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Hemotypology of a Chinese population
Fran~oise Leprince Jeanine Seger
Hemotypology of a Chinese Population Serum protein groups and red blood cell enzymes were studied in a
CentreNational de TransfusionSanguine, population of 125 unrelated Chinese men and women of the Macao Region9 53 bd Diderot Paris 75012,France In most systems the results were similar to those already reported, but
differencies were found in the GPT system9
Jacques
Ruffi~
Laboratoired'AnthropologiePhysiquedu Coll~gede France,53 bd Diderot, Paris 75012, France Keywords: hemotypology,Chinese,
Macao, serum protein systems, erythrocyte enzymes. Received 20 May 1980
1. Introduction We present here the results of serum protein groups and red-blood-cell enzymes for a population of 125 unrelated Chinese men and women, natives of the city of Macao or of the neighboring province of K u a n g - T u n g (southeastern China). The samples were drawn into vacutainers and sent by air in refrigerated boxes to the laboratory. On arrival, they were immediately frozen at - 8 0 ~ after separation of cells and plasma.
2. Methods (a) Serum Proteins The following systems were studied: Haptoglobin (Hp) Group-specific component (Gc) Third complement component (G3) Transferrin (T o Haptoglobin was analysed in starch gel with a discontinuous Poulic buffer. Group-specific component was tested according to the method of Teisberg (1970), immunofixation being done according to the method of Alper & Johnson (1969). Transferrin was studied with Ca according to the method of Teisberg (1970). 9(b) Erythroeyte Enzymes Eight erythrocyte enzymes were studied: acid phosphatase (PAc 1); adenosine deaminase (ADA); adenylate kinase (AK 1); D esterase (D est); soluble glutamic pyruvic transaminase (GPT); Journal of Human Evolution (1983) 12, 145-148
0047 2484/83/020145 + 04 $03.00/00
9 1983 Academic Press Inc. (London) Limited
146
F. L E P R I N C E E T A L .
phosphoglucomutase 1 and 2 ( P G M 1 and P G M 2); 6 phosphogluconate dehydrogenase (6PGD); glucose 6 phosphate dehydrogenase (G6PD). The packed red cells were lysed by freezing for GPT; by the addition of one volume of a 0'2% solution of [3 mercaptoethanol followed by freezing for P A C and ADA; by the addition of one volume of a solution containing 2'7 mM E D T A , 7 mM [3 mercaptoethanol, 10 btM N A D P followed by freezing for G6PD, the hemolysate being then diluted 1/5 in the electrophoretic buffer immediately before being tested; and by the addition of one volume of distilled water followed by freezing for the other enzymes. All the enzymes were studied by horizontal electrophoresis in starch gel. T h e techniques used are described by Hopkinson et al. (1963) for PAc 1, Fildes & Harris (1966) for AK1, Fildes & Parr (1963) for 6PGD, and the technique recommended by the W H O (1967) in E B T buffer for G6PD. For A D A we used the techniques described by Spencer et al. (1968) with a p H 6'5 phosphate buffer and a p H 7"5 buffer for the coloration. D est was analysed using the technique of Hopkinson et al. (1973) with a tris-maleate buffer p H 7'4 and 4 methyl umbelliferyl acetate for the coloration. For the study of G P T , we used the technique of G u s s m a n & R a m e s (1972) with a buffer identical to that used for P G M , but the non-diluted rbcs were applied to a W h a t m a n 17 paper; the migration was at 6 V / c m during 22 hours. T h e Phosphoglueomutase isoenzymes were studied by the technique described by Spencer et al. (1964), but the dilution was 1/20 for the gel which gave a better resolution than the 1/10 dilution originally described and the migration lasted 20 hours at 6 V/cm, using W h a t m a n 17 paper for the insertions.
3. R e s u l t s
O u r results are presented in Table l(a)-(j). Table l(a) Gc*
Gc
(b) Haptoglobin
No. subjects (n = 1 4 2 )
11 83 21 50 22 8 1 1 Allele ~eq: 1 = 0"7641, 2 = 0'2324.
Phenotype frequencies 0"585 0"352 0"056 0"007
(c) Cs
Haptoglobin
No. subjects (n = 1 4 0 )
Phermtype frequencies
11 14 0-10 2I 56 0.40 22 67 0.479 0 3 0.021 Allele freq:l = 0'3000,2 = 0"6786, X2 = 0'272
(d) Transferin
Ca
No. subjects (n = 1 4 2 )
22 140 21 2 Allele freq: 1 = 0"0070, 2 = 0"9930
Phenotype frequencies 0"986 0"014
Transferrin C CD
No. subjects (n = 142)
128 12 140 Allele freq: C = 0"9571, D = 0'0429
Phenotype frequencies 0.914 0.086
147
HEMOTYPOLOGY OF A CHINESE POPULATION
(f) PGMt
(e) PAc PAc
No. subjects (n = 125)
Phenotype frequencies
A AB B CA
6 47 71 1
0.048 0.376 O'568 0"008
PGM
No. subjects (n = 125)
Phenotype frequencies
ll 21 22
71 43 11
0.568 0.344 0.088
Allele ~eq: 1 = 0.74000, 2 = 0'2600, (X2 = 1'405).
Allele ~eq: A = 0.2400, B = 7560, C = 0-0040 (X2 = 0'262).
(h) 6PGD
(g) A D A ADA
No. subjects (n = 125)
Phenotype frequencies
6PGD
No. subjects (n = 125)
Phenotype frequencies
11 12
112 13
0"896 0'104
A AC
110 15
0"880 0"120
5 GPT
No. subjects (n = 123)
Phenotype frequencies
ll 21 22 "3" 1 "6" 1 "X" 1
20 50 50 1 1 1
0.162 0-407 0.407 0.008 0.008 0-008
Allele freq: 1 = 0.9480, 2 = 0'0520.
(j) 5 GPT:]:
(i) D est D est
No. subjects (n = 125)
Phenotype frequencies
ll 21 22
51 56 18
0"408 0'448 0"I44
Allele ~ e q : l = 6320, 2 = 0"3680 (Xz = 0"169).
Allele freq: 1 = 0"3780, 2 = 0"6098 (X2 = 1-525), " 3 " - " 6 " - " X " = 0'0122.
* O n e subject showed a rare allele. With simple immunofixation, this al,lele migrates more slowly than allele 2. The etectrophoretic mobility in relation to 2 is 0"90 which is more rapid than the Z allele described by J o h n s o n (1975). Isoelectric focusing reveals that it is a 2C type but the exact position has not yet been determined. It seems close to the 2C2 described by M. T h y m a n n (GcZ Copenhagen) and H. Cleve (GcZ Germany) (Constans & Cleve, 1979). "~ We found no rare alleles for P G M I nor for PGM2. { In this system we found three rare alleles, but the studies are not yet complete. O n e seems identical to the allele 3 described by S h i - H a n - C h e n (1972); the second, more rapid, seems to correspond to a new allele; the slow third allele may be identical to the allele 6 described by Shi-HanChen. Family studies now being carried out should allow us to confirm these results. For the G6PD system, out of 125 subjects studied, five showed almost no activity of 0"04 for the deficient allele. O n e subject showed a hemoglobin which migrates at the level of H b S but the study could not be carried further.
148
V. LEPRINCE ET AL.
4. C o n c l u s i o n s T h r e e remarks m a y be m a d e a b o u t this p r e l i m i n a r y study. Firstly, it seems that the C~ system has not yet been studied in Asiatic populations. O u r results seem to show that the C3 p o l y m o r p h i s m is almost absent in the Chinese p o p u l a t i o n of Macao, as the frequency of the allele is only 0'007. Secondly, our results in the G P T system are noticeably different from those given by Blake (1976) for the Chinese of Singapore. T h e frequency of the allele 2 which we found was m u c h higher. Thirdly, we found three rare alleles, two already described by S h i - H a n - C h e n (1972) a n d also reported by Blake (1976) in Asiatic populations. T h e third allele seems to be new. For the other serum protein systems, our results are very close to those already published for the p o p u l a t i o n s of M a c a o in Southeast Asia.
References Alper, C. A. & Johnson, A. M. (1969). Immunofixation electrophoresis: a technique for the study of protein polymorphism. Vox Sanguinis 17, 44,5-452. Blake, N. M. (1976). Glutamic pyruvie transaminase and esterase D types in the Asian-Pacific area. Human Genetics 35, 91-102. Constans, J. & Cleve, H. (1979). Group specific component report on the first international workshop. Human Genetics 48, 143-149. Fildes, R. A. & Harris, H. (1966). Genetically determined variation of adenylate kinase in man. Nature 209, 261-263. Fildes, R. A. & Parr, C. W. (1963). Human red cell phosphogluconatedehydrogfinase.Nature 200, 890-891. Gussman, S. & Rames, K. (1972). Die Darstellung der Polymorphismen Glutamat-Pyruvat-Transaminase (GPT, EC2.6.1.2.) und Phosphoglucomutase(PGMIEC2.7.5.1.) mittels horizontaler St~irkegerelektrophorese in einem Arbeitsgang. Zeitschrift ffir Rechtsmedizin; 70, 148-149. Hopkinson, D. A., Mestriner, M. A., Cortner, J. & Harris, H. (1973) Esterase D: a new human polymorphism. Annals of Human Genetics (Lond) 37, 119-137. Hopkinson, D. A., Spencer, N. & Harris, H. (1963). Red cell acid phosphatase variants: a new human polymorphism. Nature 199, 969-971. Johnson, A. M. et al. (1975). Variants of the group specifc component system as demonstrated by immunofixation electrophoresis. Report of a new variant, Gc Boston (Gc B). American Journal of Human Genetics 27, 728-736. OMS (1967) Normalisation des techniques d'~tude de la Glucose-6-phosphated~shydrog~nase. Org. Mond. Santg Ser. Rap. Techn. 1967, 366. Shi-Han-Chen et al. (1972). Genetics of glutamic-pyruvic transaminase: its inheritance, common and rare variants, population distribution, and differenciesin catalytic activity.Annals of Human Genetics (Lond) 35, 401. Spencer, N., Hopkinson, D. A. & Harris, H. (1964). Phosphoglucomutasepolymorphism in man. Nature 204, 742-745. Spencer, N., Hopkinson, D. A. & Harris, H. (1968). Adenosinedeaminase polymorphism in man. Annals of Human Genetics (Lond) 32, 9-14. Teisberg, P. (1970). High voltage agarose gel electrophoresis in the study of C3 polymorphism. Vox Sanguinis 19, 47-56.
Hemotypology of a Chinese Population Serum protein groups and red blood cell enzymes were studied in a
CentreNational de TransfusionSanguine, population of 125 unrelated Chinese men and women of the Macao Region9 53 bd Diderot Paris 75012,France In most systems the results were similar to those already reported, but
differencies were found in the GPT system9
Jacques
Ruffi~
Laboratoired'AnthropologiePhysiquedu Coll~gede France,53 bd Diderot, Paris 75012, France Keywords: hemotypology,Chinese,
Macao, serum protein systems, erythrocyte enzymes. Received 20 May 1980
1. Introduction We present here the results of serum protein groups and red-blood-cell enzymes for a population of 125 unrelated Chinese men and women, natives of the city of Macao or of the neighboring province of K u a n g - T u n g (southeastern China). The samples were drawn into vacutainers and sent by air in refrigerated boxes to the laboratory. On arrival, they were immediately frozen at - 8 0 ~ after separation of cells and plasma.
2. Methods (a) Serum Proteins The following systems were studied: Haptoglobin (Hp) Group-specific component (Gc) Third complement component (G3) Transferrin (T o Haptoglobin was analysed in starch gel with a discontinuous Poulic buffer. Group-specific component was tested according to the method of Teisberg (1970), immunofixation being done according to the method of Alper & Johnson (1969). Transferrin was studied with Ca according to the method of Teisberg (1970). 9(b) Erythroeyte Enzymes Eight erythrocyte enzymes were studied: acid phosphatase (PAc 1); adenosine deaminase (ADA); adenylate kinase (AK 1); D esterase (D est); soluble glutamic pyruvic transaminase (GPT); Journal of Human Evolution (1983) 12, 145-148
0047 2484/83/020145 + 04 $03.00/00
9 1983 Academic Press Inc. (London) Limited
146
F. L E P R I N C E E T A L .
phosphoglucomutase 1 and 2 ( P G M 1 and P G M 2); 6 phosphogluconate dehydrogenase (6PGD); glucose 6 phosphate dehydrogenase (G6PD). The packed red cells were lysed by freezing for GPT; by the addition of one volume of a 0'2% solution of [3 mercaptoethanol followed by freezing for P A C and ADA; by the addition of one volume of a solution containing 2'7 mM E D T A , 7 mM [3 mercaptoethanol, 10 btM N A D P followed by freezing for G6PD, the hemolysate being then diluted 1/5 in the electrophoretic buffer immediately before being tested; and by the addition of one volume of distilled water followed by freezing for the other enzymes. All the enzymes were studied by horizontal electrophoresis in starch gel. T h e techniques used are described by Hopkinson et al. (1963) for PAc 1, Fildes & Harris (1966) for AK1, Fildes & Parr (1963) for 6PGD, and the technique recommended by the W H O (1967) in E B T buffer for G6PD. For A D A we used the techniques described by Spencer et al. (1968) with a p H 6'5 phosphate buffer and a p H 7"5 buffer for the coloration. D est was analysed using the technique of Hopkinson et al. (1973) with a tris-maleate buffer p H 7'4 and 4 methyl umbelliferyl acetate for the coloration. For the study of G P T , we used the technique of G u s s m a n & R a m e s (1972) with a buffer identical to that used for P G M , but the non-diluted rbcs were applied to a W h a t m a n 17 paper; the migration was at 6 V / c m during 22 hours. T h e Phosphoglueomutase isoenzymes were studied by the technique described by Spencer et al. (1964), but the dilution was 1/20 for the gel which gave a better resolution than the 1/10 dilution originally described and the migration lasted 20 hours at 6 V/cm, using W h a t m a n 17 paper for the insertions.
3. R e s u l t s
O u r results are presented in Table l(a)-(j). Table l(a) Gc*
Gc
(b) Haptoglobin
No. subjects (n = 1 4 2 )
11 83 21 50 22 8 1 1 Allele ~eq: 1 = 0"7641, 2 = 0'2324.
Phenotype frequencies 0"585 0"352 0"056 0"007
(c) Cs
Haptoglobin
No. subjects (n = 1 4 0 )
Phermtype frequencies
11 14 0-10 2I 56 0.40 22 67 0.479 0 3 0.021 Allele freq:l = 0'3000,2 = 0"6786, X2 = 0'272
(d) Transferin
Ca
No. subjects (n = 1 4 2 )
22 140 21 2 Allele freq: 1 = 0"0070, 2 = 0"9930
Phenotype frequencies 0"986 0"014
Transferrin C CD
No. subjects (n = 142)
128 12 140 Allele freq: C = 0"9571, D = 0'0429
Phenotype frequencies 0.914 0.086
147
HEMOTYPOLOGY OF A CHINESE POPULATION
(f) PGMt
(e) PAc PAc
No. subjects (n = 125)
Phenotype frequencies
A AB B CA
6 47 71 1
0.048 0.376 O'568 0"008
PGM
No. subjects (n = 125)
Phenotype frequencies
ll 21 22
71 43 11
0.568 0.344 0.088
Allele ~eq: 1 = 0.74000, 2 = 0'2600, (X2 = 1'405).
Allele ~eq: A = 0.2400, B = 7560, C = 0-0040 (X2 = 0'262).
(h) 6PGD
(g) A D A ADA
No. subjects (n = 125)
Phenotype frequencies
6PGD
No. subjects (n = 125)
Phenotype frequencies
11 12
112 13
0"896 0'104
A AC
110 15
0"880 0"120
5 GPT
No. subjects (n = 123)
Phenotype frequencies
ll 21 22 "3" 1 "6" 1 "X" 1
20 50 50 1 1 1
0.162 0-407 0.407 0.008 0.008 0-008
Allele freq: 1 = 0.9480, 2 = 0'0520.
(j) 5 GPT:]:
(i) D est D est
No. subjects (n = 125)
Phenotype frequencies
ll 21 22
51 56 18
0"408 0'448 0"I44
Allele ~ e q : l = 6320, 2 = 0"3680 (Xz = 0"169).
Allele freq: 1 = 0"3780, 2 = 0"6098 (X2 = 1-525), " 3 " - " 6 " - " X " = 0'0122.
* O n e subject showed a rare allele. With simple immunofixation, this al,lele migrates more slowly than allele 2. The etectrophoretic mobility in relation to 2 is 0"90 which is more rapid than the Z allele described by J o h n s o n (1975). Isoelectric focusing reveals that it is a 2C type but the exact position has not yet been determined. It seems close to the 2C2 described by M. T h y m a n n (GcZ Copenhagen) and H. Cleve (GcZ Germany) (Constans & Cleve, 1979). "~ We found no rare alleles for P G M I nor for PGM2. { In this system we found three rare alleles, but the studies are not yet complete. O n e seems identical to the allele 3 described by S h i - H a n - C h e n (1972); the second, more rapid, seems to correspond to a new allele; the slow third allele may be identical to the allele 6 described by Shi-HanChen. Family studies now being carried out should allow us to confirm these results. For the G6PD system, out of 125 subjects studied, five showed almost no activity of 0"04 for the deficient allele. O n e subject showed a hemoglobin which migrates at the level of H b S but the study could not be carried further.
148
V. LEPRINCE ET AL.
4. C o n c l u s i o n s T h r e e remarks m a y be m a d e a b o u t this p r e l i m i n a r y study. Firstly, it seems that the C~ system has not yet been studied in Asiatic populations. O u r results seem to show that the C3 p o l y m o r p h i s m is almost absent in the Chinese p o p u l a t i o n of Macao, as the frequency of the allele is only 0'007. Secondly, our results in the G P T system are noticeably different from those given by Blake (1976) for the Chinese of Singapore. T h e frequency of the allele 2 which we found was m u c h higher. Thirdly, we found three rare alleles, two already described by S h i - H a n - C h e n (1972) a n d also reported by Blake (1976) in Asiatic populations. T h e third allele seems to be new. For the other serum protein systems, our results are very close to those already published for the p o p u l a t i o n s of M a c a o in Southeast Asia.
References Alper, C. A. & Johnson, A. M. (1969). Immunofixation electrophoresis: a technique for the study of protein polymorphism. Vox Sanguinis 17, 44,5-452. Blake, N. M. (1976). Glutamic pyruvie transaminase and esterase D types in the Asian-Pacific area. Human Genetics 35, 91-102. Constans, J. & Cleve, H. (1979). Group specific component report on the first international workshop. Human Genetics 48, 143-149. Fildes, R. A. & Harris, H. (1966). Genetically determined variation of adenylate kinase in man. Nature 209, 261-263. Fildes, R. A. & Parr, C. W. (1963). Human red cell phosphogluconatedehydrogfinase.Nature 200, 890-891. Gussman, S. & Rames, K. (1972). Die Darstellung der Polymorphismen Glutamat-Pyruvat-Transaminase (GPT, EC2.6.1.2.) und Phosphoglucomutase(PGMIEC2.7.5.1.) mittels horizontaler St~irkegerelektrophorese in einem Arbeitsgang. Zeitschrift ffir Rechtsmedizin; 70, 148-149. Hopkinson, D. A., Mestriner, M. A., Cortner, J. & Harris, H. (1973) Esterase D: a new human polymorphism. Annals of Human Genetics (Lond) 37, 119-137. Hopkinson, D. A., Spencer, N. & Harris, H. (1963). Red cell acid phosphatase variants: a new human polymorphism. Nature 199, 969-971. Johnson, A. M. et al. (1975). Variants of the group specifc component system as demonstrated by immunofixation electrophoresis. Report of a new variant, Gc Boston (Gc B). American Journal of Human Genetics 27, 728-736. OMS (1967) Normalisation des techniques d'~tude de la Glucose-6-phosphated~shydrog~nase. Org. Mond. Santg Ser. Rap. Techn. 1967, 366. Shi-Han-Chen et al. (1972). Genetics of glutamic-pyruvic transaminase: its inheritance, common and rare variants, population distribution, and differenciesin catalytic activity.Annals of Human Genetics (Lond) 35, 401. Spencer, N., Hopkinson, D. A. & Harris, H. (1964). Phosphoglucomutasepolymorphism in man. Nature 204, 742-745. Spencer, N., Hopkinson, D. A. & Harris, H. (1968). Adenosinedeaminase polymorphism in man. Annals of Human Genetics (Lond) 32, 9-14. Teisberg, P. (1970). High voltage agarose gel electrophoresis in the study of C3 polymorphism. Vox Sanguinis 19, 47-56.