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2.
BMC Nephrol ; 21(1): 117, 2020 04 06.
Artigo em Inglês | MEDLINE | ID: mdl-32252662

RESUMO

BACKGROUND: In the past urine was considered sterile. Through the introduction of next generation sequencing, it has become clear that a urinary microbiome exists. Acute kidney injury (AKI) represents a major threat to kidney transplant recipients. Remarkable changes in the urinary metabolome occur during AKI, which may influence the urinary microbiome. To our knowledge, this is the first study that examines the urinary microbiome in renal transplant recipients (RTX) and non-transplant recipients (nRTX) at time of AKI. METHODS: In this cross-sectional pilot-study the urinary microbiome of 21 RTX and 9 nRTX with AKI was examined. Clean catch morning urine samples were obtained from all patients on the first day of AKI diagnosis. AKI was defined according to KDIGO guidelines. Urinary microbiota and the urinary metabolome during AKI were assessed in one patient. 16S rRNA sequencing was performed. Sequences were processed using UPARSE-pipeline for operational taxonomic units (OTU) and taxon finding. RESULTS: We successfully extracted and sequenced bacterial DNA from 100% of the urine samples. All 30 patients revealed at least 106,138 reads. 319 OTU and 211 different genera were identified. The microbiotic diversity richness in the RTX group was no different from the nRTX group. Eighteen genera were solely present in nRTX and 7 in RTX. CONCLUSIONS: The urinary microbiome at time of AKI showed different bacterial genera in RTX compared to nRTX. The nRTX group exhibited no different diversity to the RTX group. Irrespective of the status of a previous renal transplantation, the urinary microbiome comprised > 210 different genera. An intraindividual change in microbiota diversity and richness was observed in one study patient during recovery from AKI.


Assuntos
Injúria Renal Aguda , DNA Bacteriano , Transplante de Rim/efeitos adversos , Microbiota/genética , RNA Ribossômico 16S , Infecções Urinárias , Injúria Renal Aguda/etiologia , Injúria Renal Aguda/microbiologia , Injúria Renal Aguda/urina , Estudos Transversais , DNA Bacteriano/isolamento & purificação , DNA Bacteriano/urina , Feminino , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Humanos , Falência Renal Crônica/cirurgia , Transplante de Rim/métodos , Masculino , Projetos Piloto , RNA Ribossômico 16S/isolamento & purificação , RNA Ribossômico 16S/urina , Transplantados , Infecções Urinárias/complicações , Infecções Urinárias/microbiologia , Infecções Urinárias/urina
3.
Vaccines (Basel) ; 10(4)2022 Mar 26.
Artigo em Inglês | MEDLINE | ID: mdl-35455265

RESUMO

First-generation vaccines against SARS-CoV-2 do not provide adequate immune protection. Therefore, we engineered a divalent gene construct combining the receptor-binding domain (RBD) of the spike protein and the immunodominant region of the viral nucleocapsid. This fusion protein was produced in either E. coli or a recombinant baculovirus system. Subsequently, the fusion protein was mixed with adjuvant and administered to mice in a prime-booster mode. Mice (72%) produced an IgG response against both proteins (titer: 10-4-10-5) 14 days after the first booster injection, which was increased to 100% by a second booster. Comparable IgG responses were detected against the delta, gamma and omicron variants of the RBD region. Durability testing revealed IgGs beyond 90 days. In addition, cytolytic effector cell molecules were increased in lymphocytes isolated from peripheral blood. Ex vivo stimulation of T cells by nucleocapsid and RBD peptides showed antigen-specific upregulation of CD44 among the CD4+ and CD8+ T cells of vaccinated mice. No side effect was documented in the central nervous system. Cumulatively, these data represent a proof-of-principle approach alternative to existing mRNA vaccination strategies.

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