Proteomic studies in Collagen Induced Arthritis (CIA) rats
|
|
- Brianna Wade
- 5 years ago
- Views:
Transcription
1 Proteomic studies in Collagen Induced Arthritis (CIA) rats
2 5.1. Introduction Over the last few decades, both naturally occurring and genetically engineered animal models of human diseases have been employed for evaluating new therapeutic options and for elucidating pathological pathways. The biological analysis of animal models that mimic human diseases has led to a better understanding of numerous pathological mechanisms and is extremely helpful in the initial testing of new therapeutic strategies (Glasser and Nogee, 2006; Hansen and Khanna, 2004; Monnet and Chachques, 2005; Patel and Goldstein, 2004). Although the usage of human cell cultures can give highly relevant data with respect to therapeutic dosage and toxic side effects (Findikli et al., 2006) but whole animal studies still forms a central part of pharmacological and biomedical evaluations (Contag, 2002). In pharmacological studies, cell culture models cannot be employed to mimic the complexity of adsorption, distribution, first pass hepatic elimination, modification and excretion of a drug. Also organ selective toxicity can only be studied at the whole body level (Boyd et al., 1983). Once a suitable animal model of a disease has been established, proteomic screening can be used to identify new candidate proteins with an altered expression level. While genomic data only predict the effects of changes in gene expression relating to the actual concentration of proteins or the extent of PTMs, proteomics can accurately provide the status of protein abundance, isoform expression patterns and peptide modification. Once interesting candidate proteins have been found through animal model proteomics, an informed decision can be made on the optimum design of subsequent clinical studies. Thus, the initial discovery of new biomarkers by proteomic studies in 81
3 animal model may have the potential for improving diagnostic procedures in the short term and will be helpful in the establishment of novel therapeutic targets in the long term. Collagen induced arthritis (CIA), as described in details in chapter 1, is an experimental model of arthritis induced in susceptible strains of rats by immunization with heterologous collagen type II, a joint specific protein, in complete Freund s adjuvant (Trentham et al., 1977). CIA has been extensively studied to elucidate the pathological mechanisms relevant to human RA and to identify potential therapeutic targets. The development of CIA, as of RA, is thought to depend on T cells, and disease susceptibility is linked to the MHC region (Wooley et al., 1981). Following T cell activation an inflammatory cascade is triggered that involves T cells, macrophages/monocytes, B cells and activated synoviocytes. The different leukocytes and synovial cells produce a complex array of cytokines and other soluble mediators that are thought to be responsible for cartilage destruction and bone erosion (Yang et al., 1999). Very few reports are available where the proteomic profiling of various tissues of CIA mice have been done to study disease specific changes in the protein expression (Biswas et al., 2009; Lorenz et al., 2003). In this study, we have developed the rat CIA model using porcine collagen II and investigated the plasma proteome of CIA rat. WGA affinity column was used to enrich plasma glycoproteins followed by 2 DE of WGA bound as well as unbound plasma proteins to study the differential expression of proteins. The proteins were identified by MALDI TOF MS followed by antibody validation of the most differentially expressed protein. Further, the gene expression levels of the two important proteins were checked by RT PCR analysis. 82
4 Thus, this study was conducted with the objective to ascertain the establishment of disease model of RA at proteomic level which may be important for testing various therapeutic agents. 5.2 Results Visual evaluation of arthritis in CIA Fig (A) Representative photographs of hind paws of (a) Control and (b) CIA rats. (B) Percent change in body weight of control and CIA rats with reference to day 12 (appearance of inflammation) till the day of euthanization (day 42 th ). Animal group immunized with collagen showed symptoms of disease progression like inflammation in joints within days period (Fig. 5.1 A). The visual examination showed that there was significant loss of body weight in CIA rats compared with control rats (Fig. 5.1 B). Similarly, the severity of disease as measured by arthritic index (Fig. 5.2 A) and arthritis score (Fig. 5.2 B) was found to be significantly increased in CIA rats as compared to control rats. 83
5 Radiographic evaluation Typical radiographs of the hind limbs in each group were obtained on 42 nd day. Radiographic joint damage including bone erosion and loss of joint space was detected in X rays of hind limbs from CIA rats unlike those of control rats (Fig. 5.3 A) Fig (A) % mean arthritic index and (B) mean arthritic score of control and CIA rats till the day of euthanization (day 42) Histopathological evaluation 84
6 Hematoxylin and eosin stained joint tissue slides were analyzed for cartilage destruction. Histological photomicrographs (Fig. 5.3 B) shows active synovitis and bone erosion in CIA (b) as compared to the control (a). Arthritic joints in CIA illustrate synovial hyperplasia, massive inflammatory cells infiltration (monomorphonuclear and polymorphonuclear cells), erosion of bone Fig (A) Representative radiographs of the hind limbs (showing the tibiotarsal and tibiofemoral joints) of control and CIA rats. Arrows indicate the bone damage in both tibiotarsal as well as tibiofemoral joints. (B) Representative histological figures (hematoxylin and eosin stained slides) of knee joints showing smooth and monolayer synovial linings and uniform synovial space of control rats. Hyperplastic synovial cells indicated by circle, erosion and disruption of synovial linings as indicated by arrows was observed in CIA rats. m=meniscus, f=fibula, js=joint space, t=tibila. 85
7 and cartilage. These histological analyses of hind limb joints were in agreement to the radiographic studies which demonstrate a significant joint destruction in CIA rats Proteomic analysis of plasma in CIA rats WGA affinity chromatography was used to enrich plasma glycoproteins and deplete high abundant proteins such as albumin. 2 DE profiling of WGA bound (Fig. 5.4) and unbound (Fig. 5.5) plasma proteins showed the presence of Fig A representative 2 DE image of WGA bound plasma of pooled control and CIA rats from each group. Three hundred microgram of protein was focused in ph 3 10 IPG strips followed by second dimensional separation in 12 % polyacrylamide gel. many protein spots, which were differentially present in the control and CIA rats. However, the protein spots, which were successfully identified by MS/MS analysis by MALDI TOF (Fig. 5.6), were given in Table 5.1. Most of the differentially expressed proteins were found to be positive acute phase proteins, such as orosomucoid (AGP), hemopexin, kininogen, and alpha 1 major acute 86
8 phase protein, which were found to be over expressed in the CIA rats. The changes in fold expression of each identified spot were given in Table Western blotting The expression level of spot no 16 identified as alpha 1 major acute phase protein or T kininogen 1, which was highly expressed in CIA rats, was further validated by 2 D Western blot analysis of plasma of rats. It is an Fig 5.5. A representative 2 DE image of flow through of WGA bound plasma of pooled control and CIA rat from each group. Three hundred microgram of protein was focused in ph 3 10 IPG strips followed by second dimensional separation in 12 % polyacrylamide gel. important inflammation related protein. The Western blot confirms the increased expression of T Kininogen 1 in CIA rats (Fig 5.7.) Gene expression studies in CIA Estimation of m RNA levels of three genes in liver tissues of CIA and co 87
9 Fig Mascot search result showing the significance of the result and MS/MS sequence spectra of alpha 1 major acute phase protein. ntrol rats after euthanization on 42 th day, using semi quantitative RT PCR analysis (Fig. 5.8 A). The densitometric analysis revealed the ~ 3 fold increase, with respect to housekeeping gene GAPDH, in the level of kininogen mrna with statistical significance. Similarly, the level of clusterin mrna was found to be increased by ~1.2 fold in CIA albeit statistical significance (Fig. 5.8 B) Discussion CIA is a widely used experimental model of polyarthritis that has many histopathological features in common with RA in human. It has been widely demonstrated that the disease progression in RA is reflected in the plasma concentration of acute phase proteins which are also denoted as disease biomarkers. It is considered that therapeutic agents that affect the acute phase 88
10 Fig 5.7. Validation of expression of alpha 1 major acute phase protein (T kininogen 1) in CIA rat by 2 DE Western blotting. protein profile of plasma in RA not only impart relief from the disease symptoms but can also ensure intervention in the disease progression itself. Therefore, we investigated the changes in the expression of plasma proteins in CIA rats. Since most of the plasma proteins are glycoproteins in nature, we studied both WGA bound as well as unbound plasma by 2 DE for complete proteomic profiling. WGA bound plasma showed the complete depletion of albumin which may otherwise interfere in the 2 DE profiling. Most of the differentially expressed proteins like Clusterin, AGP, hemopexin, alpha 1 major acute phase protein (T kininogen 1), are the acute phase glycoproteins. Kininogens are important precursor molecules for vasodilator peptide called bradykinin via plasma kallikrein kinin system (KKS) which participates in the pathogenesis of various inflammatory reactions like involved in cellular injury, coagulation, complement activation, cytokine secretion, release of proteases. It has been demonstrated that therapy with a specific plasma kallikrein inhibitor modulated the experimental arthritis and systemic inflammation (Isordia Salas et al., 2005). 89
11 Fig (A) Representative gel image of semi quantitative RT PCR amplified mrna of three genes namely Clusterin, Kinninogen, and GAPDH in liver tissues of control and CIA rat groups. (B) Densities of the above mrnas were calculated with respect to housekeeping gene GAPDH resulting in relative density values (IDV). Values are expressed as means ± S.D. (n=5 rats per group). Only expression of kininogen was found to be statistically significant (p<0.05). Earlier studies have shown that the total plasma level of kininiogen is increased to many folds in adjuvant induced arthritis. Analysis of the kininogens demonstrated that increase in T kininogen 1 was the major reason for the rise in total kininogen level. High molecular weight and low molecular weight kininogens showed little or no change (Barlas et al., 1985). In our study, we have also found the ~6 fold increase in the level of T kininogen 1 while very slight decrease in the level of LMW kininogen was observed in plasma of CIA. Clusterin has been reported to function in many inflammatory and 90
12 immunological processes, particularly, autoimmunity (Falgarone and Chiocchia, 2009). The expression level of clusterin, both at genomics and proteomics level has been detected in synovial tissues in RA patients and healthy controls and has been found to be decreased in RA patients (Devauchelle et al., 2006). AGP, as described in details in chapter 3, is one of the major acute phase proteins. It is synthesized in large quantities during acute phase reactions and accumulates at the vessel wall surface via binding to E selectin providing an anchoring template for plasminogen activator inhibitor type 1 (PAI 1) on the cellular surface and stabilizes its inhibitory activity toward plasminogen activators (Boncela et al., 2001). Its expression is almost twofold increased in CIA plasma which is in agreement with plasma studies in RA. Hemopexin or Beta 1B glycoprotein is an another important acute phase protein which has been reported to be increased in inflammation (Marinkovic et al., 1989) and adjuvant induced arthritic experimental animals (Saso et al., 1992). It functions as a heme scavenging molecule protecting cells from oxidative damage by free heme molecules released by hemolysis and preserves the iron level in the body. Low hemopexin level shows the degradation of hemoglobin which indicates hemolysis. Thus, low hemopexin level is one of the diagnostic features of a hemolytic anemia. 5.4 Conclusion Thus, the proteomic analysis of plasma proteins in CIA showed that this model mimics the disease i.e. RA at the proteomic level. This study can be very useful where the effects of different compounds are tested on the animal model of various diseases. Besides many biochemical and histochemical assays used for this study, the effects of these compounds can be checked on the expression levels of these plasma proteins. A study by Sahu et al has shown the effect of a 91
13 well known synthetic compound suramin on the level of few plasma acute phase proteins in CIA which were brought to normalization after treatment with suramin (Sahu et al., 2011). 92
14 Table 5.1. List of differentially expressed 2 DE protein spots in WGA bound and unbound plasma fractions of CIA as identified by MALDI TOF MS. Spot no Protein name *Accession number M.W./P.I Fold change Precursor ions Sequence of peptide matched 1 Immunoglobulin J chain precursor isoform gi ,229/ IIPSPEDPNEDIVER Immunoglobulin J chain precursor isoform gi ,229/ IIPSPEDPNEDIVER 92 3 Complement C1q subcomponent, B chain gi ,801/ , VITNVNDNYEPR, 72 precursor TVNSALRPNQAIR 4 Complement C1q subcomponent subunit A gi ,198/ , DQPRPAFSAIR, QNPPTYGNVVVFDK 5 Preprohaptoglobin gi ,563/ SCAVAEYGVYVR 54 6 Clusterin gi ,930/ LTQQYNELLHSLQSK 42 7 Orosomucoid 1 precursor (AGP) gi ,731/ DVGMDESEIVFVDWTK 77 8 Ig gamma 2A chain C region gi ,268/ VTCVVVDISQNDPEVR 43 9 Coagulation factor X gi ,322/ DTYDFDIAMLR Corticosteroid binding globulin gi ,813/ , GIWELPFSPENTR, GLAPTNVDFAFNLYQR 11 Alpha 1 major acute phase protein gi ,324/ YNAELESGNQFLLYR Leukemia inhibitory factor receptor alpha gi ,808./ , WDDIPVEELR 180 chain IASMEIPNDDITVEQAVGLGNR 13 Alpha 1 macroglobulin gi ,422/ , AISYLISGYQR ALLAYAFALAGNR 14 Beta 2 glycoprotein (Apolipoprotein H) gi ,743/ , VCPFAGILENGVVR TFYDPGEQIVYSCKPGYVSR 15 Hemopexin gi ,351/ LFQEEFPGIPYPPDAAVECHR Fibrinogen beta chain gi ,896/ LYIDETVNDNIPLNLR Complement component 3 gi ,825/ , TIYTPGSTVFYR, EPGQDLVVLSLPITPEFIPSFR 18 Transferrin precursor gi ,090/ ,2 KPVDQYEDCYLAR, LPEGTTYEEYLGAEYLQAVGNIR 19 Albumin precursor gi ,670/ , LGEYGFQNAVLVR, DVFLGTFLYEYSR 20 Transferrin gi ,512/ , LYLGHSYVTAIR, DQYELLCLDNTR 87 Ion score
15 21 Alpha 1B glycoprotein gi ,127/ VLNIQGFSPTR Complement component 3 gi ,825/ , VYSYYNLEESCTR, ILLQGTPVAQMAEDAVDGER 23 Leukemia inhibitory factor receptor gi ,394/ IASMEIPNDDITVEQAVGLGNR Murinoglobulin 1 precursor gi ,590/ , ALGCLEASWETIEQGR, SEGYLYTPQASSAEVEMSAYVVLAR 278 *NCBI accession number average M.W and p.i values as available in the NCBI database. fold expression values were calculated in CIA with respect to control and difference values 1.5 fold were considered. indicates up regulation and indicates down regulation of proteins, measured as ratio of spot densities of CIA/control.
Supporting Information: Protein Corona Analysis of Silver Nanoparticles Exposed to Fish Plasma
Supporting Information: Protein Corona Analysis of Silver Nanoparticles Exposed to Fish Plasma Jiejun Gao 1, Lu Lin 2, Alexander Wei 2,*, and Maria S. Sepúlveda 1,* 1 Department of Forestry and Natural
More informationIntroduction. Methods RESEARCH FUND FOR THE CONTROL OF INFECTIOUS DISEASES. TCW Poon *, HLY Chan, HWC Leung, A Lo, RHY Lau, AY Hui, JJY Sung
RESEARCH FUND FOR THE CONTROL OF INFECTIOUS DISEASES Liver specific glycoforms of serum proteins in chronic hepatitis B infection: identification by lectin affinity chromatography and quantitative proteomic
More informationTECHNICAL NOTE. Accurate and fast proteomics analysis of human plasma with PlasmaDive and SpectroDive
TECHNICAL NOTE Accurate and fast proteomics analysis of human plasma with PlasmaDive and SpectroDive In this technical note you will learn about: Step-by-step set-up of parallel reaction monitoring (PRM)
More informationRheumatoid Arthritis. Immunology and Inflammatory Disease. In pursuit of your success. Autoimmune Arthritis Animal Models Available:
Immunology and Inflammatory Disease Rheumatoid arthritis (RA) is a chronic debilitating autoimmune disorder characterized by synovitis that leads to cartilage and bone erosion by invading fibrovascular
More informationChapter 10apter 9. Chapter 10. Summary
Chapter 10apter 9 Chapter 10 The field of proteomics has developed rapidly in recent years. The essence of proteomics is to characterize the behavior of a group of proteins, the system rather than the
More informationIntroduction to Proteomics 1.0
Introduction to Proteomics 1.0 CMSP Workshop Pratik Jagtap Managing Director, CMSP Objectives Why are we here? For participants: Learn basics of MS-based proteomics Learn what s necessary for success using
More informationSeparation of Main Proteins in Plasma and Serum
BCH 471 Experiment (2) Separation of Main Proteins in Plasma and Serum PLASMA PROTEINS Mw The main plasma proteins are: þ Albumin (36-50 g/l), Mw 66.241kDa. þ Globulins (18-32 g/l), Mw of globulins Cover
More informationLECTURE-15. itraq Clinical Applications HANDOUT. Isobaric Tagging for Relative and Absolute quantitation (itraq) is a quantitative MS
LECTURE-15 itraq Clinical Applications HANDOUT PREAMBLE Isobaric Tagging for Relative and Absolute quantitation (itraq) is a quantitative MS based method for quantifying proteins subject to various different
More informationMultiple High-Abundant Protein Removal for Proteomics. Dr. Cory Szafranski, Product Manager
Multiple High-Abundant Protein Removal for Proteomics Dr. Cory Szafranski, Product Manager Agilent Multiple Affinity Removal System - What is it? H L High-Abundant Proteins (Albumin, IgG, IgA, Transferrin,
More informationMimi Roy, PhD Senior Director & Site Head Caprion Proteomics US LLC. ISBER, Orlando May 23, 2014
Controlled Analysis of Preanalytical Variables in CSF and Blood Sample Collection, Processing and Storage: Implications for Best Practices in Clinical Research Mimi Roy, PhD Senior Director & Site Head
More informationPlasma proteins Quantitatively, proteins are the most important part of the soluble components of the blood plasma.
Plasma proteins 42 Plasma proteins Quantitatively, proteins are the most important part of the soluble components of the blood plasma. concentrations of between 60 and 80 g L 1, they constitute approximately
More informationNBQX, An AMPA/Kainate Glutamate Receptor Antagonist, Alleviates Joint Disease In Models Of Inflammatory- And Osteo- Arthritis.
NBQX, An AMPA/Kainate Glutamate Receptor Antagonist, Alleviates Joint Disease In Models Of Inflammatory- And Osteo- Arthritis. Cleo S. Bonnet, PhD 1, Anwen S. Williams, PhD 1, Sophie J. Gilbert, PhD 1,
More informationAUTOIMMUNITY CLINICAL CORRELATES
AUTOIMMUNITY CLINICAL CORRELATES Pamela E. Prete, MD, FACP, FACR Section Chief, Rheumatology VA Healthcare System, Long Beach, CA Professor of Medicine, Emeritus University of California, Irvine Colonel
More informationAUTOIMMUNITY TOLERANCE TO SELF
AUTOIMMUNITY CLINICAL CORRELATES Pamela E. Prete, MD, FACP, FACR Section Chief, Rheumatology VA Healthcare System, Long Beach, CA Professor of Medicine, Emeritus University of California, Irvine Colonel
More informationEFFECT OF VITAMINS C AND E ON THE LEVELS OF CARBOHYDRATE COMPONENTS OF GLYCOPROTEINS IN COLLAGEN INDUCED ARTHRITIS
Chapter 4 EFFECT OF VITAMINS C AND E ON THE LEVELS OF CARBOHYDRATE COMPONENTS OF GLYCOPROTEINS IN COLLAGEN INDUCED ARTHRITIS 4.1. INTRODUCTION Rheumatoid Arthritis is a chronic inflammatory disease categorized
More informationGMI STUDY. COMPARATIVE PROTEOMIC STUDY BETWEEN GMI and STRAUMANN DENTAL IMPLANTS
GMI STUDY. COMPARATIVE PROTEOMIC STUDY BETWEEN GMI and STRAUMANN DENTAL IMPLANTS 2 Hypothesis. Proteomic study of first protein layer Post-implantation, the biomaterial becomes in contact with the blood,
More informationPart IV Antithrombotics, Anticoagulants and Fibrinolytics
Part IV Antithrombotics, Anticoagulants and Fibrinolytics "The meaning of good and bad, of better and worse, is simply helping or hurting" Emerson Chapter 16: Blood Coagulation and Fibrinolytic System
More informationSupporting Information. Synthesis of Zwitterionic Polymer Particles via Combined Distillation
Supporting Information Synthesis of Zwitterionic Polymer Particles via Combined Distillation Precipitation Polymerization and Click Chemistry for Highly Efficient Enrichment of Glycopeptide Jianxi Liu,
More informationCellular Pathology of immunological disorders
Cellular Pathology of immunological disorders SCBM344 Cellular and Molecular Pathology Witchuda Payuhakrit, Ph.D (Pathobiology) witchuda.pay@mahidol.ac.th Objectives Describe the etiology of immunological
More informationImprove Protein Analysis with the New, Mass Spectrometry- Compatible ProteasMAX Surfactant
Improve Protein Analysis with the New, Mass Spectrometry- Compatible Surfactant ABSTRACT Incomplete solubilization and digestion and poor peptide recovery are frequent limitations in protein sample preparation
More informationCell-Derived Inflammatory Mediators
Cell-Derived Inflammatory Mediators Introduction about chemical mediators in inflammation Mediators may be Cellular mediators cell-produced or cell-secreted derived from circulating inactive precursors,
More information1.0 Abstract. Title. Keywords. Rationale and Background
1.0 Abstract Title A Prospective, Multi-Center Study in Rheumatoid Arthritis Patients on Adalimumab to Evaluate its Effect on Synovitis Using Ultrasonography in an Egyptian Population Keywords Synovitis
More informationThe Cardiovascular System: Blood
C h a p t e r 11 The Cardiovascular System: Blood PowerPoint Lecture Slides prepared by Jason LaPres Lone Star College - North Harris Introduction to the Cardiovascular System A circulating transport system
More informationInflammation. (4 of 5)
Inflammation (4 of 5) What will we discuss today? Plasma protein derived mediators Anti-inflammatory mediators Morphologic patterns of acute inflammation Plasma protein derived mediators 3 systems: -Complement
More informationInnate vs Adaptive Response
General Immunology Innate vs Adaptive Response Innate- non-specific (4 types of barriers) anatomic- ato mechanical ca (skin), ph, mucous, normal flora Physiologic- temperature, ph, chemicals (lysozyme,
More informationChapter 11. Lecture and Animation Outline
Chapter 11 Lecture and Animation Outline To run the animations you must be in Slideshow View. Use the buttons on the animation to play, pause, and turn audio/text on or off. Please Note: Once you have
More information1. The scavenger receptor, CD36, functions as a coreceptor for which TLR? a. TLR ½ b. TLR 3 c. TLR 4 d. TLR 2/6
Allergy and Immunology Review Corner: Cellular and Molecular Immunology, 8th Edition By Abul K. Abbas, MBBS, Andrew H. H. Lichtman, MD, PhD and Shiv Pillai, MBBS, PhD. Chapter 4 (pages 62-74): Innate Immunity
More informationINFLAMMATION & REPAIR
INFLAMMATION & REPAIR Lecture 7 Chemical Mediators of Inflammation Winter 2013 Chelsea Martin Special thanks to Drs. Hanna and Forzan Course Outline i. Inflammation: Introduction and generalities (lecture
More informationPrinciples of Adaptive Immunity
Principles of Adaptive Immunity Chapter 3 Parham Hans de Haard 17 th of May 2010 Agenda Recognition molecules of adaptive immune system Features adaptive immune system Immunoglobulins and T-cell receptors
More informationSequence Coverage (%) Profilin-1 P UD 2
Protein Name Accession Number (Swissprot) Sequence Coverage (%) No. of MS/MS Queries Mascot Score 1 Reference Cytoskeletal proteins Beta-actin P60709 37 14 298 Alpha-actin P68032 33 10 141 20 Beta-actin-like
More informationAntigen Receptor Structures October 14, Ram Savan
Antigen Receptor Structures October 14, 2016 Ram Savan savanram@uw.edu 441 Lecture #8 Slide 1 of 28 Three lectures on antigen receptors Part 1 (Today): Structural features of the BCR and TCR Janeway Chapter
More informationOmar Alnairat. Tamer Barakat. Bahaa Abdelrahim. Dr.Nafez
1 Omar Alnairat Tamer Barakat Bahaa Abdelrahim Dr.Nafez It s the chemistry inside living cells. What is biochemistry? Biochemistry consists of the structure and function of macromolecules (in the previous
More informationPTM Discovery Method for Automated Identification and Sequencing of Phosphopeptides Using the Q TRAP LC/MS/MS System
Application Note LC/MS PTM Discovery Method for Automated Identification and Sequencing of Phosphopeptides Using the Q TRAP LC/MS/MS System Purpose This application note describes an automated workflow
More informationthe HLA complex Hanna Mustaniemi,
the HLA complex Hanna Mustaniemi, 28.11.2007 The Major Histocompatibility Complex Major histocompatibility complex (MHC) is a gene region found in nearly all vertebrates encodes proteins with important
More informationCh. 45 Blood Plasma proteins, Coagulation and Fibrinolysis Student Learning Outcomes: Describe basic components of plasma
Chapt. 45 Ch. 45 Blood Plasma proteins, Coagulation and Fibrinolysis Student Learning Outcomes: Describe basic components of plasma Inheritance of X-linked gene for Factor VIII hemophilia A Explain the
More informationThe Major Histocompatibility Complex (MHC)
The Major Histocompatibility Complex (MHC) An introduction to adaptive immune system before we discuss MHC B cells The main cells of adaptive immune system are: -B cells -T cells B cells: Recognize antigens
More informationIncreases Circulation Immune Complexes, Increased Fibrin Activation and Fibrosis
Inflammation Inflammation is a complex biological process in which the body s white blood cells and chemicals provide protection from infection and foreign substances, such as bacteria, yeast, and viruses
More informationAutoimmunity. Autoimmunity arises because of defects in central or peripheral tolerance of lymphocytes to selfantigens
Autoimmunity Autoimmunity arises because of defects in central or peripheral tolerance of lymphocytes to selfantigens Autoimmune disease can be caused to primary defects in B cells, T cells and possibly
More informationBlood. Plasma. The liquid part of blood is called plasma. 1. Pale yellow fluid; forms more than half the blood volume.
11 Blood FOCUS: Blood consists of plasma and formed elements. The plasma is 91% water with dissolved or suspended molecules, including albumin, globulins, and fibrinogen. The formed elements include erythrocytes,
More informationHYPERSENSITIVITY REACTIONS D R S H O AI B R AZ A
HYPERSENSITIVITY REACTIONS D R S H O AI B R AZ A HYPERSENSITIVITY REACTIONS Are exaggerated immune response upon antigenic stimulation Individuals who have been previously exposed to an antigen are said
More informationTable S1. CRC case Pool Control Pool Name UniProt No. FC b VIP value d Spectral Counts Spectral Counts
Table S1. page 1/4 Phase 1 Exploratory Study, List of proteins identified by LC-ESI-MS/MS after 1DE separation and their relative quantitation by spectral count UniProt Entry SwissProt- CRC case Pool Control
More informationCH 11 Blood OUTLINE: Functions of Blood Composition of Blood Blood Cell Disorders Blood Types Blood Clotting Functions of Blood Transportation
1 CH 11 Blood OUTLINE: Functions of Blood Composition of Blood Blood Cell Disorders Blood Types Functions of Blood Transportation Protection Regulation ph Temperature Composition of Blood Plasma: liquid
More information10. Which of the following immune cell is unable to phagocytose (a) neutrophils (b) eosinophils (c) macrophages (d) T-cells (e) monocytes
Chapter 2. Acute and chronic inflammation(6): 1. In acute inflammation, which events occur in the correct chronological order? (Remembered from 2000, 2004 exam.) p50 (a) transient vasoconstriction, stasis
More informationTissue and Fluid Proteomics Chao-Cheng (Sam) Wang
Tissue and Fluid Proteomics Chao-Cheng (Sam) Wang UAB-03/09/2004 What is proteomics? A snap shot of the protein pattern!! What can proteomics do? To provide information on functional networks and/or involvement
More informationDr Cédric DELPORTE Prof. Ass. Dr Pierre VAN ANTWERPEN
POLARIS CHIP FOR PROTEOMIC S: PROTEIN AND PEPTIDE PROFILING Dr Cédric DELPORTE Prof. Ass. Dr Pierre VAN ANTWERPEN Laboratory of Pharmaceutical Chemistry & Analytical Platform of the Faculty of Pharmacy
More informationRegulation of the IGF axis by TGF-b during periosteal chondrogenesis: implications for articular cartilage repair
Regulation of the IGF axis by TGF-b during periosteal chondrogenesis: implications for articular cartilage repair Chapter 04 Boek 1_Gie.indb 55 21-05-2007 12:27:33 Chapter 04 Abstract Goal: TGF-b and IGF-I
More informationESTUDIO GMI. ESTUDIO PROTEÓMICO-COMPARACIÓN IMPLANTES DENTALES
ESTUDIO GMI. ESTUDIO PROTEÓMICO-COMPARACIÓN IMPLANTES DENTALES 2 Hypothesis. Proteomic study of first protein layer Post-implantation, the biomaterial becomes in contact with the blood, resulting in protein
More informationMOLECULAR IMMUNOLOGY Manipulation of immune response Autoimmune diseases & the pathogenic mechanism
MOLECULAR IMMUNOLOGY Manipulation of immune response Autoimmune diseases & the pathogenic mechanism SCHMAIEL SHIRDEL CONTENT 2 Introduction Autoimmune diseases Classification Involved components Autoimmune
More informationTofacitinib ( Xeljanz) Marshall Porter & Lauren Ysais
Tofacitinib ( Xeljanz) Marshall Porter & Lauren Ysais Learning Objectives to Take with You A small molecule Janus Kinase (JAK) Inhibitor treatment for moderate to severe rheumatoid arthritis (RA) reduces
More informationBlood. Biol 105 Lecture 14 Chapter 11
Blood Biol 105 Lecture 14 Chapter 11 Outline I. Overview of blood II. Functions of blood III. Composition of blood IV. Composition of plasma V. Composition of formed elements VI. Platelets VII. White blood
More informationDense and Dynamic Polyethylene Glycol Shells Cloak Nanoparticles. from Uptake by Liver Endothelial Cells for Long Blood Circulation
Dense and Dynamic Polyethylene Glycol Shells Cloak Nanoparticles from Uptake by Liver Endothelial Cells for Long Blood Circulation Hao Zhou, Zhiyuan Fan, Peter Y. Li, Junjie Deng,, Dimitrios C. Arhontoulis,
More informationGlycosylation analysis of blood plasma proteins
Glycosylation analysis of blood plasma proteins Thesis booklet Eszter Tóth Doctoral School of Pharmaceutical Sciences Semmelweis University Supervisor: Károly Vékey DSc Official reviewers: Borbála Dalmadiné
More informationThe major histocompatibility complex (MHC) is a group of genes that governs tumor and tissue transplantation between individuals of a species.
Immunology Dr. John J. Haddad Chapter 7 Major Histocompatibility Complex The major histocompatibility complex (MHC) is a group of genes that governs tumor and tissue transplantation between individuals
More informationChapter 19. Openstax: Chapter 18. Blood
Chapter 19 Blood Openstax: Chapter 18 Chapter 19 Learning Outcomes After completing Chapter 19, you will be able to: 1. Describe the components and major functions of blood and list the physical characteristics
More informationMass Spectrometry. Mass spectrometer MALDI-TOF ESI/MS/MS. Basic components. Ionization source Mass analyzer Detector
Mass Spectrometry MALDI-TOF ESI/MS/MS Mass spectrometer Basic components Ionization source Mass analyzer Detector 1 Principles of Mass Spectrometry Proteins are separated by mass to charge ratio (limit
More informationChapter 3. Protein Structure and Function
Chapter 3 Protein Structure and Function Broad functional classes So Proteins have structure and function... Fine! -Why do we care to know more???? Understanding functional architechture gives us POWER
More informationDisease-Modifying Activity in a Model of Rheumatoid Arthritis with an Orally Available Inhibitor of Methionine Aminopeptidase Type-2,
Disease-Modifying Activity in a Model of Rheumatoid Arthritis with an rally Available Inhibitor of Methionine Aminopeptidase Type-2, PPI-2458 William Westlin, Ph.D. Praecis Pharmaceuticals Waltham, Massachusetts
More informationPlease check the slides
Quick review of main concepts: The major plasma proteins are : albumin, globulin and fibrenogen globulin consists of 3 types: α, β and γ α globulin is divided into 2 types : α1 (includes α1 antitrypsin
More informationFor questions 1-5, match the following with their correct descriptions. (24-39) A. Class I B. Class II C. Class III D. TH1 E. TH2
Questions Made by SI ATTENDEES!! :) Page 1 of 6 Student-Made Practice Exam Activity All questions, answers, and slide numbers are based off of Monday s SI activity, where students/attendees created possible
More informationSupplemental Methods: Histopathology scoring of individual components of Valentino
Supplementary Materials Online: Supplemental Methods: Histopathology scoring of individual components of Valentino synovitis grade and Mankin cartilage pathology scale Hemophilic synovitis was graded 0-10
More informationPrimary Immunodeficiency
Primary Immunodeficiency DiGeorge Syndrome Severe Combined Immunodeficiency SCID X-Linked Agammaglobulinemia Common variable immunodeficiency (CVID) IgA deficiency Hyper- IgM Syndrome Wiskott-Aldrich syndrome
More informationBIOL 2458 CHAPTER 19 Part 1 SI 1. List the types of extracellular fluids. 2. Intracellular fluid makes up of the body fluids. Where is it found?
BIOL 2458 CHAPTER 19 Part 1 SI 1 1. Extracellular fluid makes up of the body fluids. List the types of extracellular fluids. 2. Intracellular fluid makes up of the body fluids. Where is it found? 3. In
More informationNeutrophils contribute to fracture healing by synthesizing fibronectin+ extracellular matrix rapidly after injury
Neutrophils contribute to fracture healing by synthesizing fibronectin+ extracellular matrix rapidly after injury Bastian OW, Koenderman L, Alblas J, Leenen LPH, Blokhuis TJ. Neutrophils contribute to
More informationTissue and Fluid Proteomics
Tissue and Fluid Proteomics Chao-Cheng (Sam) Wang, Ph.D. 4-6919 ccwang@uab.edu UAB BMG 744-02/28/06 Sample Sources for Proteomic Analysis Cell lines. Tissue sections. Body Fluids: Blood and urine. Fluids
More information11/25/2017. THE IMMUNE SYSTEM Chapter 43 IMMUNITY INNATE IMMUNITY EXAMPLE IN INSECTS BARRIER DEFENSES INNATE IMMUNITY OF VERTEBRATES
THE IMMUNE SYSTEM Chapter 43 IMMUNITY INNATE IMMUNITY EXAMPLE IN INSECTS Exoskeleton made of chitin forms the first barrier to pathogens Digestive system is protected by a chitin-based barrier and lysozyme,
More informationMedical Virology Immunology. Dr. Sameer Naji, MB, BCh, PhD (UK) Head of Basic Medical Sciences Dept. Faculty of Medicine The Hashemite University
Medical Virology Immunology Dr. Sameer Naji, MB, BCh, PhD (UK) Head of Basic Medical Sciences Dept. Faculty of Medicine The Hashemite University Human blood cells Phases of immune responses Microbe Naïve
More informationAutoimmune Diseases. Betsy Kirchner CNP The Cleveland Clinic
Autoimmune Diseases Betsy Kirchner CNP The Cleveland Clinic Disclosures (financial) No relevant disclosures Learning Objectives Explain the pathophysiology of autoimmune disease Discuss safe administration
More informationHelen Kim, Ph.D. and John Cutts. Dept of Pharmacology & Toxicology University of Alabama at Birmingham
Understanding the actions of a dietary anti-oxidant at the protein and small molecule level using top-down proteomics, enzyme assays and mass spectrometry elen Kim, Ph.D. and John Cutts Mar 9, 2012 UAB
More informationmirna Biomarkers Seena K. Ajit PhD Pharmacology & Physiology Drexel University College of Medicine October 12, 2017
mirna Biomarkers Seena K. Ajit PhD Pharmacology & Physiology Drexel University College of Medicine October 12, 2017 Outline Introduction Circulating mirnas as potential biomarkers for pain Complex regional
More informationEtiology: Pathogenesis Clinical manifestation Investigation Treatment Prognosis
Etiology: Pathogenesis Clinical manifestation Investigation Treatment Prognosis JIA is the most common rheumatic disease in childhood and a major cause of chronic disability. Etiology: Unknown, but may
More informationTissue-specific exosome biomarkers for noninvasively monitoring immunologic rejection of transplanted tissue
Tissue-specific exosome biomarkers for noninvasively monitoring immunologic rejection of transplanted tissue Prashanth Vallabhajosyula,, Michael R. Rickels, Ali Naji J Clin Invest. 2017;127(4):1375-1391.
More informationImmunology. Lecture- 8
Immunology Lecture- 8 Immunological Disorders Immunodeficiency Autoimmune Disease Hypersensitivities Immunodeficiency 1. Immunodeficiency --> abnormal production or function of immune cells, phagocytes,
More informationBlood. BIOLOGY OF HUMANS Concepts, Applications, and Issues. Judith Goodenough Betty McGuire
BIOLOGY OF HUMANS Concepts, Applications, and Issues Fifth Edition Judith Goodenough Betty McGuire 11 Blood Lecture Presentation Anne Gasc Hawaii Pacific University and University of Hawaii Honolulu Community
More informationLYMPHOCYTES & IMMUNOGLOBULINS. Dr Mere Kende, Lecturer SMHS
LYMPHOCYTES & IMMUNOGLOBULINS Dr Mere Kende, Lecturer SMHS Immunity Immune- protection against dangers of non-self/invader eg organism 3 components of immune system 1 st line: skin/mucosa/cilia/hair/saliva/fatty
More informationFunctions of Blood. Transport. Transport. Defense. Regulation. Unit 6 Cardiovascular System: Blood
Unit 6 Cardiovascular System: Blood Functions of Blood With each beat of the heart, approximately 75 ml of blood is pumped On average, the heart beats 70 times per minute Every minute, the heart pumps
More informationChapter 11. B cell generation, Activation, and Differentiation. Pro-B cells. - B cells mature in the bone marrow.
Chapter B cell generation, Activation, and Differentiation - B cells mature in the bone marrow. - B cells proceed through a number of distinct maturational stages: ) Pro-B cell ) Pre-B cell ) Immature
More informationGLUCOSE CONCENTRATION INCREASES IGF EXPRESSION FROM SYNOVIAL MEMBRANE
GLUCOSE CONCENTRATION INCREASES IGF EXPRESSION FROM SYNOVIAL MEMBRANE Final Report Aug 17 2009 Darryl D'Lima, MD, PhD Shiley Center for Orthopaedic Research and Education at Scripps Clinic La Jolla, California
More informationTarget cell lysis Opsonization Activation of the inflammatory response (e.g. degranulation, extravasation) Clearance of immune complexes
Immunology Dr. John J. Haddad Chapter 13 Complement Major roles of complement (Figure 13-1): Target cell lysis Opsonization Activation of the inflammatory response (e.g. degranulation, extravasation) Clearance
More information4/5/17. Blood. Blood. Outline. Blood: An Overview. Functions of Blood
Outline Blood Biol 105 Chapter 11 I. Overview of blood II. Functions of blood III. Composition of blood IV. Composition of plasma V. Composition of formed elements VI. Platelets VII. White blood cells
More informationSUPPLEMENTAL TABLE I. Identified Proteins in Bovine Testicular Hyaluronidase Type I-S via LC-MS/MS
SUPPLEMENTAL TABLE I. Identified Proteins in Bovine Testicular Hyaluronidase Type I-S via LC-MS/MS No. Protein 1 serum albumin precursor gi 30794280 2 annexin A2 gi 27807289 3 Phosphatidylethanolamine-binding
More informationG. Types of White Blood Cells
1. White blood cells are also called leukocytes. G. Types of White Blood Cells 2. White blood cells function to protect against diseases. 3. Two hormones that stimulate white blood cell production are
More informationProtein Structure and Function
Protein Structure and Function Protein Structure Classification of Proteins Based on Components Simple proteins - Proteins containing only polypeptides Conjugated proteins - Proteins containing nonpolypeptide
More informationProteomics of body liquids as a source for potential methods for medical diagnostics Prof. Dr. Evgeny Nikolaev
Proteomics of body liquids as a source for potential methods for medical diagnostics Prof. Dr. Evgeny Nikolaev Institute for Biochemical Physics, Rus. Acad. Sci., Moscow, Russia. Institute for Energy Problems
More informationChapter 13. Plasma Proteins and Enzymes. Lecturer: Dr Abeer Shnoudeh. Clinical Chemistry William Marshall
Chapter 13 Plasma Proteins and Enzymes Lecturer: Dr Abeer Shnoudeh Clinical Chemistry William Marshall Introduction Measurement of Plasma Proteins Total Plasma Protein concentration of total protein in
More informationUnder the Radar Screen: How Bugs Trick Our Immune Defenses
Under the Radar Screen: How Bugs Trick Our Immune Defenses Session 7: Cytokines Marie-Eve Paquet and Gijsbert Grotenbreg Whitehead Institute for Biomedical Research HHV-8 Discovered in the 1980 s at the
More informationA. Blood is considered connective tissue. RBC. A. Blood volume and composition 1. Volume varies - average adult has 5 liters
A. Blood is considered connective tissue. RBC A. Blood volume and composition 1. Volume varies - average adult has 5 liters 2. 45% cells by volume called hematocrit (HCT) a. red blood cells (RBC) mostly
More information2402 : Anatomy/Physiology
Dr. Chris Doumen Lecture 2 2402 : Anatomy/Physiology The Endocrine System G proteins and Adenylate Cyclase /camp TextBook Readings Pages 405 and 599 through 603. Make use of the figures in your textbook
More informationWhat is Autoimmunity?
Autoimmunity What is Autoimmunity? Robert Beatty MCB150 Autoimmunity is an immune response to self antigens that results in disease. The immune response to self is a result of a breakdown in immune tolerance.
More informationWhat is Autoimmunity?
Autoimmunity What is Autoimmunity? Robert Beatty MCB150 Autoimmunity is an immune response to self antigens that results in disease. The immune response to self is a result of a breakdown in immune tolerance.
More informationIndex. Index 439. Aequorin, 84, 94 Affinity precipitation, 372, AP-1, 100 Asthma, 170, 305
Index 439 Index A Aequorin, 84, 94 Affinity precipitation, 372, 376 381 AP-1, 100 Asthma, 170, 305 B Bioassay, 185, comparison with ELISA, 318 GM-CSF bioassay, 351 IL-2 bioassay, 185 192, 300 IL-3 IL-6
More informationOverview of the immune system
Overview of the immune system Immune system Innate (nonspecific) 1 st line of defense Adaptive (specific) 2 nd line of defense Cellular components Humoral components Cellular components Humoral components
More informationAnaphylaxis: The Atypical Varieties
Anaphylaxis: The Atypical Varieties John Johnson, D.O., PGY-4 Allergy/Immunology Fellow University Hospitals of Cleveland Case Western Reserve University School of Medicine Disclosures: None What is Anaphylaxis?
More informationOverview of the Lymphoid System
Overview of the Lymphoid System The Lymphoid System Protects us against disease Lymphoid system cells respond to Environmental pathogens Toxins Abnormal body cells, such as cancers Overview of the Lymphoid
More informationIntroduction to Immunology Part 2 September 30, Dan Stetson
Introduction to Immunology Part 2 September 30, 2016 Dan Stetson stetson@uw.edu 441 Lecture #2 Slide 1 of 26 CLASS ANNOUNCEMENT PLEASE NO TREE NUTS IN CLASS!!! (Peanuts, walnuts, almonds, cashews, etc)
More informationBIOCHEMISTRY & MEDICINE:
BIOCHEMISTRY & MEDICINE: INTRODUCTION Biochemistry can be defined as the science of the chemical basis of life (Gk bios "life"). The cell is the structural unit of living systems. Thus, biochemistry can
More informationMales- Western Diet WT KO Age (wks) Females- Western Diet WT KO Age (wks)
Relative Arv1 mrna Adrenal 33.48 +/- 6.2 Skeletal Muscle 22.4 +/- 4.93 Liver 6.41 +/- 1.48 Heart 5.1 +/- 2.3 Brain 4.98 +/- 2.11 Ovary 4.68 +/- 2.21 Kidney 3.98 +/-.39 Lung 2.15 +/-.6 Inguinal Subcutaneous
More informationAround million aged erythrocytes/hour are broken down.
Anemia Degradation ofheme Around 100 200 million aged erythrocytes/hour are broken down. The degradation process starts in reticuloendothelial cells in the spleen, liver, and bone marrow. [1] The tetrapyrrole
More informationImmunology - Lecture 2 Adaptive Immune System 1
Immunology - Lecture 2 Adaptive Immune System 1 Book chapters: Molecules of the Adaptive Immunity 6 Adaptive Cells and Organs 7 Generation of Immune Diversity Lymphocyte Antigen Receptors - 8 CD markers
More informationRheumatoid arthritis: A heterogeneous disease with a heterogeneous response to treatment
Rheumatoid arthritis: A heterogeneous disease with a heterogeneous response to treatment Pr Pierre Miossec MD PhD Clinical Immunology Unit Hôpital Edouard Herriot Lyon miossec@univ-lyon1.fr Pathogenesis
More informationInternational Graduate Research Programme in Cardiovascular Science
1 International Graduate Research Programme in Cardiovascular Science This work has been supported by the European Community s Sixth Framework Programme under grant agreement n LSHM-CT-2005-01883 EUGeneHeart.
More information