OUTLINE

Introduction Mediated Neutralization Cytolytic/Cytotoxic Immune Complex Anaphylactic

Cell Mediated T-cell Cytotoxic (Killer T-cells) Delayed Hypersensitivity

Antibody or Cell Mediated Granulomatous Reactions IMMUNE EFFECTOR MECHANISMS LEVELS OF REACTIONS OF ANTIGEN WITH ANTIBODY OR CELLS

PRIMARY SECONDARY TERTIARY REACTION REACTION REACTION in vitro in vivo ANTIBODY MEDIATED INACTIVATION NEUTRALIZATION

AGGLUTINATION, LYSIS CYTOLYTIC REACTIONS OPSONIZATION Ag+Ab AgAb IMMUNE COMPLEX PRECIPITATION REACTIONS

MAST CELL ANAPHYLACTIC DEGRANULATION REACTIONS

CELL MEDIATED DELAYED T-DTH HYPERSENSITIVITY LYMPHOKINES REACTIONS +Ag-> MACROPHAGE ACTIVATION

BLAST CELL TRANSFORMATION T-CTL TARGET-CELL LYSIS DESTRUCTION

Ab or + INSOLUBLE ANTIGEN GRANULOMA NEUTRALIZATION/INACTIVATION REACTIONS

PROTECTIVE REACTIONS

TOXIN NEUTRALIZATION DIPHTHERIA,, TETANUS, ANTHRAX, CHOLERA

RECEPTOR BLOCKADE PERTUSSIS VIRUSES MEASLES. FLU, POLIO, HEPATITIS, PAPILLOMA, ETC.

PASSIVE ANTIBODY TREATMENT – CHOLERA, EBOLA VIRUS

DESTRUCTIVE REACTIONS

DIABETES, HEMOPHILIA, APLASTIC ANEMIA, MYASTHENIA GRAVIS, GRAVE’S DISEASE, BULLOUS SKIN DISEASES EVIDENCE FOR NEUTRILIZING

DICK AND SCHICK TESTS

PEOPLE WHO RECOVER FROM STREPTOCOCCAL OR DIPHTHERIA INFECTION HAVE NO REACTION TO SKIN INJECTION OF TOXIN

DUE TO PRODUCTION OF NEUTRALIZING ANTIBODY SCARLET FEVER GROUP A STREPTOCOCCUS TOXIN

Sequella: Glomerulonephritis; Rheumatic heart disease (IMMUNE COMPLEX REACTION)

VACCINE UNDER DEVELOPMENT TREATED WITH ANTIBOTICS PENICILLIN OR AMOXICILLIN Dale JB, et al, Current approaches to Group A streptoccal Vaccine development. In Streptoccus pyogenes ED. Ferretti, Steverns, Fichetti. U. Oklahoma Press, 2016. DICK TEST FOR SUSCEPTIBILITY TO SCARLET FEVER 1924 GEORGE AND GLADYS DICK

INJECTION OF 0.1 CC OF SCARLET FEVER TOXIN INTO THE SKIN:

INFLAMMATORY REACTION OF 10MM WITHIN 24 HOURS INDICATES LACK OF IMMUNITY.

STAPH STREP TOXIN

NO REACTION -- INDICATES IMMUNITY ANTIBODY NEUTRALIZES TOXIN Schick Test Diphtheria Toxin Bela Schick New York 1925 0.1 ml of diphtheria toxin injected into skin of one arm 0.1 ml of heat inactivated toxin into the other

Individuals with toxin neutralizing antibodies will have no reaction at either site;

Those without will have reaction at toxin site, but not inactivated site. VACCINES INACTIVATION / NEUTRALIZATION

TOXIN NEUTRALIZATION BACTERIA TOXINS TETANUS DIPHTHERIA PERTUSSIS CHOLERA ANTHRAX

RECEPTOR BLOCKADE VIRUSES MEASLES FLU EBOLA HEPATITIS HERPES POLIO PAPILLOMA ETC.

DPT DIPHTHERIA - LARYNGEAL MEMBRANE PNEUMONIA TOXIN – Enters cell through receptors (EGF) and blocks protein synthesis

VACCINE – HEAT INACTIVATED TOXIN PERTUSSIS WHOOPING COUGH - TOXIN AND RECEPTOR

PERTUSSIS VACCINE Acellular pertussis antigens [10 µg detoxified pertussis toxin (PT), 5 µg filamentous (FHA), 3 µg pertactin, and 5 µg fimbriae types 2 and 3 (FIM)]. TETANUS

LOCKJAW

NEONATALTETANUS OPISTHOTONOS OPISTHO – BACKWARD; TONO - TENSION PROPHYLACTIC IMMUNIZATION USING TETANUS TOXOID

BACTERIA LIVE IN DEAD TISSUE

TOXIN BLOCKS INHIBITORY NEURONS WHICH MODULATE ACTIVITY OF STIMULATORY NEURONS AND PREVENT THE CONTINUOUS MUSCLE CONTRACTION OF TETANUS

ANTIBODY TO TETANUS TOXIN PREVENTS UPTAKE FROM DEAD TISSUES TO LIVING NEURONS CHOLERA

JOHN SNOW LONDON 1854

FIRST APPLICATION OF PUBLIC HEALTH TO PREVENT DISEASE

GIVEN ORALLY RECEPTOR AND TOXIN STIMULATES IgA ANTIBODY ANTHRAX VACCINE 1881 LOUIS PASTEUR

Mab TREATMENT VACCINATE COWS “HERD IMMUNITY” PREVENTS HUMAN INFECTION

ANTIBODY BLOCKS AT MULTIPLE STAGES OF PROCESS

BINDING OF TOXIN TO RECEPTOR

ASSEMBLY OF TOXIN COMPONENTS

PORE FORMATION

ENDOCYTOSIS

TOXIN TRANSLOCATION

Froude JS, Thullier P, Pelat T. Antibodies against anthrax: mechanism of action and clinical applications. Toxins 3:1433-1452, 2011 HUANIZED MONOCLONAL ANTIBODY VIRUS VACCINATION PREVENTION MOSTLY BY RECEPTOR BLOCKADE*

Measles Rabies Mumps Hepatitis Rubella Chaga disease Varicella Yellow Fever Influenza Japanese Encephalitis Coronavirus Dengue Polio West Nile Zika HIV Rotavirus Ebola Etc.

* ALSO T-CELL CYTOXICITY TO BE PRESENTED LATER Virus

Receptor Blockade MMRV VACCINE MEASLES, MUMPS, RUBELLA, VARICELLA ATTENUATED LIVE VIRUS VARICELLA TARGET MUCOSA OF RESPIRATORY TRACT MEASLES, MUMPS , REBULLA VIRUS INFECTS MUCOSA Vaccine induced IgA antibody blocks protein receptor

IgAantibody

However, with direct contact with skin some skin infections ( ie. Smallpox) are not blocked by antibody and require T-CTL (more later)

DISEASE RETURNS WHEN FAMLIES REFUSE VACINE

POLIO VACCINES POLIO BOTH IgG and IgA ANTIBODY

INCLUDES INFLUENZA, CORONAVIRUS PATHOGENESIS OF ACUTE INFLUENZA PNEUMONIA H&E-stained section of the lung from a 1918 influenza victim showing necrotizing bronchiolitis. There is necrosis of the bronchiolar wall, with submucosal edema and vascular congestion. The epithelial layer is desquamating, and necrotic epithelial cells are present in the lumen. A mixed inflammatory cell infiltrate is present throughout (original magnification400×). Taubenberger JK, Morens DM. The pathology of influenza virus infections. Annu. Rev. Pathol. Mech. Dis. 3:499-522, 2008. INFLUENZA VACCINES THREE LEVELS OF PROTECTIVE IMMUNITY

1. ANTI-VIRUS 2. ANTI-CELL MEMBRANE 3. T-CTL

T-CTL TO BE PRESENTED IN DETAIL IN NEXT LECTURE INFLUENZA VACCINE

Why is a “new” vaccine required every year?

Antigenic drift

Mutation produces “new” surface HA (hemaglutinin) INFLUENZA VACCINE KILLED OR WEAKENED VIRUS

HEMAGGLUTININ ANTIGENIC COMPONENTS OF HEMGGLUTININ CHANGE EVERY FLU SEASON, NEW VARIENTS ARE SELECTED FOR EACH YEAR,

•egg-based flu vaccine, •cell-based flu vaccine, and •recombinant flu vaccine.

Most common

Egg-based vaccine manufacturing is used to make both inactivated (killed) vaccine (usually called the “flu shot”) and live attenuated (weakened) vaccine (usually called the “nasal spray” flu vaccine MUTIPLE IMMUNE MECHANISMS IN RESPONSE TO FLU VIRUS

IgA BLOCKING AB IgG NEUTRALIZING AB

CYTOTOXIC T-CELLS DTH T-CELL More later

SARS CoV-2 POTENTIAL IMMUNOGENS CORNAVIRUS VACCINE MODERNA ASTRZENKA mRNA-1273 Spike protein

DA approved for Phase 3 clinical trials HUMAN PAPPILOMA VIRUS AND HUMAN PAPILLOMA VIRUS ANTIBODY AND T-CTL

HIV NO VACCINE - INFECTS CD 4 HELPER T-CELLS

AZT AZIDOTHYMIDINE THYMIDINE ANALOG BLOCKS REVERSE TRANSCRIPTASE VACCINATION PREVENTION BY RECEPTOR BLOCKADE

H. Influenza HIV Meningococcus Rabies Pneumococcus Hepatitis Viral Exanthems Polio Smallpox Chagas disease Measles Yellow Fever Rubella Japanese Encephalitis Varicella Dengue Influenza West Nile Mumps Zika Ebola Typhoid Rotavirus MONOCLONAL ANTIBODIES

TREATMENT WITH INACTIVATING ANTIBODIES PASSIVE TRANSFER OF MONOCLONAL ANTIBODIES FOR

ANTHRAX

EBOLA SNAKE VENOM EBOLA VIRUS

Rial P, Elias SC. et al. (28 Co-authors) Alan R. Townsend. Therapeutic monoclonal antibodies for Ebola virus infection derived from vaccinated humans. Cell Reports 27:172-`86, 2019 NY times August 13, 2019

2018/19 Epidemic

%died

Untreated 70

Anti-viral 33 (Giliad)

Z-Mapp 24 (Mapp Biopharm.

REGN-EB3 6 (Regeneron) mAb-114 (Ridgeback Biother.) 11 ANTI-VENOM TREATMENT

Kills 100,000 annually and permanently disfigures >300,000

• May 16, 2019 AVI (International AIDS Vaccine Initative)and the Liverpool School of Tropical Medicine (LSTM) have formed a research consortium to apply antibody discovery technologies to develop affordable, accessible, and effective monoclonal antibodies (mAbs) for snakebite treatment. The consortium, the Scientific Research Partnership for Neglected Tropical Snakebite (SRPNTS), also includes • The Nigeria Snakebite Research & Intervention Centre (Bayero University, Kano), • The Kenya Snakebite Research & Intervention Centre (Institute of Primate Research, Nairobi), • The Indian Institute of Science (Bangalore), • The Scripps Research (La Jolla).

• The consortium is funded with UK aid from the UK government through the Department for International Development (DFID). NEUTRALIZATION DISEASES

ANTIBODIES BLOCK NORMAL BIOLOGICALLY ACTIVE MOLECULES OR RECEPTORS

DIABETES INSULIN, INSULIN RECEPTOR ISLET CELLS

HEMOPHILIA BLOOD CLOTTING FACTORS

PERNICIOUS ANEMIA PARIETAL CELLS

APLASTIC ANEMIA ERYTHROID TANSCRIPTION FACTOR

MYASTHENIA GRAVIS ACTEYLCHOLINE RECEPTOR

GRAVE’S DISEASE HYPERTHYROIDISM THYROID HORMONE RECEPTOR LATS

BULLOUS SKIN DISEASES EPIDERMAL CELLS/BASEMENT MEMB. IMMUNE FACTORS IN DIABETES

. NORMAL ISLET LYMPHOCYTIC INFILTRATE

HYLINIZED ISLET IMMUNOLABELING TISSUE TEST FOR ISLET CELL AUTOANTIBODIES 1. MICROSCOPIC SLIDE OF PANCREAS 2. ADD PATIENT’S SERUM, INCUBATE AND WASH 3. ADD PEROXIDASE LABELED ANTI-HUMAN IgG

ROLE OF ANTI-ISLET CELL ANTIBODIES NOT CLEAR

INFILTRATING LYMPHOCYTES ARE CD 8 (AUTO-REACTIVE CYTOTOXIC T-CELLS)

La Torre D, Lenmark A. of beta-cell destruction. Adv. Exp. Med. Biold2010:654:537-583 doi: 10.1007/978-90-481-3271-3_24. Veld, PI. Insulinitis inhuman type 1 diabetes. Islets 3:131-138, 2011 soi: 10.4616/isl.3.4.15728 INSULIN ANTIBODIES

INSULIN- NOBLE PRIZE FOR DISCOVERY – 1923 Frederick Banting and John Macleod

ANTIBODIES TO EXOGENOUS ADMINISTERED INSULIN ARE COMMON WITH TREATMENT

USUALLY IgG – SEVERE INSULIN RESISTENCE (AUTOIMMUNE HYPOGLYCEMIA) ALSO IgE – INSULIN ALLERGY

HIGHER FREQUENCY WITH BEEF OR PORK INSULIN, MUCH LESS WITH RECOMBINANT HUMAN INSULIN

MEASURED BY RIA (NOBLE PRIZE IN 1977; Rosalyn Yalow)

TREATMENT – SWITCH INSULIN SOURCES GLUCOCORTICOIDS PERNICIOUS ANEMIA – Megaloblastic Anemia

Failure to absorb Vitamin B12 (poor diet, gastrectomy, H. pylori infection, congenital deficiency of intrinsic factor)

Immune Atrophic gastritis Auto antibodies to parietal cells

NORMAL STOMACH AUTOIMMUNE ATROPHIC GASTRITIS ANTIBODIES TO PARIENTAL CELLS HEMOPHILIA HEREDITARY Congenital lack of a blood clotting factor, most often Factor VIII Bleed into joints (hemarthrosis) ACQUIRED AUTOANTI-FACTOR 8 (Post-partum, cancer, infections, etc.)

FACTOR VIII EPITOPES

Oh, J, Lim, Y, Jang MJ, Huh JY, Shima M, Oh D. Characterization of anti-factor VIII antibody in a patient with acquired hemophilia A. Blood Res 48:58-62, 2013 MYASTHENIA GRAVIS - SEVERE MUSCLE WEAKNESS

EXOPTHALMOS – HYPERTHYROIDISM (GRAVE’S DISEASE) AUTOANTIBODY MIMICS EFFECT OF THYROID STIMULATING HORMONE (TSH) ACTIVATIOIN LATS – LONG ACTING THYROID STIMULATOR NORMAL THYROID GRAVE’S DISEASE

BLISTERING SKIN DISEASES Normal skin Epidermolysis bulosa acqusita

EPIDERMOLYIS BULLOSA AQUISITA

AQUIRED SKIN BLISTERS

SEPARATION OF EPIDERMIS SERUM FROM PATIENT FROM DERMIS AT BASEMENT BINDS TO BASEMENT MEMBRANE MEMBRANE OF NORMAL SKIN PEMPHIGUS ANTIBODY TO SKIN EPITHELIAL CELLS

NEUTRALIZATION/INACTIVATION REACTIONS

PROTECTIVE REACTIONS

TOXIN NEUTRALIZATION TETANUS, DICK TEST, DIPHTHERIA, PERTUSSIS, ANTHRAX, CHOLERA

RECEPTOR BLOCKADE VIRUSES MEASLES. FLU, POLIO, HEPATITIS, PAPILLOMA, ETC.

PASSIVE ANTIBODY TREATMENT – CHOLERA, EBOLA VIRUS

DESTRUCTIVE REACTIONS

DIABETES, POMPE’S DISEASE, HEMOPHILIA, APLASTIC ANEMIA, MYASTHENIA GRAVIS, GRAVE’S DISEASE, BULLOUS SKIN DISEASES CYTOLYTIC REACTIONS

PROTECTIVE REACTIONS

LYSIS AND PHAGOCYTOSIS OF VIRUSES AND BACTERIA

DESTRUCTIVE REACTIONS

LYSIS AND PHAGOCYTOSIS OF BLOOD CELLS

) ANTIBODY DIRECTED COMPLEMEMT MEDIATD LYSIS OF BACTERIA PNEUMOVAX STREPTOCCUS PNEUMONIA COMPLEMENT SYSTEM - CLEAVAGES AND AGGREGATIONS

Immune complex reactions CYTOLYTIC REACTIONS BLOOD CELLS

RED CELLS HEMOLYTIC ANEMIA HEMOLYTIC DISEASE OF NEWBORN TRANSFUSION REACTIONS

PLATELETS IDIOPATHIC THROMBO-CYTOPENIC PURPURA

WHITE BLOOD CELLS (PMNS) AGRANULOCYTOSIS - INFECTIONS NORMAL RBC + ANTIBODY TO RBC +C

AGGLUTINATION LYSIS

HEMAGGLUTINATION AND LYSIS TRANSFUSION REACTION

+ COMPLEMENT

ERYTHROBLASTOSIS FETALIS

ERYTHRO- BLASTOSIS FETALIS

BLOOD FORMING CELLS IN LIVER COOMB’S TEST

TEST FOR ANTIBODY COATED CELLS

TEST FOR ANTIBODIES

NEUTROPENIA NEXT LECTURE PURPURA IN INDIOPATHIC THRMBOCYTOPENIC PURPURA

CYTOLYTIC REACTIONS BLOOD CELLS

RED CELLS HEMOLYTIC ANEMIA HEMOLYTIC DISEASE OF NEWBORN TRANSFUSION REACTIONS

PLATELETS IDIOPATHIC THROMBO-CYTOPENIC PURPURA

WHITE BLOOD CELLS (PMNS) AGRANULOCYTOSIS - INFECTIONS

) IMMUNE COMPLEX REACTIONS

ARTHUS REACTION

SERUM SICKNESS

LEUKOCLASTIC VASCULITIS

POLYARTERITIS NODOSA

GLOMERULONEPHRITIS

RHEUMATOID ARTHRITIS

SCLERODERMA (PREGRESSIVE SYSTEMIC SCLEROSIS)

COLLAGEN DISEASES