2. PATHOLOGY + PRACTICALS IMMUNOLOGY COURSE 11 SZEMINARS/PRACTICALS 1 class/week Weeks 1-11 26 LECTURES 2 lectures/week Weeks 1-13 BASIC IMMUNOLOGY and PATHOLOGY DEMO 1. BASIC + SEMINARS 2. PATHOLOGY + PRACTICALS

BOOKS Peter Parham: The immune system (Garland Science) 2nd Edition 2005 Janeway C.A. Jr., Travers P., Walport M., Shlomchik M.: Immunbiology (Garland Publishing) 5th Edition 2001 Rosen F., Geha R.: Case Studies in Immunology (Garland Publishing) 5th Edition 2001 Abbas A.K., Lichtman A.H., Pober J.S.: Cellular and Molecular Immunology (W.B. Saunders Company) 4th Edition 2000 www.immunology.unideb.hu Username: student PASSWORD: download TESTS

ENVIRONMENTAL GENOMICS IMMUNOLOGY MICROBIOLOGY EPIDEMIOLOGY CELL BIOLOGY GENETICS BIOCHEMISTRY BIOPHYSICS MOLECULAR BIOLOGY BASIC IMMUNE DEFICIENCIES HYPERSENSITIVITY REACTIONS INFECTIOUS DISEASES AUTOIMMUNITY TUMOR IMMUNOLOGY TRANSPLANTATION IMMUNOLOGY CLINICAL ALLERGOLOGY IMMUNOGENETICS IMMUNOGENOMICS ENVIRONMENTAL GENOMICS

HISTORY OF IMMUNOLOGY Babylonian Epic of Gilgamesh (2000 B.C.) diseases, pestilence Egypt older dynasties severe epidemics Phobus Apolló plague in the Greek army Old testament God’s punishment 1880 – First World War study of disases, vaccines 1920 – 1960 scientific revolution, chemistry and biology Immunological memory Thucydides, historian, Athen 430 i.e. plague „yet it was with those who recovered from the disease that the sick and the dying found most compassion……. No fear for themselves; as no man was never attacked twice – never at least fatally” Immunity Immunitas (latin) – excemption from taxes Depletion theories First infection depletes nutrients required for the propagation of the pathogen Variolation Smallpox live, wild-type virus inoculated (from China) but practiced in Europe too

FIRST VACCINATION Edward Jenner 1796 Immunity (protection) can be induced (cowpox - smallpox)

Louis Pasteur 1880 rabies, 1888 Pastuer Institute 1884 Ilya Mechnikoff Phagocytosis Immunization with attenuated pathogens CELLULAR IMMUNOLOGY

Koch Laboratory Berlin 1890, Diphteria and Tetanus toxin Antibodies in serum – bound to relevant pathogens Humoral factors HUMORAL IMMUNE RESPONSE Emil Behring Shimbasaru Kitasato Many disease occurs only once (natural protection) Some diseases can be prevented by vaccination The blood contains anti-bacterial activity (anti-toxins, serum therapy)

MILESTONES OF IMMUNOLOGY RESEARCH I. YEAR NAME DISCOVERY NOBEL PRIZE 1890 Emil von Behring Anti-toxins Serotherapy (diphteria) 1901 Robert Koch Tuberculosis, anthrax Cellular immunity, tuberculin reaction 1905 1883 1900 Elie Mecsnyikov Paul Ehrlich Phagocytosis, inflammation Cellular protection Side chain theory 1908 1902 Charles Richet (Paul Portier) Anaphylaxis 1913 1894 Jules Bordet Complement Antibodies/bacteriolysis 1919 Karl Landsteiner A/B/0 blood groups - serology 1930 1940 Max Theiler Vaccine against yellow fever 1951 Daniel Bovet Anti-histamines, treatment of allergy 1957

MILE STONES OF IMMUNOLOGY RESEARCH II. 1944 Peter Medawar Macfarlane Burnet Acquired tolerance Clonal selection theory 1960 1959 Rodney Porter Gerald Edelman Antibody structure 1972 Rosalyn Yalow Roger Guillemin Andrew Schally Radioimmunoassay Peptide hormon production in brain 1977 1958 Baruj Benacerraf Jean Dausset, George Snell Histocompatibility antigens 1980 1975 George Köhler Cesar Milstein Niels Jerne Monoclonal antibody Network theory 1984 1979 Susumi Tonegawa Gene rearrangement 1987 E. Donnall Thomas Joseph Murray Transplantation immunology 1990 1974 Rolf Zinkernagel, Peter Doherty MHC restriction 1996

GENERAL CHARACTERIZATION OF THE IMMUNE SYSTEM

GENERAL FEATURES OF THE IMMUNE SYSTEM STRUCTURE – various cell types, diffuse Cell communication Partners Mode – direct – soluble factors Cell – to – cell communication Th macrophage extracellular matrix Adhesion Homing Migration B pathogen macrophage 2. ACTION – dynamic Homeostasis – environmental factors Replacement vs death Activation vs differentiation neutrophil Inflammed tissue Endothelial cell 3. FUNCTION Defense against pathogens Recognize, prevent spread, clear from the body Protection of self 4. SPECIAL FEATURES Recognition – self - antigen - danger Signal processing and transduction Signal storage – learning, memory SIMILARITIES TO THE NERVOUS SYSTEM

WHY IS THE IMMUNE SYSTEM SO IMPORTANT? Species have been evolved in the presence of pathogens Bacteria PATHOGENS Virus 3 hours Viruses DIVERSITY VARIABILITY Multicellular parazites (helminths) Monocellular parazites Cells of human body: 90% microbes, 10% human 1012 (1.5kg) bacteria in the gut Human population: 7x109 (7 billion) Biomass: 90% microbes Animal mass < 5 – 25x microbes

TWO LINES OF IMMUNE DEFENSE TWO TYPES OF IMMUNE RESPONSES ACQUIRED/ADAPTIVE IMMUNITY Requires the activation and clonal expansion of cell to protect against pathogens INNATE/NATURAL IMMUNITY Innate immunity constitutes those components that protect against infection without any requirement for prior activation or clonal expansion First line of defense Inherited It is always present Rapid response Short term protection

PHYSICAL BARRIERS PROTECTING OUR BODY FROM HAIR SKIN NAILS PHYSICAL BARRIERS PROTECTING OUR BODY FROM THE ENVIRONMENT GASTROINTESTINAL SYSTEM Sinuses Trachea Lungs BRONCHIAL TRACT EYES Oral cavity esophagus WALDEYER RING Tonsils, adenoids Palatinal, pharyngeal lingual and tubar tonsils Stomach Intestines Kidney Bladder Vagina UROGENITAL SYSTEM Damage Infection Mucus glycoproteins, proteoglycanes, enzymes

EPITELIAL SURFACES ARE IMPORTANT IN THE FIRST LINE OF DEFENSE

DEFENSE LINES OF NATURAL IMMUNITY ANATOMIC BORDERS Skin Inhibits entry of pathogens, pH3 – 5 inhibits growth Mucosa Normal bacterial flora competes for binding sites and nutrients Mucus keeps away pathogens from the surface Cilia remove pathogens PHYSIOLOGICAL BORDERS Temperature Physiological body temperature and fever inhibits growth of certain pathogens Low pH Most pathogens are destroyed in the stomach Chemical Lysosym degrades bacterial cell wall Type I interferons induce anti-viral resistance The complement system is able to lyse bacteria and promotes phagocytosis PHAGOCYTOSIS/ENDOCYTOSIS Many cells can take up microorganisms by receptor-mediated internalization Special professional phagocytes (monocyte, neutrophil, macrophage) are able to internalize, kill and degrade microorganisms INFLAMMATION Tissue damage and infection results in the leakage of anti-bacterial proteins and peptides to the affected tissue Phagocytic cells leave the blood stream and enter inflammed tissues

PHAGOCYTES ARE ABLE TO RECOGNIZE PATHOGENS Toll receptor CR3 (LPS) Toll receptor (fungi) PHAGOCYTES (macrophages, dendritic cells, neutrophil granulocytes) RECOGNIZE PATHOGENS BY PATTERN RECOGNITION RECEPTORS RECOGNITION IS UNAVOIDABLE Macrophage, dendritic cell – ACT AS TISSUE SENSORS Neutrophil granulocytes – MIGRATE FROM THE BLOOD TO THE SITE OF INFLAMMATION

WHAT IS RECOGNIZED BY INNATE AND ACQUIRED IMMUNITY WHAT IS RECOGNIZED BY INNATE AND ACQUIRED IMMUNITY? HOW DO THEY RECOGNIZE PATHOGENS? RECEPTORS Common pattern of groups of pathogens Pathogen Associated Molecular Pattern PAMP Recognition by receptors Pattern Recognition Receptor PRR Unique structural elements Antigenic determinant Recognition by highly specific antigen receptors B cell receptor BCR (sIg) T cell receptor TCR 9-13 various Toll-receptors TLR family Several millions antigen receptors Innate immunity Ancient 450 million years Acquired immunity

TOLL RECEPTORS RECOGNIZE VARIOUS MICROBIAL STRUCTURES Bacteria CpG DNA Virus TLR3 dsRNA TLR9 TLR7 TLR8 ssRNS TLR2 Peptidoglycane Gram+ TLR5 Flagellin TLR4 LPS TLR6 Gram- Interferon producing cell pDC IFN Macrophage/Dendritic cell ALL STRUCTURES ARE ESSENTIAL FOR THE SURVIVAL OR REPLICATION OF THE PATHOGEN

GLYCOSYLATION OF PROTEINS IS DIFFERENT IN VARIOUS SPECIES Eukariotic cells Prokariotic cells Mannose Galactose Glucoseamin Mannose Neuraminidase

PATTERN RECOGNITION BY MANNAN BINDING LECTIN Bacterium lysis Complement activation Macrophage Phagocytosis CR3 LECTIN PATHWAY Strong binding No binding

IL- 6 THE ACUTE PHASE RESPONSE Mannose binding lectin/protein MBL/MBP Complement C-reactive protein Complement Liver Serum Amyloid Protein (SAP) Phagocytosis, ECM stability Fibrinogen IL-6 induces the production of acute phase protiens

PHAGOCYTOSIS MACROPHAGES ACTIVATE OTHER MECHANISMS OF INNATE IMMUNITY Macrophages ingest and degrade particulate antigens through the use of long pseudopodia that bind and engulf bacteria. The engulfed bacteria are degraded when the phagosome fuses with a vesicle containing proteolytic enzymes (lysosome), forming the phagolysosome. Specialized compartments also exist in the macrophage to promote antigen processing for presentation to antigen-specific T cells. MACROPHAGES ACTIVATE OTHER MECHANISMS OF INNATE IMMUNITY

CELLULAR AND HUMORAL MECHANISMS OF INNATE IMMUNITY Phagocytosis Intracellular killing PHAGOCYTOSIS Phagocyte Bacterium Bacterium Complement proteins Lysis of bacteria Inflammation Complement-dependent phagocytosis COMPLEMENT INFLAMMATION Bacterium LPS Cytokines Neutrophil NK-cell Macrophage TNF IL-12 IFN NK-CELLS Virus-infected cell NK-cell Lysis of infected cell

TWO LINES OF IMMUNE DEFENSE TWO TYPES OF IMMUNE RESPONSES INNATE/NATURAL IMMUNITY ACQUIRED/ADAPTIVE IMMUNITY Phagocytes (monocyte/macrophage, neutrophil, dendritic cell) Killer cells (NK cell, δ T cell) B1 lymphocytes (CD5+) CELLS HUMORAL FACTORS B lymphocytes (B2) T lymphocytes helper T cell cytotoxic T cell Enzymes (lysozyme,transferrin, lactoferrin, spermin, trypsin) Antibacterial peptides Complement system Cytokines, chemokines Antibodies

Rapid, prompt response (hours) No variable receptors NATURAL/INNATE Rapid, prompt response (hours) No variable receptors Limited number of specificities No improvement during the response No memory Not transferable Can be exhausted, saturated CHARACTERISTICS OF INNATE IMMUNITY ADAPTIVE/ACQUIRED Time consuming Variable antigen receptors Many very selective specificities Efficacy is improving during the response Memory Can be transferred Regulated, limited Protects self tissues COMMON EFFECTOR MECHANISMS FOR THE ELIMINATION OF PATHOGENS

TWO LINES OF IMMUNE DEFENSE INNATE/NATURAL IMMUNITY Innate immunity constitutes components that protect against infection without any requirement for prior activation or clonal expansion ACQUIRED/ADAPTIVE IMMUNITY Requires the activation and clonal expansion of cells to protect against pathogens Induced by antigen Response is under genetic control Depends on environmental stimuli First line of defense Inherited Always present

FUNCTIONAL ATTRIBUTES OF INNATE AND ADAPTIVE IMMUNITY

The innate immune response causes inflammation at sites of infection Chapter 1 Elements of the Immune System and their Roles in Defense The innate immune response causes inflammation at sites of infection The adaptive immune response adds to an ongoing innate immune response Potent immune responses require the collaboration of innate and adaptive immune responses © Garland Science 2009 29

FIRST LINE OF DEFENSE BY INNATE IMMUNITY EPITHELIAL CELLS Pattern recognition receptors (PRR) Cytokine, chemokine secretion NEUTROPHIL GRANULOCYTES Phagocytosis Intracellular cytotoxicity MONOCYTE – MACROPHAGE – DENDRITIC CELL NETWORK Internalizing receptors NATURAL KILLER CELLS Cytoxicity Cytokine production

CELLS & MECHANISMS OF INNATE IMMUNITY Soluble proteins – Defensins Enzymes - Complement system - Chemotaxis Recognition by Pattern Recognition Receptors (PRR) Macrophage & dendritic cell subsets Neutrophils Pro-inflammatory and inflammatory cytokine secretion Local effects Systemic effects Chemokine receptors & ligands – cell recruitment, other functions Cytotoxicity – NK cells