hans-martin jäck division of molecular immunology dept. of internal medicine iii...

Hans-Martin JäckDivision of Molecular ImmunologyDept. Of Internal Medicine IIINikolaus-Fiebiger-CenterUniversity of Erlangen-Nürnberg

History of Immunology

Part 3: START OF IMMUNOLOGY

Core Module ImmunologyDoctoral Training Group GK1660Erlangen 2011

Doctoral Training Group GK1660 - University of Erlangen-Nürnberg 2

TOPICS - History of Immunology

Repeat: Discovery of cells and germs (1683 - 1876)

Repeat: Prevention of Infection (1840 – today)

Start of Immunology (1796-1910)

Immunochemistry - The antibody problem (1910 - 1975)

Self-/non-self discrimination (1940 – today)

Models to explain antibody diversity (1897 and 1950s)

Discovery of B and T cells (1960s)

The molecular revolution (1974 – today)

DISCOVERY OF CELLS & GERMS(brief repeat from last lecture)

(1683 - 1876)


Germ Theory

1683 Leeuwenhoek described in a letter to the Royal Society very likely bacteria (types of animalcules) in the saliva and tooth scrapings from his mouth.

1859 Louis Pasteur – disproves spontaneous generation of cells from decayed organic matter → Germ Theory: Infectious disease are caused by germs

~1860 Louis Pasteur - Fermentation process and souring of wine is caused by the growth of different microorganisms

1876 Robert Koch isolates the first disease-causing pathogen (anthrax) and provides definitive proof of the germ theory

1884 Koch’s postulates to classify infectious agent

PREVENTION OF INFECTION


Preventions of Infections

DESINFECTION – External1840s Ignaz Semmelweis - hand washing prevents childbirth fever

1867 Joseph Lister - carbol to treat wounds for surgery

“DESINFECTION” – Internal (antimicrobial compounds)1881 E. v. Behring - unsuccessful attempts to treat infections with chemicals

1909 P. Ehrlich - First organic compound to treat syphilis (Salvarsan)

1929 Fleming - Penicillin

1935 Domagk - Sulfonamides

PREVENTIVE VACCINATION1796 Jenner - Cow pox vaccination

1880 Pasteur - cholera vaccination in chicken – generalization of Jenner’s use of cow pox vaccine. Pasteur introduces general term vaccination

1886 Pasteur - rabies vaccination


IMMUNITY – Timeline 2

Discovery of cells and germs (1683 - 1876)

Prevention of Infection (1840 – today)

Start of Immunology (1796 or 1890?? -1910)







Immunity - Etymology

Late 14c. "exempt from service or obligation" o from Latin immunitatem (nom. immunitas) "exemption

from performing public service or charge (tax)o or from Latin “immunis "exempt, free," from in - "not" +

munis "performing services“

1775 The term “Immunitas” was first used by Van Sweiten, a Dutch physician, to describe the effects induced by an early attempt at variolization.

1879 "protection from disease“o Pasteur, Koch, Ehrlich ?????

• http://www.etymonline.com/index.php?term=immunity

• Yufang Shi, Ph.D., Introduction and History of ImmunologyUMDNJ-Robert Wood Johnson Medical School


Phase I: Phenomenon: Immunity(500 B.C. - 1796)

Phase II: Introduction of Vaccination & Immunochemistry (1860 – 1945)

Phase III: Identification of cellular and molecular components (1945 – today)

IMMUNOLOGY – Timeline

START OF IMMUNOLOGY


1. Preventive Immunizationo Jenner (1789) - 1. designed immunization (1798)o Pasteur (1880) – chicken cholera generalized Jenner‘s small pox approach

2. Cellular Immunityo Methnikoff (1884) - discovers phagocytic activity

3. Humoral Immunity & Serotheraphy o Bering (1890/91) – Tetanus/Diphtheriao Ehrlich‘s Sidechain Theory (1897)

4. Cytotoxic antibodies und complemento Bordet (1899): substance sensibilisatrice + Buchner‘s Alexino Ehrlich (1899): Amboreceptor + Komplement

5. Serodiagnostic (Start of Serology)o Bordet (1901) - Complement fixation testo Wassermann (1905) - Syphilis-Nachweiso Landsteiner (1901) – Blood goups in human

6. Anaphylaxis and Related Disorders (harmless antigens make us sick)o Portier & Richet (1902) - Anaphylaxiso Arthus reaction (1903)o Von Pirquet (1906) - Serum sickness – Allergieo Wolff_Eisner (1906) - Heufiebero Meltzer (1910) - Asthma

TOPCIS: Start of Immunology

Nobel 1908

Nobel 1901Nobel 1908

Nobel 1919

Nobel 1913

Nobel 1930

START OF IMMUNOLOGY

Preventive Vaccination


Small Poxs – Variolation & Vaccination

Variolationo Oriental habit to protect children through intentional infection with lymph

or pustules from a person that recovered from a mild infection of small pox.

o Common practice before vaccination

o Worked if exposed to a weak strain of smallpox

o Wrong treatments could kill or be ineffective. o Strains of small pox differ in mortality rate (1-20%)

Vaccinationo Edward Jenner discovered that cowpox could protect against smallpox

with less complications than variolation.o Louis Pasteur coined the term vaccination (from lat, vacca = cow) as a

general procedure o immunize people against other disease


Ca. 1000 Chinese inoculated children with material from infected people

Ca. 1500 Variolation introduced into Turkish harems

1717 Lady Mary Montagu introduced smallpox inoculation to Europe.

1760 Variolation of the families of Maria Theresia und George III. popularized variolation

1776 Washington began variolating the Continental Army

Small Poxs – Timeline of Variolation

Lady Mary Montagu, wife of the British ambassodour to Turkey,


Country lore (Bauernweisheit)/Obervationo “Milkmaids who caught cowpox from their cows

could not catch smallpox”. o Milk maids infected by cowpox have on their

hands scars very similar to smallpox scars.

Hypothesiso Cowpox infection protects from small pox

The experiment (May 14, 1796) o Infected a boy with the lymph of a cowpox-

infected milkmaid. o On 1st July, Jenner infected (variolated) the

boy with small pox

Result o Boy did not get sick

http

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Cowpox pustule on the hand of the dairymaid Sarah Nelmes

Jenner infects James Phipps, the son of his gardener who had not

yet suffered smallpox with Sarah’s lymph. James became mildly ill

Small Pox – The Jenner Experiment 1796


Rabies & Cholera - Pasteur

Vaccination against rabies (1885) (The case of Johann Meister)

Vaccínation against chicken cholera (1878)

Though Pasteur knew that the vaccine worked, but no one then in the world of science knew how it worked!


aktiv passiv

Indikation Prophylaxe Prophylaxe, Therapie

Gabe von Antigen Antikörper

Gabe wie oft wenige Male immer wieder

Schutzeintritt spät sofort

Schutzdauer lange kurz

Gedächtnis ja nein

Unterschiede: Aktive Immunisierung vs. passive ImmunitätAktive und passive Immunität unterscheiden sich in einigen wesentlichen Punkten:

http://www.ilo.at/text.php?M_ID=58

Protective Vaccination - Summary

START OF IMMUNOLOGY

Cellular Immunity


1. Preventive Immunizationo Jenner (1789)-1. designed immunization (1798)o Pasteur (1880) – chicken cholera generalized Jenner‘s small pox approach


3. Humoral Immunity & Serotheraphyo Bering (1890/91) – Tetanus/Diphtheriao Ehrlich‘s Sidechain Theory (1897)




Nobel 1908


Nobel 1919

Nobel 1913

Nobel 1930

Will be covered by C. Bogdan


START OF IMMUNOLOGY

Serotherapy







Nobel 1908


Nobel 1919

Nobel 1913

Nobel 1930



aktiv passiv

Indikation Prophylaxe Prophylaxe, Therapie

Gabe von Antigen Antikörper

Gabe wie oft wenige Male immer wieder

Schutzeintritt spät sofort

Schutzdauer lange kurz

Gedächtnis ja nein

Unterschiede: Aktive Immunisierung vs. passive ImmunitätAktive und passive Immunität unterscheiden sich in einigen wesentlichen Punkten:

http://www.ilo.at/text.php?M_ID=58

Passive Vaccination - Summary


Characteriticso Transfer of serum, gammaglobulin or monoclonal antibodies from humans or animalso Immediate protection oShort duration (t/2 = 20 days) oSide effects through immune response to foreign proteins (serum disease, anaphylaxy) oContraction of hepatitis or HIV through antibody preparations from human serum

Naturally acquired passive immunityo placental transport of maternal IgG from mother in the fetus througho Transfer if maternal IgA into newborn through milk

Artifically induced passive immunizationo Injection or transfusion of gammaglobulin from other individuals or animalso Treatment of an acute infection (diphtheria, tetanus, rabies, FSME, rubella (Röteln) …)o Toxins (Insects, snake, scorpions, botulinus)oProphylactic before travel to foreign countriesoRhesus factor prophylactic

Vaccination – Passive immunization








Nobel 1908


Nobel 1919

Nobel 1913

Nobel 1930


SIDE VISIT

Bacterial toxins


Exotoxinso released by microorganismso act at the surface of host cells, for

example by binding to receptors

Endotoxinso Intrinsic components of microbial

structureo Trigger e.g., phagocytes to release

cytokines that produce local or systemic symptoms

SIDE VISIT – Bacterial toxins

Janeway, 2011 (8th edition)

microbe

exotoxin

Toxoido Coined by Ehrlich (1908) to describe a derivate of a toxin that still

bound to immunoreceptor but has lost its toxic group (toxophore)Ehrlich P (1908): Über Antigene und Antikörper. Handbuch der Technik und Methodik der Immunitätsforschung: 1-10.


Endotoxins – Structure and function

Immunogenicity Toxicity

Endotoxin is a lipopolysaccharide complex associated with the outer membrane of Gram-negative pathogens (Escherichia coli, Salmonella, Shigella, Pseudomonas, Neisseria, Haemo-philus influenzae, Bordetella pertussis and Vibrio cholerae)

http://www.textbookofbacteriology.net/endotoxin.htm

LPS induces proliferation of mouse B cells via TLR4 (TI1 antigen) → mitogen

LPS activates/induces• Phagocytosis• IL1 (fever)• TNFa (endotoxin-induced shock in

patients severely infected by gram-negative bacteria)


1894 Richard Pfeiffer Endotoxin - Heat stable toxic material from the membrane of Vibrio Cholerae that is released only after the cells are disintegrated.

1933 Andre Boivin and Lydia Mesrobeanu (Pasteur)Re-discover endotoxin and show that the partially purified toxic fraction contains polysaccharides, lipids, and proteins.

1950s Otto Westphal und Otto Lüderitz (MPI Freiburg) Prepare protein-free endotoxin

Ref: 1. JOSEPH E. ALOUF (1987). From “Diphtheritic” Poison to Molecular Toxicology ASM News 53, 1987. p.5472. Reimond Beck, A Chronology of Microbiology in Historical Context (paperback)

Endotoxin – Discovery (LPS)

Richard Pfeiffer(1858- 1945)

Germany

Otto Westphal(1913-2004)

Germany

Otto LüderitzGermany


Exotoxins – (AB toxins)


Diphtheria toxin


Diphteria Toxin – Mechanism of Action

• Diphtheria toxin (about 530 aa) is cleaved by proteolysis in disulfide-linked A and B fragment

• Fragment B facilitates entry into the cell via Heparin-binding EGF-like growth factor (through receptor-mediated endocytosis) as well as the transport of A fragment into the cytosol.

• Fragment A prevents protein synthesis by inactivting eEF2 through transfer of a ADP ribosyl moiety form NAD+ onto the unusual amino acid diphthamide in the eEF2.

NAD+ + peptide diphthamide ↔ nicotinamide + peptide N-(ADP-D-ribosyl)diphthamide


Diphtheria - ADP ribosylates translational elongation factor 2 → cells dies

Pseudomonas - ADP ribosylates translational elongation factor 2 → cells dies

Pertussis - ADP ribosylates adenylate cyclase Gi regulatory protein (blocks e.g., chemokine receptors)

Cholera toxin - ADP ribosylates eucaryotic adenylate cyclase Gs regulatory protein

- increased level of intracellular cAMP, which promotes secretion of fluid and electrolytes in intestinal epithelium leading to diarrhea

Botulinus - Zn++ dependent protease acts on synaptobrevin at motor neuron

- Inhibits acetylycholine release from peripheral cholinergic neurons resulting in flaccid paralysis

Tetanus - Zn++ dependent protease acts on synaptobrevin in CNS

- Inhibits neurotransmitter release from inhibitory neurons in the CNS resulting in spastic paralysis

Exotoxins – More examples AB toxins


Gehört zu einem genetisch veränderten Stamm der harmlosen E. coli-Bakterien

Der Keim kommt vor allem im Darm von Wiederkäuern wie Rindern, Schafen oder Ziegen vor.

Produziert Toxine, die zu wässrigem Durchfall und bis zu blutiger Diarrhoe mit Bauchkrämpfen • Spezielles Hüllenprotein (Adhäsin), das sich an die Epithelzellen der

Darmwand anheftet.

• Ein über Phageninfektion eingeschleustes Gen für das neurotoxische und nekrotisierende Shiga-Toxin oder auch Vero-Toxin (zerstört Vero-Zellen = Affennierenzellen) → Hemmt Proteinsynthese

• Plasmidkodiertes Hämolysin → blutzellenzerstörendes Toxin

(EHEC) – Entero-haemorragic E. coli


BC Homer describes Egyptian disease

~1800 Fidele Bretonneau coins “Diphtheria” for the disease (“Häutchen auf Mandeln) and introduces tracheotomy as ultima ratio in treatment

1883 Edwin Klebs (student of Rudolph Virchow) discovers bacteria in diphtheria patients

1884 Friedrich Löffler • Cultivates and identifies C. diphtheriae as the agent of the disease • hypothesize that infected people die of a toxic bacterial product since the

bacillus does not grow well in infected patients

1888 Roux and Yersin - soluble and filterable toxin in diphtheria cultures causes disease

1890 More than 50,000 children/year die in Germany of diphtheria

1890 Behring reports 1. successful vaccination with weakened C. diphtheriae in guinea pigs

Diphtheria Exotoxin – Timeline


Major Observations

• The filtrates of from old alkaline diphtheria cultures when injected into animals mimicked the symptoms of the natural disease.

• Germ-free urine of infected children contained sufficient toxin to kill guinea pigs.

• “The discovery was serendipity since high calcium tap water led to precipitation of calcium phosphate and, with it, to the lowering of free iron ions in the medium, which, as we know today, is required for optimal toxinogenesis”.

1888 Roux and Yersin discover at the newly founded Pasteur institute the first bacterial protein toxin from diphtheria,→ explains, for the first time, the mechanism of pathogenicity

of a microorganism for humans in terms of a soluble toxic substance.

Diphtheria toxin - Discovery

Emile Roux(1853- 1933)

France

Alexandre Yersin(1863- 1943)

France/Schweiz ALOUF (1987). From “Diphtheritic” Poison to Molecular Toxinology ASM News VOL.53,NO. l0


Bacterial toxins – Summary


SEROTHERAPY


Discovery of inducible soluble immunity

1889 Gameleia (Gamaleia) describes inducible humoral immunity against anthrax• Serum from sheep immunized with attenuated anthrax kills anthrax in vitro• Activity in blood vanishes after one month• But sheep remains immune for much longer time (memory !!!!???)→ soluble activity with short in vitro half-live and Memory Gamelai (1889). Sur la Destruction des Microbes dans les Corps des Animaux.

Febricitants. Ann. Inst. Pasteur, p. 229.

1890 Bouchard shows that bacteria-killing power is greater in blood serum from immunized than from naive animals → Inducible soluble activity

M. BOUCHARD (1890). ACTIONS DES PRODUITS SCREnTtS PAR LBS MICROBES PATHOGiNEsBy. Paris: Gauthier, Villars et File. (Summary published in Nov. 15. 1890. The British Medical JOURNAL. P. 1129

Nice overview in a „News and Views“ to Behring‘s Dec 1890 article about the serum therapy in animals by Hankin, EH (1890). A cure for Tetatus and Diphthria, Nature No 1101, Vol. 43, p. 121


May 1890 von Behring and Nissen show that serum from animals immunized with anthrax killed anthrax in vitro but not bacillus pyocyaneus

→ Inducibilty and Specificity von Behring and Nissen (May 1890), Ueber den bakterienfeindlichen Einfluss von verschiedenen Serumarten, Z. für Hygine vol. viii, p. 412)

Discovery of inducible soluble immunity


Weakened Tetanus

Weakened Tetanus

• Transferred immune serum protects mice against tetanus

• Works also therapeutically in sick mice !!!!!!• No data of experiments with Diphtheria bacillus were reported !!!!!

Shibasaburo Kitasato

(1853 - 1931)Japan

Emil v. Behring(1854- 1917)

Germany

Immune against pathogenic Tetanus

and tetanus toxin

ControlSerumControlSerum

ImmuneSerum

ImmuneSerum

NoTetanus

NoTetanus

10

10

PathogenicTetanus

PathogenicTetanus

20

20

BEHRING und KITASATO, 1890: Ueber das Zustandekom-men der Diphtherie-Immunität und der Tetanus-Immunität bei Thieren. Deutsche Medicinische Wochenschrift, 16. Jahrgang, Nr. 49, 4. December, S. 1113 / 1114.

Serum Therapy - Tetanus (December 1890)


PathogenicDiphtheria


Weakened Diphtheria

(Jodtrichloride)

Weakened Diphtheria

(Jodtrichloride)

• Induced immunity with chemically ‚weakened‘ germs.

• Did not transfer serum of immunized animals (as cited in many text books)

• However, tried “internal” desinfection (chemotherapy) using Jodtrichloride or Peroxide (not convincing)

• 1891 – Successful serum therapy of diphtheria in guinea pigs Behring, Emil. 1891. “Ueber Desinfection am lebenden Organismus.” Deutsche Medicinische Wochenschrift 17(52):1393–1397.


Germany

Non-immunized

Non-immunized

BEHRING 1890: “Untersuchungen über das Zustandekommen der Diphtherie-Immunität bei Thieren.” Deut. Med. Wochenschr. 16(50):1145–1147.

Protection against Diphtheria (December 1890)


• Behring, Emil. 1891. “Ueber Desinfection am lebenden Organismus.” Deutsche Medicinische Wochenschrift 17(52):1393–1397.(Summary of experiment)

• Behring, Emil, and Erich Wernicke. 1892. “Ueber Immunisierung und Heilung von Versuchsthieren bei der Diphtherie.” Zeitschrift für Hygiene und Infections-krankheiten 12:10–44.(Description of experiments)

Weakened (JCL3)

Diphtheria

Weakened (JCL3)

Diphtheria

Immune against pathogenic diphtheria

or diphtheria toxin

ControlSerumControlSerum

ImmuneSerum

ImmuneSerum

NoDiphtheria

NoDiphtheria

10

10

Wernicke, Frosch and Behring



20

20

Serum Therapy – Diphtheria (1891)


p. 1396



Zeitschrift für Hygiene und Infectionskrankheiten (1992). 12:10–44.

p.16

p.12

p.13

p.10

Wernicke, Frosch and Behring

Visibilty of serum therapy in humans

Inducible immunity

Weakening of diphtheria toxin

Sucessful serum therapy of diphtheria



Behring‘s Goals

• “ . . . may also be of use for the treatment of humans suffering from diphtheria or tetanus”. … „Das Blut ist ein ganz besonderer Saft“ (Goethe)Behring (1890): Behring E. A. and Kitasato S. (1890) Uber das Zustandekommen der Diphtherie-Immunitat und der Tetanus-Immunitlt bei Thieren. Dtsch. med. Wochenschr. 49, 1113.

• “The final aim of our experiments remains the production of the substance in such amounts and with such effectivity that humans, too, may be treated for diphtheria with it” Behring (1893): „Behring E. A. (1893). Die Geschichte der Diphtherie, p. 186. Thieme, Leipzig. Behring E. A. (1894)

1. CAUSATIVE TREATMENT OF INFECTIOUS DISEASES

2. PROMOTE FIRST IMMUNOLOGICAL PARADIGM • „Specific immunity induced by antigens is associated with the

formation of soluble antibodies“


1890 More than 50,000 children in Germany die every year of diphtheria.

1890 Behring and Kitasato discover soluble, inducible and transferable anti-tetanus activity in blood of immunized rabbits

1891 Behring successfully repeats serum therapy of diphtheria-infected guinea pigs

1891 Behring apparently treats successfully a diphtheria-diseased child with injection of anti-diphtheria serum produced in horses (report not proven)

1892 Wernicke & Behring improve immunization of rabbits, guinea pigs and goats with weakened diphtheria bacillus

Serum Therapy: From Bench to bed side


Serum Therapy: Status Quo 1892

• 1891 – Successful serum therapy of diphtheria in guinea pigs Behring, Emil. 1891. “Ueber Desinfection am lebenden Organismus.” Deutsche Medicinische Wochenschrift 17(52):1393–1397.

• 1892 – Successful serum therapy in other animals but not yet tested in humansBehring, Emil, and Erich Wernicke. 1892. “Ueber Immunisierung und Heilung von Versuchsthieren bei der Diphtherie.” Zeitschrift für Hygiene und Infectionskrankheiten 12:10–44


1892 Ehrlich & Behring develop procedures to enrich and standardize antitoxins from large animals (goats)

1992 Behring and Ehrlich set-up a lab in Berlin-Steglitz (Stadtbahnbogen, now MPI), where they could obtain large amounts of serum by using large animals – first sheep and later horses.

1892 Industrial production by Hoechst provides anti-diphtheria sera from 20 sheep for 1st clinical trial in Berlin (published by Behring and Ehrlich in 1894)



http://en.wikisource.org/wiki/Index:Popular_Science_Monthly_Volume_46.djvu

Samuel Amstrong (1895). The serum tretament of diphtheria. The popular Science Vol XLVI (46), p. 512-522)


1892 Roux succesfully produces antisera in horse that protects guinea pigs against diphtheria

START OF TRANSLATIONAL IMMUNOLOGY

Use results obtained in experiments withanimals to treat disease

in humans


Serum Therapy: 1st Clinical „Trial“ (Berlin)

• Six hospitals treated 220 patients under der supervision of Behring and Ehrlich with a serum that was standardized by Ehrlich (1893)

• Therefore, all patients received the same effective dose of antiserum

Ehrlich P, Kossel H, Wassermann A (1894): Ueber Gewinnung und Verwendung des Diphterieheilserums. Deutsche medizinische Wochenschrift 20: 353-355.

Summary of all treated patients

Treatment after onset of infection


“Since antitoxin does not neutralize toxin that is already bound to tissues, delaying its administration is associated with an increase in mortality risk. Therefore, the decision to administer diphtheria antitoxin is based on clinical diagnosis, and should not await laboratory confirmation.”

Serum Therapy: Time of application

Atkinson W, Hamborsky J, McIntyre L, Wolfe S, eds. (2007). Diphtheria. in: Epidemiology and Prevention of Vaccine-Preventable Diseases (The Pink Book) (10 ed.). Washington DC: Public Health Foundation. pp. 59–70.


p. 520

Serum Therapy: Clinical „Trial“ (Paris)


1894 After introduction of serum therapy (Roux serum) mortality of diphtheria in Paris falls from 52% to 25%

1894 Production of antitoxins in horses in US (William Hallock Park und Anna Wessels Williams in NYC)

1900 Industrial production of anti-diphtheria sera in US

http://commons.wikimedia.org/wiki/File:AmDruggist1900NoOtherSerum.jpg

22 October 190022 October 1900

Advertisement for anti-diphtheria serum by Parke Davis & Company.

One of the first bottles of diphtheria antitoxin produced at the Hygienic State Laboratory which became the NIH in 1930.

18951895

http://history.nih.gov/exhibits/history/docs/page_03.html



1904 Foundation of „Behringwerke" in Marburg (uses two Million Reichsmark from the Nobel prize)

→ 1. biotech company


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Old vials (1897 and 1906) with hand-written labels.

Behring watches diphtheria immunization of horses in Marburg


Roitt p. 321



Abb. 10 und 11: Nicht anders erging es Emil von Behring der bei seinen Forschungen beginnend mit Meerschweinchen, später erfolgreich das Pferd und in Fortsetzung auch Rinder und Schafe zur Gewinnung des Diphtherieantitoxins heranzog – er wurde als Pferdedoktor belächelt. Behring als Dompteur der Kühe mit Peitsche und Spritze und der Titel Heilserum vom Pferd, beides aus den „Lustigen Blättern 1894“.


1901 Behring uses attenuated diphtheria and successfully immunizes laboratory animals

1913 Behring & Wernicke introduce active vaccination by injection of a safe mixture of diphtheria toxin and antitoxin. Was replaced by active vaccination with inactivated diphtheria toxoid

1924 Gaston Ramon (1886-1963) inactivates tetanus and diphtheria toxins by heat/formalin treatment for active immunizations

Diphtheria – Active Immunizations


1924 Felton serum (Felton antibodies)

29 of 30 pneumonia patients were successfully treated with partially purified antibodies from anti-pneumococcal horse sera

• Felton, L. D. 1926. Bull. Johns Hopkins Hosp. 38:33-60• PARK, William H. (1924). Use of Vaccines and Pneumonia

Antibody in the Treatment and Prevention of Pneumonia and the Use of Convalescent Serum in the Prevention of Measles. Proceedings. Int. Conf. Health Probl. Trop. Amer., Kingston, Jamaica, July 22-August 1, 1924, pp. 834-849

1944 Protection against measlesStokes uses globulin fractions of pooled human plasma for passive protection against measles in 891 individuals

Other successful serum therapies


Problem Antisera were not sufficiently concentrated No Standardization to compare different anti-toxin preparations

Solution Concentration e.g., by ammonium sulfate precipation

(Behring → para-albumin, Morawitz→ pseudo-albumin, today → gamma-globulin)

Higher titers by improving immunization procedures Ehrlich (1892 – 1894)

“Immunitätseinheiten” - Develops a method to standarize anti-toxin preparations by comparig activity of antiserum against a standard anti-toxin serum in an in vivo toxin neutralization test

Coins the term “Titer”: Dilution of anti-toxin that just neutralizes completely a given amount of toxin

SERUM THERAPY – Problem 1

Ehrlich P (1894): Über die Gewinnung, Werthbestimmung und Verwerthung des Diphtherieheilserums. Hygienische Rundschau 4: 1140-1152.


Ehrlich P (1894): Über die Gewinnung, Werthbestimmung und Verwerthung des Diphtherieheilserums. Hygienische Rundschau 4: 1140-1152.

Ehrlich P, Kossel H, Wassermann A (1894): Ueber Gewinnung und Verwendung des Diphterieheilserums. Deutsche medizinische Wochenschrift 20: 353-355.

Ehrlich P, Kossel H (1894): Ueber die Anwendung des Diphtherieantitoxins.Zeitschrift fuer Hygiene und Infektionskrankheiten, medizinische Mikrobiologie, Immunologie und Virologie 17: 486-488.

Ehrlich P (1894): Ueber die Behandlung der Diphtherie mit Heilserum.Verhandlungen der Gesellschaft Deutscher Naturforscher und Aerzte : 402.

Ehrlich P, Wassermann A (1894): Ueber die Gewinnung der Diphterie-Antitoxine aus Blutserum und Milch immunisirter Thiere.Zeitschrift fuer Hygiene und Infektionskrankheiten, medizinische Mikrobiologie, Immunologie und Virologie 18: 239-250.

Serum Therapy: Improvements and Quantitation

START OF IMMUNOLOGY

Anaphylaxis and Related Disorders


SERUM THERAPY – Problem 2

1906 Inflammation (immune repones can cause disease !!!

• Freiherr Clemens von Pirquet (Wiener Kinderarzt) beobachtete bei einigen Diphteriepatienten nach wiederholter Injektion mit Antiseren gegen Diphterie eine Entzündungsreaktion

• erkannte, dass Antikörper nicht nur schützende Immunantworten vermitteln, sondern auch Überempfindlichkeitsreaktionen auslösen können.

• Pirquet führte für diese Serumkrankheit den Begriff Allergie (aus dem Griechischen „die Fremdreaktion“) ein

• Unter Allergie versteht man überschießende Reaktion des Immunsystems auf normaler-weise harmlose, fremde Stoffe


Hypersensitivity –Injection of Antigens

1902 Anaphylaxis, an acute and serious hypersensitivity reaction• Paul Portier and Charles Richet (France) report that some dogs

that had received a sublethal dose of sea anemone die after second subcutaneous injection

• Richet receives Nobel price for Medicine 1913

1903 Arthus reaction• Nicolas Maurice Arthus (a Swiss physician) observes serve local

inflammation in rabbits that had received several injections of harmless substances such as milk and horse serum

• Occurs also with repeated expose of airborne antigens such as fungi

• Cautioned that the same could happen during a serum therapy!!!!


Hypersensitivity - Today








Nobel 1908


Nobel 1919

Nobel 1913

Nobel 1930

Will be covered by S. Finotto



Naive individual can be protected and cured !!!! from diphtheria and tetanus by transfering serum from an immunized animal

► Retter der Kinder (Diphtherie)

► Retter der Soldaten (Tetanus)

1st Nobel prize in Medicine

for serum therapy (1901)


Germany

Serum Therapy: Summary

1st Immunological paradigm:

„Specific immunity induced by antigens is associated with the

formation of antibodies“


Klebs discovers

bacteria on material

from diceased diphtheria

patient

Klebs discovers

bacteria on material

from diceased diphtheria

patient

Behring-werke in Marburg

Behring-werke in Marburg

Behring&Ehrlich (Berlin)

1st serum therapy in humans

Behring&Ehrlich (Berlin)

1st serum therapy in humans

TIMELINE: Serum Therapy of Diphtheria

Löffler identifies C. diphtheriae

as the cause of

diphtheria

Löffler identifies C. diphtheriae

as the cause of

diphtheria

Roux and Yersin idenify soluble

diphtheria toxin

Roux and Yersin idenify soluble

diphtheria toxin

Hoechst(Behring)Industrial

production of antisera in sheep

Hoechst(Behring)Industrial

production of antisera in sheep

Roux develops antisera in horses

Roux develops antisera in horses

Behring1st Serum

therapy (diphtheria) in guinea

pigs

Behring1st Serum

therapy (diphtheria) in guinea

pigs

1883 18941893 190418921884 18911888 1890

Behring & Kitasato1st serum therapy

(tetanus) in mice

Behring & Kitasato1st serum therapy

(tetanus) in mice

Roux & Chaillon (Paris)Serum therapy in humans

Park & Williams (NYC)Production of antisera in

Roux & Chaillon (Paris)Serum therapy in humans

Park & Williams (NYC)Production of antisera in

1924

Safe Active Vacci-nation

Ramon

Safe Active Vacci-nation

Ramon


Serum Therapy: Today

Llewelyn et al. (1992). Monoclonal antibodies in Medicine. BMJ, 305:1269


IgG und Rhesusunverträglichkeit (Landsteiner 1940)

For further explanation, see Kuby, 4th edition, p. 414

• Mutter ist Rh-Vater ist Rh+Baby ist Rh+

• Während 1. Schwanger-schaft werden nur geringe Mengen an anti-Rh-AK produziert (IgM). Jedoch entstehen Gedächtnis-B-Zellen

• Während 2. Schwanger-schaft werden Gedächt-nis-B aktiviert und schnell hochaffine IgG-anti-Rh-AK produziert

• Anti-Rhesusfaktor-AK sind vom IgG-Typ und passie-ren die Plazenta

• Lyse von Erys

Erythroblastosis Fetalis

Hemolytic Disease of the Newborn (HDN)


Übertritt größerer fötaler Blutmengen in den mütterlichen Kreislauf

o Normalerweise nur während der Geburt

o Während der Schwangerschaft bei Amniocentese oder anderen

invasiven Eingriffen

Fehlgeburten

Abtreibung

Schweren inneren Blutungen

Behandlung der Mutter durch Gabe von Rhogam = Anti-D (Rhesusfaktor)-Antikörper vom IgG-Typ)

o Routinemäßig nach 28. Schwangerschaftswoche und kurz nach Geburt

o Spätestens 72 Stunden nach Fehlgeburten, Ab-treibungen, Amniocentese oder anderen invasi-ven Eingriffen

o Zerstören fötale Erys, die in den mütterlichen Kreislauf übertreten, und verhindern so die Aktivierung von B-Zellen und die Bildung von Gedächtnis-B-Zellen

o Nur kleine Mengen an anti-Rh-AK werden verab-reicht, diese werden im Blut der Mutter verdünnt, treten deshalbb nur langsam und in sehr geringen Mengen den fötalen Kreislauf über, zerstören aber effizient fötale Eyrs im Blut der Mutter.

Rhogam

Fötale Erys

Mütterliche Anti-Rh-B-Zelle

Anti-Rh-Antikörper (Rhogam)

Discovery of a inducible, soluble and specific activity in the blood (later termed

„antibodies“) in 1890

Discovery of a inducible, soluble and specific activity in the blood (later termed

„antibodies“) in 1890

The first paradigm in immunology„Specific immunity induced by antigens is

associated with the formation of antibodies“

The first paradigm in immunology„Specific immunity induced by antigens is

associated with the formation of antibodies“

START OF IMMUNOLOGY ??START OF IMMUNOLOGY ??

START OF IMMUNOLOGY

Antitoxins and cytotoxic antibodies





4. Cytotoxic humoral immunity and complemento Bordet (1899): substance sensibilisatrice + Buchner‘s Alexino Ehrlich (1899): Amboreceptor + Komplement

5. Serodiagnostic (Start of Serology)o Widal (1896) – Widal agglutination test for typhoid fevero Bordet (1901) - Complement fixation testo Wassermann (1905) – Syphilis testo Landsteiner (1901) – Blood goups in human


Nobel 1908


Nobel 1919

Nobel 1913

Nobel 1930


ANTITOXINS

Mechanism of action


Mechanisms of antitoxic effect of Behring‘s serum therapy?

Hypothesis 1 :Antitoxins destroys toxin. Disproved since toxins could be detected in toxin/anti-toxin mixtures

Hypothesis 2 (e.g., Roux und Buchner): „Antitoxin soll keine aktive Wirkung auf das Toxin ausüben, sondern in erster Reihe auf die Zellen einwirken und dieselben gewissermassen gegen die Giftwirkung immunisieren“.

Hypothesis 3 (Ehrlich):„Gift und Gegengift paaren in den Gewebsflüssigkeiten zu einer Art Doppelverbindung, welche nicht mehr in bestimmten Geweben fixiert wird und welche daher keine Krankheitserscheinungen mehr auslöst.“

P. Ehrlich (1897). Zur Erkenntniss der Antitoxinwirkung. Fortschritte der Medicin, Bd 15, No 2, p. 41-43

Antitoxins: Mechanism of action (Ehrlich, 1897)


Experiment:

Antitoxins: Mechanism of action (Ehrlich, 1897)

P. Ehrlich (1897). Zur Erkenntniss der Antitoxinwirkung. Fortschritte der Medicin, Bd 15, No 2, p. 41-43

Conclusion→ Cellular explanation of Roux and Buchner (hypothesis 2) disproved

→ First evidence for direct (from mix in vitro) and chemical interference (from titration) of antitoxin with toxin

Ricin

Mix of anti-ricin Serum and ricin before adition to

RBCTubes with blood from un-immunized rabbits

- +

Oberservation→ Anti-ricin toxin prevents ricin (lectin)-mediated clumping of red blood cells in a

concentration dependent manner

Ricin mediates clumbing of RBC + Ricin + + +

CYTOTOXIC HUMORAL IMMUNITY

Bacteriolysins(today:antibody + complement)


Blood plasma o yellow liquid component of blood without blood cells o 55% of the total blood volumeo contains 93% water by volume and dissolved proteins,

glucose, clotting factors, mineral ions, hormones and carbon dioxide

o Prepared by spinning a tube of fresh blood containing an anti-coagulant

Blood serum o blood plasma without fibrinogen or the other clotting factors o Produced by centrifugation of coagulated blood

Blood Plasma and Serum


1888 George Nuttall (US, working in Breslau) reports for the frist that time the bactericidal power of the blood serum from non-immunized animals in the absence of cells (macrophages) • Nuttal, G. (1888) Experimente über die bakterienfeindlichen Einflüsse

des thierischen Körpers, Z. Hygiene vol iv, p 353-393• On the Formation of Specific Anti-Bodies in the Blood, Following

Upon Treatment with the Sera of Different Animals George H. F. Nuttall The American Naturalist. Vol. 35, No. 419 (Nov., 1901), pp. 927-932

1889 Hans Buchner (1850 – 1902, Germany) was the first to demonstrate the presence of a soluble bacteria-killing, heat-labile substance (“alexin”, from the Greek alexô defend”) in normal serum Buchner, H. E. (1889). "Über die bakterientödtende Wirkung des zellenfreien Blutserums (Concerning the Bacteriological Effects of Cell-free Blood Serum), Zbl. Bakt. 5: 817)

Co-discovererd with his younger brother Eduard Buchner (1907 Nobel Prize in Chemistry for cell-free fermentation) the yeast enzyme zymase.

Bacteriolysins: The beginning

Hans Buchner(1850- 1902)

Germany

George Nuttall(1862- 1937)

San Francisco

Buchner‘sAlexin=

Ehrlich‘s complement


1894 Describes a heat labile antibactericidal activity in the blood of immunized animals• Guinea pigs were immunized with cholera bacteria• Blood was collected and added to live cholera cultures

→ Bacteria became motionless• Heating the plasma abolished the effect („destroyed“ complement)

→ Bacteriolysis or Pfeiffer Phenomenon

Other DiscoveriesVaccination of guinea pigs against cholera or typhus (1894)Initiated (in parallel with Almoth Wright) the first successful typhus vaccination (1996)Discovered endotoxin as a heat-stabile toxic activity of bacteriaIsolated in 1892 Haemophilus influenzae, which he thought was the causative agent of influenza. (Peter Olitsky and Frederick Gates, Rockefeller, could not isolate bacteria in influenza patients during the 1918 pandemic)

Bacteriolysin: Coining the name

Richard Pfeiffer(1858- 1945)

GermanyPaul Fildes Biographical Memoirs of Fellows of the Royal Society Vol. 2, (Nov., 1956), pp. 237-247


Jules Bordet (1870-1961)

Belgium

Nobel price Medicine

1919

Bacteriolysin: Labile component (1895)

Bordet verifies Buchner‘s and Pfeiffer‘s obervations of the prescence of a heat-labile factor required for bacterilysis in vitro

Fresh serum from animals immunized with bacteria lysis bacteria in vitro

However, if serum ages, bacteriolytic activity is lost

CYTOTOXIC HUMORAL IMMUNITY

Hemolysins(today:antibody + complement)


Hemolysins

1875 Landois (Greifswald), in his work on transfusion, demonstrates the hemolytic action of animal sera on heterologous erythrocytes, i.e., the ability of a serum from one animal to dissolve red cells from an animal of another species.

1898 Carbone & Belfanti (Italy) show for the first time that red cells could act as antigens and that the serum of an animal species 1 treated with injections of blood from animals of another species had a high toxicity for animal species 1BELFANTI, S. AND CARBONE, T.: Produzione di sostanze tossiche mmcl siero di animale inoculati con sangue eterogeneo. Gior. d.r. Accad. di. med. di Torino, Series 4, 46: 321, 1898.

1898 Bordet showed that the cytotoxic effect of an immune serum was due to two substances contained in the same fluid, one was Buchner’s alexine (Ehrlich’s complement), the other he name substance sensibilisatrice (Ehrlich‘s amboceptor). Bordet, J. 1898. Sur l'agglutination et la dissolution des globules rouges par le serum d'animaux injectes de sang defibrine. Ann. De l'Inst. Pasteur. xii: 688-695.

John M. POLK (1904). A CLINICAL STUDY OF THE HEMOLYTIC ACTION OF HUMAN BLOOD SERUM. Journal of Medical Research. (NEW SERIES, VOLUME VII.) VOL. XII. OCTOBER, 1904. No. 3.

Defintion: The activity in serum to clump and lyse erythrocytes


immuneserum

Lysis requires specific Immune factor + heat-labile factor

Experimental Set-up

Hemolysin – The Bordet Experiment (1898)

Fresh serum from non-immunized animals

rabbit blood

no lysis

Non-immuneserum

Guinea pig

blood no lysis

550C/30 min

specific factor

lysis

lysis

inducible factor

unspecifc heat-labile

factor

No lysis

Bordet, J. 1898. Sur l'agglutination et la dissolution des globules rouges par le serum d'animaux injectes de sang defibrine. Ann. De l'Inst. Pasteur. xii: 688-695.


RBC

Bordet: Alexin „helps“ inducible factor (1898)

ExplanationHemolysin activity of immune serum was due to two substances contained in the same serum:

1. a heat-labile unspecific factor present in blood of non-immunized and immunized animals (Buchner’s/Bordet’s alexin, today Ehrlich’s complement)

2. a heat-stable species-specific factor only present in the immune serum (Bordet’s substance of sensibilisatrice, today Ehrlich‘s antibodies)

Bordet’s mechanism of actionJules Bordet

(1870-1961)Belgium

Nobel price Medicine

1919

Substance Sensibilisatrice(specificity and sensibilsation)

senibilsation

Buchner‘s Alexin(toxic)

Cleary separeted haemolysin activity into an innate

and an adaptive humoral component


Paul Ehrlich(1854-1915)

GermanyNobel price Medicine

1908

Together with J. Morgenroth, Ehrlich verified the presence of the two factors in the immune serum (of a goat) required to lysis red blood cells from a mutton (German: Hammel).

The thermostable and inducible immunebodies were termed “amboreceptors” (today: antibodies).

and the heat-labile component was termed “complement” due to the fact that it “complemented” the activity of the amboreceptors.

Paul Ehrlich: Alexin → Complement (1899)

• Ehrlich & Morgenroth (1899). Über Haemolysie – zweite Mitteilung. Berl. Klin. Wochenschr. Bd. 22, p. 481-486. („Komplement „ mentioned on page 482, left column)

• Ehrlich (1899). Zur Theorie der Lysinwirkung. Berl. Klein. Wochenzeitschr. No. 1, p. 6 (very nice summary of the first two publications)

• Ehrlich, P. & Morgenroth, J. (1900). Ueber Hämolysine – 3. Mitteilung. Berl. Klin. Wochenchr. 37, 453-485


Mechanism of Action

Ehrlich & Morgenroth (1900). Über Haemolysie – vierte Mitteilung. Berl. Klin. Wochenschr. Juli p. 681.

a. Complement

b. Amboreceptor

c. Receptor auf RBC

toxophore

haptophore

Anti-ToxinHemolysin

• Complement

• Zwischenkörper• Immunkörper• Amboreceptor

toxophore

Haptophore 1

Haptophore 2

haptophore

2. lysis

1. recognition

• Amboceptors have bifunctional binding capacity: recognizes the specific antigen AND binds to the heat-labile antimicrobial component of fresh serum. So complement gives the amboreceptor the potential to kill

• In contrast, antitoxins posseses both binding (via haptophore grouop) and killing (via taxophore) activity

Paul Ehrlich: Hemolysin – Mode of Action


Deutsch, L. & Feistmantel, C. Die Impfstoffe und Sera. Thieme, Leipzig, 1903.

Mechanism of Hemolysis: Textbook 1903

Although Ehrlich and Bordet did not agree on the mechanism of action, their work clearly separated the haemolysin activity into an unspecific innate (complement) and an specific adaptive (antibody) component


Lysis - rupturing membranes of foreign cells

Opsonization - enhancing phagocytosis of antigens

Chemotaxis - attracting macrophages and neutrophils

Clumping of antigen-bearing agents

http://en.wikipedia.org/wiki/File:Complement_pathway.png

Complement - Today


Studies by Bordet and Ehrlich clearly separated the haemolysin activity into an innate (complement) and an adaptive (antibody) component

Established that humoral immune bodies (i.e., antibodies) can be cytotoxic

Allowed to establish sensitive tests to detect the presence of antibodies against a germ in the blood of patients – Start of serology (e.g., complement fixation test)

SUMMARY – Bordet‘s and Ehrlich‘s Impact

HUMORAL IMMUNITY

Other activities


1903 Almroth Wright (nickname ‘Almost Right’) and Douglas communicate to the Royal Society that humoral, inducible and specific substances (i.e., antibodies) in the body fluids reinforce he action of phacytosis → named these factors opsonins (today antibodies)

189? Develops anti-typhoid vaccine with weakened bugs (all British solders were immunized during the 1st WW) with Wright’s vaccine.

Properties of Antibodies (Opsonins)

Sir Almroth Edward Wright (1861–1947)

Diggins F. (2002). Who was... Almroth Wright? Biologist (London). 49(6):280-2.

Wright, A. E., and S. R. Douglas. 1904. An experimental investigation of the role of the body fluids in connection with phagocytosis. Proc. R. Soc. London 72:357-370.

See also Roitt p. 322

1908 Paul Ehrlich:

Ehrlich (1908). Über Antigene und Antikörper. Einleitung in „Handbuch der Immunitätsforschung“. P.1 -10 Very nice overview about the knowledge of antibody and antigen in 1908.

Another Paradigm in Immunology„Infections are cleared by cellular

and humoral immunity“

Another Paradigm in Immunology„Infections are cleared by cellular

and humoral immunity“


1869 Adolf Creite (Göttingen) reports accumulation (“Anhäufung”) of red blood cells when adding sera from one animal to the blood of another animal

1875 Leonard Landois (Göttingen) reports disolving (“Auflösung”) und ball formation (“Zusammen-ballung”) when he mixed blood and serum from diferent animals

Agglutinins

1847 - 1921Germany

1.12.1837 – 1902Germany

Creite, A. (1869a) Versuche über die Wirkung des Serumeiweisses nach Injection in das Blut. Zeitschrift für Rationelle Medicin, 36, 90–108.

• Landois, L. (1875) Die Transfusion des Blutes, Leipzig.• http://de.wikipedia.org/wiki/Leonard_Landois

N. C. Hughes-Jones & Brigitte Gardner (2002). Historical Review: RED CELL AGGLUTINATION: THE FIRST DESCRIPTION BY CREITE (1869) AND FURTHER OBSERVATIONS MADE BY LANDOIS (1875) AND LANDSTEINER (1901). British Journal of Haematology, 2002, 119, 889–893.


1896 Herbert E. Durham and Max von Gruber introduce the term Agglutinin for any substance that caused agglutination of cells → Gruber-Durham reaction

1896 Fernand Widal uses the Gruber-Durham reaction to develop a test for typhoid fever. o Obseravtion: Serum from a typhoid carrier caused a culture

of typhoid bacteria to clump, whereas serum from a typhoid-free person did not.

o Widal test was the first example of serum diagnosis.

1853 – 1927Ausria

1862–1929France

http://en.wikipedia.org/wiki/Agglutination_%28biology%29


Agglutinins


1900 Landsteiner's first agglutination test with blood

1901 Landsteiner discovers two types of blood antigens (blood groups)

Agglutinins – Discovery of Blood Groups

1868 -1943Austria

Nobelpreis 1930

Landsteiner (1900). Zur Kenntnis der antifermentativen, lytischen und agglutinierenden Wirkungen des Blutserums und der Lymphe.Centralblatt fur Bakteriologie, Parasitenkunde und Infektionskrankheiten, vol. 27,pp. 357-362 1900

http://www.biologieunterricht.info/pool/pdf/ab_landsteinerversuch.pdf

Landsteiner, K. (1901) Über Agglutinationserscheinungen normal menschlichen Blutes. Wiener Klinische Wochenschrift, 14, 1132–1134.


1901 Discovered that blood types of donors and recipients need to be matched before transfusionso made blood transfusions a routine o saved countless liveso Nobel Prize for Medicine1930.

1920 Introduces the term hapten

1909 discovered (with biologist Erwin Popper) the infectious nature of poliomyelitis (the polio virus).

1927 discovered with pathologist Philip Levine,the M and N agglutinogens.

1940 discovered (with pathologist Alexander Wiener the rhesus (Rh) factor in blood.

While working in his laboratory on 24 June 1943, Landsteiner suffered a heart attack, and died from its after-effects two days later.

1868 -1943Austria(US)Nobel Prize

1930

Karl Landsteiner – Discoveries


Precipitins

1897 Rudolf Kraus (1868-1932; Austria) observes precipitates between immune sera and soluble lystes of bacteria

Experiment• Filter lysates from cholera, plaque and typhus through a bacteria-

removing filter• Add sera from patients or animals

Obeservation• Precipitates are visible

Conclusion→ Not only bateria but also their soluble compounds react with

corresponding antibodies

Kraus, R. Wien. Klini Wochenzeitschrift 10:736 (1897)http://www.oegai.org/html/index.php?module=ContentExpress&func=display&btitle=CE&mid=&ceid=84

PRECIPITINS


Summary: Humoral immunity (1905)

• Eichmann, Klaus (2000). The network collective: rise and fall of a scientific paradigm • http://en.wikipedia.org/wiki/Humoral_immunity#cite_note-G.E-3• JEAN LINDENMANN (1984). Origin of the Terms 'Antibody' and 'Antigen‘ Scand. J. Immunol., 19, 281-285

In 1905 it was clear that all these humoral activities can be traced back to the same class of inducible compounds (i.e., the antibody molecule)

Today, Antikörper (Antibody) is a neutral term for the common component in all the different biological activities of immune sera


1890 Behring never uses the noun „Antitoxin“ his his two 1890 publications(Only used adj. „antitoxisch“ in a footnote in the 1890 paper with Kitasato)

1891 Tizzioni and Cattani who were looking rather for a substance than an activity introduced for the first time the noun antitoxinTizzoni, G. & Cattani, G. Ueber die Eigenschaften des Tetanus-Antitoxins. Zentralbl. Bakteriol. Mikrobiol. Hyg. [A] 9, 685, 1891.

1891 Ehrlich uses for the first time the term “Antikörper” in his second 1891 paper on page 1219 at the end of the 6. paragraph; „ If two substances give rise to two different 'Antikörper', then they themselves must be different. Ehrlich, P. Experimentelle Untersuchungen über Immunität. II. Ueber Abrin. Dtsch. med. Wochenschr. 17, 1218, 1891.

1897 Ehrlich uses again the term Antikörper in his 1897 paper: “Nachdem die ursprüngliche Annahme von einer Zerstörung des Giftes durch den Antikörper als unhaltbar sich erwiesen hatte, ….P. Ehrlich (1897). Zur Erkenntniss der Antitoxinwirkung. Fortschritte der Medicin, Bd 15, No 2, p. 41-43

Other terms: Immunkörper, Amboceptor, Zwischenkörper,, Immunisin, substance sensibilisatrice, copula, Desmon, philocytase, fixateur

Origin: Terms „Antitoxin“ & „Antikörper“


1891 1897

Deutsche Med. Wochenzeitschr. (1891) Nr. 32, p 976

Coining Term „Antikörper“ – Ehrlich


Ehrlich’s term was not accepted immediately and several other terms were used: Immunkörper, Amboceptor, Zwischenkörper, substance sensibilisatrice, copula, Desmon, philocytase, fixateur, and Immunisin.

Antibody was obviously constructed in formal analogy to 'Antitoxin', but in conceptual analogy to 'Immunkörper'. The antitoxin is something directed against a toxin; the antibody is a body directed

against something (a body ???)

This imbroglio (Verwirrung) is apparent in Ehrlich's paper (Ehrlich, P. Experimentelle Untersuchungen er Immunitat. II. Ueber Abrin. Dtsch. med. Wochenschr. 17, 1218, 1891):

“All these phenomena result from the presence, in the blood, of a body—the anti-abrin—which completely paralyses the effects of abrin—probably by destruction of this body’”

Body number 1 in this sentence is antibody; body number 2 is antigen. Antibody is actually confusing since it is itself a body that reacts against another

body. Therefore, „Immunkörper“ might have been a better term !!!????

Term Antikörper – Lindemann 1984 JEAN LINDENMANN (1984). Origin of the Terms 'Antibody' and 'Antigen‘ Scand. J. Immunol., 19, 281-285

START OF IMMUNOLOGY

Serology





4. Cytotoxic humoral immunity and complemento Bordet (1899): substance sensibilisatrice + Buchner‘s Alexino Ehrlich (1899): Amboreceptor + Komplement

5. Serodiagnostic (Start of Serology)o Widal (1896) – Widal agglutination test for typhoid fevero Landsteiner (1901) – Blood goups in humano Bordet (1901) - Complement fixation testo Wassermann (1905) – Syphilis test


Nobel 1908


Nobel 1919

Nobel 1913

Nobel 1930



1896 Herbert E. Durham and Max von Gruber introduce the term Agglutinin for any substance that caused agglutination of cells → Gruber-Durham reaction

1896 Fernand Widal uses the Gruber-Durham reaction to develop a test for typhoid fever. o Obseravtion: Serum from a typhoid carrier caused a culture

of typhoid bacteria to clump, whereas serum from a typhoid-free person did not.

o Widal test was the first example of serum diagnosis.

1853 – 1927Ausria

1862–1929France



Widal Test (Typhoid fever)


http://www.bordet.be/en/presentation/history/bordet/life.htm

Bordet/Gengou: Complement fixation (1901)

Observation (Jules Bordet and Octave Gengou 1898, Pasteur Institute)

• Antigen-antibody reaction leads to the binding (fixation) of complement to the target antigen

• This discovery allowed him to develop blood tests (serodiagnosis technique) that indicate whether a person has been in contact with any infectious agent.

Jules Bordet (1870-1961)

BelgiumNobel price Medicine

1919

Bordet, J. and O. Gengou. 1901. Sur l'existencede substances sensibilisatrices dans la plupart des serums antimicrobiens. Ann. De l'Inst. Pasteur. xv: 289-303.


Complement in patient (has anti-germ) and ctrl serum (no anti-germ) is destroyed by heating

Sheep red blood cells (sRBCs)

and anti-sRBC are added

Equal amounts of fresh C (guinea pig serum) and germ-

Ag are added

Antigen

Fixed Compl.

Anti-germ

Anti-SRBC

Free Compl.

Agglutinated sRBC

Lysed SRBC

Complement fixation test

Nice Animation: http://highered.mcgraw-hill.com/sites/0072556781/student_view0/chapter31/animation_quiz_4.html

Patient

Control

Anti-SRBC


1. A Wassermann, A. Neisser and C. Bruck. Eine serodiagnostische Reaktion bei Syphilis. Deutsche medicinische Wochenschrift, Berlin, 1906, 32: 745-746.

2. http://plumbot.com/Wassermann_test3. http://www.whonamedit.com/synd.cfm/2962.html

Wassermann Test (Syphillis)

August Paul von Wassermann

(1866- 1925)Bamberg, Germany

1904 Fritz R. Schaudinn and Paul E. Hoffmann isolate the causative organism of syphilis.

1905 Wassermann introduces complement-fixation test with blood serum or cerebro-spinal fluid to detect a syphilis infection

Procedure •Used lipid called cardiolipin from heart as "antigen", which has nothing to do with syphilis. •However, this antigen reacts with certain antibodies that are commonly present in the blood of syphilis patients•Antibodies are also present in malaria, lepra and autoimmune disorders → Not specific for syphilis•Also called Bordet-Wassermann


Complement Fixation Test - Summary

The bacterium is present, if the red blood cells (“reporter”) added to the blood of infected patients remain intact in the presence of

1. the bacterium's specific antibody,

2. complement and

3. the red blood cells' specific antibody


Bacterio- & Hemolysins: Summary

1888 George Nuttal reports that bloodserum from non-immunzed animals kills bacteria in the absence of cells

1889 Hans Buchner describes for the frist time a heat-labile substance in blood serum that was capable of destroying bacteria (“alexin”)

1894 Richard Pfeiffer shows for the frist time that blood from immunized animals can lyse bacteria (bacteriolysis) in the absence of cells

1898 Jules Bordet shows that two different ‘bodies’ (today antibody and complement) are required for lysis of erythrocytes (immune hemolysis)

1899 Paul Ehrlich & Morgenroth rename alexin in complement and show that amboreceptor (today antibody) mediates complement

1901 Bordet introduces the complement-fixation test

1905 August v. Wassermann invents antibody test for syphyilis


IMMUNOLOGY: Own Discipline

PAUL-EHRLICH-INSTITUTPaul-Ehrlich Institute für Serum-forschung und Serumprüfung (1896) bis Paul-Ehrlich Institute Bundesamt für Sera und Impfstoffe (1990)

Das Paul-Ehrlich-Institut für

Serumprüfungund Serumforschung

Jahre1896 in Steglitz bei Berlin

Paul-Ehrlich-Institut im Jahre 1990 als

Bundesamt für Sera und Impfstoffe

in Langen bei Frankfurt/Main

Königliches Institut für experimentelle Therapie + Georg-

Speyer-Haus1922. Frankfurt

Ab 1947 Paul-Ehrlich-Institut für

Exp. Therapie Georg-Speyer-Haus,

1906, Frankfurt

Königliches Institut für experimentelle

Therapie, Frankfurt1899


JOURNALSo Zeitschrift für Immunitätsforschung (1908)o J. of Immunology (1913) o Eur. J. Immunology (1970)

PROFESSIONAL SOCIETIESo American Associaten of Immunologist (1913)o Deutsche Gesellschaft für Immunologie = DGfI (1953)

IMMUNOLOGY: Own Discipline


TIME LINE - History of Immunology

Discovery of cells and germs (1683 - 1876)

Prevention of Infection (1840 – today)

Start of Immunology (1796-1910)






IMMUNOCHEMISTRY

The Antibody Problem(1910 - 1975 )

hans-martin jäck division of molecular immunology dept. of internal medicine iii...

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