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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.
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Syndrome coronavirus 2 (SARS-CoV-2) pandemic is causing a second outbreak significantly delaying the hope for the virus' complete eradication. In the absence of effective vaccines, we need effective treatments with low adverse effects that can treat hospitalized patients with COVID-19 disease. In this study, we determined the existence of SARS-CoV-2-specific T cells within CD45RA- memory T cells in the blood of convalescent donors. Memory T cells can respond quickly to infection and provide long-term immune protection to reduce the severity of COVID-19 symptoms. Also, CD45RA- memory T cells confer protection from other pathogens encountered by the donors throughout their life. It is of vital importance to resolve other secondary infections that usually develop in patients hospitalized with COVID-19. We found SARS-CoV-2-specific memory T cells in all of the CD45RA- subsets (CD3+, CD4+, and CD8+) and in the central memory and effector memory subpopulations. The procedure for obtaining these cells is feasible, easy to implement for small-scale manufacture, quick and cost-effective, involves minimal manipulation, and has no GMP requirements. This biobank of specific SARS-CoV-2 memory T cells would be immediately available "off-the-shelf" to treat moderate/severe cases of COVID-19, thereby increasing the therapeutic options available for these patients.
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Five new HLA class I alleles are described, A*30:129, B*08:195, B*51:01:62, C*01:147 and C*12:195:02.
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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.
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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
A new DRB1*07 allele, DRB1*07:83, was described in a Caucasian Spanish donor.
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Alelos , Cadeias HLA-DRB1/genética , Sequência de Bases , Éxons/genética , Teste de Histocompatibilidade , Humanos , Alinhamento de SequênciaRESUMO
Two novel HLA-B alleles, B*07:299 and B*35:350, were characterized by genomic full-length sequencing.
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Alelos , Antígenos HLA-B/genética , Sequência de Aminoácidos , Antígenos HLA-B/química , Humanos , Domínios ProteicosRESUMO
Three new HLA class I alleles with synonymous mutations were identified.
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Alelos , Antígeno HLA-A3/genética , Antígenos HLA-C/genética , Mutação Silenciosa , Feminino , Humanos , MasculinoRESUMO
A new HLA-A allele, A*30:125, was characterized in a Spanish individual.
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Antígenos HLA-A/química , Antígenos HLA-A/metabolismo , Alelos , Sequência de Aminoácidos , Sequência de Bases , Éxons/genética , Humanos , Ligação Proteica , Domínios ProteicosRESUMO
Two novel HLA-C alleles, C*07:109:02 and C*15:143, were characterized in two Spanish individuals.
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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
Four new HLA class I alleles, HLA-A*02:681, HLA-A*30:111, HLA-A*68:164 and HLA-B*35:01:46 were described in Spaniards.
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Alelos , Antígenos HLA-A/genética , Antígeno HLA-A2/genética , Antígeno HLA-B35/genética , HumanosRESUMO
The new B*50:51 allele was found in 3 Caucasians from Southern Spain.
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Alelos , Antígeno HLA-B14/genética , Recombinação Genética , Feminino , Humanos , MasculinoRESUMO
Three new HLA class I alleles, HLA-A*02:620, HLA-B*27:150 and HLA-B*07:05:01:02, were described in the Spanish Caucasoid population.
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Alelos , Antígenos HLA-A/genética , Antígenos HLA-B/genética , Genes MHC Classe I/imunologia , Antígenos HLA-A/imunologia , Antígenos HLA-B/imunologia , Humanos , Espanha , População Branca/genéticaRESUMO
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 .
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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.
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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.
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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
A novel HLA-A*30:99 allele was characterized in a Spanish individual.
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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.
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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.
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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.