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Monoclonal antibodies against human blood group related antigens in tests with non-human primate red cells
TCB 1997
1:111-114
Monoclonal antibodies against human blood group related antigens in tests with non-human primate red cells W.W. SOCHA, S.G. LASANO Laboratoryfor Experimental Medicine and Surgery in Primates (LEMSIP), New York University School of Medicine, New York, USA
Monoclonal antibodies (Mabs) directed against a single Rh epitope on human red blood cells (rbcs) were shown to frequently cross-react with the red cells of non-human primates often displaying wide individual differences in their reactivity. Observations of differences among reagents revealed rich diversity of combining sites on the Rh molecule and implied the usefulness of ape and monkey blood as a reference system for defining minute specificity variations among Mabs raised against human red blood cell antigens [1, 2]. In the present study, panels of red cells of various species of apes and Old and New World monkeys were used to compare reactivities of monoclonal antibodies against human blood group related antigens.
Material and M e t h o d s All 23 monoclonal antibodies against human blood group related antigens were tested against panels of red cells of apes (common chimpanzees, lowland gorillas, orangutans and white-handed gibbons), Old World monkeys (rhesus monkeys, crab-eating (cynomolgus) macaques, vervet monkeys and baboons) and N e w World monkeys (marmosets and tamarins). As it is customary in work with non-human primate blood [3, 4], the tests were carried out by three agglutination techniques: saline, antiglobulin (IAT) and enzyme (ficin)-treated red cell methods (ETC)_ Rabbit anti-human globulin sera were used for testing chimpanzee, gibbon and gorillla, gibbon anti-human (absorbed with orangutan rbcs) for testing orangutan
blood, and rabbit anti-rhesus serum in tests with lower primates red cells. Supplies of reagents allowing, some of the Mabs were titrated with selected, positively-reacting rbcs.
Results (Table 1) Anti-Fy3 Both Anti-Fy3 Mabs detected polymorphism in gorillas in tests by antiglobulin and enzyme treated red cell methods. They agglutinated ficinated rbcs of all chimpanzees and orangutans tested. The representative titers of Mab 2D1-1 for enzyme-treated red cells of chimpanzee, orangutans and gorillas were 1:32, 1:16 and 1:2, respectively. The reagents proved inactive, by any method, with blood of other primates.
Anti-Jk a A single reagent of this specificity agglutinated red cells of all species investigated except gibbons and marmosets. It detected polymorphisms in chimpanzees (by IAT and ETC), gorillas (saline, IAT and ETC methods), rhesus monkeys (IAT) and baboons (saline and IAT). Representative titers with positively reacting rbcs were, depending on species, 1:8-1:16 by saline and antiglobulin and 1:16-1:128 by enzymetreated red cell methods.
Anti-Jk b Correspondence to: WladyslawW. Socha,M.D., LEMSIP,New York
University Medical Center, RD#1,Long Meadow Road, Tuxedo, New York 10987,USA.
Both Mabs of this specificity reacted, in general, by all three agglutination methods, with all the Old World monkey species and tamarins' red cells. However, 111
112
W.W. S O C H A & S.G. L A S A N O
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MONOCLONAL ANTIBODIES AGAINST H U M A N BLOOD GROUP RELATED ANTIGENS
only one Mab (2D1-6) brought about agglutination of chimpanzee and orangutan red cells. Those reactions were not type-specific and occurred only with enzyme-modified cells. By contrast, reactions with the red cells of lower monkeys were produced by two or even three agglutination methods and were often type-specific.
113
Anti-K14 This reagent detected, in tests by enzyme method, individual differences among orangutans and gorillas and agglutinated marmoset and tamarin red cells. It failed to react with blood of chimpanzees and Old World monkeys.
Anti-Kpb-like Anti-Jr a Proved inactive in all tests.
Anti-Js b Detected individual differences among chimpanzees in tests on ficinated red cells. Tests with red cells of other apes and monkeys were negative b y any method.
Anti-K The four reagents tested gave negative results, by any method, with the red cells of most species. Only two (2D1-9 and 2D1-10) produced agglutination (by IAT exclusively) of blood of N e w World monkeys: marmoset and tamarin.
Anti-k-like Agglutinated red cells of gorillas (to the titers of 1:2 and 1:8 by IAT and ETC methods, respectively) and orangutans (to the titer of 1:16 by all three methods), detecting individual differences among animals of these two species_ Tests with chimpanzees, as well as all Old and N e w World monkeys were negative, if one disregards weak agglutination of ficinated tamarin red cells.
Anti-Ku-like Both reagents agglutinated red cells of all chimpanzees, gorillas and orangutans tested (up to the titers of 1:8 and 1:128 by IAT, and ETC, respectively) but only one (2D1-16) reacted with blood of some, but not all, lower monkey species, namely rhesus monkeys, crab-eating macaques and vervet monkeys (up to the titer of 1:8 by ETC), marmosets and tamarins (up to the titer of 1:32 by ETC). It detected individual differences among crab-eating macaques but failed to agglutinate baboon red cells_
Detected polymorphism in gorillas in tests by antiglobulin method, a technique which produced uniformly negative results with all Other species. The same reagent, however, agglutinated ficinated red cells of all chimpanzees, gorillas and orangutans to the same titer of 1:16. The red cells of lower monkeys remained unagglutinated by any method.
Anti-Kell The four reagents of this specificity were, by and large, inactive against red cells of non-human primates. The only positive results were produced by two Mabs: 2D1-21 and 2D1-22, in antiglobulin tests with marmoset and tamarin rbcs. Anti-Lu b
Agglutinated by IAT the red cells of orangutans but proved inactive, by any method, with blood of other primates.
Discussion Similarly to the monoclonal antibodies against Rh and Rh-related epitopes, the reagents included in this study in many instances cross-reacted with red cells of non-human primates. As expected, most positive reactions were observed with blood of species taxonomically closest to man, namely anthropoid apes and, particularly, African apes (chimpanzee and gorilla) which are known to share with man a number of immunological traits [5, 6]. Notable exceptions were Mabs of anti-Kidd family which reacted with the red cells of most, if not all, representatives of Old and N e w World monkey species here investigated. When two or more antibodies of the same specificity were tested side-by-side some differences in their reactivity were detected indicating that each reagent recognized different element (s) of the epitope. This was the case with anti-Jk b, anti-K, anti-Ku-
114
W_W. S O C H A & S_G. L A S A N O
like a n d a n t i - K e l l Mabs. It w o u l d b e of i n t e r e s t to see w h e t h e r those v a r i a t i o n s r e c o g n i z e d b y tests w i t h n o n - h u m a n p r i m a t e r e d cells c a n b e r e p l i c a t e d i n tests w i t h h u m a n r e d cells of rare t y p e s a n d v a r i a n t s .
References [1] Socha W.W., Ruffi6 J. (1990) Monoclonal antibodies directed against human Rh antigens in tests with the red cells of nonhuman primates. Rev. F~: Transfus. Hdmobiol., 33, 39-48.
[2] Blancher A., Socha W.W.,Ruffi6 J. (1992) Diversity of human anti-D monoclonal antibodies revealed by reactions with chimpanzee red blood cells. Vox Sang., 63, 112-118. [3] Erskine A.E., Socha W.W. (1978) The Principles and Practice of Blood Grouping, 2nd edition. Mosby, Saint Louis. [4] Socha W.W., Ruffi6 J. (1983) Blood Groups of Primates. Theory, Practice, EvolutionaryMeaning. A.R. Liss, New York. [5] Socha W.W., Moor-Jankowski J. (1978) Blood groups of anthropoid apes and their relationship to human blood groups. J. Hum. Evol., 8, 30-38. [6] Socha W.W., Blancher A., Moor-Jankowski J. (1995) Red cell polymorphisms in non-humanprimates. A review. J. Med. Primatol., 24, 282-305.
1:111-114
Monoclonal antibodies against human blood group related antigens in tests with non-human primate red cells W.W. SOCHA, S.G. LASANO Laboratoryfor Experimental Medicine and Surgery in Primates (LEMSIP), New York University School of Medicine, New York, USA
Monoclonal antibodies (Mabs) directed against a single Rh epitope on human red blood cells (rbcs) were shown to frequently cross-react with the red cells of non-human primates often displaying wide individual differences in their reactivity. Observations of differences among reagents revealed rich diversity of combining sites on the Rh molecule and implied the usefulness of ape and monkey blood as a reference system for defining minute specificity variations among Mabs raised against human red blood cell antigens [1, 2]. In the present study, panels of red cells of various species of apes and Old and New World monkeys were used to compare reactivities of monoclonal antibodies against human blood group related antigens.
Material and M e t h o d s All 23 monoclonal antibodies against human blood group related antigens were tested against panels of red cells of apes (common chimpanzees, lowland gorillas, orangutans and white-handed gibbons), Old World monkeys (rhesus monkeys, crab-eating (cynomolgus) macaques, vervet monkeys and baboons) and N e w World monkeys (marmosets and tamarins). As it is customary in work with non-human primate blood [3, 4], the tests were carried out by three agglutination techniques: saline, antiglobulin (IAT) and enzyme (ficin)-treated red cell methods (ETC)_ Rabbit anti-human globulin sera were used for testing chimpanzee, gibbon and gorillla, gibbon anti-human (absorbed with orangutan rbcs) for testing orangutan
blood, and rabbit anti-rhesus serum in tests with lower primates red cells. Supplies of reagents allowing, some of the Mabs were titrated with selected, positively-reacting rbcs.
Results (Table 1) Anti-Fy3 Both Anti-Fy3 Mabs detected polymorphism in gorillas in tests by antiglobulin and enzyme treated red cell methods. They agglutinated ficinated rbcs of all chimpanzees and orangutans tested. The representative titers of Mab 2D1-1 for enzyme-treated red cells of chimpanzee, orangutans and gorillas were 1:32, 1:16 and 1:2, respectively. The reagents proved inactive, by any method, with blood of other primates.
Anti-Jk a A single reagent of this specificity agglutinated red cells of all species investigated except gibbons and marmosets. It detected polymorphisms in chimpanzees (by IAT and ETC), gorillas (saline, IAT and ETC methods), rhesus monkeys (IAT) and baboons (saline and IAT). Representative titers with positively reacting rbcs were, depending on species, 1:8-1:16 by saline and antiglobulin and 1:16-1:128 by enzymetreated red cell methods.
Anti-Jk b Correspondence to: WladyslawW. Socha,M.D., LEMSIP,New York
University Medical Center, RD#1,Long Meadow Road, Tuxedo, New York 10987,USA.
Both Mabs of this specificity reacted, in general, by all three agglutination methods, with all the Old World monkey species and tamarins' red cells. However, 111
112
W.W. S O C H A & S.G. L A S A N O
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MONOCLONAL ANTIBODIES AGAINST H U M A N BLOOD GROUP RELATED ANTIGENS
only one Mab (2D1-6) brought about agglutination of chimpanzee and orangutan red cells. Those reactions were not type-specific and occurred only with enzyme-modified cells. By contrast, reactions with the red cells of lower monkeys were produced by two or even three agglutination methods and were often type-specific.
113
Anti-K14 This reagent detected, in tests by enzyme method, individual differences among orangutans and gorillas and agglutinated marmoset and tamarin red cells. It failed to react with blood of chimpanzees and Old World monkeys.
Anti-Kpb-like Anti-Jr a Proved inactive in all tests.
Anti-Js b Detected individual differences among chimpanzees in tests on ficinated red cells. Tests with red cells of other apes and monkeys were negative b y any method.
Anti-K The four reagents tested gave negative results, by any method, with the red cells of most species. Only two (2D1-9 and 2D1-10) produced agglutination (by IAT exclusively) of blood of N e w World monkeys: marmoset and tamarin.
Anti-k-like Agglutinated red cells of gorillas (to the titers of 1:2 and 1:8 by IAT and ETC methods, respectively) and orangutans (to the titer of 1:16 by all three methods), detecting individual differences among animals of these two species_ Tests with chimpanzees, as well as all Old and N e w World monkeys were negative, if one disregards weak agglutination of ficinated tamarin red cells.
Anti-Ku-like Both reagents agglutinated red cells of all chimpanzees, gorillas and orangutans tested (up to the titers of 1:8 and 1:128 by IAT, and ETC, respectively) but only one (2D1-16) reacted with blood of some, but not all, lower monkey species, namely rhesus monkeys, crab-eating macaques and vervet monkeys (up to the titer of 1:8 by ETC), marmosets and tamarins (up to the titer of 1:32 by ETC). It detected individual differences among crab-eating macaques but failed to agglutinate baboon red cells_
Detected polymorphism in gorillas in tests by antiglobulin method, a technique which produced uniformly negative results with all Other species. The same reagent, however, agglutinated ficinated red cells of all chimpanzees, gorillas and orangutans to the same titer of 1:16. The red cells of lower monkeys remained unagglutinated by any method.
Anti-Kell The four reagents of this specificity were, by and large, inactive against red cells of non-human primates. The only positive results were produced by two Mabs: 2D1-21 and 2D1-22, in antiglobulin tests with marmoset and tamarin rbcs. Anti-Lu b
Agglutinated by IAT the red cells of orangutans but proved inactive, by any method, with blood of other primates.
Discussion Similarly to the monoclonal antibodies against Rh and Rh-related epitopes, the reagents included in this study in many instances cross-reacted with red cells of non-human primates. As expected, most positive reactions were observed with blood of species taxonomically closest to man, namely anthropoid apes and, particularly, African apes (chimpanzee and gorilla) which are known to share with man a number of immunological traits [5, 6]. Notable exceptions were Mabs of anti-Kidd family which reacted with the red cells of most, if not all, representatives of Old and N e w World monkey species here investigated. When two or more antibodies of the same specificity were tested side-by-side some differences in their reactivity were detected indicating that each reagent recognized different element (s) of the epitope. This was the case with anti-Jk b, anti-K, anti-Ku-
114
W_W. S O C H A & S_G. L A S A N O
like a n d a n t i - K e l l Mabs. It w o u l d b e of i n t e r e s t to see w h e t h e r those v a r i a t i o n s r e c o g n i z e d b y tests w i t h n o n - h u m a n p r i m a t e r e d cells c a n b e r e p l i c a t e d i n tests w i t h h u m a n r e d cells of rare t y p e s a n d v a r i a n t s .
References [1] Socha W.W., Ruffi6 J. (1990) Monoclonal antibodies directed against human Rh antigens in tests with the red cells of nonhuman primates. Rev. F~: Transfus. Hdmobiol., 33, 39-48.
[2] Blancher A., Socha W.W.,Ruffi6 J. (1992) Diversity of human anti-D monoclonal antibodies revealed by reactions with chimpanzee red blood cells. Vox Sang., 63, 112-118. [3] Erskine A.E., Socha W.W. (1978) The Principles and Practice of Blood Grouping, 2nd edition. Mosby, Saint Louis. [4] Socha W.W., Ruffi6 J. (1983) Blood Groups of Primates. Theory, Practice, EvolutionaryMeaning. A.R. Liss, New York. [5] Socha W.W., Moor-Jankowski J. (1978) Blood groups of anthropoid apes and their relationship to human blood groups. J. Hum. Evol., 8, 30-38. [6] Socha W.W., Blancher A., Moor-Jankowski J. (1995) Red cell polymorphisms in non-humanprimates. A review. J. Med. Primatol., 24, 282-305.