RESUMO
BACKGROUND: Effective treatments are still needed to reduce the severity of symptoms, time of hospitalization, and mortality of COVID-19. SARS-CoV-2 specific memory T-lymphocytes obtained from convalescent donors recovered can be used as passive cell immunotherapy. METHODS: Between September and November 2020 a phase 1, dose-escalation, single centre clinical trial was conducted to evaluate the safety and feasibility of the infusion of CD45RA- memory T cells containing SARS-CoV-2 specific T cells as adoptive cell therapy against moderate/severe cases of COVID-19. Nine participants with pneumonia and/or lymphopenia and with at least one human leukocyte antigen (HLA) match with the donor were infused. The first three subjects received the lowest dose (1 × 105 cells/kg), the next three received the intermediate dose (5 × 105 cells/kg) and the last three received the highest dose (1 × 106 cells/kg) of CD45RA- memory T cells. Clinicaltrials.gov registration: NCT04578210. FINDINGS: All participants' clinical status measured by National Early Warning Score (NEWS) and 7-category point ordinal scales showed improvement six days after infusion. No serious adverse events were reported. Inflammatory parameters were stabilised post-infusion and the participants showed lymphocyte recovery two weeks after the procedure. Donor microchimerism was observed at least for three weeks after infusion in all patients. INTERPRETATION: This study provides preliminary evidence supporting the idea that treatment of COVID-19 patients with moderate/severe symptoms using convalescent CD45RA- memory T cells is feasible and safe. FUNDING: Clinical Trial supported by Spanish Clinical Research Network PT17/0017/0013. Co-funded by European Regional Development Fund/European Social Fund. CRIS CANCER Foundation Grant to AP-M and Agencia Valenciana de Innovación Grant AVI-GVA COVID-19-68 to BS.
RESUMO
Three new HLA class I alleles were described in the Spanish population. HLA-A*68:169 and -B*39:129 show one amino acid replacement at the α1-domain, compared to A*68:02 (P47 > L47) and -B*39:06 (S11 > A11), respectively. HLA-B*07:298 presents one nucleotide mutation within exon 1, resulting in a new amino acid position -14, L>Q, which has not been previously described in any HLA protein. Prediction of the B*07:298 signal peptide cleavage did not show significant differences in comparison with that obtained for the rest of HLA-B genes.
Assuntos
Alelos , Sequência de Bases , Antígenos HLA-A/genética , Antígenos HLA-B/genética , Antígeno HLA-B7/genética , Análise de Sequência de DNA , Sequência de Aminoácidos , Antígenos HLA-A/química , Antígenos HLA-B/química , Antígeno HLA-B7/química , Haplótipos , Humanos , Peptídeos/químicaRESUMO
Primary refractory or relapsed pediatric leukemia yield significant morbidity and mortality, with long-term survival rates <40%. Here we present a post-hoc analysis assessing safety and efficacy of infusing activated and expanded Natural Killer cells (NKAE) from haploidentical donors in patients from 2 clinical trials. In total, 18 children, adolescents and young adults with relapse or refractory acute leukemia were treated with two cycles of rescue chemotherapy followed by fresh NKAE cells infusions and low doses of IL-2. The overall response rate, complete remission achievement at the end of the study, was 72% (13 of 18). We infused 52 NKAE cell products containing a median of 6.76â¯×â¯106 NK cells/kg (0.7-34.16) and 0.49â¯×â¯106â¯T cells/kg (0-11). All infusions were well tolerated with no graft versus host disease nor other serious adverse events. Among the 14 patients who completed treatment, 4 of them are alive and leukemia-free more than 750 days post-transplant. We conclude that infusion of fresh NKAE cell therapy is feasible and safe in heavily pretreated pediatric population, and should be further investigated in advanced-phase clinical trials as well as a consolidation therapy to decrease relapse in patients with high-risk leukemia. TRIALS REGISTRATION: Registered at www.clinicaltrials.gov as NCT01944982 and NCT02074657.
Assuntos
Interleucina-15/genética , Células Matadoras Naturais/transplante , Leucemia Mieloide Aguda/terapia , Recidiva Local de Neoplasia/terapia , Leucemia-Linfoma Linfoblástico de Células Precursoras B/terapia , Leucemia-Linfoma Linfoblástico de Células T Precursoras/terapia , Adolescente , Criança , Pré-Escolar , Tratamento Farmacológico , Estudos de Viabilidade , Feminino , Humanos , Lactente , Células Matadoras Naturais/imunologia , Leucemia Mieloide Aguda/genética , Ligantes , Masculino , Recidiva Local de Neoplasia/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras B/genética , Leucemia-Linfoma Linfoblástico de Células T Precursoras/genética , Terapia de Salvação , Análise de Sobrevida , Transplante Haploidêntico , Resultado do Tratamento , Adulto JovemRESUMO
Two novel HLA-C alleles, C*07:109:02 and C*15:143, were characterized in two Spanish individuals.
Assuntos
Alelos , Éxons , Antígenos HLA-C/genética , Polimorfismo de Nucleotídeo Único , Doadores de Tecidos , Substituição de Aminoácidos , Sequência de Bases , Clonagem Molecular , Códon/química , Feminino , Expressão Gênica , Antígenos HLA-C/imunologia , Haplótipos , Transplante de Células-Tronco Hematopoéticas , Teste de Histocompatibilidade , Humanos , Masculino , Reação em Cadeia da Polimerase , Alinhamento de Sequência , Análise de Sequência de DNA , Irmãos , EspanhaRESUMO
BACKGROUND: The assignment of human leukocyte antigen (HLA) null alleles is clinically relevant in the setting of stem cell transplantation. Cell surface expression profiling and mRNA processing analysis of the HLA-B allele previously designated as B*07:44, have been performed. MATERIALS AND METHODS: Cell surface expression of HLA-B*07:44 was determined using flow cytometry. Genomic full-length and HLA-B*07-specific cDNA sequencing were carried out by Sanger procedure. RESULTS: Flow cytometric analysis confirmed previous serologic results and demonstrated a lack of cell membrane expression of the HLA-B protein. The mRNA processing, studied using direct HLA-B*07-specific cDNA sequencing, revealed the presence of a unique, aberrantly spliced mRNA, with a deletion of the last 43 bp on the 5'-end of exon 4. The substitution from T to G at genomic position 1799 compared to B*07:02:01 introduced a new and stronger splice donor site at exon 4. This alternative splicing produced an mRNA containing a premature stop codon at position 280, explaining the absence of mature HLA-B7 protein on the cell surface. CONCLUSION: These findings led us to consider this HLA-B variant as a HLA null allele. The World Health Organization (WHO) Nomenclature Committee has since renamed this variant B*07:44N .
Assuntos
Processamento Alternativo , Sequência de Bases , Antígeno HLA-B7/genética , RNA Mensageiro/genética , Deleção de Sequência , Alelos , Transplante de Medula Óssea , Clonagem Molecular , Códon de Terminação , DNA Complementar/genética , DNA Complementar/imunologia , Éxons , Citometria de Fluxo , Antígeno HLA-B7/imunologia , Humanos , RNA Mensageiro/imunologia , Alinhamento de Sequência , Análise de Sequência de DNA , Terminologia como Assunto , Doadores de TecidosRESUMO
Two new HLA class I alleles, HLA-A*11:01:01:04 and HLA-B*35:330, were characterized in a Spanish patient.
Assuntos
Alelos , Antígeno HLA-A11/genética , Antígeno HLA-B35/genética , Polimorfismo de Nucleotídeo Único , Doadores de Tecidos , Substituição de Aminoácidos , Sequência de Bases , Clonagem Molecular , Códon/química , Éxons , Expressão Gênica , Genótipo , Antígeno HLA-A11/imunologia , Antígeno HLA-B35/imunologia , Transplante de Células-Tronco Hematopoéticas , Heterozigoto , Teste de Histocompatibilidade , Humanos , Íntrons , Mieloma Múltiplo/genética , Mieloma Múltiplo/imunologia , Mieloma Múltiplo/patologia , Mieloma Múltiplo/terapia , Reação em Cadeia da Polimerase , Alinhamento de Sequência , Análise de Sequência de DNA , Irmãos , EspanhaRESUMO
A novel HLA-DRB1*13:216 allele was characterized in a Spanish bone marrow donor.
Assuntos
Alelos , Substituição de Aminoácidos , Éxons , Cadeias HLA-DRB1/genética , Mutação , Sequência de Bases , Transplante de Medula Óssea , Expressão Gênica , Cadeias HLA-DRB1/imunologia , Teste de Histocompatibilidade , Humanos , Análise de Sequência de DNA , Espanha , Doadores de TecidosRESUMO
An important factor affecting the success in the setting of related haploidentical hematopoietic stem cell transplantation (HSCT) is the graft-versus-leukemia effect mediated by natural killer (NK) cells when the donor displays NK alloreactivity versus the recipient. NK cell function is regulated by killer immunoglobulin-like receptors (KIR) and it has been described that donor KIR genotype influences transplantation outcome. This has led to a requirement of laboratories to have a quality assurance program for validation and control of their KIR genotyping methods. The goal of the 1st and 2nd Spanish KIR Genotyping Workshops was to provide an external proficiency testing program in KIR genotyping for Spanish immunology and transplant laboratories. These workshops were conducted during the years 2014-2016 and consisted of 17 participating laboratories typing a set of 20 samples. The presence/absence of 16 mandatory KIR loci (2DL1, 2DL2, 2DL3, 2DL4, 2DL5, 2DS1, 2DS2, 2DS3, 2DS4, 2DS5, 2DP1, 3DL1, 3DL2, 3DL3, 3DS1, and 3DP1) was evaluated per sample. Methods for KIR genotyping included polymerase chain reaction with the use of sequence-specific primers and sequence-specific oligoprobes. Consensus typing was reached in all samples, and the performance of laboratories in external proficiency testing was satisfactory in all cases. The polymorphism detected in the small sample studied in both workshops is indicative of an ample variety of KIR gene profiles in the Spanish population.
Assuntos
Seleção do Doador/métodos , Transplante de Células-Tronco Hematopoéticas/métodos , Receptores KIR/genética , Frequência do Gene , Genótipo , Humanos , Células Matadoras Naturais/imunologia , Reação em Cadeia da Polimerase/métodos , Polimorfismo Genético , Controle de QualidadeRESUMO
Two novel HLA-C alleles, C*04:239 and C*05:137, were characterized in Spanish individuals.
Assuntos
Alelos , Éxons , Antígenos HLA-C/genética , Polimorfismo de Nucleotídeo Único , Sequência de Aminoácidos , Substituição de Aminoácidos , Clonagem Molecular , Códon/química , Expressão Gênica , Antígenos HLA-C/imunologia , Haplótipos , Transplante de Células-Tronco Hematopoéticas , Teste de Histocompatibilidade , Humanos , Reação em Cadeia da Polimerase , Alinhamento de Sequência , Análise de Sequência de DNA , Espanha , Doadores de Tecidos , TransplantadosRESUMO
Characterization of four new human leukocyte antigen (HLA) class I alleles, A*32:01:23, B*18:01:24, B*18:72:02 and C*12:166.
Assuntos
Alelos , Éxons , Antígenos HLA-A/genética , Antígeno HLA-B18/genética , Antígenos HLA-C/genética , Mutação Puntual , Doadores de Tecidos , Sequência de Bases , Clonagem Molecular , Expressão Gênica , Genótipo , Antígenos HLA-A/imunologia , Antígeno HLA-B18/imunologia , Antígenos HLA-C/imunologia , Transplante de Células-Tronco Hematopoéticas , Teste de Histocompatibilidade , Humanos , Isoformas de Proteínas/genética , Isoformas de Proteínas/imunologia , Alinhamento de Sequência , Análise de Sequência de DNA , EspanhaRESUMO
In this report we describe a case of somatic mutation in the HLA-B gene in the tumour cells of a patient with acute myelogenous leukaemia (AML). Routine human leukocyte antigen (HLA) typing of patient by serology and molecular methodologies rendered discrepant results regarding the expression of a B*15:01 antigen. Sequencing-based typing of a patient's sample including a very high percentage of blast cells revealed the presence of one nucleotide insertion at exon 3, position 440, codon 123. This insertion resulted in a reading frame shift and a premature stop codon at position 152. The mutation was absent in a sample obtained when the patient was in remission. This case report points out the relevance of the sample source for HLA typing in patients with haematological malignancies.
Assuntos
Alelos , Éxons , Mutação da Fase de Leitura , Antígeno HLA-B15/genética , Leucemia Mieloide Aguda/genética , Sequência de Bases , Códon sem Sentido , Feminino , Expressão Gênica , Genótipo , Antígeno HLA-B15/imunologia , Teste de Histocompatibilidade , Humanos , Leucemia Mieloide Aguda/imunologia , Leucemia Mieloide Aguda/patologia , Pessoa de Meia-Idade , Alinhamento de Sequência , Análise de Sequência de DNARESUMO
Characterization of two new human-leukocyte antigen (HLA)-A alleles, A*02:572 and HLA-A*03:225.
Assuntos
Alelos , Anemia Aplástica/genética , Células da Medula Óssea/imunologia , Antígeno HLA-A2/genética , Antígeno HLA-A3/genética , Polimorfismo de Nucleotídeo Único , Substituição de Aminoácidos , Anemia Aplástica/imunologia , Anemia Aplástica/patologia , Sequência de Bases , Células da Medula Óssea/patologia , Códon , Éxons , Expressão Gênica , Loci Gênicos , Antígeno HLA-A2/imunologia , Antígeno HLA-A3/imunologia , Teste de Histocompatibilidade , Humanos , Dados de Sequência Molecular , Alinhamento de Sequência , Análise de Sequência de DNA , IrmãosRESUMO
Three novel HLA class II alleles, DRB1*08:70, DQA1*01:13 and DQA1*03:01:03, were characterized.
Assuntos
Alelos , Antígenos de Histocompatibilidade Classe II/genética , Sequência de Aminoácidos , Éxons , Cadeias alfa de HLA-DQ/química , Cadeias alfa de HLA-DQ/genética , Cadeias HLA-DRB1/química , Cadeias HLA-DRB1/genética , Antígenos de Histocompatibilidade Classe II/química , Humanos , Linfoma não Hodgkin/genética , Análise de Sequência de DNARESUMO
HLA-A*31:01:02:02 differs from A*31:01:02 in a single nucleotide mutation at intron 3, nucleotide position 1000 (G > A).
Assuntos
Alelos , Antígenos HLA-A/genética , Polimorfismo de Nucleotídeo Único , Doadores de Tecidos , Sequência de Bases , Transplante de Medula Óssea , Códon , Éxons , Expressão Gênica , Antígenos HLA-A/imunologia , Teste de Histocompatibilidade , Humanos , Íntrons , Dados de Sequência Molecular , Alinhamento de SequênciaRESUMO
HLA-A*03:168N differs from HLA-A*03:01:01 by a single nucleotide substitution resulting in a coding change Y27X.
Assuntos
Alelos , Antígenos HLA-A/genética , Células-Tronco Hematopoéticas/metabolismo , Teste de Histocompatibilidade , Análise de Sequência de DNA , Doadores de Tecidos , Sequência de Bases , Éxons/genética , Humanos , Masculino , Dados de Sequência Molecular , Noruega , Alinhamento de Sequência , SorotipagemRESUMO
HLA-B*08:108 shows one nucleotide difference regarding B*08:01:01 at codon 109 (CTC>TTC, L109>F109).
Assuntos
Alelos , Éxons , Antígeno HLA-B8/genética , Polimorfismo de Nucleotídeo Único , Sequência de Bases , Transplante de Medula Óssea , Códon , Antígeno HLA-B8/imunologia , Teste de Histocompatibilidade , Humanos , Dados de Sequência Molecular , Alinhamento de Sequência , Análise de Sequência de DNA , Doadores de Tecidos , População BrancaRESUMO
Two novels alleles, HLA-A*01:128 and HLA-C*05:88 are characterized.
Assuntos
Alelos , Antígenos HLA-A/genética , Antígenos HLA-C/genética , Sequência de Aminoácidos , Substituição de Aminoácidos , Transplante de Medula Óssea , Códon , Antígenos HLA-A/imunologia , Antígenos HLA-C/imunologia , Teste de Histocompatibilidade , Humanos , Dados de Sequência Molecular , Mutação , Alinhamento de Sequência , Análise de Sequência de DNA , Espanha , Doadores de TecidosRESUMO
BACKGROUND: The presence of circulating antibodies (CA) against human leukocyte antigen (HLA) and major-histocompatibility-complex class I-related chain A (MICA) antigens has been associated with worse renal function and reduced kidney allograft survival. We sought to describe the presence of donor-specific anti-HLA antibodies, non-donor specific antibodies, and antibodies against MICA antigens among a cohort of renal transplant recipients with respect to their evolution effects on renal function and occurrence of an acute rejection episode (AR) after transplantation. METHODS: This prospective study of 22 renal transplant recipients of deceased donor kidneys underwent studies of antibodies before and 3 months after grafting using Luminex technology. RESULTS: Ten patients (five men and five women) showed preexistent CA. Comparing patients with versus without preformed CA, we did not observe a significant difference in donor and recipient age or gender. Eight patients (80%) with CA had undergone induction treatment with anti-human-activated T-lymphocyte rabbit immunoglobulin and 2 (20%) with basiliximab. There were no differences between groups regarding the incidence of acute rejection episodes (ARE n = 3 each). There was one case of Banff grade IIB ARE in a patient without preexisting CA; the other episodes were low-grade cellular responses. There were no differences in other variables including cold ischemia time, HLA mismatches, panel-reactive antibody levels, number of transfusions, cytomegalovirus infection or renal function at discharge and 3 months later. Retransplantation was the only factor associated with preformed CA. Retransplantation and preformed CA were associated with CA at 3 months after transplantation. CONCLUSIONS: CA monitoring is important for highly sensitized renal transplants, although our experience failed to show a difference in graft survival or renal function in the first 3 months' follow-up.
Assuntos
Antígenos HLA/imunologia , Antígenos de Histocompatibilidade Classe I/imunologia , Isoanticorpos/sangue , Transplante de Rim/imunologia , Doença Aguda , Adulto , Idoso , Idoso de 80 Anos ou mais , Anticorpos Monoclonais/uso terapêutico , Soro Antilinfocitário/uso terapêutico , Basiliximab , Biomarcadores/sangue , Feminino , Rejeição de Enxerto/epidemiologia , Rejeição de Enxerto/imunologia , Rejeição de Enxerto/cirurgia , Humanos , Imunossupressores/uso terapêutico , Incidência , Transplante de Rim/efeitos adversos , Masculino , Pessoa de Meia-Idade , Monitorização Imunológica , Estudos Prospectivos , Proteínas Recombinantes de Fusão/uso terapêutico , Reoperação , Fatores de Risco , Espanha/epidemiologia , Fatores de Tempo , Resultado do TratamentoRESUMO
The newly detected HLA-DPB1*136:01 is distinguished from HLA-DPB1*69:01 by a single-nucleotide exchange at position 258, resulting in a coding change 57D-->57E .
Assuntos
Cadeias beta de HLA-DP/genética , Polimorfismo de Nucleotídeo Único , Adulto , Alelos , Substituição de Aminoácidos , Sequência de Bases , Códon , Éxons , Cadeias beta de HLA-DP/imunologia , Transplante de Células-Tronco Hematopoéticas , Teste de Histocompatibilidade , Humanos , Íntrons , Masculino , Dados de Sequência Molecular , Sistema de Registros , Análise de Sequência de DNA , Homologia de Sequência do Ácido Nucleico , Doadores de TecidosRESUMO
A*33:49 has one nucleotide change regarding A*33:01:01 at exon 3, producing an amino acid replacement at codon 97, M97 to I97.