse effects, Antibodies, Monoclonal therapeutic use, Antineoplastic Combined Chemotherapy Protocols, Cell Dedifferentiation physiology, Dexamethasone administration & dosage, Dexamethasone adverse effects, Dexamethasone therapeutic use, Flow Cytometry methods, Humans, Immunophenotyping methods, Male, Multiple Myeloma diagnosis, Multiple Myeloma pathology, Mutation, Myelodysplastic Syndromes chemically induced, Thalidomide administration & dosage, Thalidomide adverse effects, Thalidomide analogs & derivatives, Thalidomide therapeutic use, Treatment Failure, Tumor Suppressor Protein p53 genetics, Cell Dedifferentiation drug effects, Multiple Myeloma drug therapy, Paraproteinemias blood, Plasma Cells pathology
- Published
- 2021
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133. Utility of TRBC1 Expression in the Diagnosis of Peripheral Blood Involvement by Cutaneous T-Cell Lymphoma.
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Horna P, Shi M, Jevremovic D, Craig FE, Comfere NI, and Olteanu H
- Subjects
- Adult, Aged, Aged, 80 and over, Biomarkers, Tumor blood, CD4-Positive T-Lymphocytes immunology, Case-Control Studies, Female, Flow Cytometry, Healthy Volunteers, Humans, Immunophenotyping methods, Lymphoma, T-Cell, Cutaneous blood, Lymphoma, T-Cell, Cutaneous immunology, Male, Middle Aged, Receptors, Antigen, T-Cell, alpha-beta blood, Sensitivity and Specificity, Skin Neoplasms blood, Skin Neoplasms immunology, T-Lymphocyte Subsets immunology, Young Adult, Biomarkers, Tumor metabolism, CD4-Positive T-Lymphocytes metabolism, Lymphoma, T-Cell, Cutaneous diagnosis, Receptors, Antigen, T-Cell, alpha-beta metabolism, Skin Neoplasms diagnosis, T-Lymphocyte Subsets metabolism
- Abstract
Peripheral blood involvement by cutaneous T-cell lymphoma is typically assessed by flow cytometry and plays a critical role in diagnosis, classification, and prognosis. Simplified strategies to detect tumor cells (Sezary cells) fail to exclude reactive subsets, whereas tumor-specific abnormalities are subtle and inconsistently present. We implemented a flow cytometric strategy to detect clonal Sezary cells based on the monotypic expression of one of two mutually exclusive TCR constant β chains, TRBC1 and TRBC2. Analysis of CD4
+ T-cell subsets and TCR variable β classes from healthy donors showed polytypic TRBC1 staining. Clonal Sezary cells were identified by TRBC1 staining in 56 of 111 (50%) samples from patients with cutaneous T-cell lymphoma, accounting for 7-18,155 cells/μl and including 13 cases (23%) lacking tumor-specific immunophenotypic abnormalities. CD4+ T-cell subsets from 86 patients without T-cell lymphoma showed polytypic TRBC1 staining, except for five patients (6%) with minute T-cell clones of uncertain significance accounting for 53-136 cells/μl. Assessment of TRBC1 expression within a comprehensive single-tube flow cytometry assay effectively overcomes interpretative uncertainties in the identification of Sezary cells without the need for a separate T-cell clonality assay., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)- Published
- 2021
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134. Altered immune response to the annual influenza A vaccine in patients with myeloproliferative neoplasms.
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Alimam S, Ann Timms J, Harrison CN, Dillon R, Mare T, DeLavallade H, Radia D, Woodley C, Francis Y, Sanchez K, Kordasti S, and McLornan DP
- Subjects
- Adult, Aged, B-Lymphocytes drug effects, B-Lymphocytes immunology, CD4-Positive T-Lymphocytes drug effects, CD4-Positive T-Lymphocytes immunology, Case-Control Studies, Female, Humans, Immunity immunology, Immunologic Memory drug effects, Immunophenotyping methods, Influenza, Human immunology, Influenza, Human virology, Male, Middle Aged, Myeloproliferative Disorders pathology, Neoplasms diagnosis, Neoplasms immunology, T-Lymphocytes drug effects, T-Lymphocytes immunology, Immunity drug effects, Influenza A virus immunology, Influenza, Human prevention & control, Myeloproliferative Disorders immunology, Vaccination adverse effects
- Abstract
The seasonal influenza A vaccine is recommended for patients with myeloproliferative neoplasms (MPNs). We hypothesised that immune deregulation associated with MPNs may affect the immune response gained following vaccinations when compared to healthy controls. Using deep immunophenotyping with high-dimensional single-cell analysis and mass cytometry we could demonstrate an altered immune response in MPN patients following vaccination. We found that prior to vaccination, MPN patients had reduced numbers of naive CD4 T cells. Furthermore, at 3-weeks and 3-months post-vaccination there was evidence of both delayed and impaired B- and T-memory cells responses. Thus, although, the immune systems of MPN patients can 'recognise' the Influenza A vaccine, the response appears inferior compared to healthy controls., (© 2020 The Authors. British Journal of Haematology published by British Society for Haematology and John Wiley & Sons Ltd.)
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- 2021
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135. Gender Differences in Hemocyte Immune Parameters of Hong Kong Oyster Crassostrea hongkongensis During Immune Stress.
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Lu J, Shi Y, Yao T, Bai C, Jiang J, and Ye L
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- Animals, Crassostrea metabolism, Female, Hemocytes cytology, Histocytochemistry methods, Immunophenotyping methods, Male, Phagocytosis immunology, Sex Factors, Stress, Physiological genetics, Biomarkers, Crassostrea immunology, Hemocytes immunology, Hemocytes metabolism, Stress, Physiological immunology
- Abstract
Gender differences in individual immune responses to external stimuli have been elucidated in many invertebrates. However, it is unclear if gender differences do exist in the Hong Kong oyster Crassostrea hongkongensis , one of the most valuable marine species cultivated along the coast of South China. To clarify this, we stimulated post-spawning adult C. hongkongensis with Vibrio harveyi and lipopolysaccharide (LPS). Gender-based differences in some essential functional parameters of hemocytes were studied via flow cytometry. Obvious gender-, subpopulation-, and immune-specific alterations were found in the hemocyte immune parameters of C. hongkongensis . Three hemocyte subpopulations were identified: granulocytes, semi-granulocytes, and agranulocytes. Granulocytes, the chief phagocytes and major producers of esterase, reactive oxygen species, and nitric oxide, were the main immunocompetent hemocytes. Immune parameter alterations were notable in the accumulation of granulocyte esterase activities, lysosomal masses, nitric oxide levels, and granulocyte numbers in male oysters. These results suggest that post-spawning-phase male oysters possess a more powerful immune response than females. Gender and subpopulation differences in bivalve immune parameters should be considered in the future analysis of immune parameters when studying the impact of pathogenic or environmental factors., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Lu, Shi, Yao, Bai, Jiang and Ye.)
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- 2021
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136. Hyposialylation Must Be Considered to Develop Future Therapies in Autoimmune Diseases.
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Bordron A, Morel M, Bagacean C, Dueymes M, Pochard P, Harduin-Lepers A, Jamin C, and Pers JO
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- Animals, Autoantibodies immunology, Autoimmune Diseases therapy, Humans, Immunophenotyping methods, Precision Medicine methods, Sialic Acids immunology, Autoantibodies metabolism, Autoimmune Diseases immunology, Protein Processing, Post-Translational, Sialic Acids metabolism
- Abstract
Autoimmune disease development depends on multiple factors, including genetic and environmental. Abnormalities such as sialylation levels and/or quality have been recently highlighted. The adjunction of sialic acid at the terminal end of glycoproteins and glycolipids is essential for distinguishing between self and non-self-antigens and the control of pro- or anti-inflammatory immune reactions. In autoimmunity, hyposialylation is responsible for chronic inflammation, the anarchic activation of the immune system and organ lesions. A detailed characterization of this mechanism is a key element for improving the understanding of these diseases and the development of innovative therapies. This review focuses on the impact of sialylation in autoimmunity in order to determine future treatments based on the regulation of hyposialylation.
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- 2021
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137. Tumor microenvironment immune subtypes for classification of novel clear cell renal cell carcinoma profiles with prognostic and therapeutic implications.
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Wang Q, Hu J, Kang W, Wang J, Xiang Y, Fu M, Gao H, and Huang Z
- Subjects
- Biomarkers, Tumor immunology, Carcinoma, Renal Cell immunology, Carcinoma, Renal Cell mortality, Chemokine CX3CL1, DNA Copy Number Variations immunology, DNA-Binding Proteins, Gene Expression Regulation, Neoplastic immunology, HMGB1 Protein, Humans, Interferon-alpha, Kidney Neoplasms immunology, Kidney Neoplasms mortality, Lymphocytes, Tumor-Infiltrating immunology, Methylation, Mutation immunology, Neoplasm Grading, Neoplasm Staging, P-Selectin, Predictive Value of Tests, Prognosis, Receptors, Tumor Necrosis Factor, Member 14, Signal Transduction immunology, Survival Analysis, Toll-Like Receptor 4, Transcription Factors, Von Hippel-Lindau Tumor Suppressor Protein, Carcinoma, Renal Cell classification, Immunophenotyping methods, Kidney Neoplasms classification, Lymphocyte Subsets immunology, Tumor Microenvironment immunology
- Abstract
Abstract: Currently, no effective prognostic model of clear cell renal cell carcinoma (ccRCC) based on immune cell infiltration has been developed. Recent studies have identified 6 immune groups (IS) in 33 solid tumors. We aimed to characterize the expression pattern of IS in ccRCC and evaluate the potential in predicting patient prognosis. The clinical information, immune subgroup, somatic mutation, copy number variation, and methylation score of patients with TCGA ccRCC cohort were downloaded from UCSC Xena for further analysis. The most dominant IS in ccRCC was the inflammatory subgroup (immune C3) (86.5%), regardless of different pathological stages, pathological grades, and genders. In the C3 subgroup, stage IV (69.1%) and grade 4 (69.9%) were the least presented. Survival analysis showed that the IS could effectively predict the overall survival (OS) (P < .0001) and disease-specific survival (DSS) (P < .0001) of ccRCC alone, of which group C3 (OS, HR = 2.3, P < .001; DSS, HR = 2.84, P < .001) exhibited the best prognosis. Among the most frequently mutated ccRCC genes, only VHL and PBRM1 were found to be common in the C3 group. The homologous recombination deficiency score was also lower. High heterogeneity was observed in immune cells and immunoregulatory genes of IS. Notably, CD4+ memory resting T cells were highly infiltrating, regulatory T cells (Treg) showed low infiltration, and most immunoregulatory genes (such as CX3CL1, IFNA2, TLR4, SELP, HMGB1, and TNFRSF14) were highly expressed in the C3 subgroup than in other subgroups. Enrichment analysis showed that adipogenesis, apical junction, hypoxia, IL2 STAT5 signaling, TGF-beta signaling, and UV response DN were activated, whereas E2F targets, G2M checkpoint, and MYC targets V2 were downregulated in the C3 group. Immune classification can more accurately classify ccRCC patients and predict OS and DSS. Thus, IS-based classification may be a valuable tool that enables individualized treatment of patients with ccRCC., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. All authors have read and approved the final version of the manuscript., (Copyright © 2021 the Author(s). Published by Wolters Kluwer Health, Inc.)
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- 2021
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138. A hyperacute immune map of ischaemic stroke patients reveals alterations to circulating innate and adaptive cells.
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Krishnan S, O'Boyle C, Smith CJ, Hulme S, Allan SM, Grainger JR, and Lawrence CB
- Subjects
- Aged, Aged, 80 and over, Antigens, CD blood, Antigens, CD immunology, B-Lymphocytes metabolism, Brain Ischemia complications, CD4-Positive T-Lymphocytes metabolism, CD8-Positive T-Lymphocytes metabolism, Cohort Studies, Female, Flow Cytometry methods, Humans, Immunologic Memory immunology, Ischemic Stroke blood, Ischemic Stroke etiology, Lymphocyte Activation immunology, Male, Myeloid Cells metabolism, B-Lymphocytes immunology, CD4-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes immunology, Immunophenotyping methods, Ischemic Stroke immunology, Myeloid Cells immunology
- Abstract
Systemic immune changes following ischaemic stroke are associated with increased susceptibility to infection and poor patient outcome due to their role in exacerbating the ischaemic injury and long-term disability. Alterations to the abundance or function of almost all components of the immune system post-stroke have been identified, including lymphocytes, monocytes and granulocytes. However, subsequent infections have often confounded the identification of stroke-specific effects. Global understanding of very early changes to systemic immunity is critical to identify immune targets to improve clinical outcome. To this end, we performed a small, prospective, observational study in stroke patients with immunophenotyping at a hyperacute time point (< 3 h) to explore early changes to circulating immune cells. We report, for the first time, decreased frequencies of type 1 conventional dendritic cells (cDC1), haematopoietic stem and progenitor cells (HSPCs), unswitched memory B cells and terminally differentiated effector memory T cells re-expressing CD45RA (TEMRA). We also observed concomitant alterations to human leucocyte antigen D-related (HLA-DR), CD64 and CD14 expression in distinct myeloid subsets and a rapid activation of CD4
+ T cells based on CD69 expression. The CD69+ CD4+ T cell phenotype inversely correlated with stroke severity and was associated with naive and central memory T (TCM) cells. Our findings highlight early changes in both the innate and adaptive immune compartments for further investigation as they could have implications the development of post-stroke infection and poorer patient outcomes., (© 2020 The Authors. Clinical & Experimental Immunology published by John Wiley & Sons Ltd on behalf of British Society for Immunology.)- Published
- 2021
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139. Characterization of autoantibodies, immunophenotype and autoimmune disease in a prospective cohort of patients with idiopathic CD4 lymphocytopenia.
- Author
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Cudrici CD, Boulougoura A, Sheikh V, Freeman A, Sortino O, Katz JD, Sereti I, and Siegel RM
- Subjects
- Adult, Aged, Autoimmune Diseases complications, Cohort Studies, Communicable Diseases complications, Female, Humans, Male, Middle Aged, Prospective Studies, T-Lymphocytopenia, Idiopathic CD4-Positive complications, Young Adult, Autoantibodies analysis, Autoimmune Diseases immunology, Immunophenotyping methods, T-Lymphocytopenia, Idiopathic CD4-Positive immunology
- Abstract
Objective: Characterize autoantibodies and autoimmune diseases in a prospective cohort of patients with Idiopathic CD4 Lymphocytopenia (ICL) a rare immunodeficiency characterized by an absolute CD4
+ T count of <300 cells/μl in the absence of HIV or HTLV infection., Methods: Single-Center prospective study of 67 patients conducted over an 11-year period. Rheumatologic evaluation and measurement of autoantibodies were systematically conducted, and flow cytometry of immune cell subsets was performed in a subset of patients., Results: 54% of referred patients had clinical evidence of autoimmunity, with 34% having at least one autoimmune disease, most commonly autoimmune thyroid disease. 19%, had autoantibodies or incomplete features of autoimmune disease. Patients with autoimmune disease had more elevated serum immunoglobulins, and more effector memory T cells than those without autoimmunity., Conclusions: Evidence of autoimmunity, including autoimmune diseases, is more prevalent in ICL than the general population, and should be considered part of this syndrome., (Published by Elsevier Inc.)- Published
- 2021
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140. Perivascular epithelioid cell tumors (PEComa) of the gynecologic tract.
- Author
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Bennett JA and Oliva E
- Subjects
- Basic Helix-Loop-Helix Leucine Zipper Transcription Factors genetics, Biomarkers, Tumor, Diagnosis, Differential, Female, Gene Fusion, Gene Rearrangement, Humans, Immunophenotyping methods, MTOR Inhibitors pharmacology, Neoplasms, Connective and Soft Tissue genetics, Neoplasms, Connective and Soft Tissue immunology, Neoplasms, Connective and Soft Tissue metabolism, Tuberous Sclerosis Complex 1 Protein genetics, Tuberous Sclerosis Complex 1 Protein metabolism, Perivascular Epithelioid Cell Neoplasms genetics, Perivascular Epithelioid Cell Neoplasms immunology, Perivascular Epithelioid Cell Neoplasms metabolism
- Abstract
PEComas of the female genital tract are rare mesenchymal neoplasms that are most common in the uterus, but also may occur in other gynecologic locations. As they morphologically and immunohistochemically resemble smooth muscle tumors, distinction between the two entities is often challenging, and may be aided by molecular analysis. Thus far, two distinct molecular groups-classic PEComas with TSC mutations and TFE3-translocation associated PEComas with TFE3 fusions have been described. Recognition of the first group is imperative as these patients may benefit from targeted therapy with mTOR inhibitors, if malignant. This review will focus on recognition of the morphologic and immunophenotypic features of PEComas, as well as the role of molecular testing in their diagnosis and treatment, analysis of the different algorithms to predict behavior, and differential diagnosis., (© 2020 Wiley Periodicals LLC.)
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- 2021
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141. Apoptosis and immunophenotyping of peripheral blood lymphocytes in Iranian COVID-19 patients: Clinical and laboratory characteristics.
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Taghiloo S, Aliyali M, Abedi S, Mehravaran H, Sharifpour A, Zaboli E, Eslami-Jouybari M, Ghasemian R, Vahedi-Larijani L, Hossein-Nattaj H, Amjadi O, Rezazadeh H, Ajami A, and Asgarian-Omran H
- Subjects
- Adult, Aged, Apoptosis immunology, Biomarkers blood, COVID-19 immunology, Female, Flow Cytometry, Humans, Iran, Lymphocyte Count, Lymphopenia immunology, Male, Middle Aged, B-Lymphocytes cytology, CD4-Positive T-Lymphocytes cytology, CD8-Positive T-Lymphocytes cytology, Immunophenotyping methods, Killer Cells, Natural cytology, SARS-CoV-2 immunology
- Abstract
A novel member of human coronavirus, named severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), has been recently recognized in China and rapidly spread worldwide. Studies showed the decreasing of peripheral blood lymphocytes in a majority of patients. In this study, we have reported the clinical features, laboratory characteristics, the frequency of peripheral blood lymphocyte subpopulations, and their apoptosis pattern in Iranian coronavirus infectious disease (COVID-19) patients. Demographic and clinical data of 61 hospitalized confirmed cases with COVID-19 at Imam Khomeini Hospital were collected and analyzed. Peripheral blood mononuclear cells were isolated from all samples and the apoptosis pattern was evaluated using Annexin V/propidium iodide method. The frequency of lymphocyte subsets, including T-CD4
+ , T-CD8+ , NK, B cells, and monocytes, was measured in all patients and 31 controls by flow cytometry. Our findings demonstrated that the percentage of lymphocytes, CD4+ , and CD8+ T cells were decreased in COVID-19 patients compared with the control group. Regarding the clinical severity, the number of lymphocytes, CD4+ , CD8+ T cells, and NK cells were also decreased in severe cases when compared with mild cases. Finally, our data have also indicated the increase in apoptosis of mononuclear cells from COVID-19 patients which was more remarkable in severe clinical cases. The frequency of immune cells is a useful indicator for prediction of severity and prognosis of COVID-19 patients. These results could help to explain the immunopathogenesis of SARS-CoV-2 and introducing novel biomarkers, therapeutic strategies, and vaccine candidates., (© 2020 Wiley Periodicals LLC.)- Published
- 2021
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142. An immunostaining panel of C-reactive protein, N-cadherin, and S100 calcium binding protein P is useful for intrahepatic cholangiocarcinoma subtyping.
- Author
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Akita M, Sawada R, Komatsu M, Suleman N, Itoh T, Ajiki T, Heaton N, Fukumoto T, and Zen Y
- Subjects
- Aged, Aged, 80 and over, Bile Duct Neoplasms metabolism, Bile Ducts, Intrahepatic metabolism, Bile Ducts, Intrahepatic pathology, Biomarkers, Tumor analysis, Cholangiocarcinoma metabolism, Diagnosis, Differential, Female, Humans, Immunophenotyping methods, Liver Neoplasms metabolism, Liver Neoplasms pathology, Male, Middle Aged, Bile Duct Neoplasms pathology, C-Reactive Protein metabolism, Cadherins metabolism, Calcium-Binding Proteins metabolism, Cholangiocarcinoma pathology, Neoplasm Proteins metabolism
- Abstract
This study aimed to establish an immunohistochemical panel useful for subclassification of intrahepatic cholangiocarcinoma (iCCA) into small- and large-duct types. Fifty surgical cases of iCCA consisting of small- (n = 31) and large-duct types (n = 19) were examined. To imitate liver needle biopsies, tissue microarrays were constructed using three tissue cores (2 mm in diameter) obtained from one representative paraffin block of each case. Immunostaining for C-reactive protein (CRP), N-cadherin, tubulin beta-III (TUBB3), neural cell adhesion molecule (NCAM), and S100 calcium binding protein P (S100P) was conducted. Most cases of small-duct iCCA were immunoreactive to CRP and N-cadherin, whereas expressions of these markers were markedly less common in large-duct iCCA (CRP, 97% vs. 5%, P < 0.001; N-cadherin, 87% vs. 16%, P < 0.001). TUBB3 and NCAM were also more frequently expressed in small-duct iCCA (65% vs. 32%, P = 0.006; 58% vs. 5%, P < 0.001), but their sensitivities were lower than those of CRP and N-cadherin. S100P was more commonly expressed in large-duct iCCA than in small-duct iCCA (95% vs. 29%, P < 0.001), and diffuse expressions were observed in 17 of 19 cases of large-duct iCCA (90%). All cases with a CRP+/S100P- immunophenotype were of small-duct type, whereas all but one case with a CRP-/S100P+ immunophenotype were of large-duct type. Of 10 cases with a double-positive or double-negative immunophenotype, 7 were appropriately classified based on immunoreactivity to N-cadherin. In conclusion, CRP, N-cadherin, and S100P form a useful immunohistochemical panel for iCCA subclassification, and correct subclassification was possible in 92% of cases based on a proposed, simple algorithm., (Copyright © 2020. Published by Elsevier Inc.)
- Published
- 2021
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143. Novel tumour-infiltrating lymphocyte-related risk stratification based by flow cytometry for patients with de novo angioimmunoblastic T cell lymphoma.
- Author
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Zhu Q, Deng X, Yao W, Chen Z, Ye Y, Gao L, Zhang W, Liu W, and Zhao S
- Subjects
- Adult, Aged, Aged, 80 and over, China, Cohort Studies, Disease Progression, Female, Humans, Immunoblastic Lymphadenopathy diagnosis, Immunophenotyping methods, Lymphoma, T-Cell diagnosis, Male, Middle Aged, Neoplasm Staging, Predictive Value of Tests, Prognosis, Retrospective Studies, Risk Assessment, Risk Factors, Sensitivity and Specificity, Tumor Microenvironment immunology, Flow Cytometry, Immunoblastic Lymphadenopathy pathology, Lymphocytes, Tumor-Infiltrating pathology, Lymphoma, T-Cell pathology
- Abstract
Tumour-infiltrating lymphocytes (TILs) account for a large proportion of tumour microenvironment (TME) in angioimmunoblastic T cell lymphoma (AITL), and at present the significance of TIL in TME of AITL remains unclear. Overall, 50 de novo AITL patients undergoing lymph node flow cytometry from 2014 to 2019 were retrospectively analysed to assess the relationship between TILs and AITL prognosis. We found that high TIL-Bs (≥ 42.4%, p = 0.004) and high CD4:CD8 (≥ 0.85, p = 0.024) were independent favourable prognostic factors for de novo AITL in univariate or multivariate analyses. New TIL-related risk stratification was established based on TIL-Bs and CD4:CD8 factors. Patients in the low-risk group (TIL-Bs ≥ 42.4% and CD4:CD8 ≥ 0.85) had significantly better overall survival than the high-risk (TIL-Bs < 42.4% and CD4:CD8 < 0.85) (p < 0.001) or intermediate-risk group (TIL-Bs ≥ 42.4% and CD4:CD8 < 0.85 or TIL-Bs < 42.4% and CD4:CD8 ≥ 0.85) (p = 0.011). To our knowledge, our cohort is the largest one focusing on the TILs in de novo cases of AITL by analysing lymph node samples using flow cytometry, which is the first time to comprehensively consider humoral immunity and cellular immunity influence on AITL. Our new risk stratification was valuable and useful in evaluating prognosis of AITL and guiding immunotherapy strategies.
- Published
- 2021
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144. STAT3 Silencing and TLR7/8 Pathway Activation Repolarize and Suppress Myeloid-Derived Suppressor Cells From Breast Cancer Patients.
- Author
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Safarzadeh E, Mohammadi A, Mansoori B, Duijf PHG, Hashemzadeh S, Khaze V, Kazemi T, Derakhshani A, Silvestris N, and Baradaran B
- Subjects
- CD3 Complex immunology, Cell Differentiation immunology, Cell Proliferation physiology, Cytokines immunology, Female, HLA-DR Antigens immunology, Humans, Immune Tolerance immunology, Immunophenotyping methods, Middle Aged, Myeloid Cells immunology, T-Lymphocytes immunology, Tumor Microenvironment immunology, Breast Neoplasms immunology, Myeloid-Derived Suppressor Cells immunology, STAT3 Transcription Factor immunology, Signal Transduction immunology, Toll-Like Receptor 7 immunology, Toll-Like Receptor 8 immunology
- Abstract
Cancer cells escape immune destruction. From this perspective, myeloid-derived suppressor cells (MDSCs), which are immunosuppressive in various cancers including breast cancer (BC), are significant. However, the precise mechanisms are unknown. We isolated HLA-DR
- CD33+ MDSCs and CD3+ T cells from BC patients' peripheral blood and healthy donors through MACS and immunophenotyped by flow cytometry. Transfection of short-interfering RNAs and treatment with a TLR7/8 agonist altered pathway activities in vitro . Gene expression was analyzed using qRT-PCR, western blotting, and immunohistochemistry. Our findings showed an association between the progression of BC and increased levels of circulating HLA-DR- CD33+ MDSCs. These cells strongly suppress both autologous and analogous CD3+ T cell proliferation and enter the tumor microenvironment. We also identified increased STAT3 signaling and increased IDO and IL-10 expression in BC-derived MDSCs as immunosuppression mechanisms. Further, STAT3 inhibition and TLR7/8 pathway stimulation reduce the immunosuppressive activity of patient-derived MDSCs on T cells by inducing MDSC repolarization and differentiation into mature myeloid cells. This also alters the expression of critical cytokines and transcription factors in CD3+ T cells and, importantly, reduces breast cancer cells' proliferation. Finally, while chemotherapy is able to significantly reduce circulating MDSCs' level in patients with breast cancer, these MDSCs remained highly T cell-suppressive. We identified a novel molecular mechanism of MDSC-mediated immunosuppression. STAT3 inhibition and TLR7/8 pathway stimulation in MDSCs repolarize and suppress MDSCs from breast cancer patients. This offers new opportunities for BC immunotherapy., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Safarzadeh, Mohammadi, Mansoori, Duijf, Hashemzadeh, Khaze, Kazemi, Derakhshani, Silvestris and Baradaran.)- Published
- 2021
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145. Lymphocyte Immunophenotyping and CD4/CD8 Ratio in Cerebrospinal Fluid for the Diagnosis of Sarcoidosis-related Uveitis.
- Author
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Paule R, Denis L, Chapuis N, Rohmer J, Hadjadj J, London J, Chauvin A, Bonnet C, Mouthon L, Le Jeunne C, Monnet D, Blanche P, Brezin A, and Terrier B
- Subjects
- Adult, Biomarkers cerebrospinal fluid, CD4-CD8 Ratio, Female, Flow Cytometry, Humans, Male, Middle Aged, Retrospective Studies, Sarcoidosis cerebrospinal fluid, Sarcoidosis complications, Uveitis cerebrospinal fluid, Uveitis etiology, CD4-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes immunology, Immunophenotyping methods, Sarcoidosis immunology, Uveitis diagnosis
- Abstract
Background : This study aimed to assess the diagnostic relevance of CD4/CD8 ratio in cerebrospinal fluid (CSF) for the etiological diagnosis work-up of uveitis. Methods : We consecutively included patients who were referred to our department for the diagnostic workup of intermediate and/or posterior uveitis. Etiological diagnoses were established in a blind manner regarding CD4/CD8 ratio. Results : Fifty-two patients were included. A diagnosis of ocular sarcoidosis was made in 15 (29%) patients, 21% had another determined diagnosis while 50% remained of undetermined origin. Median CD4/CD8 ratio in CSF was 4.57 (IQR 3.39-5.47) in ocular sarcoidosis, 1.74 (1.60-3.18) in uveitis due to other determined cause ( P = .008), and 2.83 (2.34-3.54) in those with uveitis of undetermined origin ( P = .007). CD4/CD8 ratio >3.23 was associated with a diagnosis of ocular sarcoidosis. Conclusion : Determination of CD4/CD8 ratio in CSF can be useful for diagnosis work-up since a CD4/CD8 ratio >3.23 in CSF is associated with ocular sarcoidosis.
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- 2021
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146. Double remission of chronic lymphocytic leukemia and secondary acute myeloid leukemia after venetoclax monotherapy: A case report.
- Author
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Wang L and Lin N
- Subjects
- Antibiotics, Antineoplastic administration & dosage, Antibiotics, Antineoplastic therapeutic use, Antimetabolites, Antineoplastic administration & dosage, Antimetabolites, Antineoplastic therapeutic use, Antineoplastic Agents administration & dosage, Antineoplastic Agents therapeutic use, Antineoplastic Combined Chemotherapy Protocols therapeutic use, Bridged Bicyclo Compounds, Heterocyclic administration & dosage, Bridged Bicyclo Compounds, Heterocyclic therapeutic use, Cytarabine administration & dosage, Cytarabine therapeutic use, Daunorubicin administration & dosage, Daunorubicin therapeutic use, Disease Progression, Hematopoietic Stem Cell Transplantation standards, Humans, Immunophenotyping methods, Leukemia, Lymphocytic, Chronic, B-Cell pathology, Leukemia, Myeloid, Acute pathology, Leukocyte Count, Lymphocyte Count, Male, Middle Aged, Remission Induction, Sulfonamides administration & dosage, Sulfonamides therapeutic use, Treatment Outcome, Leukemia, Lymphocytic, Chronic, B-Cell complications, Leukemia, Lymphocytic, Chronic, B-Cell drug therapy, Leukemia, Myeloid, Acute drug therapy, Leukemia, Myeloid, Acute etiology
- Abstract
Rationale: The abnormal expression of B-cell lymphoma-2 (Bcl-2) family members is often associated with the progression of the disease. Bcl-2 inhibitors (eg, venetoclax) were first reported to inhibit the proliferation of malignant lymphocytes and have a significant effect on patients with chronic lymphoblastic leukemia, but research on myeloid tumors is relatively delayed. Venetoclax was approved in 2018 for the treatment of acute myeloid leukemia (AML) patients who were not suitable for high-dose chemotherapy. The approval of venetoclax is an advance in the treatment of hematological tumors., Patient Concerns: Here we report a 64-year-old male with an increased white blood cell (WBC) count (39.0 × 109/L) and lymphocyte count (30.6 × 109/L) on physical examination in July 2014. The patients were diagnosed with chronic lymphocytic leukemia (CLL) through bone marrow (BM) smears and immunophenotyping without any cytogenetic or molecular abnormalities. Chlorambucil was prescribed, WBC was stable between 15 × 109/L and 25 × 109/L in the past 6 years. He came to the hospital again in May 2020 and complained of fatigue for 2 weeks. WBC (16.7 × 109/L) and lymphocyte (14.76 × 109/L) counts were increased, hemoglobin (HGB) and platelet (PLT) were decreased in peripheral blood, which indicated the progression of the disease., Diagnoses: The patient was diagnosed as secondary AML after CLL based on the clinical and laboratory findings., Interventions: He achieved a morphological complete remission in both AML and CLL without any adverse reactions after one course of venetoclax monotherapy., Outcomes: He received standard daunorubicin and cytarabine combined with venetoclax as consolidation therapy and is now ready for allogeneic-hematopoietic stem cell transplantation., Lessons: Our case presents a challenge to traditional treatment. New drugs such as venetoclax have shown outstanding effects in this respect. High expression of Bcl-2 can identify the responders of venetoclax. These findings should be validated in future clinical trials. We fully believe that in the near future, the comprehensive use of targeted drugs with different mechanisms will not only improve the quality of life of patients, but also completely change the prognosis of patients with recurrent and refractory hematological malignancies., Competing Interests: The authors have no conflicts of interest to disclose., (Copyright © 2021 the Author(s). Published by Wolters Kluwer Health, Inc.)
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- 2021
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147. Utility of adjuvant whole abdominal radiation therapy in ovarian clear cell cancer (OCCC): a pragmatic cohort study of women with classic immuno-phenotypic signature.
- Author
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Stevens MJ, West S, Gard G, Renaud C, Nevell D, Roderick S, and Le A
- Subjects
- Adenocarcinoma, Clear Cell immunology, Adenocarcinoma, Clear Cell pathology, Adenocarcinoma, Clear Cell radiotherapy, Aged, Female, Humans, Middle Aged, Ovarian Neoplasms immunology, Ovarian Neoplasms pathology, Ovarian Neoplasms radiotherapy, Positron Emission Tomography Computed Tomography, Prognosis, Radiotherapy Dosage, Survival Rate, Abdomen radiation effects, Adenocarcinoma, Clear Cell mortality, Immunophenotyping methods, Ovarian Neoplasms mortality, Radiotherapy, Adjuvant mortality, Radiotherapy, Intensity-Modulated mortality
- Abstract
Background: To evaluate the initial experience and clinical utility of first-line adjuvant intensity-modulated whole abdominal radiation therapy (WART) in women with ovarian clear cell cancer (OCCC) referred to an academic center., Methods: Progression-free and overall survival was analyzed in a pragmatic observational cohort study of histologically pure OCCC patients over-expressing HNF-1ß treated between 2013 and end-December 2018. An in-house intensity-modulated WART program was developed from a published pre-clinical model. Radiation dose-volume data was curated to American Association of Physics in Medicine (AAPM) Task Group 263 recommendations. A dedicated database prospectively recorded presenting characteristics and outcomes in a standardized fashion., Results: Five women with FIGO (2018) stage IA to IIIA2 OCCC were treated with first-line WART. Median age was 58 years (range 47-68 years). At diagnosis CA-125 was elevated in 4 cases (median 56 kU/L: range 18.4-370 kU/L) before primary de-bulking surgery. Severe premorbid endometriosis was documented in 3 patients. At a median follow-up of 77 months (range 16-83 mo.), all patients remain alive and progression-free on clinical, biochemical (CA-125), and
18 Fluoro-deoxyglucose (FDG) PET/CT re-evaluation. Late radiation toxicity was significant (G3) in 1 case who required a limited bowel resection and chronic nutritional support at 9 months post-WART; 2 further patients had asymptomatic (G2) osteoporotic fragility fractures of axial skeleton at 12 months post-radiation treated with anti-resorptive agents (denosumab)., Conclusions: The clinical utility of intensity-modulated WART in OCCC over-expressing HNF-1β was suggested in this small observational cohort study. The hypothesis that HNF-1β is a portent of platinum-resistance and an important predictive biomarker in OCCC needs further confirmation. Curating multi-institutional cohort studies utilizing WART by means of "Big Data" may improve OCCC care standards in the future.- Published
- 2021
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148. From the archives of MD Anderson Cancer Center: Untreated leukemic non-nodal mantle cell lymphoma with relapse as pleomorphic variant mantle cell lymphoma 21 years later.
- Author
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Fang H, Medeiros LJ, Tang Z, Wang W, Ok CY, Patel KP, Khoury JD, and Thakral B
- Subjects
- Adult, Antineoplastic Combined Chemotherapy Protocols therapeutic use, Biopsy, Follow-Up Studies, Humans, Immunophenotyping methods, Leukemia, Lymphocytic, Chronic, B-Cell diagnosis, Leukemia, Lymphocytic, Chronic, B-Cell drug therapy, Lymphadenopathy pathology, Lymphocytosis etiology, Lymphocytosis pathology, Lymphoma, Mantle-Cell diagnosis, Lymphoma, Mantle-Cell metabolism, Lymphoma, Mantle-Cell therapy, Male, Recurrence, Remission Induction, Splenectomy methods, Stem Cell Transplantation methods, Transplantation, Homologous methods, Cyclin D1 metabolism, Leukemia, Lymphocytic, Chronic, B-Cell pathology, Lymphoma, Mantle-Cell pathology, Lymphoma, Non-Hodgkin pathology
- Abstract
Leukemic, non-nodal mantle cell lymphoma (MCL) is a distinct, rare, indolent variant of mantle cell lymphoma, but can relapse aggressively. It can present with lymphocytosis with chronic lymphocytic leukemia (CLL)-like morphologic and immunophenotypic features as was initially considered in the index case. However, at time of splenectomy, two years later cyclin D1 overexpression was shown and the disease was realized to be leukemic non-nodal MCL. The patient was followed for 21 years, without therapy, before he developed clinically aggressive MCL with lymphadenopathy. Lymph node biopsy showed MCL, pleomorphic variant. We review the literature and discuss the features of leukemic non-nodal MCL as well as the potential pitfalls in diagnosis. Furthermore, we are not aware of another cases reported with a 21 year interval from initial diagnosis of leukemic non-nodal MCL to aggressive MCL., (Published by Elsevier Inc.)
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- 2021
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149. Reliable measurements of extracellular vesicles by clinical flow cytometry.
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Kuiper M, van de Nes A, Nieuwland R, Varga Z, and van der Pol E
- Subjects
- Animals, Extracellular Vesicles pathology, Flow Cytometry standards, Humans, Immunophenotyping standards, Observer Variation, Organelle Size, Reference Standards, Biomarkers metabolism, Extracellular Vesicles metabolism, Flow Cytometry methods, Immunophenotyping methods
- Abstract
Extracellular vesicles (EVs) are cell-derived particles with a phospholipid membrane present in all body fluids. Because EV properties change in health and disease, EVs have excellent potential to become biomarkers for diagnosis, prognosis, or monitoring of disease. The only technique capable of detecting, sizing, and phenotyping a million of EVs within minutes is (clinical) flow cytometry. A flow cytometer measures light scattering and fluorescence signals of single EVs. Although these signals contain valuable information about the presence and composition of EVs, the signals are expressed in arbitrary units, which make the comparison of measurement results impossible between instruments and laboratories. Additionally, unintended and undocumented variations in the source, preparation, and analysis of the sample lead to orders of magnitude variations in the measured EV concentrations. Here, we will explain the basics, challenges, and common misconceptions of EV flow cytometry. In addition, we provide an overview of recent standardization initiatives, which are a prerequisite for comparison of clinical data and thus for clinical biomarker exploration of EVs., (© 2020 The Authors. American Journal of Reproductive Immunology published by John Wiley & Sons Ltd.)
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- 2021
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150. Switched and unswitched memory B cells detected during SARS-CoV-2 convalescence correlate with limited symptom duration.
- Author
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Newell KL, Clemmer DC, Cox JB, Kayode YI, Zoccoli-Rodriguez V, Taylor HE, Endy TP, Wilmore JR, and Winslow GM
- Subjects
- Adult, Aged, Antibodies, Neutralizing immunology, Antibodies, Viral immunology, Antibody Formation, B-Lymphocytes immunology, Convalescence, Disease Progression, Female, Humans, Immunity immunology, Immunoglobulin G immunology, Immunoglobulin M metabolism, Immunophenotyping methods, Male, Middle Aged, Recovery of Function immunology, SARS-CoV-2 immunology, SARS-CoV-2 isolation & purification, B-Lymphocyte Subsets immunology, COVID-19 immunology
- Abstract
Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), the causative agent of the pandemic human respiratory illness COVID-19, is a global health emergency. While severe acute disease has been linked to an expansion of antibody-secreting plasmablasts, we sought to identify B cell responses that correlated with positive clinical outcomes in convalescent patients. We characterized the peripheral blood B cell immunophenotype and plasma antibody responses in 40 recovered non-hospitalized COVID-19 subjects that were enrolled as donors in a convalescent plasma treatment study. We observed a significant negative correlation between the frequency of peripheral blood memory B cells and the duration of symptoms for convalescent subjects. Memory B cell subsets in convalescent subjects were composed of classical CD24+ class-switched memory B cells, but also activated CD24-negative and natural unswitched CD27+ IgD+ IgM+ subsets. Memory B cell frequency was significantly correlated with both IgG1 and IgM responses to the SARS-CoV-2 spike protein receptor binding domain (RBD) in most seropositive subjects. IgM+ memory, but not switched memory, directly correlated with virus-specific antibody responses, and remained stable over 3 months. Our findings suggest that the frequency of memory B cells is a critical indicator of disease resolution, and that IgM+ memory B cells may play an important role in SARS-CoV-2 immunity., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2021
- Full Text
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