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along with environmental factors&#46;<a class="elsevierStyleCrossRefs" href="#bib0115"><span class="elsevierStyleSup">3&#44;4</span></a> Various genes of the immune system have been reported to contribute to the development of this disease&#44; such as the autoimmune regulator &#40;AIRE&#41; gene&#44; human leukocyte antigen &#40;HLA&#41;&#44; interleukin &#40;IL&#41; genes&#44; cytotoxic T lymphocyte-associated antigen 4 &#40;CTLA4&#41;&#44; protein tyrosine phosphatase non-receptor type 22 &#40;PTPN22&#41;&#44; tumor necrosis factor superfamily member 6 or CD95 &#40;FAS&#41; and FAS ligand &#40;FASL&#41; among others&#46;<a class="elsevierStyleCrossRefs" href="#bib0125"><span class="elsevierStyleSup">5&#44;6</span></a></p><p id="par0010" class="elsevierStylePara elsevierViewall">The cytotoxic T-lymphocyte antigen 4 &#40;CTLA4&#41; gene &#40;2q34&#41; is a member of the immunoglobulin superfamily encoding a protein whose function is that of a potent negative regulator of T-cell response&#46; Also involved in the maintenance of immune tolerance&#44; it has been implicated as a susceptibility gene in human autoimmune diseases&#44; presenting several polymorphisms that have been associated with its development&#46;<a class="elsevierStyleCrossRef" href="#bib0135"><span class="elsevierStyleSup">7</span></a> Among the latter&#44; two genetic variants may contribute to AA susceptibility&#58; &#43;49AG &#40;rs231775&#41;&#44; a missense variation leading to a threonine to alanine aminoacid substitution at codon 17 in the leader peptide &#40;T17A&#41;&#59; and CT60 &#40;rs3087243&#41;&#44; located 236 bp downstream of CTLA4 gene &#40;3&#8242;-UTR&#41;&#46;<a class="elsevierStyleCrossRef" href="#bib0140"><span class="elsevierStyleSup">8</span></a> However&#44; recent studies conducted in different populations have shown that the relationship of these genetic variants in the development of AA is controversial&#46;<a class="elsevierStyleCrossRefs" href="#bib0115"><span class="elsevierStyleSup">3&#44;9</span></a></p><p id="par0015" class="elsevierStylePara elsevierViewall">The current study sought to determine whether the CTLA4 &#43;49AG &#40;rs231775&#41; 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The patients were recruited from the Dermatology Department after the acceptance and signature of an informed consent&#46; Ethical approval was given by the Institutional Review Board and this study was registered under the code DE09-001&#46; Further details regarding the clinical and demographic characteristics&#44; AA type and distribution of each patient were obtained during the clinical evaluation&#46;</p></span><span id="sec0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0055">DNA isolation and genotyping</span><p id="par0025" class="elsevierStylePara elsevierViewall">Genomic DNA was isolated from peripheral venous blood using a &#8220;salting-out&#8221; method and suspended in Tris&#8211;EDTA &#40;pH 7&#46;8&#41; at a final concentration 0&#46;1&#8211;1&#46;0<span class="elsevierStyleHsp" style=""></span>&#956;g&#47;&#956;L before use&#46; Genotype and allele frequencies of the CTLA4 &#43;49AG &#40;rs231775&#41; andCT60 &#40;rs3087243&#41; gene variants were characterized by PCR-RFLP using a MJ Mini PTC1148 thermal cycler &#40;Bio-Rad&#44; Hercules&#59; CA&#44; USA&#41;&#44; specific oligonucleotide primers &#40;IDT&#44; Coralville&#44; IA&#44; USA&#41; and restriction enzymes &#40;New England Biolabs&#44; Ipswich&#44; MA&#44; USA&#41; according to a previously published protocol&#46;<a class="elsevierStyleCrossRef" href="#bib0150"><span class="elsevierStyleSup">10</span></a> The fragments obtained were analyzed by electrophoresis in a 2&#46;5&#37; agarose gel containing ethidium bromide&#44; visualized in UVP model 2UV High Performance Transilluminator &#40;Upland&#44; CA&#44; USA&#41; and documented&#46;</p></span><span id="sec0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0060">Statistical analysis</span><p id="par0030" class="elsevierStylePara elsevierViewall">For genetic analysis&#44; the sample size was calculated based on 2&#37; worldwide alopecia areata incidence&#44;<a class="elsevierStyleCrossRef" href="#bib0155"><span class="elsevierStyleSup">11</span></a> assuming a power of 99&#46;0&#37; &#40;a <span class="elsevierStyleItalic">Z</span> value of 2&#46;33&#41;&#44; a minimum number of 43 subjects is sufficient&#46;</p><p id="par0035" class="elsevierStylePara elsevierViewall">Data analysis was performed with the SPSS v21&#46;0 software for windows &#40;SPSS&#44; Inc&#46;&#44; Chicago&#44; IL&#44; USA&#41; and the Epi-INFO TM 7 statistical program &#40;Stone&#46; Mountain&#44; GA&#44; USD Inc&#41;&#46; Hardy&#8211;Weinberg equilibrium for the alleles was obtained using a goodness-of-fit test and the genotypic dependence between patients and controls was determined with a <span class="elsevierStyleItalic">&#967;</span><span class="elsevierStyleSup">2</span> test&#59; OR was calculated from 2<span class="elsevierStyleHsp" style=""></span>&#215;<span class="elsevierStyleHsp" style=""></span>2 contingency tables &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05&#41;&#46;</p></span></span><span id="sec0030" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0065">Results</span><span id="sec0035" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0070">Clinical characteristics of the participants</span><p id="par0040" class="elsevierStylePara elsevierViewall">The present study analyzed the genotype distribution and allele frequency of CTLA4 &#43;49AG and CT60 gene variants&#46; Our universe &#40;<span class="elsevierStyleItalic">n</span>&#41; consisted of 50 AA patients and 100 control participants&#46; The clinical presentation of AA in the patients was&#58; 41 Patchy AA subjects &#40;single patch <span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>14&#44; 8 females and 6 males&#59; multiple patches <span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>27&#44; 19 females and 8 males&#41;&#44; 4 Ophiasis AA subjects &#40;3 females and 1 male&#41;&#44; 1 female with Totalis AA and 4 Universalis AA subjects &#40;3 males and 1 female&#41;&#46; Ten patients &#40;20&#37;&#41; had a personal history of autoimmune diseases &#40;Vitiligo&#44; type 2 DM&#44; among others&#41; and 36 subjects &#40;72&#37;&#41; have family history of immune disease&#46;</p></span><span id="sec0040" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0075">Allele and genotype analysis</span><p id="par0045" class="elsevierStylePara elsevierViewall">Several tests were performed for both genetic variants in order to assess the Hardy&#8211;Weinberg equilibrium &#40;HWE&#41; &#40;<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>&#41; all of which had a <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#62;<span class="elsevierStyleHsp" style=""></span>0&#46;05&#44; indicating that rs231775 and rs3087243 gene variants were in HWE in both AA patients and controls&#46;</p><elsevierMultimedia ident="tbl0005"></elsevierMultimedia><p id="par0050" class="elsevierStylePara elsevierViewall">We compared the CTLA4 rs231775 and the rs3087243 alleles and genotypes of the control group with AA subjects &#40;all AA types&#41;&#44; and with the different types of AA&#44; detailed results are shown in <a class="elsevierStyleCrossRef" href="#tbl0005"><span class="elsevierStyleBold">table 1</span></a>&#46; For CTLA4 rs231775&#44; 30&#37; of AA patients showed an homozygous wild type AA genotype&#44; 48&#37; showed the heterozygous AG genotype&#44; and the remaining 22&#37; were variant homozygous GG genotype&#46; For CTLA4 rs3087243&#44; 44&#37; of the AA patients showed the homozygous wild type GG genotype&#44; 44&#37; showed the heterozygous AG genotype and the remaining 12&#37; were variant homozygous AA genotype&#46;</p></span><span id="sec0045" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0080">Association of the genotypes in patients con alopecia areata</span><p id="par0055" class="elsevierStylePara elsevierViewall">When CTLA4 rs231775 and rs3087243 genotypes and allele frequencies were compared between the whole cohort &#40;all AA types&#41; and controls&#44; we observed that the heterozygous AG genotype for CTLA4 rs231775 and rs3087243 was more frequent in controls than across all AA patient groups&#46; Further&#44; no evidence was observed concerning the association of the allele variants and AA in the analyzed patients &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#62;<span class="elsevierStyleHsp" style=""></span>0&#46;05&#41; &#40;<a class="elsevierStyleCrossRef" href="#tbl0010">Table 2</a>&#41;&#46; Furthermore&#44; no relationship was found between genotype and type of AA with the personal&#47;familiar history of autoimmune diseases or the sex of the analyzed patients &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#62;<span class="elsevierStyleHsp" style=""></span>0&#46;05&#41; &#40;<a class="elsevierStyleCrossRef" href="#tbl0015">Table 3</a>&#41;&#46;</p><elsevierMultimedia ident="tbl0010"></elsevierMultimedia><elsevierMultimedia ident="tbl0015"></elsevierMultimedia></span></span><span id="sec0050" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0085">Discussion</span><p id="par0060" class="elsevierStylePara elsevierViewall">AA is an autoimmune hair-loss condition that has been associated with multiple gene variants&#44; several of which participate in immune response pathways related to HLA&#46;<a class="elsevierStyleCrossRef" href="#bib0160"><span class="elsevierStyleSup">12</span></a> The CTLA4 gene product is a receptor expressed by CD4&#43; and CD8&#43; T-cells&#46; Experiments in knockout mice have demonstrated the important role that CTLA4 plays in protection against autoimmunity&#46; CTLA-4 is a critical negative regulator of T cell responses&#44; and the action of CTLA4 on Treg cells can control the activity of other cells and controlling fatal autoimmunity&#46;<a class="elsevierStyleCrossRef" href="#bib0165"><span class="elsevierStyleSup">13</span></a></p><p id="par0065" class="elsevierStylePara elsevierViewall">The association between the presence of the CTLA4 gene variants and the development of different autoimmune diseases has been suggested&#46;<a class="elsevierStyleCrossRefs" href="#bib0170"><span class="elsevierStyleSup">14&#44;15</span></a> However in immune dermatological diseases&#44; the results have been controversial&#59; in vitiligo their participation or lack of influence<a class="elsevierStyleCrossRef" href="#bib0180"><span class="elsevierStyleSup">16</span></a> has been seen&#46; A similar situation has been observed with the development of psoriasis&#46;<a class="elsevierStyleCrossRefs" href="#bib0185"><span class="elsevierStyleSup">17&#44;18</span></a></p><p id="par0070" class="elsevierStylePara elsevierViewall">In this regard&#44; considering the possible contribution of the autoimmune function in alopecia&#44; genetic variant studies of CTLA4 gene have been explored in diverse populations&#46; A previous association analysis has suggested the plausible involvement of CTLA4 gene variants as a risk factor in the development of AA in a European population&#44;<a class="elsevierStyleCrossRef" href="#bib0140"><span class="elsevierStyleSup">8</span></a> with a stronger effect in patients with severe forms of the disorder&#46;</p><p id="par0075" class="elsevierStylePara elsevierViewall">Another study conducted on an Italian population&#44; where rs231775 and rs3087243 gene variants were analyzed&#44; observed the latter as a risk factor in the development of patchy AA&#46;<a class="elsevierStyleCrossRef" href="#bib0145"><span class="elsevierStyleSup">9</span></a> On the other hand&#44; a study conducted on Iran nationals failed to prove the association of the rs3087243 gene variant in the development of AA&#46;<a class="elsevierStyleCrossRef" href="#bib0115"><span class="elsevierStyleSup">3</span></a></p><p id="par0080" class="elsevierStylePara elsevierViewall">Previously&#44; we observed the participation of genetic variants of TNF&#945; and PTPN22<a class="elsevierStyleCrossRef" href="#bib0195"><span class="elsevierStyleSup">19</span></a> as a risk factor for AA in Mexican patients&#59; both of these genes&#8217; function is related to the regulation of immune mechanisms&#46; However&#44; the participation of CTLA4 genetic variants in AA has not been analyzed in this population&#46;</p><p id="par0085" class="elsevierStylePara elsevierViewall">Some SNPs within the CTLA-4 gene have already been analyzed for susceptibility to rheumatoid arthritis &#40;RA&#41; in the Mexican population&#59; among them&#44; rs5742909&#44; rs231775 and rs3087243&#44; suggesting that the &#8722;319C&#47;&#43;49G&#47;CT60G haplotype of the CTLA-4 gene is a risk factor for RA&#44; whereas that the rs3087243 SNP could be a protective factor against this type of autoimmune disease&#46;<a class="elsevierStyleCrossRef" href="#bib0200"><span class="elsevierStyleSup">20</span></a></p><p id="par0090" class="elsevierStylePara elsevierViewall">However&#44; no significant differences were observed for rs231775 and rs3087243 in our study population &#40;<a class="elsevierStyleCrossRef" href="#tbl0010">Table 2</a>&#41;&#44; suggesting that these variants are not a risk factor in the development of AA or Patchy AA in the Mexican population&#46;</p></span><span id="sec0055" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0090">Conclusion</span><p id="par0095" class="elsevierStylePara elsevierViewall">In conclusion&#44; the genetic variants rs231775 and rs3087243 of the CTLA4 gene do not constitute a risk factor in the development of AA in the Mexican population of Monterrey&#44; Mexico&#46; Further&#44; these genetic variants have no association between family&#47;personal backgrounds of autoimmune diseases or the gender of the study subjects&#46;</p></span><span id="sec0060" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0095">Financial support</span><p id="par0100" class="elsevierStylePara elsevierViewall">None declared&#46;</p></span><span id="sec0065" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0100">Authors&#39; contributions</span><p id="par0105" class="elsevierStylePara elsevierViewall">Mauricio Andr&#233;s Salinas Santander&#58; Statistic analysis&#59; approval of the final version of the manuscript&#59; conception and planning of the study&#59; elaboration and writing of the manuscript&#59; obtaining&#44; analysis&#44; and interpretation of the data&#59; effective participation in research orientation&#59; critical review of the literature&#59; critical review of the manuscript&#46;</p><p id="par0110" class="elsevierStylePara elsevierViewall">Cristina Susana Cantu-Salinas&#58; Approval of the final version of the manuscript&#59; conception and planning of the study&#59; elaboration and writing of the manuscript&#59; obtaining&#44; analysis&#44; and interpretation of the data&#59; effective participation in research orientation&#59; critical review of the literature&#59; critical review of the manuscript&#46;</p><p id="par0115" class="elsevierStylePara elsevierViewall">Jorge Ocampo-Candiani&#58; Approval of the final version of the manuscript&#59; conception and planning of the study&#59; effective participation in research orientation&#59; critical review of the literature&#59; critical review of the manuscript&#46;</p><p id="par0120" class="elsevierStylePara elsevierViewall">Victor de Jesus Suarez-Valencia&#58; Approval of the final version of the manuscript&#59; elaboration and writing of the manuscript&#59; obtaining&#44; analysis&#44; and interpretation of the data&#59; critical review of the literature&#59; critical review of the manuscript&#46;</p><p id="par0125" class="elsevierStylePara elsevierViewall">Jennifer Guadalupe Ramirez-Guerrero&#58; Approval of the final version of the manuscript&#59; elaboration and writing of the manuscript&#59; obtaining&#44; analysis&#44; and interpretation of the data&#59; critical review of the literature&#46;</p><p id="par0130" class="elsevierStylePara elsevierViewall">Celia Nohemi Sanchez-Dominguez&#58; Approval of the final version of the manuscript&#59; elaboration and writing of the manuscript&#59; critical review of the literature&#59; critical review of the manuscript&#46;</p></span><span id="sec0070" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0105">Conflicts of interest</span><p id="par0135" class="elsevierStylePara elsevierViewall">None declared&#46;</p></span></span>"
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    "fechaRecibido" => "2019-06-06"
    "fechaAceptado" => "2019-09-24"
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            0 => "Alopecia areata"
            1 => "Autoimmune diseases"
            2 => "Polymorphism"
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        "resumen" => "<span id="abst0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0010">Background</span><p id="spar0005" class="elsevierStyleSimplePara elsevierViewall">Alopecia areata is an autoimmune disease that produces non-scarring hair loss around the body&#46; Gene variants of the cytotoxic T-lymphocyte antigen 4 &#40;CTLA4&#41; gene&#44; a negative regulator of T-cell response&#44; have been associated with a predisposition to autoimmune diseases in different populations&#59; however&#44; the involvement of these genetic variants in the development of AA is controversial&#46;</p></span> <span id="abst0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0015">Objective</span><p id="spar0010" class="elsevierStyleSimplePara elsevierViewall">The present study evaluated the potential association of two CTLA4 gene variants with alopecia areata in a Mexican population&#46;</p></span> <span id="abst0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0020">Methods</span><p id="spar0015" class="elsevierStyleSimplePara elsevierViewall">We genotyped &#43;49AG &#40;rs231775&#41; and CT60 &#40;rs3087243&#41; variants in 50 AA patients and 100 healthy control participants through PCR-RFLP&#46;</p></span> <span id="abst0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0025">Results</span><p id="spar0020" class="elsevierStyleSimplePara elsevierViewall">No statistical difference was observed for either of the gene variants regarding allele or genotype frequencies between AA patients and the controls when the parameters of family&#47;personal history of autoimmune diseases or gender were considered &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#62;<span class="elsevierStyleHsp" style=""></span>0&#46;05&#41;&#46; <span class="elsevierStyleItalic">Study limitations&#58;</span> Small sample size of patients and the data were obtained from Northeast Mexico population&#46;</p></span> <span id="abst0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0030">Conclusion</span><p id="spar0025" class="elsevierStyleSimplePara elsevierViewall">The genetic variants rs231775 and rs3087243 of the CTLA4 gene are not a risk factor for the development of alopecia areata in the analyzed Mexican population&#46;</p></span>"
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        "nota" => "<p class="elsevierStyleNotepara" id="npar0005">How to cite this article&#58; Salinas-Santander MA&#44; Cantu-Salinas CS&#44; Ocampo-Candiani J&#44; Suarez-Valencia VJ&#44; Ramirez-Guerrero JG&#44; Sanchez-Dominguez CN&#46; CTLA4 &#43;49AG &#40;rs231775&#41; and CT60 &#40;rs3087243&#41; gene variants are not associated with alopecia areata in a Mexican population from Monterrey Mexico&#46; An Bras Dermatol&#46; 2020&#59;95&#58;283&#8211;8&#46;</p>"
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        "nota" => "<p class="elsevierStyleNotepara" id="npar0010">Study conducted at the Department of Investigation&#44; Facultad de Medicina Unidad Saltillo&#44; Universidad Aut&#243;noma de Coahuila&#44; Coahuila&#44; M&#233;xico&#44; and Dermatology Service&#44; Hospital Universitario Dr&#46; Jos&#233; Eleuterio Gonz&#225;lez&#44; Facultad de Medicina&#44; Universidad Aut&#243;noma de Nuevo Le&#243;n&#44; Nuevo Le&#243;n&#44; M&#233;xico&#46;</p>"
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          "en" => "<p id="spar0040" class="elsevierStyleSimplePara elsevierViewall">Analysis of the gene variants CTLA4 &#43;49AG &#40;rs231775&#41; and CT60 &#40;rs3087243&#41; frequencies with the sex and personal&#47;familiar history of autoimmune diseases&#46;</p>"
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                            0 => "T&#46; Simakou"
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