DOCSLIB.ORG

B Cell Development the Stages of B Cell Development Ordered Gene Rearrangements a Model for Allelic Exclusion the Role of the Pr

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
B Cell Development the Stages of B Cell Development Ordered Gene Rearrangements a Model for Allelic Exclusion the Role of the Pr Regular Office Hours: Tuesdays 11-12 B Cell Development Extra office hours: Wed, Feb 7 12-1pm Thurs, Feb 8 11am-12 Fri, Feb 9 2-4pm Antigen-independent Antigen- phase dependent phase I WILL NOT BE HOLDING OFFICE HOURS ON TUESDAY Feb 13!! (bone marrow, fetal (spleen, lymph liver) node) Dina, Tim, and I encourage all confused students to come to our office hours and discussion sections so we can try to help un-confuse you. Molecular V(D)J rearrangement Class switch, events Somatic The GSIs will conduct a review session in our regular class period on Tues hypermutation Feb 13. Cellular proB > preB > B cell events mature B cell activation, First midterm: Thurs Feb 15 at 6pm in 155 Dwinelle (not 2050 VLSB as listed development Memory and in the original schedule). plasma B cell Midterm will focus on material covered in lectures and will be designed to differentiation be taken in 90 min. (We have the room till 8pm.) 1 2 Bone marrow stromal cells provide secreted and cell surface factors that promote B cell maturation. B cell development The stages of B cell development In vitro cultures of bone marrow Ordered gene rearrangements stromal cells and A model for allelic exclusion progenitor B cells The role of the preBCR in B cell development can accurately B cell tolerance recapitulate the normal steps of B cell development. 3 4 1 Mixture of cells labeled with fluorescent antibodies Flow Cytometry Stages in B cell development: proB cells: no HC or LC expression preB cell: HC (cytoplasmic), but no LC expression B cell: surface HC and LC expression 5 6 1 parameter histograms: Ultrasonic nozzle vibrator Flow cytometric Cell suspension Cell < 1 % Sheath fluid Isolating Unstained cells analysis number positive (negative control) of cells stained with 2 defined cell Drop-charging different labeled signal populations Fluorescence intensity antibodies laser using Cell 30% 2 parameter dot plot: ) number positive n Fluorescence Stained e e cells r g ( 20% 20% y t Green Activated Cell i “double s only n Positive” e Fluorescence intensity (red) t n i Sorting e c n e c s FACS Cell 40% e r 50% 10% number positive o u red Stained l “double F only cells negative” Cell collector Cell collector Fluorescence intensity (red) 7 8 Fluorescence intensity (green) Flask for undeflected droplets 2 Successive stages of B cell development can be distinguished •B220+ CD43+ cells are “pro-B” (Fractions A-C) by correlated expression of various cell surface markers •B220+ CD43- IgM- cells are “pre-B” (Fraction D) •B220+ IgM+ IgD- cells are “immature B” (Fraction E) •B220+ IgM+ IgD+ cells are “mature B” (Fraction F) Randy Hardy’s scheme for fractionating bone marrow B cells. Different populations were FACS sorted, cultured in vitro, and reanalyzed to establish developmental order. 9 10 V(D)J recombination can be B cell development detected by as simple PCR assay The preB cell Ordered gene rearrangements A model for allelic exclusion The role of the preBCR in B cell development B cell tolerance 11 12 3 C F Sequential gene reAarrangemD ent and regulated PCR assay for gene rearrangement E gene expression B Gene rearrangement Gene expression A B C D E F D-to-JH actin V-to-DJ H TdT λ5 V-to-DJH RAG-1 RAG-2 V-to-Jκ 13 14 Sequence of Ig gene rearrangement Sequence of Ig gene rearrangement as determined by FACS sorted developing B cells (and as determined by FACS sorted developing B cells (and from studies using transformed “preB cell-like” cell from studies using transformed “preB cell-like” cell lines). lines). DJH , VDJH , VJk , VJλ DJH , VDJH , VJk , VJλ (proB cell), (preBcell), (B cell) 15 16 4 B cell development Allelic exclusion The preB cell of Ig genes Ordered gene rearrangements A model for allelic exclusion The role of the preBCR in B cell development B cell tolerance 17 18 Ordered Gene Rearrangement and Allelic Exclusion Flexibility in joining of gene segments contributes to junctional diversity. (Note most rearrangements are µo = germline heavy-chain locus non-productive!) κo = germline light-chain locus Some B cell clones have VDJ rearrangements on both HC alleles. In these clones, only one allele is productively rearranged!! 19 20 5 A Model for Allelic Exclusion 21 22 Heavy-chain expression signals to A puzzle: developing B cells: Antibody including the surface version of Ig (BCR) is composed of paired Heavy and Light chains. • Proliferate (3 to 4 cycles) • Shut off allelic heavy-chain V-to-DJ rearrangement Heavy chain expression effects B cell development at a stage (the preB cell) in which light chain is not • Start V-to-Jκ rearrangement yet expressed. • Alterations in gene expression (e.g., shut off TdT) What is the form of the B cell antigen receptor on • Don’t die preB cells? 23 24 6 Ig HC (µ) in pre B cells is paired with a surrogate light chain complex The BCR versus the pre-BCR consisting of VpreB, λ5: whole complex known as the “preBCR” Figure 7. Heavy-chain mu/ surrogate VpreB light-chain complex and genes. !5 5' 3' Mu VpreB !5 1 kb •Surrogate light chains: •Homologous to Vλ and Cλ exons •Genes DO NOT rearrange •Expressed only in developing B cells. 25 26 Transgenesis & Targeted Mutation: Collect fertilized eggs Techniques for altering the mouse Transgenesis-- Inject cloned DNA introduction of genome Into one of the pronuclei Implant injected eggs into oviduct cloned DNA of pseudo-pregnant female Transgenic mice are engineered to express foreign DNA into germline inserted randomly into genome. Technique usually Pseudo-pregnant female produces dominant mutations. (random insertion into host chromosome; Mice with targeted mutations (gene “knock-out” mice) different in each founder) have engineered mutations in endogenous genes. About 10%-30% of offspring contain transgene Technique usually produces recessive mutations. Test for presence of transgene 27 28 Breed transgenics 7 Engineering mice to express a “pre- rearranged” Heavy chain transgene. Ig heavy-chain transgenic mouse VDJH exons CH exons Promoter Rearranged HC chain cloned from a mature B cell. pre pro Mice expressing a rearranged HC transgene have reduced endogenous V-to-DJH rearrangement (allelic exclusion). Evidence that presence of a rearranged HC gene feeds back to shut down rearrangement of other HC loci. Evidence that presence of a rearranged HC genes promotes 29 development from the proB to the preB stage. 30 Ig heavy-chain expression is sufficient to inhibit The origin of Embryonic Stem (ES) cells endogenous heavy chain rearrangement (allelic exclusion) and to promote developmental Fertilized 2 cells Morula, 8 cells 16 cells Section of blastocyst progression in mice that cannot rearrange their mouse egg 1.5 days 2.5 days 3 days 4 days endogneous HC genes (rag-). Is the membrane form of Heavy Chain necessary for Inner cell allelic exclusion and developmental progression? mass Gene targeting approach to delete the membrane exons of Heavy chain. ES cells can be grown indefinately in culture. ES cells are pluripotent: can give rise to all tissues when 31 transplanted into a new embryo. 32 8 Gene targeting using homologous Blastocyst recombination in ES cells neoR Injection TK Targeting construct •Start with correctly targeted ES cell Target locus •Hold blastocyst with suction pipette -Transfect ES cells with targeting construct DNA. •Inject ES cells into blastocyst •Injected ES cells become part of host -Select for neoR ES cells (transfected DNA has integrated into blastocyst genomic DNA of ES cells) •Implant embryo into foster mother Select against TK expression (correctly targeted insertion deletes TK gene). •Embryo develops into chimeric mouse •Breed to achieve germline transmission neoR Targeted allele 33 34 Membrane associated vs. secreted Ig: Differential mRNA splicing 35 36 9 Targeted mutagenesis in mice used to delete the exons The pre-BCR is required for normal B cell encoding the membrane associated version of HC. development The membrane- λ5− deficient mice show impaired B cell development associated form of Ig Wild type µ : λ5T/+ λ5T/ λ5T 5 r HC is required for: 4 e 5511%% 31% 2% D k r C a regulation the rearrangement of LC genes; s m a i l l l a e B220 inactivate rearrangement of heavy-chain genes (allelic exclusion); c 0 2 B 2 cause proliferation of pre-B cells, selecting for cells with B “productive” rearrangements; Thy1: T cell marker prevent the apoptosis of developing cells. 37 38 B1 B cells, a “non conventional” B cell population preBCR (HC and surrogate light chains) Characteristic B-2 B-1 ____________ signals lead to: surface markers IgDhi IgMlo IgMhi IgDlo (CD5+) development throughout life fetal & perinatal period lifetime weeks entire lifetime • Shut off heavy-chain V-to-DJ rearrangement (allelic exclusion) location blood, spleen, lymph nodes body cavities (peritoneum) • Pro B cell > pre B cell transition antibody all Ig classes; very diverse IgM, limited diversity role in immunity adaptive responses programmed response – Start V-to-Jκ rearrangement – Alterations in gene expression (e.g., shut off TdT) – Don’t die – Proliferation 39 40 10 AB individuals do not make antibodies to A or B carbohydrate structures B cell development because they are tolerant to The preB cell their own red blood cells. Ordered gene rearrangements A model for allelic exclusion The role of the preBCR in B cell development B cell tolerance 41 42 Tolerance to self involves both B and T cells and operates at early and late stages of B cell development B cell tolerance • There are many overlapping mechanisms that ensure self- • Clonal deletion-- the removal, by apoptosis, of B cells tolerance. with self-specific antigen receptors • Self-reactive B and T cells are eliminated or inactivated during • Anergy-- the biochemical inactivation of self-specific their development B cells • Most B cell responses depend on T cell help, so T cell tolerance • Receptor editing-- ongoing V(D)J recombination helps to ensure that antibodies against self are not generated.
Load more

Recommended publications
  • CD137 Microbead Kit CD137 Microbead + Cells
    CD137 MicroBead Kit human Order no. 130-093-476 Contents 1.2 Background information 1. Description The activation-induced antigen CD137 (4-1BB) is a 30 kDa glycoprotein of the tumor necrosis factor (TNF) receptor 1.1 Principle of the MACS® Separation + + superfamily. It is mainly expressed on activated CD4 and CD8 1.2 Background information T cells, activated B cells, and natural killer cells, but can also be 1.3 Applications found on resting monocytes and dendritic cells. As a costimulatory molecule, CD137 is involved in the activation 1.4 Reagent and instrument requirements and survival of CD4, CD8, and NK cells. Its engagement enhances 2. Protocol expansion of T cells and activates them to secrete cytokines. CD137 has been described to be a suitable marker for antigen- 2.1 Sample preparation specific activation of human CD8+ T cells, as CD137 is not expressed 2.2 Magnetic labeling on resting CD8+ T cells and its expression is reliably induced after 2.3 Magnetic separation 24 hours of stimulation.¹,² 3. Example of a separation using the CD137 MicroBead Kit 1.3 Applications 4. References ● Enrichment of CD137+ T cells for phenotypical and functional 5. Appendix characterization. ● Enrichment of activated antigen-specific T cells after antigen- Warnings specific stimulation. Reagents contain sodium azide. Under acidic conditions sodium 1.4 Reagent and instrument requirements azide yields hydrazoic acid, which is extremely toxic. Azide ● compounds should be diluted with running water before discarding. Buffer: Prepare a solution containing phosphate-buffered These precautions are recommended to avoid deposits in plumbing saline (PBS), pH 7.2, 0.5% bovine serum albumin (BSA), where explosive conditions may develop.
    [Show full text]
  • Antagonist Antibodies Against Various Forms of BAFF: Trimer, 60-Mer, and Membrane-Bound S
    Supplemental material to this article can be found at: http://jpet.aspetjournals.org/content/suppl/2016/07/19/jpet.116.236075.DC1 1521-0103/359/1/37–44$25.00 http://dx.doi.org/10.1124/jpet.116.236075 THE JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS J Pharmacol Exp Ther 359:37–44, October 2016 Copyright ª 2016 by The American Society for Pharmacology and Experimental Therapeutics Unexpected Potency Differences between B-Cell–Activating Factor (BAFF) Antagonist Antibodies against Various Forms of BAFF: Trimer, 60-Mer, and Membrane-Bound s Amy M. Nicoletti, Cynthia Hess Kenny, Ashraf M. Khalil, Qi Pan, Kerry L. M. Ralph, Julie Ritchie, Sathyadevi Venkataramani, David H. Presky, Scott M. DeWire, and Scott R. Brodeur Immune Modulation and Biotherapeutics Discovery, Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, Connecticut Received June 20, 2016; accepted July 18, 2016 Downloaded from ABSTRACT Therapeutic agents antagonizing B-cell–activating factor/B- human B-cell proliferation assay and in nuclear factor kB reporter lymphocyte stimulator (BAFF/BLyS) are currently in clinical assay systems in Chinese hamster ovary cells expressing BAFF development for autoimmune diseases; belimumab is the first receptors and transmembrane activator and calcium-modulator Food and Drug Administration–approved drug in more than and cyclophilin ligand interactor (TACI). In contrast to the mouse jpet.aspetjournals.org 50 years for the treatment of lupus. As a member of the tumor system, we find that BAFF trimer activates the human TACI necrosis factor superfamily, BAFF promotes B-cell survival and receptor. Further, we profiled the activities of two clinically ad- homeostasis and is overexpressed in patients with systemic vanced BAFF antagonist antibodies, belimumab and tabalumab.
    [Show full text]
  • IHC) Outreach Services
    IIImmunohistochemistry (IHC) Outreach Services Note type of fixative used if not neutral buffered formalin. Note type of tissue/specimen Unless specified otherwise, positive and negative controls react satisfactorily. Available Chromogen – All markers have been validated with 3,3’-Diaminobenzidine Tetrahydrochloride (DAB) which results in a brown/black precipitate. DAB is the routine chromogen. In addition, some markers have also been validated using the Fast Red (RED), which results in a red precipitate. If available with both chromogens and one is not selected, the def ault will be the DAB chromogen. Antibody Common Applications St aining Charac teristi cs Actin (muscl e sp eci fic ) Smoo th, ske letal & ca rdiac musc le Cytoplasm ic Actin (smoo th muscle ) Smoo th muscl e an d myoep itheli al cel ls Cytoplasm ic and memb rane ALK Pr otein ALK1 posi tive lymp homa s Cytoplasm ic and/or nuclea r Alph a-1-Antitr ypsi n Demo nstr ates A-1-AT in liver Cytoplasm ic (A-1-AT) Bcl-2 Oncop rotein Foll icula r lymph oma an d so ft tiss ue Cytoplasm ic tumors Bcl-6 Foll icula r lymph oma Nucl ear Ber-EP 4, Epithelia l Antige n Aden oca rcin oma vs. meso the liom a Memb rane and cytoplasm ic. The and epithelial tumors membrane staining is preferentially basolateral. Beta-Amyloid Pos t mo rt em dia gnos is of demen tia E xtr acel lular de pos ition (am yloid plaques), vascular deposition (amyloid angiopathy) BRS T-3, (B72.3) Aden oca rcin oma vs.
    [Show full text]
  • TACI:Fc Scavenging B Cell Activating Factor (BAFF) Alleviates Ovalbumin-Induced Bronchial Asthma in Mice
    EXPERIMENTAL and MOLECULAR MEDICINE, Vol. 39, No. 3, 343-352, June 2007 TACI:Fc scavenging B cell activating factor (BAFF) alleviates ovalbumin-induced bronchial asthma in mice 1,2,3 2 Eun-Yi Moon and Sook-Kyung Ryu the percentage of non-lymphoid cells and no changes were detected in lymphoid cell population. 1 Department of Bioscience and Biotechnology Hypodiploid cell formation in BALF was decreased Sejong University by OVA-challenge but it was recovered by TACI:Fc Seoul 143-747, Korea treatment. Collectively, data suggest that asthmatic 2 Laboratory of Human Genomics symptom could be alleviated by scavenging BAFF Korea Research Institute of Bioscience and Biotechnology (KRIBB) and then BAFF could be a novel target for the Daejeon 305-806, Korea develpoment of anti-asthmatic agents. 3 Corresponding author: Tel, 82-2-3408-3768; Fax, 82-2-466-8768; E-mail, [email protected] Keywords: asthma; B-cell activating factor; ovalbu- and [email protected] min; transmembrane activator and CAML interactor protein Accepted 28 March 2007 Introduction Abbreviations: BAFF, B cell activating factor belonging to TNF- family; BALF, bronchoalveolar lavage fluid; OVA, ovalbumin; PAS, Mature B cell generation and maintenance are regu- periodic acid-Schiff; Prx, peroxiredoxin; TACI, transmembrane lated by B-cell activating factor (BAFF). BAFF is pro- activator and calcium modulator and cyclophilin ligand interactor duced by macrophages or dendritic cells upon stim- ulation with LPS or IFN- . BAFF belongs to the TNF family. Its biological role is mediated by the specific Abstract receptors, B-cell maturation antigen (BCMA), trans- membrane activator and calcium modulator and cy- Asthma was induced by the sensitization and chal- clophilin ligand interactor (TACI) and BAFF receptor, lenge with ovalbumin (OVA) in mice.
    [Show full text]
  • CD Markers Are Routinely Used for the Immunophenotyping of Cells
    ptglab.com 1 CD MARKER ANTIBODIES www.ptglab.com Introduction The cluster of differentiation (abbreviated as CD) is a protocol used for the identification and investigation of cell surface molecules. So-called CD markers are routinely used for the immunophenotyping of cells. Despite this use, they are not limited to roles in the immune system and perform a variety of roles in cell differentiation, adhesion, migration, blood clotting, gamete fertilization, amino acid transport and apoptosis, among many others. As such, Proteintech’s mini catalog featuring its antibodies targeting CD markers is applicable to a wide range of research disciplines. PRODUCT FOCUS PECAM1 Platelet endothelial cell adhesion of blood vessels – making up a large portion molecule-1 (PECAM1), also known as cluster of its intracellular junctions. PECAM-1 is also CD Number of differentiation 31 (CD31), is a member of present on the surface of hematopoietic the immunoglobulin gene superfamily of cell cells and immune cells including platelets, CD31 adhesion molecules. It is highly expressed monocytes, neutrophils, natural killer cells, on the surface of the endothelium – the thin megakaryocytes and some types of T-cell. Catalog Number layer of endothelial cells lining the interior 11256-1-AP Type Rabbit Polyclonal Applications ELISA, FC, IF, IHC, IP, WB 16 Publications Immunohistochemical of paraffin-embedded Figure 1: Immunofluorescence staining human hepatocirrhosis using PECAM1, CD31 of PECAM1 (11256-1-AP), Alexa 488 goat antibody (11265-1-AP) at a dilution of 1:50 anti-rabbit (green), and smooth muscle KD/KO Validated (40x objective). alpha-actin (red), courtesy of Nicola Smart. PECAM1: Customer Testimonial Nicola Smart, a cardiovascular researcher “As you can see [the immunostaining] is and a group leader at the University of extremely clean and specific [and] displays Oxford, has said of the PECAM1 antibody strong intercellular junction expression, (11265-1-AP) that it “worked beautifully as expected for a cell adhesion molecule.” on every occasion I’ve tried it.” Proteintech thanks Dr.
    [Show full text]
  • CD134 (OX40) Antibodies, Human for Research Use Only
    CD134 (OX40) antibodies, human For research use only One test corresponds to labeling of up to 107 cells in a total volume of 100 µL. Product Content Order no. CD134 (OX40)­VioBright FITC for 30 tests 130­109­664 CD134 (OX40)­VioBright FITC for 100 tests 130­109­605 CD134 (OX40)­PE for 30 tests 130­109­660 CD134 (OX40)­PE for 100 tests 130­109­601 CD134 (OX40)­APC for 30 tests 130­109­661 CD134 (OX40)­APC for 100 tests 130­109­602 CD134 (OX40)­PE­Vio770 for 30 tests 130­109­662 CD134 (OX40)­PE­Vio770 for 100 tests 130­109­603 CD134 (OX40)­APC­Vio770 for 30 tests 130­109­663 CD134 (OX40)­APC­Vio770 for 100 tests 130­109­604 CD134 (OX40)­Biotin for 30 tests 130­109­659 CD134 (OX40)­Biotin for 100 tests 130­109­600 Warnings Reagents contain sodium azide. Under acidic conditions sodium azide yields hydrazoic acid, which is extremely toxic. Azide compounds should be diluted with running water before discarding. These precautions are recommended to avoid deposits in plumbing where explosive conditions may develop. Technical data and background information Antigen CD134 (OX40) Clone REA621 Isotype recombinant human IgG1 Isotype control REA Control (S) antibodies Alternative names of antigen OX­40, OX40 Molecular mass of antigen [kDa] 27 Distribution of antigen B cells, endothelial cells, fibroblasts, lymphocytes, T cells Product format Reagents are supplied in buffer containing stabilizer and 0.05% sodium azide. Fixation Cells should be stained prior to fixation, if formaldehyde is used as a fixative. Storage Store protected from light at 2–8 °C.
    [Show full text]
  • The Thrombopoietin Receptor : Revisiting the Master Regulator of Platelet Production
    This is a repository copy of The thrombopoietin receptor : revisiting the master regulator of platelet production. White Rose Research Online URL for this paper: https://eprints.whiterose.ac.uk/175234/ Version: Published Version Article: Hitchcock, Ian S orcid.org/0000-0001-7170-6703, Hafer, Maximillian, Sangkhae, Veena et al. (1 more author) (2021) The thrombopoietin receptor : revisiting the master regulator of platelet production. Platelets. pp. 1-9. ISSN 0953-7104 https://doi.org/10.1080/09537104.2021.1925102 Reuse This article is distributed under the terms of the Creative Commons Attribution (CC BY) licence. This licence allows you to distribute, remix, tweak, and build upon the work, even commercially, as long as you credit the authors for the original work. More information and the full terms of the licence here: https://creativecommons.org/licenses/ Takedown If you consider content in White Rose Research Online to be in breach of UK law, please notify us by emailing [email protected] including the URL of the record and the reason for the withdrawal request. [email protected] https://eprints.whiterose.ac.uk/ Platelets ISSN: (Print) (Online) Journal homepage: https://www.tandfonline.com/loi/iplt20 The thrombopoietin receptor: revisiting the master regulator of platelet production Ian S. Hitchcock, Maximillian Hafer, Veena Sangkhae & Julie A. Tucker To cite this article: Ian S. Hitchcock, Maximillian Hafer, Veena Sangkhae & Julie A. Tucker (2021): The thrombopoietin receptor: revisiting the master regulator of platelet production, Platelets, DOI: 10.1080/09537104.2021.1925102 To link to this article: https://doi.org/10.1080/09537104.2021.1925102 © 2021 The Author(s).
    [Show full text]
  • Allelic Exclusion of the Immunoglobulin Heavy Chain Locus Is Independent of Its Nuclear Localization in Mature B Cells Sjoerd J
    Published online 7 June 2013 Nucleic Acids Research, 2013, Vol. 41, No. 14 6905–6916 doi:10.1093/nar/gkt491 Allelic exclusion of the immunoglobulin heavy chain locus is independent of its nuclear localization in mature B cells Sjoerd J. B. Holwerda1, Harmen J. G. van de Werken1, Claudia Ribeiro de Almeida2, Ingrid M. Bergen2, Marjolein J. W. de Bruijn2, Marjon J. A. M. Verstegen1, Marieke Simonis1, Erik Splinter1, Patrick J. Wijchers1, Rudi W. Hendriks2,* and Wouter de Laat1,* 1Hubrecht Institute-KNAW & University Medical Center Utrecht, Utrecht 3584 CT, The Netherlands and 2Department of Pulmonary Medicine, Erasmus MC Rotterdam, Rotterdam, Box 2040, 3000 CA, The Netherlands Received April 7, 2013; Revised May 6, 2013; Accepted May 11, 2013 ABSTRACT INTRODUCTION In developing B cells, the immunoglobulin heavy B and T lymphocytes express a large repertoire of antigen chain (IgH) locus is thought to move from repressive receptors that safeguard the robustness of our adaptive to permissive chromatin compartments to facili- immune response. Lymphocyte development uniquely tate its scheduled rearrangement. In mature B relies on scheduled genomic rearrangement of V (variable), D (diversity) and J (joining) gene segments in cells, maintenance of allelic exclusion has been the antigen receptor loci (1–3). proposed to involve recruitment of the non-product- The murine IgH locus spans nearly 3Mb, with ive IgH allele to pericentromeric heterochromatin. upstream 150 functional VH segments spread over Here, we used an allele-specific chromosome con- 2.4 Mb, followed by DH and JH segments and a 200 kb formation capture combined with sequencing constant (CH) gene region.
    [Show full text]
  • Antigen-Receptor Genes & B Cell Development Learning
    Katherine L. Knight, Ph.D. Host Defense 2011 Antigen-Receptor Genes and B Cell Development ANTIGEN-RECEPTOR GENES & B CELL DEVELOPMENT Date: Friday, March 18, 2011 Monday, March 21, 2011 10:30 AM – 11:30 AM 10:30 AM – 11:30 AM LEARNING GOAL You will be able to diagram and describe the stages of B cell development including somatic DNA rearrangements, and explain how a large antibody repertoire is developed. You will also be able to diagram the structure and rearrangement of T cell receptor (TCR) genes and explain how a large TCR repertoire develops. OBJECTIVES To attain the goal for these lectures you will be able to: • State the approximate size of antibody repertoire needed for immune protection. • Diagram the order of H and L chain gene rearrangements that occur during B cell development. • Describe the mechanism by which individual B cells make only one antibody (allelic exclusion) • Diagram the sequence of events leading from rearrangements of germline Ig genes to production of an Ig molecule. • Describe how antibody diversity is generated through combinatorial joining, junctional diversity, and N segment addition. • Describe how antibody diversity is generated in secondary lymphoid tissues • Describe the maturation of B cells from stem cells, including the pre-B cell receptor. • State the cell origin of CLL, Burkitt’s lymphoma, Hodgkin’s lymphoma, ALL, and multiple myeloma • Draw the structure of αβ and γδ T cell receptors. • Diagram the mechanism by which T cells express a single antigen receptor READING ASSIGNMENT Janeway’s Immunobiology (2008), Chapter 4; Chapter 7, pp 262-272; 308-312 LECTURER Katherine L.
    [Show full text]
  • Induces Antigen Presentation in B Cells Cell-Activating Factor of The
    B Cell Maturation Antigen, the Receptor for a Proliferation-Inducing Ligand and B Cell-Activating Factor of the TNF Family, Induces Antigen Presentation in B Cells This information is current as of September 27, 2021. Min Yang, Hidenori Hase, Diana Legarda-Addison, Leena Varughese, Brian Seed and Adrian T. Ting J Immunol 2005; 175:2814-2824; ; doi: 10.4049/jimmunol.175.5.2814 http://www.jimmunol.org/content/175/5/2814 Downloaded from References This article cites 54 articles, 36 of which you can access for free at: http://www.jimmunol.org/content/175/5/2814.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • 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 by guest on September 27, 2021 *average 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 © 2005 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology B Cell Maturation Antigen, the Receptor for a Proliferation-Inducing Ligand and B Cell-Activating Factor of the TNF Family, Induces Antigen Presentation in B Cells1 Min Yang,* Hidenori Hase,* Diana Legarda-Addison,* Leena Varughese,* Brian Seed,† and Adrian T.
    [Show full text]
  • CD43 Expression in Malignant Melanoma: a Diagnostic Pitfall
    Asian Archives of Pathology 2014; Vol. 10 No.4, 97-109 Case Report CD43 Expression in Malignant Melanoma: A Diagnostic Pitfall Sanya Sukpanichnant Received: 24 September 2014; accepted 30 October 2014. ABSTRACT CD43 is commonly concerned as a marker for hematopoietic cells and, at times, a preferential T-cell marker. However, a number of non-hematologic cancers, mostly carcinoma, can express CD43. Recently, a case of malignant melanoma mimicking malignant lymphoma by cytomorphology was reported to express CD43 as well as melanoma markers but the other hematologic markers [Buehler et al. Diagn Cytopathol 2012;40:619- 23]. This is another case report. A 52-year-old man suffered from a rapidly growing left tonsillar mass for one month causing slurred speech. An incisional biopsy showed gray tissue and, beneath the epithelium, a sheet of - lastic lymphoma, as the original workup showed focal and faint CD138 expression but negative keratins and CD45. On consultation, hematologic malignancy and malignant melanoma were considered. The tumor cells express CD43, CD68 (focal), S-100, HMB45, Melan A, and vimentin. The negative markers included CD3, CD20, CD30, CD79a, MUM1, PAX5, kappa, lambda, and CK5/6. The proliferation index was high (70-75%) another case report of malignant melanoma with CD43 expression, mimicking malignant lymphoma, raises the awareness of diagnostic pitfall when dealing with undifferentiated tumors showing only CD43 expression among various hematologic markers. A panel for keratins, vimentin, S-100, CD45, CD30, and CD138 will be helpful when dealing with undifferentiated large cell tumor. Keywords: melanoma *This case report has been presented partly in a poster session in the 12th Japanese-Korean Lymphoreticular- Workshop 2014 in conjunction with the 5th Asian Hematopathology Symposium on January 25, 2014 at Nagoya University Hospital Lecture Hall.
    [Show full text]
  • Poor Quality Vβ Recombination Signal Sequences Stochastically Enforce Tcrβ Allelic Exclusion
    ARTICLE Poor quality Vβ recombination signal sequences stochastically enforce TCRβ allelic exclusion Glendon S. Wu1,2, Katherine S. Yang-Iott2, Morgann A. Klink2, Katharina E. Hayer2, Kyutae D. Lee2, and Craig H. Bassing1,2 The monoallelic expression of antigen receptor (AgR) genes, called allelic exclusion, is fundamental for highly specific immune responses to pathogens. This cardinal feature of adaptive immunity is achieved by the assembly of a functional AgR gene on one allele, with subsequent feedback inhibition of V(D)J recombination on the other allele. A range of epigenetic mechanisms have been implicated in sequential recombination of AgR alleles; however, we now demonstrate that a genetic mechanism controls this process for Tcrb. Replacement of V(D)J recombinase targets at two different mouse Vβ gene segments with a higher quality target elevates Vβ rearrangement frequency before feedback inhibition, dramatically increasing the frequency of T cells with TCRβ chains derived from both Tcrb alleles. Thus, TCRβ allelic exclusion is enforced genetically by the low quality of Vβ recombinase targets that stochastically restrict the production of two functional rearrangements before feedback inhibition silences one allele. Introduction Monoallelic expression is an essential process that limits the regulatory mechanisms, the frequent assembly of out-of-frame dosage of numerous genes. Important examples include genetic rearrangements and the requirement for AgR protein expression imprinting and X-chromosome inactivation, as well as the to drive T and B cell development dictate that biallelic expression tissue-specific allelic exclusion of olfactory neuron receptors of any TCR or Ig gene should occur in 20% of lymphocytes (Brady and lymphocyte antigen receptors (AgR).
    [Show full text]