Karl Landsteiner and the
Discovery of Blood Groups
Core Module Immunology
Doctoral Training Group GK1660
Erlangen  2011
Lena Krzyzak
Outline
• Life of Karl Landsteiner
• History of the Discovery of Blood Groups
• Landsteiner and his Discovery of Blood Groups
• Discovery of Rhesus Factor
• Haemolytic Disease of the New-born
• Blood Group System Today
Life of Karl Landsteiner
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
• 1896 Vienna General Hospital - Assistant under Max
von Gruber, Institute for Microbiology and Hygiene
• 1897 Vienna Institute of Pathology - Performing
autopsies 3639 dissection
Speiser, 1961 (Hollinek)
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
• 1939 Emeritus Professor at the Rockefeller Institute
• Died in 1943 at the age of 75 in New York, USA
Speiser, 1961
(Hollinek)
Work of Karl Landsteiner
Human blood groups ABO
Pathogenesis of paroxysmal haemoglobinuria
Introduction of dark-field microscopy to
visualize the spirochetes of syphilis
Amongst the first to
purify antibodies
Introduction of
Haptens
Polio research: Polio belongs to a
group of filterable microorganisms
Research work on Syphilis  Detection of Treponema pallidum
 360 Publications
Most of the time, Landsteiner was quite poor
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.’
scienceheroes.com
History of the Discovery of
Blood Groups
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 für Rationelle Medizin’
•
‘Versuche über die Wirkung des
Serumeiweisses nach Injection in das Blut’
– Serum proteins could „dissolving“ (lysis) and
„cluster“(agglutination)
– Inspired by Stokvis (1867) and Bernard (1856)
Hughes-Jones, 2002 (Brit. Jour. Haem.)
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 für Rationelle Medizin’
•
‘Versuche über die Wirkung des
Serumeiweisses nach Injection in das Blut’
– Serum proteins could „dissolving“ (lysis) and
„cluster“(agglutination)
– Inspired by Stokvis (1867) and Bernard (1856)
Hughes-Jones, 2002 (Brit. Jour. Haem.)
Creite‘s Experiments
• calf
• pig
• dog
Sera from
•
•
•
•
•
•
•
•
calf
pig
dog
sheep
cat
chicken
duck
goat
Injection of
8 ml serum
•
•
•
•
•
sheep
cat
chicken
duck
goat
No Effect
• “blood-stained
urine“
• general malaise
• death
 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
affected foreign red cells directly (‘dissolve red cells’)
 Active ingredient could be serum protein
Hughes-Jones, 2002 (Brit. Jour. Haem.)
Creite: 1st Description of Agglutination
In vitro Experiments
Sera from
•
•
•
•
•
•
dog
sheep
cat
chicken
duck
goat
• rabbit
Add to a drop
of fresh blood
Microscopy
No Effect
‘cells suddenly flow
together forming
different-shaped droplike clusters’
Possibility that some
blood 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
Hughes-Jones, 2002 (Brit. Jour. Haem.)
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 of agglutination
•
1875 published: ‘Die Transfusion des Blutes’
•
did not cite Creite
• died 1902 in Greifswald
Hughes-Jones, 2002 (Brit. Jour. Haem.)
Lehrbuch der
Physiologie des
Menschen
(1880)
Landois‘ Experiments
Question: Is lysis of red cells are those of the recipient or the donor?
Incubation at 37°C
Add fresh
blood
Sera from
Observation of
the initiation
of red cell lysis
eight different
animals
Transparent
Sera from
dogs
Non-transparent
Add fresh
blood
Transparent
Non-transparent
e.g. Sera from dogs
Incubation at
room temperature
Incubation
Transparent,
cells are no
longer visible
• Nearly all foreign
red cells lyse
within minutes
• cells in close
proximity to each
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
Sera from
Add fresh
blood
Observation of
the initiation
of red cell lysis
eight different
animals
Transparent
Non-transparent
Incubation at
room temperature
Transparent,
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
interaction between the serum and the red cells
Hughes-Jones, 2002 (Brit. Jour. Haem.)
Landsteiner and his Discovery of
Blood Groups
The Way to Landsteiner
1890
1898-1901
1900
Discovery of the antibodies specific for tetanus by Emil von
Behring und Kitasato
37 Papers on the subject of red cell agglutinins and lysins
published
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
human serum is called ‘Isoagglutination’ (Berliner klinische Wochenschrift)
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
wikipedia.org
1900
 First suggestion of the existence of serum agglutinins and red cell antigens
(footnote):
Landsteiner, 1900 (Wiener Klin. Wochenschrift)
1901
Landsteiner‘s 17th publication
Classification of three different groups
according to its agglutination properties
Landsteiner, 1901 (Wiener Klin. Wochenschrift)
Crosstesting sera and red cells from six healty 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 selftolerance
Landsteiner, 1901 (Wiener Klin. Wochenschrift)
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 & antiB 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)
Landsteiner, 1901 (Wiener Klin. Wochenschrift)
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
?
• linked to diseases from which people had recovered
 Exact triggers responsible for the 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
determinants on microorganisms present in the normal flora and the gut
1902: Discovery of 4th group (AB)
(von Decastello and Sturli)
Alfred von Decastello-Rechtwehr (1872–1960) and Adriano Sturli (1873–1964),
two kollegues 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
History of the Discovery of Blood Types
1869
1875
1901
Adolf Creite
Leonard Landois
Karl Landsteiner
1st
description of
agglutination
Research on
agglutination
Discovery of the
blood groups A,
B and C (0)
1902
1911
von Decastello
and Sturli
von Dungern
and Hirszfeld
Discovery of the
blood group AB
Discovery of
mendelian
inheritance
of A and B
Kuby 5th edition
The Rhesus factor
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 for
Experimental Biology and Medicine)
1941
Blood from rhesus
monkeys
Immunization
Clinical importance:
• Patients with haemolytic reactions after receiving repeated
blood transfusion of the proper blood group
 Serum of these patients contained anti-Rh antibodies
(but cell factor was lacking)
• Appearance of immune isoantibodies in pregnancies
Obtaining serum
Agglutinable factor
different from A and
B was detected
Rhesus factor
Goal: Develop a practical method of testing for the presence of Rh factor and
investigate heredity
Test System for the Presence of Rh
Washed red blood cells
from rhesus monkeys
Injection of
cells i.p.
After 7 days
Day 1 and 5
Results
Obtaining serum
Using sera for
practical diagnosis
Sera + washed blood
suspension
In narrow tube
wait 30 min
Sedimentation
 Typing of patients and prospective blood donors
Inspection of
bottom of
tube
 Examination of families showed Rh factor is inherited Mendelian dominant
Haemolytic Disease of the New-born
within 24-48h
after 1st
delivery
Kuby 5th edition
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
 Rise in antibody titer  Coombs test
Isolated fetal red blood cells + Coombs reagent
(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
(elevation 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
diseases
• McLeod syndrome
Goeff, 2001 (Wiener Klin. Wochenschrift)
Nomenclature and updated tables regarding new blood groups and their
antigens  International Society for Blood Transfusion
Distribution World Wide
wikipedia.org
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‘)
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.
scienceheroes.com
“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
Thank you for your attention!
scienceheroes.com