Ligand for Bovine CD5 the Identification and Characterization of A

Total Page:16

File Type:pdf, Size:1020Kb

Ligand for Bovine CD5 the Identification and Characterization of A The Identification and Characterization of a Ligand for Bovine CD5 Karen M. Haas and D. Mark Estes This information is current as J Immunol 2001; 166:3158-3166; ; of September 27, 2021. doi: 10.4049/jimmunol.166.5.3158 http://www.jimmunol.org/content/166/5/3158 Downloaded from References This article cites 46 articles, 18 of which you can access for free at: http://www.jimmunol.org/content/166/5/3158.full#ref-list-1 Why The JI? Submit online. http://www.jimmunol.org/ • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication *average by guest on September 27, 2021 Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2001 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Identification and Characterization of a Ligand for Bovine CD51 Karen M. Haas* and D. Mark Estes2*† CD5, a type I glycoprotein expressed by T cells and a subset of B cells, is thought to play a significant role in modulating Ag receptor signaling. Previously, our laboratory has shown that bovine B cells are induced to express this key regulatory molecule upon Ag receptor cross-linking. To date, a ligand has not been described for bovine CD5. Given the importance ligand binding presumably plays in the functioning of CD5 on this B cell subset and on T cells, we sought to characterize the ligand for this protein using a bovine CD5-human IgG1 (CD5Ig) fusion protein produced by both mammalian and yeast cells. As determined by CD5Ig binding, expression of this ligand is negative to low on freshly isolated lymphocytes, with low-density expression being limited to activated B cells. Activation with LPS, PMA, and calcium ionophore, or ligation of CD40 alone or in combination with anti-IgM, resulted in B cell-specific expression of this ligand. Interestingly, activation through B cell Ag receptor cross-linking alone, Downloaded from although able to induce CD5 expression, did not result in expression of CD5 ligand (CD5L). In addition, we demonstrate a functional role for CD5L as a costimulatory molecule that augments CD40L-stimulated B cell proliferation. Finally, immuno- precipitation with CD5Ig suggests that the ligand characterized in this study has a molecular mass of ϳ200 kDa. The data reported herein, as well as future studies aimed at further characterizing this newly identified bovine CD5L, will undoubtedly aid in understanding the role that the CD5-CD5L interaction plays in immune responses. The Journal of Immunology, 2001, 166: 3158–3166. http://www.jimmunol.org/ D5 is a 67-kDa type I monomeric glycoprotein belonging (22), and HIV infection (23) is evidenced by reports demonstrating to the ancient family of scavenger receptor cysteine-rich disease-related CD5ϩ B cell-specific expansion. CD5ϩ B cells C proteins (1). It has remained highly conserved throughout have also been implicated in the development of certain autoim- evolution and, therefore, is thought to play a conserved role in both mune conditions, such as Sjogren’s syndrome and rheumatoid ar- lymphocyte development and function among species (2–5). CD5 thritis (24), as well as various B cell lymphomas and leukemias, is found to be expressed by all T cells and a subset of B cells, such as B cell chronic lymphocytic leukemia, which are often of a previously termed B-1 cells (6). Although the origination of this CD5ϩ phenotype (25). ϩ CD5 B cell subset remains controversial, it has been clearly es- Recent studies have suggested that the function of CD5 may by guest on September 27, 2021 tablished by our laboratory and others that CD5Ϫ B cells, or con- differ with respect to specific cell population. For example, on ventional B (B-2) cells, from multiple species can be activated to peripheral T cells, CD5 appears to play a costimulatory role (26– express this marker (7–12). However, whether a B-1 cell is devel- 29), yet on Jurkat cells, CD5 was recently shown to negatively opmentally distinct from a B-2 cell induced to express CD5 as a regulate TCR signaling (30). Although it has been suggested that result of activation (such as Ag receptor cross-linking) remains to CD5 may function to provide continual stimulation through the B 3 be clearly established. cell Ag receptor (BCR) via its interaction with VH framework CD5ϩ B cells have been reported to exist in a number of spe- regions (31), studies involving CD5 knockout mice have suggested cies, although the proportion of B cells expressing CD5, as well as that CD5 negatively regulates Ag receptor signaling in both thy- their anatomical localization, varies among species. CD5 is ex- mocytes and B cells (32, 33). In addition, a recent study has dem- pressed by subpopulations of human (hu), mouse, and bovine B onstrated that ligation of CD5 on tonsillar B cells results in apo- cells (6, 13–16), while in other species, such as the rabbit (17) and ptosis, while ligation on resting peripheral T cells does not (34). chicken (2), nearly all B cells express CD5. The importance of this That CD5 negatively regulates signaling through the BCR is fur- B cell population during the immune response to a variety of ther supported by a recent biochemical study that has shown CD5 pathogenic infections, such as trypanosomosis (16, 18), schistoso- to recruit SHP-1, a protein tyrosine phosphatase, in B cells (35). In miasis (19, 20), bovine leukemia virus (21), chronic hepatitis C further support of the negative regulatory role of CD5 on B cells, a recent study by Hippen et al. (36) has suggested that CD5 plays a role in maintaining B cell anergy. Although the role CD5 ex- *Department of Molecular Microbiology and Immunology, School of Medicine, and pression plays on various cell populations remains to be com- † Department of Veterinary Pathobiology, College of Veterinary Medicine, University pletely understood, the interaction between CD5 and its ligand(s) of Missouri, Columbia, MO 65211 in all likelihood plays a critical role in regulating the function of Received for publication August 14, 2000. Accepted for publication December 28, 2000. CD5. Thus, the identification of the ligand(s) for CD5 is key to The costs of publication of this article were defrayed in part by the payment of page further elucidating the function of CD5 on T and B cell charges. This article must therefore be hereby marked advertisement in accordance populations. with 18 U.S.C. Section 1734 solely to indicate this fact. 1 This work was supported by the U.S. Department of Agriculture National Research Initiative Competitive Grants Program Project 96-35204-3584. 3 Abbreviations used in this paper: BCR, B cell Ag receptor; L, ligand; sIgM, surface 2 Address correspondence and reprint requests to Dr. D. Mark Estes, Department of IgM; TD, T cell-dependent; TI, T cell-independent; CD5Ig, bovine CD5-huIgG1; Veterinary Pathobiology, University of Missouri, 201 Connaway Hall, Columbia, MO mCD5Ig, MOP-8-expressed bovine CD5Ig; yCD5Ig, yeast-expressed bovine CD5Ig; 65211. E-mail address: [email protected] DTSSP, 3,3Јdithiobis-(sulfosuccinimidylpropionate); bo, bovine; hu, human. Copyright © 2001 by The American Association of Immunologists 0022-1767/01/$02.00 The Journal of Immunology 3159 The constitutive B cell marker, CD72, was described by Van de ture, transfected cells and nontransfected DAP3 cells were treated with 50 Velde et al. (37) as the first ligand for CD5. However, recent stud- ␮g/ml mitomycin C for 30 min at 37°C, washed three times in HBSS, and ies have suggested that CD5 may bind to one or more alternative allowed to adhere to culture dishes for 1 h and residual unbound fibroblasts were removed. PMA and calcium ionophore A23187 (Sigma, St. Louis, ligand(s). Using a CD5-huIgG1 fusion protein, Biancone et al. (38) MO) stimulation of B cells was performed at final concentrations of 10 described an activation-induced CD5 ligand (CD5L) expressed by ng/ml and 1 ␮g/ml, respectively. Endotoxin (from Escherichia coli O55: activated murine B cells, as well as T cell clones. Similarly, Bikah B5; Sigma) and pokeweed mitogen (Phytolacca americana lectin; Sigma) et al. (39) have described a ligand expressed by murine peritoneal were used at concentrations of 20 EU/ml and 10 ␮g/ml, respectively. B cells, activated splenic B cells, and B lymphoma cell lines. More Proliferation assay recently, Calvo et al. (40) described a CD5L expressed by cell types of lymphoid, myeloid, and epithelial origin. In addition to B cell proliferation assays were conducted on B cells cultured in triplicate at a concentration of 2 ϫ 105 B cells per well in a 96-well plate. Twenty- these studies, which suggest that CD5 may potentially interact with four hours after culture, CD5Ig or isotype control (huIgG1) was added to more than ligand, CD5 has been reported to interact with VH Ig cultures at concentrations of 25, 12.5, or 6.25 ␮g/ml. B cells were pulsed framework regions in both rabbit and human studies (41, 42). with 1 ␮Ci [3H]thymidine (DuPont/NEN, Boston, MA) after 72 h culture Given the variation in function CD5 appears to serve with respect and harvested 18 h later onto Skatron filter mats (Skatron Instruments, Lier, to cell-specific expression, as well its structural similarity to scav- Norway) using a cell harvester (Skatron Instruments).
Recommended publications
  • Human and Mouse CD Marker Handbook Human and Mouse CD Marker Key Markers - Human Key Markers - Mouse
    Welcome to More Choice CD Marker Handbook For more information, please visit: Human bdbiosciences.com/eu/go/humancdmarkers Mouse bdbiosciences.com/eu/go/mousecdmarkers Human and Mouse CD Marker Handbook Human and Mouse CD Marker Key Markers - Human Key Markers - Mouse CD3 CD3 CD (cluster of differentiation) molecules are cell surface markers T Cell CD4 CD4 useful for the identification and characterization of leukocytes. The CD CD8 CD8 nomenclature was developed and is maintained through the HLDA (Human Leukocyte Differentiation Antigens) workshop started in 1982. CD45R/B220 CD19 CD19 The goal is to provide standardization of monoclonal antibodies to B Cell CD20 CD22 (B cell activation marker) human antigens across laboratories. To characterize or “workshop” the antibodies, multiple laboratories carry out blind analyses of antibodies. These results independently validate antibody specificity. CD11c CD11c Dendritic Cell CD123 CD123 While the CD nomenclature has been developed for use with human antigens, it is applied to corresponding mouse antigens as well as antigens from other species. However, the mouse and other species NK Cell CD56 CD335 (NKp46) antibodies are not tested by HLDA. Human CD markers were reviewed by the HLDA. New CD markers Stem Cell/ CD34 CD34 were established at the HLDA9 meeting held in Barcelona in 2010. For Precursor hematopoetic stem cell only hematopoetic stem cell only additional information and CD markers please visit www.hcdm.org. Macrophage/ CD14 CD11b/ Mac-1 Monocyte CD33 Ly-71 (F4/80) CD66b Granulocyte CD66b Gr-1/Ly6G Ly6C CD41 CD41 CD61 (Integrin b3) CD61 Platelet CD9 CD62 CD62P (activated platelets) CD235a CD235a Erythrocyte Ter-119 CD146 MECA-32 CD106 CD146 Endothelial Cell CD31 CD62E (activated endothelial cells) Epithelial Cell CD236 CD326 (EPCAM1) For Research Use Only.
    [Show full text]
  • T Cells the Usual Subsets
    T cells: the usual subsets Chen Dong and Gustavo J. Martinez T cells have important roles in immune responses and function by directly secreting soluble mediators or important for adaptation of immune responses in different microenvironments and might be particularly through cell contact-dependent mechanisms. Many T cell subsets have been characterized. Although relevant for host defence against pathogens that colonize different tissues. Distinct T cell subsets, or effector T cells were originally considered to be terminally differentiated, a growing body of evidence has differentiation states, can be identified based on the cell surface markers expressed and/or the effector challenged this view and suggested that the phenotype of effector T cells is not completely fixed but is molecules produced by a particular T cell population. This Poster summarizes our current understanding of more flexible or plastic. T cells can have ‘mixed’ phenotypes (that is, have characteristics usually the surface markers, transcriptional regulators, effector molecules and functions of the different T cell associated with more than one T cell subset) and can interconvert from one subset phenotype to another, subsets that participate in immune responses. Further knowledge of how these T cell subsets are regulated IMMUNOLOGY although instructive signalling can lead to long-term fixation of cytokine memory. T cell plasticity can be and cooperate with each other will provide us with better tools to treat immune-related diseases. Cytotoxic T cell Exhausted T cell
    [Show full text]
  • Optimal Minimal Panels of Immunohistochemistry for Diagnosis of B-Cell Lymphoma for Application in Countries with Limited Resources and for Triaging Cases Before
    AJCP /ORIGINAL ARTICLE Optimal Minimal Panels of Immunohistochemistry for Diagnosis of B-Cell Lymphoma for Application in Countries With Limited Resources and for Triaging Cases Before Referral to Specialist Centers Downloaded from https://academic.oup.com/ajcp/article-abstract/145/5/687/2195691 by World Health Organization user on 09 January 2019 Maria Giulia Disanto, MD,1 Maria Raffaella Ambrosio, MD, PhD,2 Bruno Jim Rocca, MD, PhD,2 Hazem A. H. Ibrahim, FRCPath, PhD,1,3 Lorenzo Leoncini, MD, PhD,2 and Kikkeri N. Naresh, MD, FRCPath1 From the 1Department of Histopathology, Imperial College Healthcare NHS Trust & Imperial College, London, United Kingdom; 2Department of Medical Biotechnologies, Section of Pathology, University of Siena, Siena, Italy; and 3Department of Histopathology, Faculty of Medicine, Mansoura University, Mansoura, Egypt. Key Words: Lymphoma; B-cell lymphoma; Immunohistochemistry; Diagnosis; Classification; Developing countries Am J Clin Pathol May 2016;145:687-695 DOI: 10.1093/AJCP/AQW060 ABSTRACT Lymphomas are a collection of different malignancies “arising” from lymphoid cells. They include about 49 entities, Objectives: Establish and validate optimal minimal and over 19 provisional entities and subsets.1 About 85% of immunohistochemistry panels for usage in a staged lymphomas are of B-cell origin. Precision in lymphoma diag- algorithmic manner for precise diagnosis of B-cell nosis requires expertise and infrastructure. The entities are lymphomas in countries with limited resources. Suggest defined based on morphology, immunohistochemistry (on short panels of immunostains to be used in referring units some occasions in situ hybridization), cytogenetics/fluores- that refer suspected lymphomas to specialist diagnostic cent in situ hybridization (FISH), molecular genetics and clin- centers in resourceful countries.
    [Show full text]
  • RA0358-C.5-IFU-RUO CD57 / B3GAT1 (Natural Killer Cell
    Instructions For Use RA0 35 8-C.5 -IFU -RUO Revision: 1 Rev. Date: Dec. 19, 2014 Page 1 of 2 P.O. Box 3286 - Logan, Utah 84323, U.S.A. - Tel. (800) 729-8350 – Tel. (435) 755-9848 - Fax (435) 755-0015 - www.scytek.com CD57 / B3GAT1 (Natural Killer Cell Marker); Clone NK/804 (Concentrate) Availability/Contents: Item # Volume RA0358-C.5 0.5 ml Description: Species: Mouse Immunogen: Recombinant human B3GAT1 protein Clone: NK/804 Isotype: IgM, kappa Entrez Gene ID: 27087 (Human) Hu Chromosome Loc.: 11q25 Synonyms: 3-Glucuronyltransferase 1; B3GAT1; Galactosylgalactosylxylosylprotein 3-beta- Glucuronosyltransferase 1; GLCATP; GlcUAT-P; Glucuronosyltransferase P; UDP GlcUA Glycoprotein beta 1, 3 Glucuronyltransferase. Mol. Weight of Antigen: ~110kDa Format: Bioreactor Concentrate with 0.05% Azide. Specificity: Anti-CD57 marks a subset of lymphocytes known as natural killer (NK) cells. Anti-CD57 also stains neuroendocrine cells and their derived tumors, including carcinoid tumors and medulloblastoma. Background: Follicular center cell lymphomas often contain many NK cells within the neoplastic follicles. Anti- CD57 can be useful in separating type B3 thymoma from thymic carcinoma when combined with a panel that includes antibodies against GLUT1, CD5, and CEA. Species Reactivity: Human. Does not react with Rat. Others not known. Positive Control: Lymph node or tonsil. Cellular Localization: Cell surface Titer/ Working Dilution: Immunohistochemistry (Frozen and Formalin-fixed): 1:50-1:100 Flow Cytometry: 5-10 µl/million cells Immunofluorescence: 1:50-1:100 Western Blotting: 1:100-1:200 Microbiological State: This product is not sterile. 8° C Storage: 2° C C ScyTek Laboratories, Inc. 205 South 600 West P EmergoEurope (31)(0) 70 345-8570 Logan, UT 84321 Molsnstraat 15 Doc: IFU-Template2-8rev2 U.S.A.
    [Show full text]
  • Flow Cytometry CPT: 88182, 88184, 88185, 88187, 88188, 88189, 86355, 86356, 86357, 86359, 86360, 86361, 86367
    Medicare Local Coverage Determination Policy Flow Cytometry CPT: 88182, 88184, 88185, 88187, 88188, 88189, 86355, 86356, 86357, 86359, 86360, 86361, 86367 CMS Policy for Alaska, Arizona, Idaho, Montana, North Dakota, Medically Supportive Oregon, South Dakota, Utah, Washington, and Wyoming ICD Codes are listed Local policies are determined by the performing test location. This is determined by the state on subsequent page(s) in which your performing laboratory resides and where your testing is commonly performed. of this document. Coverage Indications, Limitations, and/or Medical Necessity Flow cytometry (FCM) is a complex process to examine blood, body fluids, CSF, bone marrow, lymph node, tonsil, spleen and other solid tissues. The use of peripheral blood and fine needle aspirate material avoids more invasive procedures for diagnosis. A flow cytometer evaluates the physical and/or chemical characteristics of single cells as the cells pass individually in a fluid stream through a measuring device. Surface receptors, intracellular molecules, and DNA bind with fluorescent dyes that allow detection and evaluation. When light of one wave length excites electrons of certain chemicals to energy levels above their ground state and upon return to ground state emits light of a longer wavelength, fluorescence is produced. A flow cytometer detects cell characteristics by measuring the fluorescence produced by fluorochromes conjugated either directly with cell components or conjugated to antibodies directed against cell components. Indications • Cytopenias and Hypercellular Hematolymphoid Disorders Hematolymphoid neoplasia can present with cytopenias (anemia, leucopenia and/or thrombocytopenia) or elevated leukocyte counts. If medical review and preliminary laboratory testing fails to reveal a cause, bone marrow aspiration and biopsy are indicated to rule out an infiltrative process or a stem cell disorder.
    [Show full text]
  • MGD011, a CD19 X CD3 Dual-Affinity Retargeting Bi-Specific Molecule Incorporating Extended Circulating Half-Life for the Treatment of B-Cell Malignancies
    Published OnlineFirst September 23, 2016; DOI: 10.1158/1078-0432.CCR-16-0666 Cancer Therapy: Preclinical Clinical Cancer Research MGD011, A CD19 x CD3 Dual-Affinity Retargeting Bi-specific Molecule Incorporating Extended Circulating Half-life for the Treatment of B-Cell Malignancies Liqin Liu, Chia-Ying K. Lam, Vatana Long, Lusiana Widjaja, Yinhua Yang, Hua Li, Linda Jin, Steve Burke, Sergey Gorlatov, Jennifer Brown, Ralph Alderson, Margaret D. Lewis, Jeffrey L. Nordstrom, Scott Koenig, Paul A. Moore, Syd Johnson, and Ezio Bonvini Abstract Purpose: CD19, a B-cell lineage-specific marker, is highly autologous B-cell depletion in PBMCs from both species. represented in B-cell malignancies and an attractive target for MGD011-mediated killing was accompanied by target-depen- therapeutic interventions. MGD011 is a CD19 x CD3 DART dent T-cell activation and expansion, cytokine release and bispecific protein designed to redirect T lymphocytes to eliminate upregulation of perforin and granzyme B. MGD011 demon- CD19-expressing cells. MGD011 has been engineered with a strated antitumor activity against localized and disseminated modified human Fc domain for improved pharmacokinetic (PK) lymphoma xenografts reconstituted with human PBMCs. In properties and designed to cross-react with the corresponding cynomolgus monkeys, MGD011 displayed a terminal half-life antigens in cynomolgus monkeys. Here, we report on the preclin- of 6.7 days; once weekly intravenous infusion of MGD011 at ical activity, safety and PK properties of MGD011. dosesupto100mg/kg, the highest dose tested, was well Experimental Design: The activity of MGD011 was evaluated tolerated and resulted in dose-dependent, durable decreases in several in vitro and in vivo models.
    [Show full text]
  • CD3+CD5+CD4-CD8-) Alpha/Beta T Cell Receptor-Bearing Cells
    Mlsa generated suppressor cells. I. Suppression is mediated by double-negative (CD3+CD5+CD4-CD8-) alpha/beta T cell receptor-bearing cells. This information is current as of October 2, 2021. M Bruley-Rosset, I Miconnet, C Canon and O Halle-Pannenko J Immunol 1990; 145:4046-4052; ; http://www.jimmunol.org/content/145/12/4046 Downloaded from Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision http://www.jimmunol.org/ • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription by guest on October 2, 2021 Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 1990 by American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. 0022-1767/90/14512-4046$02.00/0 THEJOURNAL OF IMMUNOLCGY Vol. 145.4046-4052. No. 12. December 15. 1990 Copyright 0 1990 by The American Association of lmmunologists Printed In U.S.A. Mls" GENERATED SUPPRESSORCELLS I. Suppression is Mediated by Double-Negative (CD3+CDVCD4-CD8-) a/@ T Cell Receptor-Bearing Cells' Grafting of cells from B10.D2 (H-2d)donors into H- The GVHR3 remains a major problem after bone mar- 2 compatible lethally irradiated (DBA/2 x B1O.DZ)Fl row transplantation, even in MHC-compatible donor-re- hostsresults in a severe graft-vs-host reaction cipient combinations.
    [Show full text]
  • B-Cell Activation by Crosslinking of Surface Igm Or Ligation of CD40 Involves Alternative Signal Pathways and Results in Different B-Cell Phenotypes HENRY H
    Proc. Natl. Acad. Sci. USA Vol. 92, pp. 3348-3352, April 1995 Immunology B-cell activation by crosslinking of surface IgM or ligation of CD40 involves alternative signal pathways and results in different B-cell phenotypes HENRY H. WORTIS*t, MARK TEUTSCH*, MINDY HIGER*, JENNY ZHENG*, AND DAVID C. PARKERt§ *Department of Pathology, Tufts University School of Medicine, and Graduate Program in Immunology, Sackler School of Graduate Biomedical Sciences, Boston, MA 02111; and tDepartment of Molecular Genetics and Microbiology, University of Massachusetts Medical School, Worcester, MA 01655 Communicated by Salome G. Waelsch, Albert Einstein College of Medicine of Yeshiva University, Bronx, NY December 7, 1994 ABSTRACT Treatment of small resting B cells with sol- direct contact with an activated T-helper cell such that the uble F(ab')2 fragments of anti-IgM, an analogue of T-inde- surface molecule gp39 [CD40 ligand (CD40L)] of the T cell pendent type 2 antigens, induced activation characterized by ligates the CD40 of the B cell (10). proliferation and the expression of surface CD5. In contrast, Our strategy to determine if minimal TD and TI-2 signals B cells induced to proliferate in response to thymus-dependent were sufficient to induce phenotypic differences in B cells was inductive signals provided by either fixed activated T-helper 2 to use an antigen that could be modified so as to initiate either cells or soluble CD40 ligand-CD8 (CD40L) recombinant pro- a TD or a TI-2 response. To produce a TD response we used tein displayed elevated levels of CD23 (FcJII receptor) and no monovalent Fab of rabbit anti-IgM and provided help in the surface CD5.
    [Show full text]
  • Flow Cytometric Analysis of B-Cell Lymphoproliferative Disorders
    Flow cytometric analysis of B-cell lymphoproliferative disorders David M. Dorfman, M.D., Ph.D. Department of Pathology Brigham and Women’s Hospital and Harvard Medical School Boston, MA Objectives • Review basic principles of flow cytometric immunophenotypic analysis of B cell lymphoproliferative disorders • Discuss recent studies to overcome limitations and shortcomings – New markers – New methods Incidence of B-cell neoplasms, United States Subtype Incidence rate 2011-2012 New cases, 2016 per 100,000 Lymphoid neoplasms 34.4 136,960 Lymphoid neoplasms, B 29.0 93.3% 117,470 B-LL/L 1.4 82.2% 4,930 CLL/SLL 5.1 20,980 FL 3.4 13,960 DLBCL 6.3 27,650 MM 5.9 24,280 Lymphoid neoplasms, T/NK 2.1 8,380 T-LL/L 0.3 1,070 T-PLL <0.1 160 T-LGL 0.2 670 ATL/L <0.1 180 Teras et al. CA Cancer J Clin 2016; 66:443-459 (North American AssociationTeras of et Central al. CA Cancer Cancer Registries) J Clin 2016; 66:443-459 (North American Association of Central Cancer Registries) SS <0.1 Teras et al. CA Cancer J Clin70 2016; 66:443-459 (North American Association of Central Cancer Registries) 94% WHO revised 4th ed., 2017 Flow cytometric analysis of B-cell lymphoproliferative disorders • B-cell antigen expression (CD19, CD20, CD22) • Monoclonal surface immunoglobulin κ or λ light chain expression (or absence of surface immunoglobulin) • Expression of additional B-cell antigens or other antigens, including abnormal expression levels • Presence of cells with abnormal light scatter characteristics ( high forward scatter or side scatter) B-ALL MCL FL, HL MZL, CLL, MM LPL DLBCL DLBCL WHO revised 4th ed.
    [Show full text]
  • Functional Studies of Chronic Lymphocytic Leukemia B Cells Expressing Β2-Integrin Type Complement Receptors CR3 and CR4
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Repository of the Academy's Library Functional studies of chronic lymphocytic leukemia B cells expressing β2-integrin type complement receptors CR3 and CR4 Barbara Uzonyia,1, Bernadett Mácsik-Valentb,1, Szilvia Lukácsib, Richárd Kissc, Katalin Törökb, Mariann Kremlitzkaa,2, Zsuzsa Bajtayb, Judit Demeterd, Csaba Bödörc, Anna Erdeia,b,* a MTA-ELTE Immunology Research Group, Department of Immunology, Eötvös Loránd University, Budapest, Hungary b Department of Immunology, Eötvös Loránd University, Budapest, Hungary c MTA-SE “Lendület” Molecular Oncohematology Research Group, 1st Department of Pathology, and Experimental Cancer Research, Semmelweis University, Budapest, Hungary d 1st Department of Internal Medicine, Semmelweis University, Budapest, Hungary 1 These authors contributed equally to this work 2 Present address: Division of Medical Protein Chemistry, Department of Translational Medicine, Lund University, Malmö, Sweden * Corresponding author Anna Erdei Department of Immunology, Eötvös Loránd University, Pázmány Péter s. 1/C, Budapest H-1117, Hungary Phone: (+36)-1-3812-175 Fax: (+36)-1-3812-176 e-mail: [email protected], [email protected] Abbreviations BCR: B cell receptor, CLL: chronic lymphocytic leukemia, CR: complement receptor, IL: interleukin Keywords CLL B cells, IL-10 production, expression of CR3 and CR4, adhesion and spreading 1 Abstract The expression and role of CR3 (CD11b/CD18) and CR4 (CD11c/CD18) in B cells are not yet explored in contrast to myeloid cells, where these β2-integrin type receptors are known to participate in various cellular functions, including phagocytosis, adherence and migration. Here we aimed to reveal the expression and role of CR3 and CR4 in human B cells.
    [Show full text]
  • Significant Enlargement of a Specific Subset of CD3+CD8+ Peripheral Blood Leukocytes Mediating Cytotoxic T-Lymphocyte Activity D
    Proc. Natl. Acad. Sci. USA Vol. 90, pp. 9427-9430, October 1993 Immunology Significant enlargement of a specific subset of CD3+CD8+ peripheral blood leukocytes mediating cytotoxic T-lymphocyte activity during human immunodeficiency virus infection (T lymphocyte subset/monoclonal antibody/AIDS) A. BENSUSSAN*t, C. RABIANt, V. SCHIAVON*, D. BENGOUFAt, G. LECA*, AND L. BOUMSELL* Institut Nationale de la Sant6 et de la Recherche Medicale U93, Association Claude Bernard, and tLaboratoire d'Immunologie et d'Histocompatibilite, H6pital Saint-Louis, 1 Avenue Claude Veliefaux, 75475 Paris cedex 10, France Communicated by Jean Dausset, July 20, 1993 ABSTRACT We have obtained a monoclonal antibody studies have shown that human immunodeficiency virus termed BY55 that defines an 80-kDa cell-surface structure on (HIV) infection significantly changes the phenotype of cir- a subset ofcirculating peripheral blood mononucleocytes. This culating lymphocytes and increases CD8 T lymphocytes (19), structure, which was not detected on most cell lines or activated it was interesting to look for this cytotoxic CD3+CD8+BY55+ lymphocytes, was expressed exclusively on 15-25% of CD2+ circulating cell subset in asymptomatic HIV-positive individ- circulating lymphocytes, including a major subset within the uals with various numbers of CD4 cells. Results indicated CD3- and the T-cell receptor y6+ lymphocytes and a small that peripheral blood CD3+CD8+BY55+ cells significantly percentage of the CD3+CD8+ cells. Moreover, we have shown increased in HIV-positive individuals. that the BY55 molecule delineated the competent killer circu- lating lymphocytes. In the present report, additional two- and MATERIALS AND METHODS three-color immunofluorescence studies of peripheral blood lymphocytes were done to precisely determine BY55 expression mAbs.
    [Show full text]
  • Immunological Synapse Formation Licenses CD40-CD40L Accumulations at T-APC Contact Sites1
    The Journal of Immunology Immunological Synapse Formation Licenses CD40-CD40L Accumulations at T-APC Contact Sites1 Judie Boisvert, Samuel Edmondson, and Matthew F. Krummel2 The maintenance of tolerance is likely to rely on the ability of a T cell to polarize surface molecules providing “help” to only specific APCs. The formation of a mature immunological synapse leads to concentration of the TCR at the APC interface. In this study, we show that the CD40-CD154 receptor-ligand pair is also highly concentrated into a central region of the synapse on mouse lymphocytes only after the formation of the TCR/CD3 c-SMAC. Concentration of this ligand was strictly dependent on TCR recognition, the binding of ICAM-1 to T cell integrins and the presence of an intact cytoskeleton in the T cells. This may provide a novel explanation for the specificity of T cell help directing the help signal to the site of Ag receptor signal. It may also serve as a site for these molecular aggregates to coassociate and/or internalize alongside other signaling receptors. The Journal of Im- munology, 2004, 173: 3647–3652. uring the onset and propagation of immune responses, T Interactions of T cells with Ag-bearing APCs is associated with cells send and receive signals via Ag-dependent and Ag- a coalescence of TCRs, other related receptors such as CD2, CD4, independent ligands. Whereas peptide-class II MHC CD28, and signaling molecules such as p56lck, fyn, linker for ac- D ϩ complexes on APC are recognized by the TCR on CD4 helper T tivation of T cells, and protein kinase C␪ into the central portion of cells, separate cell surface receptors modulate T cell-APC inter- an “immunological synapse” (15–23).
    [Show full text]