RESUMEN

Sclerotic graft-versus-host disease (GVHD) is a distinctive phenotype of chronic GVHD after allogeneic hematopoietic cell transplantation, characterized by fibrosis of skin or fascia. Sclerotic GVHD has clinical and histopathological similarities with systemic sclerosis, an autoimmune disease whose risk is influenced by genetic polymorphisms. We examined 13 candidate single-nucleotide polymorphisms (SNPs) that have a well-documented association with systemic sclerosis to determine whether these SNPs are also associated with the risk of sclerotic GVHD. The study cohort included 847 consecutive patients who were diagnosed with chronic GVHD. Genotyping was performed using microarrays, followed by imputation of unobserved SNPs. The donor rs10516487 (BANK1: B-cell scaffold protein with ankyrin repeats 1) TT genotype was associated with lower risk of sclerotic GVHD (hazard ratio [HR], 0.43; 95% confidence interval [CI], 0.21-0.87; P = .02). Donor and recipient rs2056626 (CD247: T-cell receptor ζ subunit) GG or GT genotypes were associated with higher risk of sclerotic GVHD (HR, 1.57; 95% CI, 1.13-2.18; P = .007 and HR, 1.66; 95% CI, 1.19-2.32; P = .003, respectively). Donor and recipient rs987870 (5'-flanking region of HLA-DPA1) CC genotypes were associated with higher risk of sclerotic GVHD (HR, 2.50; 95% CI, 1.22-5.11; P = .01 and HR, 2.13; 95% CI, 1.00-4.54; P = .05, respectively). In further analyses, the recipient DPA1*01:03∼DPB1*04:01 haplotype and certain amino acid substitutions in the recipient P1 peptide-binding pocket of the HLA-DP heterodimer were associated with risk of sclerotic GVHD. Genetic components associated with systemic sclerosis are also associated with sclerotic GVHD. HLA-DP-mediated antigen presentation, T-cell response, and B-cell activation have important roles in the pathogenic mechanisms of both diseases.

Asunto(s)

Proteínas Adaptadoras Transductoras de Señales/genética , Antígenos CD/genética , Antígenos de Neoplasias/genética , Enfermedad Injerto contra Huésped/diagnóstico , Cadenas alfa de HLA-DP/genética , Trasplante de Células Madre Hematopoyéticas/efectos adversos , Proteínas de la Membrana/genética , Polimorfismo de Nucleótido Simple/genética , Esclerosis/diagnóstico , Enfermedades de la Piel/diagnóstico , Adolescente , Adulto , Anciano , Linfocitos B/metabolismo , Linfocitos B/patología , Niño , Preescolar , Estudios de Cohortes , Femenino , Estudios de Seguimiento , Genotipo , Enfermedad Injerto contra Huésped/etiología , Enfermedad Injerto contra Huésped/patología , Cadenas alfa de HLA-DP/química , Haplotipos , Prueba de Histocompatibilidad , Humanos , Lactante , Masculino , Persona de Mediana Edad , Pronóstico , Conformación Proteica , Esclerosis/etiología , Esclerosis/patología , Enfermedades de la Piel/etiología , Enfermedades de la Piel/patología , Trasplante Homólogo , Adulto Joven

RESUMEN

The major allergen, Cry j 1, was isolated from Japanese cedar Cryptomeria japonica (Cry j) pollen and was shown to react with immunoglobulin E antibodies in the sera from pollinosis patients. We previously reported that the frequency of HLA-DP5 was significantly higher in pollinosis patients and the immunodominant peptides from Cry j 1 bound to HLA-DP5 to activate Th2 cells. In the present study, we determined the crystal structure of the HLA-DP5 heterodimer in complex with a Cry j 1-derived nine-residue peptide, at 2.4Å resolution. The peptide-binding groove recognizes the minimal peptide with 10 hydrogen bonds, including those between the negatively charged P1 pocket and the Lys side chain at the first position in the peptide sequence. We confirmed that HLA-DP5 exhibits the same Cry j 1-binding mode in solution, through pull-down experiments using structure-based mutations of Cry j 1. We also identified the characteristic residues of HLA-DP5 that are responsible for the distinct properties of the groove, by comparing the structure of HLA-DP5 and the previously reported structures of HLA-DP2 in complexes with pDRA of the self-antigen. The comparison revealed that the HLA-DP5·pCry j 1 complex forms several hydrogen bond/salt bridge networks between the receptor and the antigen that were not observed in the HLA-DP2·pDRA complex. Evolutionary considerations have led us to conclude that HLA-DP5 and HLA-DP2 represent two major groups of the HLA-DP family, in which the properties of the P1 and P4 pockets have evolved and acquired the present ranges of epitope peptide-binding specificities.

Asunto(s)

Antígenos de Plantas/química , Cadenas alfa de HLA-DP/química , Cadenas beta de HLA-DP/química , Fragmentos de Péptidos/química , Proteínas de Plantas/química , Animales , Sitios de Unión , Cryptomeria/química , Cristalografía por Rayos X , Cadenas alfa de HLA-DP/genética , Cadenas beta de HLA-DP/genética , Humanos , Enlace de Hidrógeno , Modelos Moleculares , Filogenia , Unión Proteica , Dominios y Motivos de Interacción de Proteínas , Estructura Secundaria de Proteína , Células Sf9 , Spodoptera

RESUMEN

HLA class II α and ß chains form receptors for antigen presentation to CD4(+) T cells. Numerous pairings of class II α and ß subunits from the wide range of haplotypes and isotypes may form, but most of these combinations, in particular those produced by isotype mixing, yielded mismatched dimers. It is unclear how selection of functional receptors is achieved. At the atomic level, it is not known which interactions of class II residues regulate selection of matched αß heterodimers and the evolutionary origin of matched isotype mixed dimer formation. In this study we investigated assembly of isotype-mixed HLA class II α and ß heterodimers. Assembly and carbohydrate maturation of various HLA-class II isotype-mixed α and ß subunits was dependent on the groove binding section of the invariant chain (Ii). By mutation of polymorphic DPß sequences, we identified two motifs, Lys-69 and GGPM-(84-87), that are engaged in Ii-dependent assembly of DPß with DRα. We identified five members of a family of DPß chains containing Lys-69 and GGPM 84-87, which assemble with DRα. The Lys/GGPM motif is present in the DPß sequence of the Neanderthal genome, and this ancient sequence is related to the human allele DPB1*0401. By site-directed mutagenesis, we inspected Neanderthal amino acid residues that differ from the DPB1*0401 allele and aimed to determine whether matched heterodimers are formed by assembly of DPß mutants with DRα. Because the *0401 allele is rare in the sub-Saharan population but frequent in the European population, it may have arisen in modern humans by admixture with Neanderthals in Europe.

Asunto(s)

Evolución Molecular , Antígenos HLA-DP/genética , Antígenos HLA-DR/genética , Immunoblotting , Mutación , Secuencia de Aminoácidos , Animales , Antígenos de Diferenciación de Linfocitos B/química , Antígenos de Diferenciación de Linfocitos B/genética , Antígenos de Diferenciación de Linfocitos B/metabolismo , Sitios de Unión/genética , Células COS , Línea Celular , Chlorocebus aethiops , Antígenos HLA-DP/química , Antígenos HLA-DP/metabolismo , Cadenas alfa de HLA-DP/química , Cadenas alfa de HLA-DP/genética , Cadenas alfa de HLA-DP/metabolismo , Cadenas beta de HLA-DP/química , Cadenas beta de HLA-DP/genética , Cadenas beta de HLA-DP/metabolismo , Antígenos HLA-DR/química , Antígenos HLA-DR/metabolismo , Cadenas alfa de HLA-DR/química , Cadenas alfa de HLA-DR/genética , Cadenas alfa de HLA-DR/metabolismo , Cadenas beta de HLA-DR/química , Cadenas beta de HLA-DR/genética , Cadenas beta de HLA-DR/metabolismo , Antígenos de Histocompatibilidad Clase II/química , Antígenos de Histocompatibilidad Clase II/genética , Antígenos de Histocompatibilidad Clase II/metabolismo , Humanos , Modelos Moleculares , Hombre de Neandertal , Unión Proteica , Multimerización de Proteína , Estructura Terciaria de Proteína