RESUMO
BACKGROUND: The Djallonke sheep is well adapted to harsh environmental conditions, and is relatively resistant to Haemonchosis and resilient to animal trypanosomiasis. The larger Sahelian sheep, which cohabit the same region, is less well adapted to these disease challenges. Haemonchosis and Trypanosomiasis collectively cost the worldwide animal industry billions of dollars in production losses annually. RESULTS: Here, we separately sequenced and then pooled according to breed the genomes from five unrelated individuals from each of the Djallonke and Sahelian sheep breeds (sourced from Ghana), at greater than 22-fold combined coverage for each breed. A total of approximately 404 million (97%) and 343 million (97%) sequence reads from the Djallonke and Sahelian breeds respectively, were successfully mapped to the sheep reference genome Oar v3.1. We identified approximately 11.1 million and 10.9 million single nucleotide polymorphisms (SNPs) in the Djallonke and Sahelian breeds, with approximately 15 and 16% respectively of these not previously reported in sheep. Multiple regions of reduced heterozygosity were also found; 70 co-localised within genomic regions harbouring genes that mediate disease resistance, immune response and adaptation in sheep or cattle. Thirty- three of the regions of reduced heterozygosity co-localised with previously reported genes for resistance to haemonchosis and trypanosomiasis. CONCLUSIONS: Our analyses suggest that these regions of reduced heterozygosity may be signatures of selection for these economically important diseases.
Assuntos
Adaptação Fisiológica/genética , Resistência à Doença/genética , Genômica , Heterozigoto , Ovinos/genética , Ovinos/fisiologia , Clima Tropical , Animais , Cruzamento , Cromossomos de Mamíferos/genética , Feminino , Masculino , Polimorfismo de Nucleotídeo Único , Ovinos/imunologia , Ovinos/microbiologia , Tripanossomíase/imunologiaRESUMO
BACKGROUND: Carcinoma of unknown primary (CUP) is a metastatic epithelial malignancy in the absence of an identifiable primary tumour. Prognosis for patients with CUP is poor because treatment options are generally limited to broad spectrum chemotherapy. A shift towards personalised cancer management based on mutation profiling offers the possibility of new treatment paradigms. This study has explored whether actionable, oncogenic driver mutations are present in CUP that have potential to better inform treatment decisions. METHODS: Carcinoma of unknown primary cases (n = 21) were selected and DNA was isolated from formalin-fixed paraffin embedded sections prior to amplification and sequencing. Two distinct yet complementary targeted gene panels were used to assess variants in up to 76 known cancer-related genes for the identification of biologically relevant and actionable mutations. RESULTS: Variants were detected in 17/21 cases (81%) of which 11 (52%) were potentially actionable with drugs currently approved for use in known primary cancer types or undergoing clinical trials. The most common variants detected were in TP53 (47%), KRAS (12%), MET (12%) and MYC (12%). Differences at the molecular level were seen between common CUP histological subtypes. CUP adenocarcinomas and poorly differentiated carcinomas harboured the highest frequency of variants in genes involved in signal transduction pathways (e.g. MET, EGFR, HRAS, KRAS, and BRAF). In contrast, squamous cell carcinoma exhibited a higher frequency of variants in cell cycle control and DNA repair genes (e.g. TP53, CDKN2A and MLH1). CONCLUSION: Taken together, mutations in biologically relevant genes were detected in the vast majority of CUP tumours, of which half provided a potentially novel treatment option not generally considered in CUP.
Assuntos
Terapia de Alvo Molecular , Neoplasias Primárias Desconhecidas/genética , Adulto , Idoso , Feminino , Variação Genética , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias Primárias Desconhecidas/patologiaRESUMO
Human leucocyte antigen (HLA) genes play an important role in the success of organ transplantation and are associated with autoimmune and infectious diseases. Current DNA-based genotyping methods, including Sanger sequence-based typing (SSBT), have identified a high degree of polymorphism. This level of polymorphism makes high-resolution HLA genotyping challenging, resulting in ambiguous typing results due to an inability to resolve phase and/or defining polymorphisms lying outside the region amplified. Next-generation sequencing (NGS) may resolve the issue through the combination of clonal amplification, which provides phase information, and the ability to sequence larger regions of genes, including introns, without the additional effort or cost associated with current methods. The NGS HLA sequencing project of the 16IHIW aimed to discuss the different approaches to (i) template preparation including short- and long-range PCR amplicons, exome capture and whole genome; (ii) sequencing platforms, including GS 454 FLX, Ion Torrent PGM, Illumina MiSeq/HiSeq and Pacific Biosciences SMRT; (iii) data analysis, specifically allele-calling software. The pilot studies presented at the workshop demonstrated that although individual sequencers have very different performance characteristics, all produced sequence data suitable for the resolution of HLA genotyping ambiguities. The developments presented at this workshop clearly highlight the potential benefits of NGS in the HLA laboratory.
Assuntos
DNA/genética , Antígenos HLA , Sequenciamento de Nucleotídeos em Larga Escala , Transplante de Órgãos , Alelos , Genótipo , Antígenos HLA/classificação , Antígenos HLA/genética , Antígenos HLA/imunologia , Sequenciamento de Nucleotídeos em Larga Escala/instrumentação , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Teste de Histocompatibilidade , Humanos , Polimorfismo Genético , Análise de Sequência de DNA , SoftwareRESUMO
The region spanning the tumour necrosis factor (TNF) cluster in the human major histocompatibility complex is implicated in susceptibility to immunopathological disease, but ethnic differences and linkage disequilibrium have hampered identification of critical polymorphisms. Here, we investigate Europeans, Asians (Bidayuh, Chinese, Indian, Jehai, Malay, Temuan) and Australian Aborigines to provide a framework for disease-association studies. DNA from 999 unrelated healthy donors was genotyped at 38 loci, primarily in coding and promoter regions over a 60-kb region spanning seven genes near TNF. The PHASE algorithm was used to statistically infer TNF block haplotypes and estimate their frequencies in each population. The TNF block is carried as 31 haplotypes in all populations combined, with <19 in any single population. Only six haplotypes have a unique tag single nucleotide polymorphism (SNP) valid for all populations, but seven haplotypes could be tagged with individual SNPs in selected populations. Four to eight TNF block haplotypes exist across all ethnicities, and hence must pre-date the divergence of these populations from a common ancestor >160,000 years ago. Some haplotypes are unique to isolated populations, but they do not contain unique SNP. Hence, they reflect restricted migration and/or extinction of some families rather than de novo mutation.
Assuntos
Povo Asiático/genética , Frequência do Gene/genética , Haplótipos/genética , Havaiano Nativo ou Outro Ilhéu do Pacífico/genética , Fatores de Necrose Tumoral/genética , População Branca/genética , Alelos , Cromossomos Humanos Par 6/genética , Evolução Molecular , Variação Genética , Humanos , Filogenia , Polimorfismo de Nucleotídeo Único/genética , Regiões Promotoras GenéticasRESUMO
The implementation and popularity of next generation sequencing (NGS) has led to the development of various rapid whole mitochondrial genome sequencing techniques. We summarise an efficient and cost-effective NGS approach for mitochondrial genomic DNA in humans using the Ion Torrent platform, and further discuss our bioinformatics pipeline for streamlined variant calling. Ion 316 chips were utilised with the Ion Torrent semi-conductor platform Personal Genome Machine (PGM) to perform tandem sequencing of mitochondrial genomes from the core pedigree (n = 315) of the Norfolk Island Health Study. Key improvements from commercial methods focus on the initial PCR step, which currently requires extensive optimisation to ensure the accurate and reproducible elongation of each section of the complete mitochondrial genome. Dual-platform barcodes were incorporated into our protocol thereby extending its potential application onto Illumina-based systems. Our bioinformatics pipeline consists of a modified version of GATK best practices tailored for mitochondrial genomic data. When compared with current commercial methods, our method, termed high throughput mitochondrial genome sequencing (HTMGS), allows high multiplexing of samples and the use of alternate library preparation reagents at a lower cost per sample (~1.7 times) when compared to current commercial methodologies. Our HTMGS methodology also provides robust mitochondrial sequencing data (>450X average coverage) that can be applied and modified to suit various study designs. On average, we were able to identify ~30 variants per sample with 572 variants observed across 315 samples. We have developed a high throughput sequencing and analysis method targeting complete mitochondrial genomes; with the potential to be platform agnostic with analysis options that adhere to current best practices.
Assuntos
Genoma Mitocondrial , Sequenciamento de Nucleotídeos em Larga Escala , DNA Mitocondrial/genética , Variação Genética , Humanos , Controle de QualidadeRESUMO
We have shown that normal C57BL/6J mice are moderately resistant to infection with murine cytomegalovirus (MCMV) and that this resistance is impaired by prior infection with LP-BM5 MuLV, which causes a disease (MAIDS) similar to early HIV-induced disease. This study investigates macrophage function in MAIDS+ mice challenged with MCMV. MAIDS reduces the influx of cells into the peritoneal cavity seen in normal C57BL/6J mice 6 days after MCMV infection. The infiltrates contained cells that resembled activated macrophages, as they took up colloidal gold, expressed the macrophage marker Mac-1, had high levels of acid phosphatase activity, and were lymphocytostatic when co-cultured with activated T cells. MAIDS+ mice had a higher percentage of cells able to take up colloidal gold and higher acid phosphatase activity per cell. The cells were also more lymphocytostatic and produced higher levels of interleukin-1 and tumor necrosis factor-alpha on days 4 and 6 after MCMV infection. Hence, MAIDS enhances baseline and induced macrophage activity, but depresses infiltration into the site of inflammation.
Assuntos
Infecções por Citomegalovirus/complicações , Macrófagos Peritoneais/imunologia , Síndrome de Imunodeficiência Adquirida Murina/complicações , Síndrome de Imunodeficiência Adquirida Murina/imunologia , Fosfatase Ácida/análise , Animais , Células Cultivadas , Técnicas de Cocultura , Citomegalovirus/isolamento & purificação , Infecções por Citomegalovirus/imunologia , Feminino , Citometria de Fluxo , Inflamação , Interleucina-1/biossíntese , Ativação Linfocitária , Ativação de Macrófagos , Antígeno de Macrófago 1/análise , Camundongos , Camundongos Endogâmicos C57BL , Linfócitos T/imunologia , Fatores de Tempo , Fator de Necrose Tumoral alfa/biossínteseRESUMO
The contribution of MHC class II haplotypes to susceptibility to type I diabetes has been clearly established, and interest has now focused on the effects of additional genes in the MHC region. We have investigated the central MHC alleles on 8.1 ancestral haplotype (HLA-A1, B8, DR3, DQ2), as it is well conserved in Caucasian populations. The HLA-DR3-DQ2 genotype is a recognized risk factor for type I diabetes. Single nucleotide polymorphisms and microsatellites in the MHC were used to map segments of the 8.1 ancestral haplotype carried by type I diabetic and control subjects expressing either HLA-B8 or DR3, but not both these markers. In this way we controlled for the diabetogenic effect of carriage of DR3. Alleles of the 8.1 ancestral haplotype between TNFA-308/D6STNFa and HLA-B were carried with significantly greater frequency in B8(-), DR3(+) type I diabetic patients compared with B8(-), DR3(+) controls. This interval was marked by a BAT1 gene polymorphism and a MIB microsatellite allele.
Assuntos
Diabetes Mellitus Tipo 1/genética , Antígenos HLA/genética , Complexo Principal de Histocompatibilidade/genética , Polimorfismo de Nucleotídeo Único/genética , Alelos , Diabetes Mellitus Tipo 1/imunologia , Antígeno HLA-B8/genética , Antígeno HLA-DR3/genética , Haplótipos , Humanos , Complexo Principal de Histocompatibilidade/imunologia , Repetições de MicrossatélitesRESUMO
Susceptibility to multiple sclerosis (MS) is clearly associated with human leukocyte antigen (HLA)-DRB1*1501, but some studies show associations with HLA-B7 and -B18. These are often co-expressed with DRB1*1501 in the ancestral haplotypes (AH) denoted 7.1 (HLA-A3, B7, tumor necrosis factor [TNF]a11b4, DRB1*1501) and 18.1 (HLA-A25, B18, TNFa10b4, DRB 1*1501). Here we present a systematic study of 218 patients and 274 controls typed at all standard class II and TNF microsatellite loci, and a novel non-synonymous polymorphism in the central major histocompatibility complex gene, inhibitor of kappa B-like protein (IKBL). The C allele at IKBL+738 is only found on the 7.1 haplotype. HLA-DRB1*1501 was associated with disease, as expected. When subjects expressing DRB 1*501 were analyzed separately, TNFa11b4 and IKBL+738C were less common in the patients and, hence, mark an allele that mediates resistance which lies telomeric of IKBL. TNFa10b4 and TNFa1b5 were more common in DRB1*1501 patients than in controls. These alleles have been associated with the 18.1 and 18.2 AH, respectively. Since no component of these haplotypes was an independent risk factor in this study, it appears likely that a gene linked to TNFa10b4 and TNFa1b5 modifies the effect of the susceptibility locus marked by HLA-DRB1*1501. Potential candidate genes telomeric of the TNF cluster are discussed.
Assuntos
Antígenos HLA-DR/genética , Família Multigênica , Esclerose Múltipla/genética , Telômero , Fator de Necrose Tumoral alfa/genética , Proteínas Adaptadoras de Transdução de Sinal , Adulto , Alelos , Estudos de Coortes , Feminino , Frequência do Gene , Predisposição Genética para Doença , Cadeias HLA-DRB1 , Antígenos de Histocompatibilidade Classe II/genética , Humanos , Masculino , Pessoa de Meia-Idade , Esclerose Múltipla/imunologiaRESUMO
As MHC genes are potent determinants of susceptibility to immunopathological diseases, the mapping of SAPK2a (CSBP) and SAPK4 to chromosome 6p 21.2-21.3 suggested that these genes may mediate the effects of the MHC on disease. Here we describe the genomic structure and localisation of both genes approximately 2.3Mb centromeric of HLA-DP. Examination of the complete coding region and selected intronic regions of SAPK2a and SAPK4 from 22 human EBV-transformed B-cell lines of different MHC haplotypes and racial background revealed complete sequence conservation. There were no notable differences in levels of expression of SAPK2a and SAPK4 mRNA in cell lines of different MHC haplotypes or racial origin. Examination of the SAPK2a and SAPK4 sequences from two chimpanzees revealed 3 nucleotide differences between human and chimpanzee in each gene resulting in only one amino acid change in SAPK4, and 6 nucleotide substitutions plus 2 deletions in 600bp of intronic sequence from SAPK4. This highlights the selective pressure placed on these genes to maintain their protein sequence, but does not favour a role in genetic regulation of disease or provide evidence of linkage disequilibrium with the MHC.
Assuntos
Proteínas Quinases Dependentes de Cálcio-Calmodulina/genética , Centrômero , Éxons , Íntrons , Complexo Principal de Histocompatibilidade/genética , Proteínas Quinases Ativadas por Mitógeno/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Linhagem Celular , DNA Complementar , Haplótipos , Humanos , Proteína Quinase 13 Ativada por Mitógeno , Dados de Sequência Molecular , Pan troglodytes , Proteínas Quinases p38 Ativadas por MitógenoRESUMO
Susceptibility to several disorders, including insulin-dependent diabetes mellitus and multiple sclerosis, has been associated with alleles of HLA class II genes and loci in the TNF cluster in the central major histocompatibility complex (MHC) region. As recombination within this region is rare, it is difficult to determine which genes are important. This will be facilitated by the identification of functional polymorphisms. Hence we are sequencing reverse transcription-polymerase chain reaction products derived from central MHC genes in well characterized and conserved ancestral haplotypes. Here we address the IKBL gene, which lies near the TNF cluster at the telomeric end of the central MHC. Although the IKBL cDNA sequence was conserved between most ancestral haplotypes, a synonymous nucleotide substitution, a 3' untranslated region substitution, and a single nonsynonymous substitution were identified. The latter (IKBL+738) was present in multiple examples of the 7.1 haplotype [HLA-A3, B7, DR2 (DR15)] and resulted in a cysteine to arginine substitution in a predicted protein kinase C phosphorylation site. This polymorphism did not occur in 18 other common haplotypes from the 10th International Histocompatibility Workshop and thus appears haplospecific. A role for IKBL+738 in the association between HLA-A3,B7,DR2(DR15) and susceptibility to multiple sclerosis is discussed.
Assuntos
Antígeno HLA-A3/genética , Antígeno HLA-B7/genética , Antígenos HLA-DR/genética , Antígenos de Histocompatibilidade Classe II/genética , NF-kappa B/genética , Polimorfismo Genético , Proteínas Adaptadoras de Transdução de Sinal , Sequência de Aminoácidos , Sequência de Bases , Linhagem Celular Transformada , DNA Complementar , Subtipos Sorológicos de HLA-DR , Haplótipos , Humanos , Dados de Sequência MolecularRESUMO
BACKGROUND: BAT1 belongs to the DEAD-box family of RNA-binding proteins and is encoded in the central MHC. To determine whether it affects immune responses and hence diseases influenced by MHC haplotypes, U937, THP1 and Jurkat cells were stably transfected with anti-sense DNA corresponding to exons 2-5 of BAT1 using a retroviral vector. RESULTS: Anti-sense transfectants carried anti-sense DNA and expressed anti-sense mRNA. After mitogenic stimulation, they produced higher levels of TNFalpha, IL-1 and IL-6 than equivalent cells carrying the vector alone, suggesting that BAT1 may down-regulate acute phase cytokine production. Polyclonal antibodies raised against a peptide in exon 8 of BAT1 recognized approximately 50 kDa and approximately 38 kDa proteins in all cell lines tested, including the anti-sense transfectants. Expression was localized to the nucleolus in dividing fibroblasts. However the immunochemistry may be confounded by a recently described gene, DDXL, on chromosome 19, which shares a 89% amino acid identity with BAT1. RT-PCR analyses established that BAT1 and DDXL mRNA are expressed in resting U937, THP1 and Jurkat cells. BAT1 and DDXL are divergent in the exons selected for the anti-sense study. CONCLUSIONS: BAT1 is a negative regulator of inflammation. Future studies should address how its functions relate to those of DDXL.
Assuntos
Transportadores de Cassetes de Ligação de ATP/genética , Citocinas/biossíntese , Complexo Principal de Histocompatibilidade/genética , Transportadores de Cassetes de Ligação de ATP/fisiologia , Sequência de Bases , Western Blotting , Linhagem Celular , Nucléolo Celular , RNA Helicases DEAD-box , DNA Antissenso , Regulação para Baixo , Fibroblastos/citologia , Haplótipos , Hematopoese , Humanos , Imuno-Histoquímica , Células Jurkat , Ligantes , RNA Helicases , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , TransfecçãoRESUMO
As mice are often used to model human major histocompatibility complex (MHC)-associated diseases, it is important to understand how their MHC regions differ at the DNA level. The sequencing of the mouse MHC (H2 region) has enabled a detailed map of this region to be assembled for comparison with the human MHC. Here, Richard Allcock and colleagues outline the similarities between the human and mouse MHC regions and discuss notable differences that might affect disease models.
Assuntos
Modelos Animais de Doenças , Complexo Principal de Histocompatibilidade/genética , Animais , Humanos , Camundongos , Especificidade da EspécieRESUMO
The onset of pulmonary hypertension in patients with systemic sclerosis carries a poor prognosis. Atrial septostomy has been used successfully to palliate endstage primary pulmonary hypertension but has not been attempted in other forms of pulmonary vascular disease. We report substantial clinical improvement following atrial septostomy in a patient with systemic sclerosis complicated by severe, isolated pulmonary hypertension. After the procedure, exercise capacity was improved and exertional syncope abolished. We suggest that this procedure should be considered for other patients with this diagnosis.
Assuntos
Septos Cardíacos/cirurgia , Hipertensão Pulmonar/etiologia , Hipertensão Pulmonar/cirurgia , Escleroderma Sistêmico/complicações , Função Atrial , Débito Cardíaco , Feminino , Humanos , Pessoa de Meia-Idade , Cuidados PaliativosRESUMO
OBJECTIVES: We aimed to obtain an estimate of the prevalence and demographics of systemic sclerosis (SSc) and its subtypes at the turn of the millennium. METHODS: Case finding from multiple sources from a defined geographical area. Diagnosis confirmed by clinical examination. RESULTS: The crude prevalence of SSc in northeast England was 8.8 (95% CI: 6.8-10.8) per 100,000. The prevalence when adjusted for the entire UK is 8.2 (95% CI: 6.2-9.8) per 100,000. The ratio of women to men was 5.2:1. The median age of patients was 57.1 yr. The ratio of limited cutaneous SSc to diffuse cutaneous SSc was 4.7:1. Limited cutaneous SSc is associated with the presence of anticentromere antibodies; diffuse cutaneous SSc is associated with anti-Scl 70 antibodies, but either antibody was found in either form of SSc. CONCLUSIONS: SSc appears to be more common in northeast England than was found in the West Midlands in 1986. This may reflect changes in the diagnostic definition of SSc.
Assuntos
Escleroderma Sistêmico/epidemiologia , Adolescente , Adulto , Distribuição por Idade , Idade de Início , Idoso , Idoso de 80 Anos ou mais , Autoanticorpos/análise , Viés , Inglaterra/epidemiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Prevalência , Escleroderma Sistêmico/diagnóstico , Escleroderma Sistêmico/imunologia , Distribuição por SexoRESUMO
Factors influencing MCMV infection mediated by MHC class 1 molecules were analysed further as previous studies showed that the effects of the MHC genotype on sensitivity to this virus are important in vivo. Here we show that H-2d, H-2b, H-2r and H-2v macrophages are highly sensitive to MCMV. Moreover, transfection of H-2k L-cells with Kb or Dd conferred sensitivity to MCMV. This was not affected by amino acid substitutions in Kb alpha 1 or alpha 2, although previous studies demonstrated that exchange of the alpha 1 domain of Dd with Ld alpha 1 compromised sensitivity. Here replacement of Kb alpha 3 with Ld alpha 3 reduced susceptibility to low doses of MCMV. In addition, extracellular beta 2-microglobulin (beta 2m) promoted infection of beta 2m-negative RIE/TL8X.1 cells transfected with Db with or without a beta 2m gene. Hence MCMV infection can involve beta 2m and the alpha 1 and alpha 3 domains of MHC heavy chains. MCMV infection of L-cells expressing Dd or Kb was also inhibited by heparin, but infection of the parental L-cell line was not reproducibly affected. A role for heparan sulphate proteoglycan in MHC-mediated MCMV infection was confirmed using cells pre-treated with heparinase I or III, or propagated in chlorate to inhibit the sulphation of the glycosaminoglycan chains.
Assuntos
Genes MHC Classe I/imunologia , Glicosaminoglicanos/farmacologia , Antígenos H-2/fisiologia , Infecções por Herpesviridae/imunologia , Muromegalovirus/imunologia , Animais , Pré-Escolar , Citomegalovirus/efeitos dos fármacos , Citomegalovirus/imunologia , Infecções por Citomegalovirus/genética , Antígenos H-2/genética , Heparina/farmacologia , Heparitina Sulfato/farmacologia , Infecções por Herpesviridae/tratamento farmacológico , Infecções por Herpesviridae/etiologia , Humanos , Imunidade Inata/genética , Células L , Lipídeos de Membrana/farmacologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Muromegalovirus/efeitos dos fármacos , Muromegalovirus/genética , Mutação/imunologia , Polilisina/farmacologiaRESUMO
The BAT1 gene (D6S81E) encodes a member of the DEAD-box family of RNA-binding proteins, and lies in the central MHC. This region contains genes which affect susceptibility to immunopathological diseases. A 14-kb section of the human MHC containing the BAT1 gene and a further 5-kb telomeric of BAT1 was sequenced using DNA from individuals homozygous for HLA-A1, B8, DR3 and HLA- A1, B57, DR7. Analysis of our sequences and the previously reported human cDNA sequence showed that the expressed sequence of the 8.1 and 57.1 haplotypes is identical with only minor substitutions in the introns. Phylogenetic analysis suggests BAT1 may be a translation initiation factor. Screening of cells and tissues for BAT1 mRNA suggests an abundant member of a family of proteins expressed in multiple cell types, notably macrophages and hepatocytes. Expression was independent of MHC haplotype, consistent with the lack of sequence polymorphism.
Assuntos
Éxons , Expressão Gênica , Íntrons , Complexo Principal de Histocompatibilidade/genética , RNA Helicases/química , RNA Helicases/genética , Northern Blotting , Fator de Iniciação 4A em Eucariotos , Haplótipos , Humanos , Dados de Sequência Molecular , Família Multigênica , Fatores de Iniciação de Peptídeos/química , Fatores de Iniciação de Peptídeos/genética , Polimorfismo Genético , RNA Helicases/biossíntese , RNA Helicases/fisiologia , RNA Mensageiro/biossíntese , Homologia de Sequência de Aminoácidos , Transcrição Gênica , Células Tumorais CultivadasRESUMO
Numerous studies have associated carriage of HLA-DRB1*1501, DQA1*0102 and DQB1*0602 (DR15, DQ6) with dominant resistance to type 1 diabetes and have concluded that one or more of the component HLA class II molecules mediate this effect. Mechanisms for MHC class II-mediated resistance to diabetes have been proposed from studies of transgenic mice, usually using the diabetes-prone non-obese diabetic (NOD) strain. However, these studies have not reached any consensus on a plausible mechanism. In this study we question why the role of central MHC genes in resistance to diabetes has not been addressed, as the central MHC carries markers of susceptibility to diabetes in linkage disequilibrium with several genes with known or putative immunoregulatory functions. To illustrate the type of studies required to address this issue, we selected diabetes patients and control subjects for carriage of HLA-DR15 and the C allele at position +738 in the inhibitor of kappa B-like gene (IKBL). These alleles mark the 7.1 haplotype (HLA-A3, B7, IKBL738*C, DR15, DQ6). HLA-DR15 was the most effective marker of resistance, but an effect may be evident with IKBL738*C in a larger study. Moreover, carriage of the entire haplotype was particularly rare in patients. The best explanation for this is that the critical gene lies between IKBL and HLA-DRB1, and is more closely linked to HLA-DRB1. Candidate genes at the centromeric end of the central MHC are reviewed, highlighting the need for further study.
Assuntos
Diabetes Mellitus Tipo 1/imunologia , Genes MHC da Classe II/imunologia , Imunidade Inata/genética , Adulto , Criança , Diabetes Mellitus Tipo 1/genética , Genes MHC da Classe II/genética , Predisposição Genética para Doença , Humanos , Modelos Genéticos , Modelos ImunológicosRESUMO
BACKGROUND & AIMS: IKBL gene lies telomeric of the tumor necrosis factor cluster in the central major histocompatibility complex and carries a structural polymorphism at position +738. In the Spanish white population, we found the IKBL+738(C) allele in haplotypes carrying either HLA-DRB1(*)1501 or -DRB1(*)0103. Because these HLA class II alleles may confer susceptibility to ulcerative colitis, we investigated an association between IKBL+738(C) and this disease. METHODS: DNA-based techniques were used to type individual alleles of HLA-DRB1 and IKBL+738. The frequencies of these alleles were compared among ethnically matched populations comprising 155 patients and 298 controls. RESULTS: IKBL+738(C) allele was exclusively increased in patients with extensive and/or intractable disease. HLA-DRB1(*)0103 was the only HLA-DRB1 allele to be significantly increased in frequency in patients with UC compared with controls. It was found in patients with extensive and distal disease. In the HLA-DRB1(*)0103-negative population, patients with extensive disease still had a significant association with IKBL+738(C). The difference between the 2 groups of patients was statistically significant (13.7% vs. 1.7% in patients with distal disease; odds ratio, 9.25; P = 0.01). CONCLUSIONS: HLA-DRB1(*)0103 is associated with susceptibility to ulcerative colitis, and IKBL+738(C) marks a propensity to extensive and more severe disease.
Assuntos
Colite Ulcerativa/genética , Colite Ulcerativa/fisiopatologia , Predisposição Genética para Doença/genética , Antígenos de Histocompatibilidade Classe II/genética , Complexo Principal de Histocompatibilidade/genética , Polimorfismo Genético/genética , Proteínas Adaptadoras de Transdução de Sinal , Adulto , Alelos , Frequência do Gene , Antígenos HLA-DR/genética , Cadeias HLA-DRB1 , Humanos , Valores de Referência , Índice de Gravidade de DoençaRESUMO
Periodontal attachment loss is mediated by overproduction of tumour necrosis factor (TNF) and interleukin (IL)-1, and appears to have a genetic component. The 8.1 major histocompatibility complex (MHC) ancestral haplotype (HLA-A1,B8,TNFA-308(2),DR3) is associated with elevated TNF production and predisposes carriers to several autoimmune/immunopathological disorders, including rapid progression of HIV disease, but not early onset periodontal disease in healthy individuals. Rather a high proportion of subjects with severe periodontal disease carry allele 2 at IL-1A-889 and IL-1B+3953. We predicted that genetic associations may be different or clearer in HIV patients, as they often show elevated production of TNF and IL-1 and periodontal attachment loss. Hence periodontal parameters and IL-1 polymorphisms were assessed in HIV-positive subjects expressing HLA-B8 with or without other markers of the 8.1 haplotype. Of 16 HLA-B8 subjects, 13 demonstrated elevated probing pocket depth and clinical attachment loss. The difference was statistically significant and did not correlate with smoking, age, CD4 T-cell counts, HIV viral load or levels of dental plaque. As TNFA-308 (allele 2) was present in four non-B8 subjects who had minimal attachment loss, it may not mediate the effect of the 8.1 haplotype. Moreover, polymorphisms at IL-1A-889 and IL-1B+3953 did not significantly affect periodontal parameters. Thus a central MHC gene characteristic of the 8.1 haplotype was the clearest determinant of periodontal attachment loss in HIV-infected individuals.