Karl Landsteiner and the

Discovery of Blood Groups

David Ntiamoah Ofosu

Prin BMS & Adjunct Lecturer

13.10.2015 1
 Outline
• Life of Karl Landsteiner

• History of the Discovery of Blood Groups

• Landsteiner and his Discovery of Blood Groups

• Discovery of Rhesus Factor

•• Haemolytic
Blood GroupDisease
System of
Today
the New-born
2
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 Life of Karl Landsteiner

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 Karl Landsteiner (1868-1943)
• born June 14th, 1868 in Baden (near Vienna)

• 1885 Study of medicine at the University of Vienna

(Austria), graduated 1891

• Studies in organic chemistry at the

✻ University Zürich – Arthur Hantzsch
✻ University Würzburg – Emil Fischer
✻ University of Munich – Eugen Bamberger

• 1891 published 1st paper on the influence of diet on

the composition of blood ash

• 1897Vienna Institute of Pathology - Performing

• 1896 Vienna General Hospital - Assistant under Max
autopsies 3639 dissection Speiser, 1961 (Hollinek)
4
von Gruber, Institute for Microbiology and Hygiene
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Karl Landsteiner (1868-1943)
• 1900 Description of the agglutination of human blood

• 1901 “Agglutination phenomena of normal human blood”

• 1908 Department of Pathology – Wilhelmina Hospital Vienna (Head)

• 1911 Professor of Pathological Anatomy in the University of Vienna

• 1919 Hospital The Hague, Netherlands

• 1922 The Rockefeller Institute, New York

• 1929 Landsteiner became US citizen

• 1930 Nobel price for medicine Speiser, 1961

5
(Hollinek)

• 1939 Emeritus Professor at the Rockefeller Institute

 Work of Karl Landsteiner
Pathogenesis of paroxysmal haemoglobinuria
Human blood groups ABO
Amongst the first to Introduction of
Introduction of dark-field microscopy to purify antibodies Haptens
visualize the spirochetes of syphilis

Polio research: Polio belongs to a

group of filterable microorganisms

Research work on Syphilis Detection of Treponema pallidum

3 6 0 Pu
13 .1 0 .20 1 5 Most of the time, Landsteiner was quite poor6
blications
 Nobel price for Medicine 1930
"for his discovery of human blood groups"
‘Beginning in 1923, fourteen
different nominators had put him
up for the award for three
different discoveries - his polio
research, his immune system
work, and his discovery of the
blood groups. After 29 years, the
committee finally granted him
the award for his blood group
work.’

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 History of the Discovery of
Blood Groups

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 Adolf Creite (1869)
• born 1847 in Helmstedt, Lower Saxony (died 1921)

• Medical student at the University of Göttingen

• Physiological Institute, Prof. Georg Meissner

– Serum proteins could „dissolving“ (lysis) and

„cluster“(agglutination)

– Inspired by Stokvis (1867) and Bernard (1856)

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Hughes-Jones, 2002 (Brit. Jour. Haem.)
9
 Adolf Creite (1869)
• born 1847 in Helmstedt, Lower Saxony (died
1921)

• Medical student at the University of Göttingen

• Physiological Institute, Prof. Georg Meissner

•• 1869 ‘Zeitschrift
‘Versuche überfür
dieRationelle Medizin’
Wirkung des
Serumeiweisses nach Injection in das Blut’

– Serum proteins could „dissolving“ (lysis) and

„cluster“(agglutination)

– Inspired by Stokvis (1867) and Bernard (1856)

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Hughes-Jones, 2002 (Brit. Jour. Haem.) 10
 Creite‘s Experiments
Sera from • calf
• calf • pig No Effect
Injection of
• pig • dog
•
8 ml serum
dog
• sheep • sheep • “blood-stained
• cat • cat urine“
• chicken • chicken • general malaise
• duck
• duck • death
• goat
• goat
Blood-stained urine: similar observations in literature (injection of whole blood rather than
serum) Panum (1863)
Sheel (1803) – Ippolito Magnani as well as Denis and Gaspard de Gurye

Creite‘s conclusion:
Constituents of serum had certain ‘chemical properties’ (chemische Eigenschaften) which
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Active red cells
ingredient directly
could (‘dissolve
be serum red cells’)
protein 11
Hughes-Jones, 2002 (Brit. Jour. Haem.)
Creite: 1st Description of Agglutination
In vitro Experiments

Sera from
• dog Add to a drop ‘cells suddenly flow
• sheep Microscopy together forming different-
of fresh blood
• cat
shaped drop- like clusters’
• chicken
• duck Possibility that some blood
•• rabbit No Effect
goat cells has dissolved

Creite described his observation but did not reach the conclusion that the
formation of ‚drop-like clusters‘ could be a true agglutination reaction

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Haem.)
Hughes-Jones, 2002 (Brit. Jour. 12
 Leonard Landois (1875)
• born 1837 in Münster

• Medical student at the University of

Greifswald

• Professor and Director of the Institute of

Physiology at Greifswald

• Study of blood transfusions and the

• phenomena
1875 of ‘Die
published: agglutination
Transfusion des Blutes’
• did not cite Creite

• died 1902 in Greifswald Lehrbuch der

Physiologie des
Menschen
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(1880) 13
Hughes-Jones, 2002 (Brit. Jour. Haem.)
 Landois‘ Experiments
Question: Is lysis of red cells are those of the recipient or the donor?

Incubation at 37°C
Add fresh
blood Observation of
Sera from
the initiation of
eight different
animals
red cell lysis

Transparent Non-transparent Incubation at room Transparent,

temperature cells are no
longer visible

Add fresh
Incubation • Nearly all foreign
Sera from blood
dogs red cells lyse
Transparent within minutes
Non-transparent • cells in close
e.g. Sera from dogs
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proximity to each 14
other form clumps
Hughes-Jones, 2002 (Brit. Jour. Haem.)
 Landois‘ Experiments
Question: Is lysis of red cells are those of the recipient or the donor?

Incubation at 37°C
Add fresh
blood Observation of
Sera from
the initiation of
eight different
animals
red cell lysis

Transparent Non-transparent Incubation at room Transparent,

temperature cells are no
longer visible
High concentration of red cells clumping of red cells occurred rather than lysis
substance in the serum which acted on red cells and made the membrane soft and sticky,
when cells touched each other they aggregate
Landois was stuck with the great variation in the activity shown by different sera on
different cells, but considered that lysis and aggregation resulted from some type of
inte1r3a.1c0.t2i0o1n5 between the serum and the red cells 15
Hughes-Jones, 2002 (Brit. Jour. Haem.)
 Landsteiner and his Discovery of
Blood Groups

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 The Way to Landsteiner
1890 Discovery of the antibodies specific for tetanus by Emil von
Behring und Kitasato

1898-1901 37 Papers on the subject of red cell agglutinins and lysins

published

1900 Shattock and Grünbaum appearance of serum agglutinins were

a manifestation of certain diseases (esp. infections)

Landsteiner entered the field

1900 Landsteiner: ‘Zur Kenntnis der antifermentativen, lytischen und
agglutinierenden Wirkungen des Blutserums und der Lymphe’

1900 Ehrlich und Morgenroth Agglutination of human blood with

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human serum is called ‘Isoagglutination’ (Berliner klinische Wochenschrift) 17
 Rouleaux
Rouleaux are stacks of red blood cells,
which form because of the unique discoid
shape

• Occur, when plasma protein

concentration is high
erythrocyte sedimentation rate
is also increased

• Non-specific indicator of the presence

of diseases

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wikipedia.org
 1900

First suggestion of the existence of serum agglutinins and red cell antigens
(footnote):

13.10.2015 Landsteiner, 1900 (Wiener Klin. Wochenschr1i9ft)

 1901

Landsteiner‘s 17th publication

Classification of three different groups

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Landsteiner, 1901 (Wiener Klin. Wochenschrift)
according to its agglutination properties 20
Crosstesting sera and red cells from six healthy scientists including himself

Serum of Dr. Störck and

Landsteiner reacted with no
other red cells
None of the sera reacted with
their own red cells

First description of self-

tolerance

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Crosstesting sera and red cells from six healty scientists including himself

Serum of Dr. Pletsching

reacted with Dr. Sturli cells

Landsteiner suggested at least

two classes of antibodies:
anti-A and anti-B

Dr. Pletsching A antigen on red cells & anti- B

antibodies in his serum

Dr. Sturli B antigen on red cells & anti-A

antibodies in his serum
Cells of Dr. Störck and Landsteiner contained
neither A antigen nor B antigen,
but anti-A and anti-B antibodies in their sera
3rd type: C (today: 0)

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Landsteiner, 1901 (Wiener Klin. Wochensc hrift)
 Mechanism?
1) Eisenberg: Resorption of red cell particles?
Landsteiner rejected this idea he was not able to produce

?
autoagglutinins after injection of their own red blood cells
in animals
2) Agglutinins forming as a result of physiological decay of
organ tissue:

• phenomenon of autoimmunization induced by red cell

particles

• Exact
linked to diseases
triggers from which
responsible for thepeople had recovered
production of anti-A and anti-B in humans
have yet to be identified (Historical Review, 2002)
Today: Antibodies induced due to natural exposure to similar antigen
1 3 e.10 . inants on microorganisms present in the normal flora and the gu 2 t3
d t
20 1 5
 1902: Discovery of th
4 group (AB)
(von Decastello and Sturli)

Alfred von Decastello-Rechtwehr (1872–1960) and Adriano Sturli (1873–1964),

two colleagues of Landsteiner, discovered one year later the 4th group and called
it AB.
von Decastelo, A. & Sturli, A. (1902) Über die Isoagglutinine im Serum gesunder
und kranker Menschen. Münchner Medizinische Wochenschrift, 49, 1090-1095.

1910-1911 von Dungern and Hirszfeld

Discovery of mendelian inheritance of A and B
von Dungern, E. & Hirszfeld, L. (1911) Über gruppenspezifische Strukturen des
Blutes III. Zeitschrift für Immunologische Forschung, 8, 526-562.

1910 von Dungern and Hirszfeld

Nomenclature for the AB0-System 1928 International accepted

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History of the Discovery of Blood Types
1869 1875 1901 1902 1911

Adolf Creite Leonard Landois Karl Landsteiner von Decastello von Dungern
and Sturli and Hirszfeld
1 st Research on Discovery of the
description of agglutination blood groups A, B Discovery of the Discovery of
agglutination and C (0) blood group AB mendelian
inheritance
of A and B

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ed2i5tion
 The Rhesus factor

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 Rhesus factor – Discovery
1921 Unger: Report of intragroup transfusion reactions Additional tests should be
carried out for saver blood transfusions
1939 Levine: Report of an unusual case of intragroup agglutination (Levine and
Stetson, 1939)
• Woman (blood type 0) with a stillbirth needed a blood transfusion from her
husband (0) within 10 min she developed severe symptoms
• Cross-matched: her serum agglutinated with her husband‘s cell
104 group 0 samples were tested only 21 were compatible Levine
suggested: Isosensitization caused by ‘products‘ from the fetus
1940 Landsteiner and Wiener: Discovery of Rh factor ‘An agglutinable factor of human
blood recognizable by immune sera for rhesus blood‘
Human alloantibody was
renamed ‘anti-D‘
Landsteiner & Wiener, 1940
(Proceedings of the Society 2fo 7r
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Experimental Biology and Medicine)
 1941

Blood from rhesus Immunization Obtaining serum

monkeys Agglutinable factor
different from A and
Clinical importance: B was detected
• Patients with haemolytic reactions after receiving repeated
blood transfusion of the proper blood group Rhesus factor
Serum of these patients contained anti-Rh antibodies (but
cell factor was lacking)
• Appearance of immune isoantibodies in pregnancies
Goal: Develop a practical method of testing for the presence of Rh factor and
inve13s.1t0ig.2a01t5e heredity 28
 Test System for the Presence of Rh
Injection of
cells i.p. After 7 days
Washed red blood cells Obtaining serum
from rhesus monkeys
Using sera for
Day 1 and 5 practical diagnosis

Results Sera + washed blood

suspension

In narrow tube
wait 30 min

Sedimentation

Inspection of
Typing of patients and prospective blood donors bottom of
tube
E x amination of families showed Rh factor is inherited Mendelian
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domin29ant
Haemolytic Disease of the New-born

within 24-48h
after 1st
delivery

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 Haemolytic Disease of the New-born
• Caused by destruction of red blood cells due to alloantibodies (IgG) against Rh
antigens

• Maternal IgG antibody passes through the placenta binding the foetal Rh-positive
red blood cells and destroying them
leading to anaemia foetal brain damage death (erythoblastosis fetalis)
different forms: from mild to severe anaemia
• Development of haemolytic disease of the new-born can be detected by testing
maternal serum during the pregnancy for antibodies of the Rh antigen
Isolated fetal red blood cells + Coombs reagent
Rise in antibody titer Coombs test
(goat anti-human IgG)
Maternal IgG bound Agglutination
Treatment during pregancy: intrauterine blood transfusions,
plasmapheresis of the mother‘s blood Kuby 5th edition
Haemolytic disease (65%) due to AB0 incompatibility minor consequences
31
(ele1v3.a10t.i2o01n5 of bilirubin exposure of infant to low levels of UV light)
 Human blood groups – Today (2010)
308 blood group determinants
30 blood group systems
last one: RHAG (≈2008)
MNS system
• 46 antigens
• described by Landsteiner and
Levine (1927)
Kell-Cellano system
• Kell protein
• Alloimmune or autoimmune Goeff, 2001 (Wiener Klin. Wochenschrift)

diseases
• McLeod syndrome
Nomenclature and updated tables regarding new blood groups and their
32
ant1i3g.1e0.n20s15 International Society for Blood Transfusion
 Distribution World Wide

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g
 Blood Transfusions - History
1666 Report on a dog-dog blood transfusion from physiologist R. Lower
most experiments were successful

1667 1st successful blood transfusion to a human from a lamb by two

french physicians Denis and Emmerez
4th transfusion unsuccessfull (patient got 2nd transfusion):
haemolytic reaction due to immunization of the patient by 1st
transfusion
Transfusion were prohibited
1907 1st successful transfusion achieved by Dr. Reuben Ottenberg (Mt.
Sinai Hospital New York
Today Extensive blood donor screening Reduced risk for HIV, HBV and
HCV infection as well as for AB0 incompatibility
Packed red blood cells (‚Erythrozytenkonzentrate‘)
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Schwarz, 2003 (Brit. Jour. Haem.)
Importance of Landsteiner’s Discovery

Landsteiner (1901)

Superman of Science Makes Landmark

Discovery - Over 1 Billion Lives Saved So Far!
Almost 4 million American receive a blood
transfusion each year
Worldwide, there are about 890 million units of
blood donated each year.
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 “Wherever a blood transfusion
is performed in the world today,
wherever a worried mother’ s
threatened child is saved, Karl
Landsteiner is virtually present”

Herrmann Chiari, 1961

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 Thank you for your attention!

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 The End

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