RESUMEN
BACKGROUND: Brain metastases (BM) are common in patients with small cell lung cancer (SCLC). In recent years, the role of whole brain radiotherapy (WBRT) for brain metastases in lung cancer is being reevaluated, especially in the context of new systemic treatments available for SCLC. With this analysis, we investigate decision-making in SCLC patients with BM among European experts in medical oncology and radiation oncology. METHODS: We analyzed decision-making from 13 medical oncologists (selected by IASLC) and 13 radiation oncologists (selected by ESTRO) specialized in SCLC. Management strategies of individual experts were converted into decision trees and analyzed for consensus. RESULTS AND CONCLUSION: In asymptomatic patients, chemotherapy alone is the most commonly recommended first line treatment. In asymptomatic patients with limited volume of brain metastases, a higher preference for chemotherapy without WBRT among medical oncologists compared to radiation oncologists was observed. For symptomatic patients, WBRT followed by chemotherapy was recommended most commonly. For limited extent of BM in symptomatic patients, some experts chose stereotactic radiotherapy as an alternative to WBRT. Significant variation in clinical decision-making was observed among European SCLC experts for the first line treatment of patients with SCLC and BM.
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Neoplasias Encefálicas , Neoplasias Pulmonares , Radiocirugia , Carcinoma Pulmonar de Células Pequeñas , Neoplasias Encefálicas/radioterapia , Irradiación Craneana , Humanos , Carcinoma Pulmonar de Células Pequeñas/radioterapiaRESUMEN
We reported previously on the widespread occurrence of anti-HLA alloantibodies of the IgA isotype (anti-HLA IgA) in the sera of solid-organ re-transplantation (re-tx) candidates (Arnold et al., ). Specifically focussing on kidney re-tx patients, we now extended our earlier findings by examining the impact of the presence and donor specificity of anti-HLA IgA on graft survival. We observed frequent concurrence of anti-HLA IgA and anti-HLA IgG in 27% of our multicenter collective of 694 kidney re-tx patients. This subgroup displayed significantly reduced graft survival as evidenced by the median time to first dialysis after transplantation (TTD 77 months) compared to patients carrying either anti-HLA IgG or IgA (TTD 102 and 94 months, respectively). In addition, donor specificity of anti-HLA IgA had a significant negative impact on graft survival (TTD 74 months) in our study. Taken together, our data strongly indicate that presence of anti-HLA IgA, in particular in conjunction with anti-HLA-IgG, in sera of kidney re-tx patients is associated with negative transplantation outcome.
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Supervivencia de Injerto/inmunología , Antígenos HLA/inmunología , Inmunoglobulina A/inmunología , Isoanticuerpos/inmunología , Trasplante de Órganos , Receptores de Trasplantes , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Alelos , Especificidad de Anticuerpos/inmunología , Niño , Preescolar , Femenino , Antígenos HLA/genética , Humanos , Inmunoglobulina A/sangre , Inmunoglobulina G/sangre , Inmunoglobulina G/inmunología , Isoanticuerpos/sangre , Trasplante de Riñón , Masculino , Persona de Mediana Edad , Trasplante de Órganos/efectos adversos , Pronóstico , Retratamiento , Adulto JovenRESUMEN
Antibody identification by a bead array assay in a kidney patient revealed several HLA-specific antibodies including one directed against the HLA-B7 antigen. Low-resolution typing of the patient indicated the presence of an HLA-B*07 allele. To rule out an HLA-specific autoantibody the HLA-typing of the patient was further refined by nucleotide sequencing on a next-generation sequencing platform and eventually showed an HLA-B*39:01:01:03 and HLA-B*07:181N genotype. Thereby the allospecific nature of the antibody was proven. The HLA-B7-specific antibody could be explained by an immunization during the first kidney-transplantation in 1996 with an HLA-B*07 positive donor. When assessing the plausibility of antibodies, the presence of nonexpressed alleles should be taken into consideration.
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Alelos , Antígeno HLA-B39/genética , Antígeno HLA-B7/genética , Isoanticuerpos/genética , Trasplante de Riñón/métodos , Receptores de Trasplantes , Adulto , Secuencia de Bases , Femenino , Expresión Génica , Genotipo , Antígeno HLA-B39/inmunología , Antígeno HLA-B7/inmunología , Secuenciación de Nucleótidos de Alto Rendimiento , Prueba de Histocompatibilidad , Humanos , Reacción en Cadena de la Polimerasa , Isoformas de Proteínas/genética , Isoformas de Proteínas/inmunología , Alineación de Secuencia , Análisis de Secuencia de ADN , Donantes de Tejidos , Trasplante HomólogoRESUMEN
The classic pathway (CP) of complement is believed to significantly contribute to alloantibody-mediated transplant injury, and targeted complement inhibition is currently considered to be a promising approach for preventing rejection. Here, we investigated the mode of action and efficacy of the humanized anti-C1s monoclonal antibody TNT009 and its parental mouse variant, TNT003, in preclinical in vitro models of HLA antibody-triggered CP activation. In flow cytometric assays, we measured the attachment of C1 subcomponents and C4/C3 split products (C4b/d, C3b/d) to HLA antigen-coated flow beads or HLA-mismatched aortic endothelial cells and splenic lymphocytes. Anti-C1s antibodies profoundly inhibited C3 activation at concentrations >20 µg/mL, in both solid phase and cellular assays. While C4 activation was also prevented, this was not the case for C1 subcomponent attachment. Analysis of serum samples obtained from 68 sensitized transplant candidates revealed that the potency of inhibition was related to the extent of baseline CP activation. This study demonstrates that anti-C1s antibodies TNT009 and TNT003 are highly effective in blocking HLA antibody-triggered complement activation downstream of C1. Our results provide the foundation for clinical studies designed to investigate the potential of TNT009 in the treatment or prevention of complement-mediated tissue injury in sensitized transplant recipients.
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Anticuerpos Monoclonales/farmacología , Activación de Complemento/inmunología , Complemento C1s/inmunología , Rechazo de Injerto/tratamiento farmacológico , Antígenos HLA/inmunología , Isoanticuerpos/efectos adversos , Trasplante de Riñón/efectos adversos , Animales , Estudios de Seguimiento , Tasa de Filtración Glomerular , Rechazo de Injerto/etiología , Supervivencia de Injerto , Humanos , Fallo Renal Crónico/inmunología , Fallo Renal Crónico/cirugía , Pruebas de Función Renal , Ratones , PronósticoRESUMEN
BACKGROUND: To avert the differentiation of allergen-specific Th2 cells in atopic individuals is a major goal in the prevention and therapy of IgE-mediated allergy. We aimed to compare different toll-like receptor (TLR) agonists regarding their effects on antigen-presenting cells and the differentiation of naïve T cells from allergic patients. METHODS: Monocytes and monocyte-derived dendritic cells (mdDC) from allergic patients were stimulated with Pam3CSK4 (TLR1/2 ligand), FSL-1 (TLR2/6 ligand), monophosphoryl lipid (MPL)-A, lipopolysaccharide (LPS, both TLR4 ligands), and flagellin (TLR5 ligand). Allergen uptake and upregulation of CD40, CD80, CD83, CD86, CD58, CCR7 and PD-L1 were analyzed by flow cytometry. Functional maturation of mdDC was tested in mixed leukocyte reactions, and the synthesis of proinflammatory cytokines, IL-10 and members of the IL-12 family was assessed. TLR-ligand-activated mdDC were used to stimulate naïve CD4(+) T cells, and cytokine responses were assessed in supernatants and intracellularly. RESULTS: All TLR ligands except flagellin enhanced allergen uptake. All TLR ligands induced functional maturation of mdDC with differential expression of surface molecules and cytokines and promoted the differentiation of IFN-γ-producing T cells. LPS-matured mdDC exclusively induced Th1-like responses, whereas mdDC stimulated with the other TLR ligands induced both Th1- and Th0-like cells. Pam3CSK4 and flagellin additionally induced Th2-like cells. Th1-like responses were associated with higher expression levels of co-stimulatory molecules, PD-L1, IL-6, TNF-α, and IL-12p70. None of the TLR-ligand-stimulated mdDC induced IL-10- or IL-17-producing T cells. CONCLUSION: Different TLR ligands differently influence T-cell responses due to varying activation of the three signals relevant for T-cell activation, that is, antigen presentation, co-stimulation and cytokine milieu.
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Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/metabolismo , Células Dendríticas/inmunología , Células Dendríticas/metabolismo , Hipersensibilidad/inmunología , Hipersensibilidad/metabolismo , Receptores Toll-Like/metabolismo , Alérgenos/inmunología , Células Presentadoras de Antígenos/inmunología , Células Presentadoras de Antígenos/metabolismo , Antígenos de Superficie/metabolismo , Diferenciación Celular , Citocinas/metabolismo , Humanos , Inmunofenotipificación , Ligandos , Activación de Linfocitos/inmunología , FenotipoRESUMEN
Next generation sequencing (NGS) denotes novel sequencing technologies that enable the generation of a large number of clonal sequences in a single sequencing run. NGS was initially introduced for whole genome sequencing and for quantitation of viral variants or genetic mutations in tumor tissues; more recently, the potential for high resolution HLA typing and high throughput analyses has been explored. It became clear that the complexity of the HLA system implicates new challenges, especially for bioinformatics. From an economical point of view, NGS is becoming increasingly attractive for HLA typing laboratories currently relying on Sanger based sequencing. Realizing the full potential of NGS will require the development of specifically adapted typing strategies and software algorithms. In the present review, three laboratories that were among the first to perform HLA-typing using different NGS platforms, the Roche 454, the Illumina Miseq and the Ion Torrent system, respectively, give an overview of these applications and point out advantages and limitations.
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Antígenos HLA/clasificación , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Prueba de Histocompatibilidad/métodos , Programas Informáticos , Algoritmos , Antígenos HLA/genética , Secuenciación de Nucleótidos de Alto Rendimiento/instrumentación , Secuenciación de Nucleótidos de Alto Rendimiento/normas , Prueba de Histocompatibilidad/instrumentación , Prueba de Histocompatibilidad/normas , Humanos , Proyectos de Investigación , Análisis de Secuencia de ADNRESUMEN
We have updated the catalogue of common and well-documented (CWD) human leukocyte antigen (HLA) alleles to reflect current understanding of the prevalence of specific allele sequences. The original CWD catalogue designated 721 alleles at the HLA-A, -B, -C, -DRB1, -DRB3/4/5, -DQA1, -DQB1, and -DPB1 loci in IMGT (IMmunoGeneTics)/HLA Database release 2.15.0 as being CWD. The updated CWD catalogue designates 1122 alleles at the HLA-A, -B, -C, -DRB1, -DRB3/4/5, -DQA1, -DQB1, -DPA1 and -DPB1 loci as being CWD, and represents 14.3% of the HLA alleles in IMGT/HLA Database release 3.9.0. In particular, we identified 415 of these alleles as being 'common' (having known frequencies) and 707 as being 'well-documented' on the basis of ~140,000 sequence-based typing observations and available HLA haplotype data. Using these allele prevalence data, we have also assigned CWD status to specific G and P designations. We identified 147/151 G groups and 290/415 P groups as being CWD. The CWD catalogue will be updated on a regular basis moving forward, and will incorporate changes to the IMGT/HLA Database as well as empirical data from the histocompatibility and immunogenetics community. This version 2.0.0 of the CWD catalogue is available online at cwd.immunogenomics.org, and will be integrated into the Allele Frequencies Net Database, the IMGT/HLA Database and National Marrow Donor Program's bioinformatics web pages.
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Alelos , Antígenos HLA/clasificación , Antígenos HLA/inmunología , Histocompatibilidad/inmunología , Bases de Datos Genéticas , Frecuencia de los Genes , Sitios Genéticos/inmunología , Genética de Población , Antígenos HLA/genética , Histocompatibilidad/genética , Prueba de Histocompatibilidad , Humanos , Terminología como AsuntoRESUMEN
In this multicentre study, sera from 803 retransplant candidates, including 775 kidney transplant recipients, were analysed with regard to the presence and specificity of anti-HLA alloantibodies of the IgA isotype using a modified microsphere-based platform. Of the kidney recipients, nearly one-third (n = 237, 31%) had IgA alloantibodies. Mostly, these antibodies were found in sera that also harboured IgG alloantibodies that could be found in a total of 572 (74%) of patients. Interestingly, IgA anti-HLA antibodies were preferentially targeting HLA class I antigens in contrast to those of the IgG isotype, which targeted mostly both HLA class I and II antigens. Donor specificity of the IgA alloantibodies could be established for over half of the 237 patients with IgA alloantibodies (n = 124, 52%). A further 58 patients had specificities against HLA-C or HLA-DP, for which no information regarding donor typing was available. In summary, these data showed in a large cohort of retransplant candidates that IgA alloantibodies occur in about one-third of patients, about half of these antibodies being donor specific.
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Anticuerpos Antiidiotipos , Inmunoglobulina A , Isoanticuerpos , Trasplante de Riñón , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Anticuerpos Antiidiotipos/genética , Anticuerpos Antiidiotipos/inmunología , Especificidad de Anticuerpos , Niño , Preescolar , Femenino , Rechazo de Injerto , Antígenos HLA/genética , Antígenos HLA/inmunología , Antígenos de Histocompatibilidad Clase I , Prueba de Histocompatibilidad , Humanos , Inmunoglobulina A/sangre , Inmunoglobulina A/genética , Inmunoglobulina G/sangre , Inmunoglobulina G/genética , Lactante , Isoanticuerpos/genética , Isoanticuerpos/inmunología , Persona de Mediana Edad , Donantes de TejidosRESUMEN
Between 1988 and 2007, international searches for matched unrelated donors (MUDs) were performed for 1586 Austrian patients. Between 2004 and 2007, a MUD was identified for 76.7% of the patients. Between 1996 and 2003, a donor was identified for 71.3% of the patients, and between 1988 and 1995, only for 53.4% of the patients. Search times of successful searches decreased from 7.7 months in the first period to 1.7 months in the period from 2004 to 2007. However, transplants were not performed in all cases in which a donor was found: only in 61.6% of the patients between 2004 and 2007, in 53.4% between 1996 and 2003 and in 29.6% between 1988 and 1995. Multivariate analysis determined that having a common HLA type was the most important variable impacting on finding a MUD for a patient. Factors that most strongly influence a patient's access to transplant were the patient's European origin and a short time between diagnosis and start of donor search. The strongest factor for both finding a donor and being transplanted was a search being performed during more recent years: patients' chances increased from year to year.
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Trasplante de Células Madre Hematopoyéticas/métodos , Donadores Vivos/provisión & distribución , Obtención de Tejidos y Órganos/métodos , Donante no Emparentado/provisión & distribución , Adulto , Austria , Niño , Femenino , Antígenos HLA/genética , Antígenos HLA/inmunología , Humanos , Donadores Vivos/estadística & datos numéricos , Masculino , Persona de Mediana Edad , Fenotipo , Donante no Emparentado/estadística & datos numéricosRESUMEN
BACKGROUND: Nonspecific lipid transfer proteins (nsLTPs) are important food allergens. Often, patients allergic to the nsLTP in peach suffer from allergy to hazelnuts. We aimed to analyse the T-cell response to Cor a 8, the nsLTP in hazelnut and its immunological cross-reactivity with the nsLTP in peach, Pru p 3. METHODS: Cor a 8-reactive T-cell lines (TCL) established from patients allergic to hazelnut and peach were stimulated with 12-mer peptides representing the complete amino acid sequence of Cor a 8 to identify its T-cell-activating regions and with Pru p 3 to investigate cellular cross-reactivity. T-cell clones specific for different major T-cell-activating regions of Pru p 3 were stimulated with Cor a 8. Both nsLTPs were subjected to endolysosomal degradation assays. Immunoglobulin E (IgE) cross-reactivity between Cor a 8 and Pru p 3 was assessed in inhibition enzyme-linked immunosorbent assay. RESULTS: No major T-cell-activating region was found among 26 T-cell-reactive peptides identified in Cor a 8. Although generated with Cor a 8, 62% of the TCL responded more strongly to Pru p 3. This cross-reactivity was mediated by T cells specific for the immunodominant region Pru p 3(61-75) . Peptide clusters encompassing this region were generated during lysosomal degradation of both nsLTPs. Cor a 8 was more rapidly degraded by lysosomal proteases than Pru p 3. Pre-incubation of sera with Pru p 3 completely abolished IgE binding to Cor a 8, which was not the case vice versa. CONCLUSIONS: T-cell reactivity to Cor a 8 is predominantly based on cross-reactivity with Pru p 3, indicating that the latter initiates sensitisation to its homolog in hazelnut. The limited allergenic potential of Cor a 8 seems to be associated with rapid lysosomal degradation during allergen processing and the lack of major T-cell-activating regions.
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Alérgenos/inmunología , Antígenos de Plantas/inmunología , Corylus/inmunología , Reacciones Cruzadas/inmunología , Hipersensibilidad a los Alimentos/inmunología , Prunus/inmunología , Humanos , Lisosomas/inmunología , Proteínas de Plantas , Linfocitos T/inmunologíaRESUMEN
A novel human leukocyte antigen (HLA)-DRB3*01 allele carrying a HLA-DRB1-specific sequence motive in exon 2 is described.
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Alelos , Técnicas de Laboratorio Clínico , Antígenos HLA-DR/química , Secuencia de Bases , Electroforesis/métodos , Exones , Geles , Antígenos HLA-DR/genética , Cadenas HLA-DRB1 , Cadenas HLA-DRB3 , Prueba de Histocompatibilidad , Humanos , Datos de Secuencia Molecular , TemperaturaRESUMEN
CONTEXT: Rare haplotypes with Q318X mutations and duplicated CYP21A2 genes have been reported to occur in different populations to a varying extent. Discrimination between a normal (Q318X mutation on one of the duplicated CYP21A2 genes) and a congenital adrenal hyperplasia (CAH, Q318X mutation without duplicated functional gene) allele is of importance, particularly for prenatal diagnosis and the respective genetic counseling. Although methods to differentiate between such alleles have been published only recently, it remains unclear with which frequency Q318X mutations are associated with duplicated CYP21A2 genes and whether these haplotypes have a common ancestry. SUBJECTS AND METHODS: Human leukocyte antigen (HLA) typing has been performed in 38 unrelated individuals and in 11 family members detected to carry a Q318X mutation in the course of CYP21 genotyping using sequence, multiplex ligation-dependent probe amplification, and Southern blot analyses. RESULTS: The majority (n = 32, 84.2%) of the 38 unrelated individuals carrying the Q318X mutation had the trimodular RCCX haplotype, carrying the Q318X mutation on a duplicated CYP21A2 gene. Twenty-two individuals of these 32 (68.8%) were of the rare HLA-B*50-Cw*06 haplotype, suggesting a common ancestry of this haplotype. In five (13.2%) of the 38 subjects, the Q318X mutation was not associated with a duplicated CYP21A2 gene and thus represents a CAH allele. None of these five patients had the above mentioned HLA haplotype. CONCLUSION: The majority of individuals in whom Q318X mutations are detected carry a duplicated functional CYP21A2 gene and the rare HLA-B*50-Cw*06 haplotype.
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Efecto Fundador , Duplicación de Gen , Antígenos HLA-B/genética , Heterocigoto , Esteroide 21-Hidroxilasa/genética , Hiperplasia Suprarrenal Congénita/diagnóstico , Hiperplasia Suprarrenal Congénita/genética , Adulto , Alelos , Southern Blotting , Femenino , Amplificación de Genes , Genotipo , Glutamina , Haplotipos , Humanos , Masculino , Persona de Mediana Edad , MutaciónRESUMEN
Interactions of polymorphic killer Ig-like receptor (KIR) receptors with KIR ligands have been shown to modify the outcome of hematopoietic SCT (HSCT). The association of these genetic factors with different transplantation endpoints, however, varies substantially, depending on clinical and study setup variables. We aimed to assess whether KIR ligands, KIR genes and KIR haplotypes are associated with HSCT outcome of 124 patients with various hematological malignancies, transplanted with 12/12 HLA matched grafts from unrelated donors. For this purpose, patient and donor KIR gene and KIR ligand polymorphisms were determined and correlated with clinical data in simple and multiple models. We found that a missing HLA-C2 ligand for donor inhibitory KIR2DL1 was significantly associated with an increased risk of acute GVHD (aGVHD) (II-IV) (hazard ratio (HR)=2.23, 95% confidence interval (95% CI): 1.21-4.10, P=0.010), as were the AA KIR haplotypes in patients and donors in HLA-C1CX (HR=2.37, 95% CI: 1.16-4.84, P=0.018) and in HLA-Bw4(-) (HR=3.20, 95% CI: 1.35-7.60, P=0.008) patients. On the contrary, transplantation of HLA-C1C2 patients with KIR2DS2 positive grafts were associated with a decreased risk of aGVHD (II-IV) (HR=0.24, 95% CI: 0.07-0.85, P=0.027). Thus, our single center study provides evidence for the modification of aGVHD risk by KIRs and their ligands.
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Enfermedad Injerto contra Huésped/genética , Antígenos HLA/genética , Trasplante de Células Madre Hematopoyéticas , Prueba de Histocompatibilidad , Histocompatibilidad/genética , Receptores KIR/genética , Enfermedad Aguda , Adolescente , Adulto , Alelos , Femenino , Estudios de Seguimiento , Genotipo , Trasplante de Células Madre Hematopoyéticas/efectos adversos , Humanos , Ligandos , Donadores Vivos , Masculino , Persona de Mediana Edad , Recurrencia , Análisis de Regresión , Factores de Riesgo , Tasa de Supervivencia , Trasplante HomólogoRESUMEN
Sequencing analysis of exons 1-3 of the human leukocyte antigen (HLA)-C gene showed a novel allele, HLA-Cw*0617. While the amino acid sequence is identical with the HLA-Cw*060201 allele, the leader peptide differs in three amino acids.
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Exones/genética , Antígenos HLA-C/genética , Alelos , Secuencia de Bases , Humanos , Datos de Secuencia Molecular , Alineación de SecuenciaRESUMEN
A novel allele HLA-B*2730 and the haplotype HLA-A*03-B*2730-Cw*02-DRB1*16-DRB5 have been found in a Slovene patient and his mother. The exon 2 sequence is identical to that of HLA-B*2702, and the exon 3 sequence is identical to that of HLA-B*2719. A possible mechanism for the generation of B*2730 is a gene conversion event.
Asunto(s)
Glomerulonefritis por IGA/genética , Antígenos HLA-B/genética , Adulto , Alelos , Secuencia de Aminoácidos , Femenino , Glomerulonefritis por IGA/cirugía , Antígeno HLA-B27 , Haplotipos , Prueba de Histocompatibilidad , Homocigoto , Humanos , Masculino , Datos de Secuencia Molecular , Alineación de Secuencia , Inmunología del Trasplante , Listas de EsperaRESUMEN
A novel HLA-B (human leukocyte antigen-B) allele, HLA-B*4442, was identified both in a Czech patient with leukaemia and in his mother. The presence of a novel allele was initially suspected because conflicting results were obtained by serological and DNA typing techniques. The HLA typing using the polymerase chain reaction-sequence-specific primers (PCR-SSP) at the two-digit level indicated an allele belonging to the HLA-B*44 group, whereas serological typing indicated HLA-B21. Typing with PCR-sequence-specific oligonucleotides (PCR-SSO) resulted in a unique reaction pattern that could not be assigned to a known allele, PCR-SSP typing at the four-digit level did not match any known B*44 allele, either. The sequencing-based typing of the HLA-B locus then revealed the novel B*4442 allele that is identical with B*4405 except a single C-->G nucleotide exchange at position 572. This exchange results in an amino acid substitution from serine to tryptophan at position 167 of the expressed HLA-B protein. The B21 serological reactivity of the novel B*4442 allele product was confirmed by employing an additional serological panel of typing sera. Our findings support previous reports claiming that serine at the position 167 in the alpha-2 domain of the HLA-B protein is a major determinant of the HLA-B44(12) serological epitope.
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Alelos , Sustitución de Aminoácidos , Antígenos HLA-B/genética , Mutación Puntual , Sustitución de Aminoácidos/inmunología , Secuencia de Bases , República Checa , Femenino , Regulación Leucémica de la Expresión Génica/genética , Regulación Leucémica de la Expresión Génica/inmunología , Antígenos HLA-B/inmunología , Antígeno HLA-B44 , Humanos , Leucemia/genética , Leucemia/inmunología , Masculino , Datos de Secuencia Molecular , Linaje , Mutación Puntual/inmunologíaAsunto(s)
Alelos , Antígenos HLA-D/química , Antígenos HLA-D/genética , Secuencia de Bases , Clonación Molecular , Codón , Escherichia coli/genética , Exones , Antígenos HLA-D/aislamiento & purificación , Humanos , Masculino , Datos de Secuencia Molecular , Técnicas de Amplificación de Ácido Nucleico , Reacción en Cadena de la Polimerasa , Homología de Secuencia de Ácido NucleicoRESUMEN
BACKGROUND: The T cell reactivity to the major allergen of bee venom, phospholipase A2, has been thoroughly characterized. In contrast, only little is known about the human cellular response to major allergens from wasp venom. OBJECTIVE: To characterize the human T cell response to antigen 5 from Vespula vulgaris, Ves v 5. METHODS: Recombinant Ves v 5 was used to establish allergen-specific T cell lines (TCL) and T cell clones (TCC) from the peripheral blood of vespid-allergic and non-allergic individuals. Ves v 5-specific TCL were mapped for T cell epitopes using overlapping synthetic peptides representing the complete amino acid sequence of Ves v 5. Ves v 5-specific TCC were analysed for antigen-induced secretion of IL-4, IFN-gamma and IL-10. RESULTS: Seventeen distinct T cell epitopes were recognized by allergic individuals among which Ves v 5(181-192) was identified as a dominant T cell epitope. Partially different epitopes were observed in TCL from non-allergic subjects and the dominant epitope Ves v 5(181-192) was not prevalent in these cultures. Ves v 5-specific TCC isolated from allergic individuals did not show the typical T helper type 2 (Th2)-like cytokine profile in response to specific stimulation, i.e. high amounts of IL-4 and low IFN-gamma. TCC from non-allergic individuals showed a Th1-like cytokine pattern. CONCLUSIONS: Our findings provide evidence that the allergic T cell response to Ves v 5 is not Th2-dominated and that different immunogenic sites on this major wasp venom allergen are recognized by allergic and non-allergic individuals.
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Alérgenos/inmunología , Antígenos/administración & dosificación , Epítopos/análisis , Hipersensibilidad/inmunología , Linfocitos T/inmunología , Venenos de Avispas/inmunología , Antígenos/inmunología , Estudios de Casos y Controles , Línea Celular , Células Clonales , Epítopos/inmunología , Humanos , Immunoblotting/métodos , Inmunoglobulina E/análisis , Proteínas Recombinantes/inmunologíaRESUMEN
A novel human leukocyte antigen-B (HLA-B) allele, B*420502, was identified in a patient with leukemia (Caucasoid, Czech ancestry) and his mother during intrafamily search for the hematopoietic stem cell donor. The novel allele was initially detected by HLA typing at low resolution using both sequence specific primers and sequence specific oligonucleotides techniques that resulted in unique reaction patterns. The alleles of the HLA-B locus were separated by the haplotype-specific extraction technique. Sequencing of those alleles revealed a novel allele, B*420502, that is identical with B*420501 except a T-->G exchange (synonymous mutation) at position 618.