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Frequency of CCR5∆32 in Brazilian Populations

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Frequency of CCR5∆32 in Brazilian Populations Brazilian Journal of Medical and Biological Research (2006) 39: 321-325 CCR5∆32 in Brazilian populations 321 ISSN 0100-879X Short Communication Frequency of CCR5∆32 in Brazilian populations A.E. Vargas, A.R. Marrero, Departamento de Genética, Instituto de Biociências, F.M. Salzano, M.C. Bortolini Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brasil and J.A.B. Chies Abstract Correspondence A sample of 103 randomly chosen healthy individuals from Alegrete, Key words J.A.B. Chies RS, Brazil, was tested for the CCR5∆32 allele, which is known to • CCR5 Departamento de Genética, UFRGS influence susceptibility to HIV-1 infection. The CCR5∆32 allele was • Chemokine receptors Caixa Postal 15053 identified by PCR amplification using specific primers flanking the • Brazilian population 91501-970 Porto Alegre, RS region of deletion, followed by electrophoresis on a 3% agarose gel. • Gene flow Brasil Fax: +55-51-3316-7311 The data obtained were compared to those reported for other popula- E-mail: [email protected] tions and interpreted in terms of Brazilian history. The individuals studied came from a highly admixed population. Most of them were Research supported by PRONEX identified as white (N = 59), while blacks and browns (mulattoes) (No. 66.1002/1997.9), CNPq, were N = 13 and N = 31, respectively. The observed frequencies, FAPERGS, and PROPESQ/UFRGS. considering the white, black and brown samples (6.8, 3.8, and 6.4%, respectively), suggest an important European parental contribution, even in populations identified as black and brown. However, in Brazil Received April 29, 2005 as a whole, this allele shows gradients indicating a relatively good Accepted October 14, 2005 correlation with the classification based on skin color and other physical traits, used here to define major Brazilian population groups. One of the most interesting characteris- grants was unequal in the various Brazilian tics of the Brazilian population is its hetero- regions. In the North, the populations were geneity. When Brazil was “discovered” by formed mainly by Europeans and Amerindi- the Portuguese in 1500, the land was already ans; in the Southeast and Northeast, Europe- inhabited by Amerindians (estimated at 2 ans, Africans and Amerindians had different million people). Since then, emigration of degrees of influence, while in the South, the individuals from different countries and con- European heritage prevails (1). tinents with diverse ethnic backgrounds has CCR5 is a chemokine receptor present contributed to the establishment of the ge- mainly in cells of the immune system, such as netic pool of the contemporary Brazilian macrophages and T lymphocytes, playing a population. These parental contributions in- major role in the migration of these cells to cluded a constant influx of Portuguese, 4 sites of inflammation. The gene encoding million Africans (mainly from West-Central CCR5 (CKR5) is located in the p21.3 region of Africa) and 3.9 million Europeans (other the human chromosome 3, forming a cluster than Portuguese), who arrived here in the with other chemokine receptor genes (2). Deng 19th and 20th centuries (1). et al. (3) demonstrated that CCR5 serves as a However, the distribution of these immi- co-receptor for human immunodeficiency vi- Braz J Med Biol Res 39(3) 2006 322 A.E. Vargas et al. rus-1 (HIV-1). The variant allele CCR5∆32 The investigation was approved by the Bra- described by Liu et al. (2) contains a 32-bp zilian National Ethics Committee (CONEP deletion that generates a truncated protein, No. 1333/2002) and all donors were informed which confers relative resistance to HIV-1 about the aims of this study and signed a infection. written consent. The study of the allelic frequency of DNA was extracted from saliva or blood CCR5∆32 in 18 European populations re- samples using the Nucleon DNA Extraction vealed an interesting North-South gradient, kit (Nucleon Bioscience, Coatbridge, UK) with the highest frequencies of the variant or a salting-out method, respectively. allele being observed in Finnish and Mord- Genotyping was performed by PCR am- vinian populations (16%) and the lowest in plification with specific primers. PCR sam- Sardinia (4%) (4). The last investigators also ples were prepared to a final volume of 25 proposed that the CCR5∆32 allele origi- µL as follows: 1 µL DNA (0.2-0.5 µg), 2.5 nated from a single mutation event in North- µL 10X PCR buffer (200 mM Tris-HCl, pH eastern Europe a few thousand years ago. 8.4, 500 mM KCl), 1 µL 50 mM MgCl2, 1 µL The high frequencies of CCR5∆32 found in 3 mM dNTP mix, 1 µL 10 pmol primer mix, Europeans have been attributed to a strong and 0.2 µL Taq DNA polymerase, 5 U/µL selective pressure, possibly exerted by patho- (Invitrogen Corporation, San Diego, CA, gens such as Yersinia pestis (the bubonic USA). Samples were submitted to 40 cycles plague agent), Shigella, Salmonella, and of 1 min at 94ºC, 1 min at 55ºC, and 1 min at Mycobacterium tuberculosis, all of which 72ºC. The set of specific primers used to target macrophages, or by some other infec- amplify the CCR5 gene segment was de- tious diseases such as syphilis, smallpox and scribed by Chies and Hutz (6). It yields a influenza (5). 137-bp fragment for the wild-type allele and Thus, the prevalence of this allele is of a 105-bp fragment for the CCR5∆32 variant. obvious medical importance. We have in- PCR products were plotted on 3% agarose vestigated its distribution in a random sample gel containing ethidium bromide and sub- of individuals from Alegrete, a town located mitted to electrophoresis. Fragments were in the western region of Rio Grande do Sul visualized under UV irradiation. (29º53' S; 55º57' W) where the population Skin color is used in Brazil as the equiva- was basically established from a mixture of lent of race, and is based on a complex and Spanish, Portuguese and African individu- subjective phenotypic evaluation. In Brazil, als and native Amerindians, and compared it the emphasis is on physical appearance rather to those already reported for other popula- than ancestry, which is in contrast to the tions, interpreting the data in terms of Bra- situation in the United States. The Brazilian zilian history. Institute of Geography and Statistics (IBGE) A sample of 103 randomly chosen unre- adopts the criterion of classification of indi- lated healthy individuals from Alegrete, RS, viduals into the following categories: white Brazil, was analyzed in the present study. (in Portuguese, branco), black (preto), brown Most of the individuals studied were identi- (pardo), yellow (amarelo), and Amerindian fied as white (N = 59), while blacks and (indígena). Accordingly, in Brazil as a whole, browns (mulattoes) were N = 13 and N = 31, 53, 6, and 38% of the persons are identified respectively. This classification was based as white, black, and brown, respectively, the on physical appearance as judged by the remaining 3% being distributed among yel- researcher at the time of blood collection, low and Amerindian persons. In Rio Grande and on data about the ethnicity of parents/ do Sul (~10 million inhabitants), the num- grandparents reported by the participants. bers are 87.5, 5, 7, and 0.5% for white, black, Braz J Med Biol Res 39(3) 2006 CCR5∆32 in Brazilian populations 323 brown, and yellow + Amerindian individu- do not report admixture with non-Europe- als, respectively (7). More recently, the ex- ans. Brown will be used to refer to individu- pression Afro-descendent has been incorpo- als with intermediate physical appearance rated into this ethnic semantic definition (8). between white and black. However, the last investigators have esti- It has been widely observed that most mated that about 148 million Brazilians pres- populations share alleles at any given locus ent more than 10% of African nuclear ge- and that those alleles that are most frequent nome ancestry, and that at least 89 millions in one population are also found at high of individuals have mtDNA lineages of Af- frequency in others, reflecting the recent rican origin (8). This illustrates the exten- dispersion of Homo sapiens into continental sion of admixture in Brazil and supports the groups (9). Due to this fact, there are few suggestion that skin color and other pheno- classical or DNA markers that have been typic traits can be poor predictors of genom- demonstrated either to be population-specif- ic ancestry. These results reinforce the idea ic or to have large frequency disparities that, independently of the chosen criteria, it among geographically and ethnically defined is problematic to classify people. To facili- populations (9). tate reading and comprehension, the word In the present study, no CCR5∆32/ “black” will be used here to refer to any CCR5∆32 homozygotes were detected. The person (or population) identified and/or self- presence of the CCR5/CCR5∆32 genotype identified with some term that reports Afri- among whites, blacks and browns was 14, 8, can ancestry according to physical appear- and 13%, indicating a CCR5∆32 allele fre- ance, whereas “white” will be used to define quency of 6.8, 3.8, and 6.4%, respectively. those that, according to their physical traits, The CCR5∆32 distributions observed in Table 1. CCR5 genotype and CCR5∆32 allele frequencies in Brazilian populations and in their putative parental groups. Brazilian populations No. Genotype ∆32 allele References individuals frequencies (%) frequency (%)a CCR5/∆32 ∆32/∆32 Brown/unclassified urban/semi-urban North 203 15 1 4.2 14 Southeast 539 57 0 5.3 17-19 South 31 4 0 6.4 Present study Black urban/semi-urban Northeast 549 29 0 2.6 6 Southeast 54 4 0 1.9 6 South 71 3 0 0.7 6, present study Black Rural 296 11 0 1.9 14,15 White urban/semi-urban South 158 19 1 6.6 10, present study Parental groups Amerindians 1071 5 0 0.2 10,12,14,20 Europeans 2668 492 23 10.1 4 Africans 251 0 0 0 11 aWeighted average allele frequencies were obtained when more than one study was considered.
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