HLA antigens in amerindian groups of two different linguistic briceno1996

European Journal of Immunogenetics (1996), 23, 21-28
H L A A N T I G E N S IN A M E R I N D I A N G R O U P S O F
TWO DIFFERENT LINGUISTIC FAMILIES FROM
COLOMBIA
I . B R I C E N O ,J*. E . B E R N A LC, . D U R A NJ, . O R T E G AA
, . U M A N AA
, . GOMEZ
& S.S. P A P I H A *
lnstituto de Genetica Humana, Facultad de Medicina, Universidad Javeriana, Bogota,
Colombia and *Department of Human Genetics, University of Newcastle upon Tyne, Newcastle
upon Tyne, U . K .
(Received 24 October 1994; revised 25 June 1995; accepted 31 July 1995)
SUMMARY
Serological HLA types ( A , B, C , DR and DQ loci) were studied in five different Indian
tribes (Cubeo, Tucano, Coreguaje, Embera and Noanama) belonging to two distinct
linguistic families. For all the MHC loci, the range of variation among the five tribes
was enormous. Two tribes, Cubeo and Tucano, showed a wide spectrum of antigenic
specificities which seemed to be due to admixture from non-tribal groups, while in the
other three tribes the polymorphisms of various HLA loci showed restricted
distributions. The gene frequency data, when converted to a kinship matrix and a
two-dimensional eigenvector plot, indicated that members of the same linguistic
family tend to have greater genetic affinity.
INTRODUCTION
Some of the major factors that influence human population structure are geographical distance,
population movements, linguistic and cultural factors, demographic effects and random events.
The customary rationale for using language to interpret gene frequency patterns is a simplifying
assumption that common language frequently indicates a common origin. Cavalli-Sforza et al.
(1988) have presented empirical evidence for the correspondence between biological and
linguistic phylogenies on a worldwide basis.
The populations that show the greatest linguistic diversity are the Indian groups of North and
South America. In Colombia there are some 80 groups speaking about 60 well-defined languages
(Patino, 1991). Recently, Black etal. (1994, cited in Callegari-Jacquesef al., 1994), examining the
distribution of Class I HLA antigens in 1690South Amerindians, did not find any geographical or
linguistic clusters. Studies of HLA antigens from Colombia are limited, and no multivariate
analysis has been carried out to elucidate the genetic differentiation or affinity among different
Indian groups belonging to the same or different linguistic families. In 1988, Javeriana
Correspondence: Dr S.S. Papiha, Department of Human Genetics, 19/20 Claremont Place, Newcastle
Upon Tyne NE2 4AA. U.K.
0 1996 Blackwell Science Ltd
21
I . Briceno et al.
22
I
I
I
I
2W
400
600
4
800h
FIG.1. Map of Colombia showing sample collection sites.
University, Bogota, launched a 5-year programme, ‘Expedicion Humana’, 3 collect essential
biomedical data from different Amerindian groups. Several groups were examined for the
HLA-A, B, C, D R and DQ loci. This paper examines genetic variation and affinities derived from
HLA data on five Amerindian groups (Fig. 1) belonging to two distinct linguistic families.
Area, language and populations
Of the five Indian groups studied, Cubeo, Tucano and Coreguaje are of Macro-Tucano linguistic
stock, which is further divided into three linguistic families. The Cubeo belong to the Central
Tucano family, while the Tucano and Coreguaje belong to the Oriental and Occidental Tucano
families, respectively. The other two Indian groups, Embera-Epena and Noanama (Waunana),
speak the Noanama language. A brief demographic description of each Indian group is given
below.
Cubeo. The Cubeo Indians number nearly 4OOO individuals living near the banks of the rivers
Vaupes, Quduyari and Querari in the tropical rain forest of South-east Colombia. The Cubeo are
mainly endogamous, but intertribal marriages with neighbouring groups also occur.
Tucano. The Tucano Indians also inhabit the Vaupes region and are greater in number than the
Cubeo (population >6000). Their dialect is widespread along the Vaupes river. Like many other
0 I996 Blackwell Science Ltd, European Journal of Immunogenelics 23.21-28
HLA antigens in Amerindians
23
groups of this area, the Tucano are relatively exogamous. For both the Cubeo and Tucano
Indians, samples for HLA typing were obtained from Mandi and San Javier, near the banks of the
Quduyari river.
Coreguaje. The Coreguaje are 1400 in number and distributed in 14 settlements along the rivers
Orteguaza, Peneya and Caqueta. For the purpose of this investigation, samples were collected
from the Coreguaje living near the Orteguaza river.
Embera-Epena. The Embera-Epena Indians are part of the wider Embera family that also
includes the Chami and Catio tribes. The Embera-Epena, numbering nearly 1500 individuals,
speak Epena-Pedee, a language related to Noanama. For the purpose of this study the
Embera-Epena living in Guangui, a small settlement on the Saija river, were investigated.
Noanama (Waunana). The population size of this group is 2000 in Colombia, although additional
members of the group live in Panama. Both of these Noanama-speaking tribes live in an area near
the Pacific coast of the south-west of Colombia (Fig. 1).
MATERIAL AND METHODS
On two different field trips, blood samples were collected from Mandi, San Javier, Guangui and
settlements along the rivers Orteguaza and San Juan (Fig. 1). Collection involved random
sampling of both males and females from different villages, except for the Embera sample which
was mainly collected in Guangui. The Embera samples cannot be regarded as a collection from a
single village because Embera from neighbouring villages travelled to our camp for medical help.
Most individuals included in this survey were apparently healthy, although some with minor
medical complaints were also included. 307 individuals (Cubeo = 39, Tucano = 129, Coreguaje =
70, Noanama = 37 and Embera = 32) were tested for the number of specificitiesgiven below. Due
to field conditions and the non-availability of antisera, uniform analysis of all samples of the
different groups was not possible. Although all population groups were tested with broad
specificities such as A9, A10, A19, B5, B16, B17 and B40, in the present study the split
specificitieswere used in gene frequency (g.f.)calculations wherever the split antisera were used.
The samples were processed and HLA-typed locally using commercially available antisera
(Boehringwerke A.G.). Details of the typing have been described previously (Bernal el al.,
1988). In all, 14 specificities were examined at the HLA-A locus (21 for Coreguaje and Tucano),
23 at the B locus (44 for Coreguaje and Tucano), four at the C locus (10 for Coreguaje and
Tucano) and nine in the D region (eight at the DR locus and one at the DQ locus). In all, 77
antisera were used to type the HLA region. All specimens could not be analysed uniformly due to
difficulties in obtaining sufficient antisera and some further technical problems.
Gene frequencies and Hardy-Weinberg equilibrium were calculated using a computer
program based on the method described by Nam & Grat (1976). Genetic affinity and
differentiation among populations were computed by the method of Harpending & Jenkins
(1973), while the two-dimensional plot was constructed with the first two eigenvectors using
principal component analysis.
RESULTS A N D DISCUSSION
The observed antigen numbers and gene frequencies of the HLA-A, B, C, DR and DQ loci in the
five Amerindian groups are given in Table 1. Significant deviations from Hardy-Weinberg
0 1996 Blackwell Science Ltd, European Journal of Immunogmetics 23,21-28
N
6
93
3
9
f
2'
fi
CD
;
'D
5-
8
@
B44
B45
B51
Blocus
B7
B8
B14
B13
B18
B27
B35
B37
B38
B39
B41
B42
A1
A2
A3
A1 1
A23
A24
A25
A26
A28
A29
A30
~ 3 1
Blank
A locus
Specificity
0
10
0
NT
NT
NT
0
0
6
NT
0
1
4
0
6
2
10
0
0
29
0
1
3
10
6
4
4
1
Antigen
GF
O.oo00
O.oo00
O.oo00
O.oo00
0.1394
O.oo00
0.0131
0.0533
O.oo00
0.081 1
O.oo00
O.oo00
0.081 1
0.0126
0.4827
O.oo00
0.0126
0.0384
0.1346
0.0784
0.0515
0.0515
O.oo00
0.0254
0.1346
-0.0223
(n= 39)
cube0
44
1
3
0
7
4
4
14
1
2
6
0
0
0
1
1
109
0
4
8
32
7
7
7
3
12
22
5
Antieen
GF
0.0030
0.01 14
O.oo00
0.0268
0.0152
0.0152
0.0545
0.0030
0.0076
0.0230
O.oo00
O.oo00
O.oo00
0.0032
0.1830
0.0033
0.5185
O.oo00
0.0134
0.0269
0.1136
0.0235
0.0235
0.0235
0.0100
0.0407
0.0763
0.1268
Tucano*
(n = 129)
0
0
0
0
0
0
0
0
0
0
0
0
0
11
0
0
6
1
30
0
0
36
4
0
1
1
0
0
.
.
.
.
m
m
m
m
0 . m
0 . m
0 . m
0 . m
0 . m
0 . m
0 . m
0 . m
0 . m
0 . m
0 . m
0 . m
0 . m
0 . m
0.0806
0.0784
0.0069
0.5833
0
0
0
0
0.0069
0.0069
0 . m
0 . m
0.2900
0.0276
Antigen GF
Coreguaje'
(n = 70)
4
1
0
NT
0
0
0
0
0
0
0
0
15
NT
0
1
6
1
9
7
1
0
19
0
NT
0
20
0
Antigen
GF
O.oo00
0.0136
0.0555
O.oo00
O.oo00
O.oo00
0.2285
O.oo00
O.oo00
O.oo00
O.oo00
O.oo00
O.oo00
O.oo00
0.3024
O.oo00
0.0128
0.0795
0.0128
0.1221
0.0934
0.0930
O.oo00
0.2840
O.oo00
Noanama
(n= 37)
0
0
0
3
NT
0
NT
0
0
0
0
0
0
0
0
7
0
13
0
NT
0
4
0
0
10
0
1
7
Antigen
Embera
( n = 32)
TABLE1. Observed numbers and gene frequencies for the HLA-A locus in five Amerindian populations of Colombia
~
O.oo00
O.oo00
O.oo00
O.oo00
O.oo00
O.oo00
O.oo00
0.0598
O.oo00
O.oo00
O.oo00
O.oo00
O.oo00
O.oo00
0.0626
O.oo00
O.oo00
0.1655
O.oo00
0.0153
0.1125
0.4218
O.oo00
0.2223
O.oo00
GF
E
2
30
B
g.
9
!i2
10
5
7
8
NT
22
16
7
5
3
5
14
NT
NT
NT
11
10
NT
8
14
7
3
0
4
4
3
4
0.1305
0.1466
0.2881
0.1977
0.0806
0.0568
0.0928
0.0806
0.0568
0.2600
0. I850
0.1417
0.1277
0.1850
0.1006
0.0954
0.0533
O.oo00
0.0533
0.0533
0.0399
0.3368
20
3
17
9
NT
30
32
7
9
32
29
68
34
20
26
12
2
0
8
11
2
21
0
6
19
18
24
0.0808
0.1282
0.1374
0.0285
0.0367
0.5498
0.0121
0.0706
0.0367
0.1845
O.oo00
0.0452
0.0074
0.1009
0.0766
0.1345
0.0413
0.1134
0.2%2
0.0076
0.0822
O.oo00
0.0230
0.0747
0.0706
0.3960
NT
NT
NT
NT
NT
NT
NT
NT
NT
NT
0 . m
0.7439
0.0071
1
0 . m
0 . m
0 . m
0.0964
0.0286
0.0507
0.0733
0.0579
0.1777
0.66%
0 . m
0 . m
0.0142
0 . m
0
41
0
0
0
13
10
4
7
0
2
0
0
9
26
34
7
9
7
5
14
15
2
2
9
1
NT
NT
NT
NT
16
0
NT
4
7
12
0
8
0
0
6
3
8
0.0995
0.0228
0.0228
0. 1078
0.0113
0.1883
0.1743
0.0582
0.1078
0.3067
0.6653
0.0559
0.1OoO
0.1788
O.oo00
8
1
0
0
5
8
6
7
11
8
NT
NT
NT
NT
18
1
11
NT
2
4
5
10
0.0413
0.4621
0.0845
2
6
0
2
3
O.oo00
0.1145
O.oo00
O.oo00
0.1340
O.oo00
O.oo00
0.0892
0.1472
0.1081
0.1274
0.2096
0.1472
0.1713
0.7058
0.0309
0.0629
0.1851
0.0153
0.0394
0.1238
O.oo00
0.0394
0.0598
0.1019
0.5759
'Tucano and Coreguaje were also tested with A32, A33, A34, A36, A66, A68, A69, B46, B47, B48, BSO, B52, B53, B54, B57, B59, B63, B67 and B70antisera, but
no positive reaction was observed.
DQ 1
DQ locus
DRI
DR2
DR3
DR4
DR7
DRI 1
DR52
DR53
Blank
DR locus
c locus
CWI
cw2
cw3
cw4
cw5
CW6
cw7
CW8
CWll
Blank
B55
B60
B62
B73
Bw4
Bw6
Blank
26
I. Briceno et al.
equilibrium were observed in Coreguaje for the HLA-B and C loci, in Noanama for the HLA-C
locus and in Tucano for the HLA-D locus. Considering the large number of loci and antigens
tested, deviation in those loci may be a result of chance. However, there is a suggestion that
selection due to heterosis in South American Indians has operated (Black & Salzano, 1981),
producing fewer homozygote alleledhaplotypes, which may be responsible for the present
deviation from the null hypothesis.
For the HLA-A locus, both the Noanama-speaking tribes and the Coreguaje showed a very
restricted distribution pattern of antigens compared to the Cubeo and Tucano. The most frequent
specificity found in all five groups was A2, and its frequency ranged from 23 to 61%. Other
frequent antigens found at the A locus were A24 (split of A9) and A31 (split of A19). The genetic
variation in Tucano-speaking tribes was statistically significant for antigens A2, A24 and A25,
while differences among Noanama speakers were significant for A24. In the Coreguaje tribe the
A3 antigen, which is generally absent in American Indians (Salzano & Callegari-Jacques, 1988).
was present. At the B locus, there was a predominance of the B35, B51 and B60 antigens, but of
interest is the presence of B8 and B17 (broad specificity) in Cubeo and Tucano and B27 in
Tucano; these are rarely seen in American Indians (Black, 1980a; Belich et al., 1992). The
presence of these antigens may suggest admixture in Cubeo and Tucano. In the Coreguaje, the B
locus antigens were restricted to only a few of the commonly observed specificities in
Amerindians (B51, B60, BW4 and BW6) while in Noanama and Embera the additional
specificities B35 and B45 were also observed. These specificities are typical of those found in
north and central Amerindians (Cavalli-Sforza et al., 1994).
At the C locus, all five groups were consistently tested for Cw2, Cw3, Cw4 and Cw5 antigens.
Specificity Cw3 was the most common in all five tribes (range 6-30%). Cw4 was found in four
tribes and showed a narrow range of gene frequency (13-19y0). The Cw6 frequencies in Cubeo
andTucano were significantly different (P<O.OOl). DR and DQ typing could not be performed in
the Coreguaje, but all specificities tested were found in all four tribes and DR7 and D R l l were
the most common. The Embera tribe showed the highest frequency for DR4 (ISYO), DR52
(21%), and DR53 (14%) specificities. DQ1 was present in all the four tribes tested, with
frequencies ranging between 8 and 13%.
For all the MHC loci and for several specificities tested, the variation among the five tribes was
enormous. For analysis of genetic affinity among these five tribes belonging to two different
linguistic families, locus-by-locus comparison of gene frequencies not only proved to be difficult,
but could also be misleading. The gene frequency data consistently analysed in five groups were
converted to a single value, giving the kinship between the different groups. The kinship matrix
(R) was computed using the method of Harpending & Jenkins (1973). The R matrix is based on
the mean standard deviation of gene frequencies in each group from the total mean, and on the
average covariance of the gene frequencies in a pair of subgroups relative to the mean.
The R matrix based on the HLA data of the five Amerindian groups is given in Table 2. The
highest positive value of 0.027 was found between the Cubeo and Tucano, suggesting an
above-average affinity between these two groups. The R estimate between the Noanama
speakers was also found to be positive (0.012). However, the Coreguaje showed low positive
values with Noanama speakers, but high negative values with members of its own linguistic
family, Cubeo and Tucano. The Coreguaje tribe therefore shows below-average relationship and
is genetically distinct from the other four groups. This multidimensional R matrix was reduced to
a two-dimensional eigenvector plot (Fig. 2). This shows a relatively close relationship between
the Noanama-speaking tribes and between the Cubeo and Tucano, but the Coreguaje, who speak
oriental and occidental Tucano, as expected are genetically distinct and are separated from the
other groups in the lower part of the plot. The diagonal element of the R matrix indicates the
0 1996 Blackwell Science Lld. European Journal of Immunogenelics 23.21-28
27
HLA antigens in Amerindians
TABLE
2. R Matrix for five Amerindian groups
Population
Cubeo
Cubeo
0.0159
Tucano
0.0268
Tucano
Coreguaje
Noanama
Embera
0.0554
Coreguaje
-0.0186
-0.0433
Noanama
-0.0104
-0.0238
0.0064
0.0122
Embera
-0.021 1
-0.0410
0.0046
0.0232
0.0185
0.0232
Mean diagonal element = Rs.1. = 0.0246.
O€MBERA
WOANAMA
OTUCANO
,CUBE0
eigenvectorl
+3
-3
0
COREGUAJE
eigenvector2
FIG.2. Eigenvector plot of the R matrix for the five Colombian Amerindian tribes
degree of isolation characteristic of a particular subgroup. The highest diagonal value is for the
Tucano (0.055) and lowest for the Noanama (0.012). The mean diagonal element (RsT) provides
an index of genetic differentiation among populations. The Rsr value is 0.025, which shows a
moderately high level of genetic differentiation among these groups.
In summary, the genetic diversity at the HLA loci in the Amerindian groups studied is vast.
One possible reason for this diversity is admixture in some groups, which tends to provide a wide
spectrum of antigenic specificities, as observed in the Cubeo and Tucano, in both of which there
seems to be admixture from non-Indian groups. Unlike the findings of Black et al. (1994, cited in
Callegari-Jacques et al. 1994), our results suggest that members of the same linguistic family tend
0 1996 Blackwell Science Ltd. European Journal of lmmunogenerics 23,2 1-28
I. Briceno et al.
28
to share greater affinity, while other groups, such as the Coreguaje, who speak a slightly different
version of the Tucano language and are geographically distant from other members of the same
linguistic group, still retain the restricted HLA distribution pattern seen in most of the Indian
groups.
ACKNOWLEDGMENTS
Thanks are due to the Instituto Colombiano de Cultura Hispanica, Colciencias and to the
Academic de Ciencias Exactas, Fiscias Y Naturates de Madrid for their financial support and to
the Hospital and authorities of Mitu and the Sisters of Mama Bwe in the Orteguaza River for their
help. We are grateful to Dr D.J. Pritchard for his critical perusal of the manuscript.
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