Adenovirus vaccines, particularly the COVID-19 Ad5-nCoV adenovirus vaccine, have emerged as promising tools in the fight against infectious diseases. In this study, we investigated the structure of the T cell response to the Spike protein of the SARS-CoV-2 virus used in the COVID-19 Ad5-nCoV adenoviral vaccine in a phase 3 clinical trial (NCT04540419). In 69 participants, we collected peripheral blood samples at four time points after vaccination or placebo injection. Sequencing of T cell receptor repertoires from Spike-stimulated T cell cultures at day 14 from 17 vaccinated revealed a more diverse CD4+ T cell repertoire compared to CD8+. Nevertheless, CD8+ clonotypes accounted for more than half of the Spike-specific repertoire. Our longitudinal analysis showed a peak T cell response at day 14, followed by a decline until month 6. Remarkably, multiple T cell clonotypes persisted for at least 6 months after vaccination, as demonstrated by ex vivo stimulation. Examination of CDR3 regions revealed homologous sequences in both CD4+ and CD8+ clonotypes, with major CD8+ clonotypes sharing high similarity with annotated sequences specific for the NYNYLYRLF peptide, suggesting potential immunodominance. In conclusion, our study demonstrates the immunogenicity of the Ad5-nCoV adenoviral vaccine and highlights its ability to induce robust and durable T cell responses. These findings provide valuable insight into the efficacy of the vaccine against COVID-19 and provide critical information for ongoing efforts to control infectious diseases.

• MeSH
• Adenoviridae genetika   MeSH
• COVID-19 * prevence a kontrola   MeSH
• glykoprotein S, koronavirus MeSH
• infekční nemoci * MeSH
• lidé MeSH
• SARS-CoV-2 MeSH
• T-lymfocyty MeSH
• vakcíny proti COVID-19 MeSH
• vakcíny * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: The emergence of new SARS-CoV-2 variants of concern B.1.1.7 (Alpha), B.1.351 (Beta), P.1 (Gamma) and B.1.617.2 (Delta) that harbor mutations in the viral S protein raised concern about activity of current vaccines and therapeutic antibodies. Independent studies have shown that mutant variants are partially or completely resistant against some of the therapeutic antibodies authorized for emergency use. METHODS: We employed hybridoma technology, ELISA-based and cell-based S-ACE2 interaction assays combined with authentic virus neutralization assays to develop second-generation antibodies, which were specifically selected for their ability to neutralize the new variants of SARS-CoV-2. FINDINGS: AX290 and AX677, two monoclonal antibodies with non-overlapping epitopes, exhibit subnanomolar or nanomolar affinities to the receptor binding domain of the viral Spike protein carrying amino acid substitutions N501Y, N439K, E484K, K417N, and a combination N501Y/E484K/K417N found in the circulating virus variants. The antibodies showed excellent neutralization of an authentic SARS-CoV-2 virus representing strains circulating in Europe in spring 2020 and also the variants of concern B.1.1.7 (Alpha), B.1.351 (Beta) and B.1.617.2 (Delta). In addition, AX677 is able to bind Omicron Spike protein just like the wild type Spike. The combination of the two antibodies prevented the appearance of escape mutations of the authentic SARS-CoV-2 virus. Prophylactic administration of AX290 and AX677, either individually or in combination, effectively reduced viral burden and inflammation in the lungs, and prevented disease in a mouse model of SARS-CoV-2 infection. INTERPRETATION: The virus-neutralizing properties were fully reproduced in chimeric mouse-human versions of the antibodies, which may represent a promising tool for COVID-19 therapy. FUNDING: The study was funded by AXON Neuroscience SE and AXON COVIDAX a.s.

• MeSH
• angiotensin konvertující enzym 2 chemie   genetika   metabolismus   MeSH
• antigenní drift a shift MeSH
• COVID-19 virologie   MeSH
• farmakoterapie COVID-19 MeSH
• glykoprotein S, koronavirus genetika   imunologie   metabolismus   MeSH
• imunodominantní epitopy imunologie   MeSH
• kinetika MeSH
• lidé MeSH
• modely nemocí na zvířatech MeSH
• monoklonální protilátky imunologie   terapeutické užití   MeSH
• mutace MeSH
• myši MeSH
• neutralizační testy MeSH
• plíce patologie   MeSH
• protinádorové látky imunologicky aktivní imunologie   terapeutické užití   MeSH
• SARS-CoV-2 genetika   imunologie   izolace a purifikace   MeSH
• vazba proteinů MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Viruses rapidly co-evolve with their hosts. The 9 million sequenced SARS-CoV-2 genomes by March 2022 provide a detailed account of viral evolution, showing that all amino acids have been mutated many times. However, only a few became prominent in the viral population. Here, we investigated the emergence of the same mutations in unrelated parallel lineages and the extent of such convergent evolution on the molecular level in the spike (S) protein. We found that during the first phase of the pandemic (until mid 2021, before mass vaccination) 31 mutations evolved independently ≥3-times within separated lineages. These included all the key mutations in SARS-CoV-2 variants of concern (VOC) at that time, indicating their fundamental adaptive advantage. The omicron added many more mutations not frequently seen before, which can be attributed to the synergistic nature of these mutations, which is more difficult to evolve. The great majority (24/31) of S-protein mutations under convergent evolution tightly cluster in three functional domains; N-terminal domain, receptor-binding domain, and Furin cleavage site. Furthermore, among the S-protein receptor-binding motif mutations, ACE2 affinity-improving substitutions are favoured. Next, we determined the mutation space in the S protein that has been covered by SARS-CoV-2. We found that all amino acids that are reachable by single nucleotide changes have been probed multiple times in early 2021. The substitutions requiring two nucleotide changes have recently (late 2021) gained momentum and their numbers are increasing rapidly. These provide a large mutation landscape for SARS-CoV-2 future evolution, on which research should focus now.

• MeSH
• aminokyseliny MeSH
• glykoprotein S, koronavirus * genetika   MeSH
• mutace MeSH
• nukleotidy MeSH
• SARS-CoV-2 * genetika   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The emergence of the novel ß-coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has resulted in a global pandemic of coronavirus disease 2019 (COVID-19). Clinical studies have documented that potentially severe neurological symptoms are associated with SARS-CoV-2 infection, thereby suggesting direct CNS penetration by the virus. Prior studies have demonstrated that the destructive neurological effects of rabies virus (RABV) infections are mediated by CNS transport of the virus tightly bound to the nicotinic acetylcholine receptor (nAChR). By comparison, it has been hypothesized that a similar mechanism exists to explain the multiple neurological effects of SARS-CoV-2 via binding to peripheral nAChRs followed by orthograde or retrograde transport into the CNS. Genetic engineering of the RABV has been employed to generate novel vaccines consisting of non-replicating RABV particles expressing chimeric capsid proteins containing human immunodeficiency virus 1 (HIV-1), Middle East respiratory syndrome (MERS-CoV), Ebolavirus, and hepatitis C virus (HCV) sequences. Accordingly, we present a critical discussion that integrates lessons learned from prior RABV research and vaccine development into a working model of a SARS-CoV-2 vaccine that selectively targets and neutralizes CNS penetration of a tightly bound viral nAChR complex.

• MeSH
• Betacoronavirus chemie   imunologie   MeSH
• glykoprotein S, koronavirus chemie   genetika   imunologie   MeSH
• koronavirové infekce imunologie   metabolismus   prevence a kontrola   virologie   MeSH
• lidé MeSH
• nikotinové receptory metabolismus   MeSH
• pandemie MeSH
• proteinové domény MeSH
• replikace viru * MeSH
• virová pneumonie imunologie   virologie   MeSH
• virové vakcíny chemie   imunologie   metabolismus   MeSH
• virus rabies genetika   fyziologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

INTRODUCTION AND OBJECTIVE: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) enters the nasal cavity, penetrates the nasal epithelial cells through the interaction of its spike protein with the host cell receptor angiotensin-converting enzyme 2 (ACE2) and then triggers a cytokine storm. We aimed to assess the biocompatibility of fullerenol nanoparticles C60(OH)40 and ectoine, and to document their effect on the protection of primary human nasal epithelial cells (HNEpCs) against the effects of interaction with the fragment of virus - spike protein. This preliminary research is the first step towards the construction of a intranasal medical device with a protective, mechanical function against SARS-CoV-2 similar to that of personal protective equipment (eg masks). METHODS: We used HNEpCs and the full-length spike protein from SARS-CoV-2 to mimic the first stage of virus infection. We assessed cell viability with the XTT assay and a spectrophotometer. May-Grünwald Giemsa and periodic acid-Schiff staining served to evaluate HNEpC morphology. We assessed reactive oxygen species (ROS) production by using 2',7'-dichlorofluorescin diacetate and commercial kit. Finally, we employed reverse transcription polymerase chain reaction, Western blotting and confocal microscopy to determine the expression of angiotensin-converting enzyme 2 (ACE2) and inflammatory cytokines. RESULTS: There was normal morphology and unchanged viability of HNEpCs after incubation with 10 mg/L C60(OH)40, 0.2% ectoine or their composite for 24 h. The spike protein exerted cytotoxicity via ROS production. Preincubation with the composite protected HNEpCs against the interaction between the spike protein and the host membrane and prevented the production of key cytokines characteristic of severe coronavirus disease 2019, including interleukin 6 and 8, monocyte chemotactic protein 1 and 2, tissue inhibitor of metalloproteinases 2 and macrophage colony-stimulating factor. CONCLUSION: In the future, the combination of fullerenol and ectoine may be used to prevent viral infections as an intranasal medical device for people with reduced immunity and damaged mucous membrane.

• MeSH
• aminokyseliny diaminové MeSH
• angiotensin konvertující enzym 2 metabolismus   MeSH
• COVID-19 * prevence a kontrola   MeSH
• cytokiny metabolismus   MeSH
• epitelové buňky * účinky léků   virologie   MeSH
• fullereny * farmakologie   chemie   MeSH
• glykoprotein S, koronavirus * metabolismus   MeSH
• kultivované buňky MeSH
• lidé MeSH
• nanočástice * chemie   MeSH
• nosní sliznice účinky léků   cytologie   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• SARS-CoV-2 * účinky léků   MeSH
• syndrom uvolnění cytokinů * prevence a kontrola   MeSH
• viabilita buněk * účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Cíl studie: Cílem tohoto sdělení je přinést informaci o porovnání stanovení protilátek anti-SARS-CoV-2 dvěma různě postavenými metodami, ECLIA (Roche Diagnostics) a ELISA (Euroimmun). Název a sídlo pracoviště: Oddělení klinické biochemie a Imunologická laboratoř Fakultní Thomayerova nemocnice Praha. Materiál a metody: Vybrali jsme 134 vzorků séra od 90 pacientů, zaměstnanců Fakultní Thomayerova nemocnice a dárců rekonvalescentní plazmy (33 PCR-negativních a 57 PCR-pozitivních) s různým odstupem od začátku klinických symptomů (1–136 dní). Vyšetřili jsme protilátky anti-SARS-CoV-2 simultánně metodou ELISA ve třídě IgG a IgA a (DSX) a ECLIA (Cobas e602) a výsledky statisticky zpracovali. Měřením kontrolních materiálů, pozitivních a negativních poolů jsme vyhodnotili vybrané analytické charakteristiky. Výsledky: Zjistili jsme vysokou míru shody mezi ELISA a ECLIA. Specificita i senzitivita byla velmi vysoká u obou testů, senzitivita narůstala s odstupem od začátku symptomů s maximem po více než 20 dnech. U ECLIA lze senzitivitu ještě zvýšit aplikací optimalizovaného cut-off. Ve vysoké míře přetrvávaly protilátky ještě po 60 dnech od začátku symptomů, především ve třídě IgG. Mezilehlá preciznost a opakovatelnost klinickým účelům vyhovovaly. Závěr: Obě metody jsou velmi dobře využitelné pro sledování stavu protilátkové odpovědi na infekci SARS-CoV-2. Pro každodenní rutinu a potřebný rychlý screening je vhodnější ECLIA metoda využívající vysoce imunogenní nukleokapsidový antigen, která zachytí vysoce afinitní protilátky všech tříd. Ke konfirmaci a následnému rozlišení jednotlivých imunoglobulinových tříd je pak možné využít ELISA testy s konzervativním spike proteinem.

Objective: The aim of this study is to provide information on the comparison of the determination anti-SARS-CoV-2 antibodies by two differently designed assays, ECLIA (Roche Diagnostics) and ELISA (Euroimmun). Settings: Department of Clinical Biochemistry and Immunology Laboratory, Thomayer University Hospital Materials and Methods: We selected 134 serum samples from 90 patients, Thomayer Hospital staff, and convalescent plasma donors (33 PCR-negative and 57 PCR-positive) at various intervals from the onset of clinical symptoms (1-136 days). We examined anti-SARS-CoV-2 antibodies simultaneously by ELISA in the IgG and IgA classes (DSX) and ECLIA (Cobas e602) and statistically processed the results. By measuring control materials, positive and negative pools we evaluated selected analytical characteristics. Results: We found a high degree of agreement between ELISA and ECLIA. Specificity and sensitivity were very high in both tests, the sensitivity increasing with the distance from the beginning of the symptoms with a maximum of more than 20 days. For ECLIA, the sensitivity may be further increased by application an optimized cut-off. Antibodies persisted to a high degree after 60 days from the onset of symptoms, especially in the IgG class. Intermediate precision and repeatability suited clinical purposes. Conclusion: Both methods are very useful serological monitoring of the antibody response status to SARS-CoV-2 infection. ECLIA method using a highly immunogenic nucleocapsid antigen that captures high affinity antibodies of all classes is preferable for daily routine and the rapid screening. ELISA tests with a conservative spike protein can then be used to confirm and distinguish individual immunoglobulin classes.

• MeSH
• COVID-19 * diagnóza   MeSH
• klinické laboratorní techniky metody   MeSH
• lidé MeSH
• protilátky virové analýza   MeSH
• SARS-CoV-2 * MeSH
• senzitivita a specificita MeSH
• séroepidemiologické studie MeSH
• Check Tag
• lidé MeSH

Emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants diminishes the efficacy of vaccines and antiviral monoclonal antibodies. Continued development of immunotherapies and vaccine immunogens resilient to viral evolution is therefore necessary. Using coldspot-guided antibody discovery, a screening approach that focuses on portions of the virus spike glycoprotein that are both functionally relevant and averse to change, we identified human neutralizing antibodies to highly conserved viral epitopes. Antibody fp.006 binds the fusion peptide and cross-reacts against coronaviruses of the four genera, including the nine human coronaviruses, through recognition of a conserved motif that includes the S2' site of proteolytic cleavage. Antibody hr2.016 targets the stem helix and neutralizes SARS-CoV-2 variants. Antibody sd1.040 binds to subdomain 1, synergizes with antibody rbd.042 for neutralization, and, similar to fp.006 and hr2.016, protects mice expressing human angiotensin-converting enzyme 2 against infection when present as a bispecific antibody. Thus, coldspot-guided antibody discovery reveals donor-derived neutralizing antibodies that are cross-reactive with Orthocoronavirinae, including SARS-CoV-2 variants.

• MeSH
• COVID-19 * MeSH
• epitopy MeSH
• glykoprotein S, koronavirus MeSH
• lidé MeSH
• myši MeSH
• neutralizační testy MeSH
• neutralizující protilátky * MeSH
• protilátky virové MeSH
• SARS-CoV-2 MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH

• MeSH
• COVID-19 * prevence a kontrola   MeSH
• glykoprotein S, koronavirus MeSH
• lidé MeSH
• messenger RNA MeSH
• plíce MeSH
• příjemce transplantátu * MeSH
• protilátky virové MeSH
• SARS-CoV-2 MeSH
• T-lymfocyty MeSH
• vakcína BNT162 MeSH
• vakcíny proti COVID-19 MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The adaptive immune response to severe acute respiratory coronavirus 2 (SARS-CoV-2) is important for vaccine development and in the recovery from coronavirus disease 2019 (COVID-19). Men and cancer patients have been reported to be at higher risks of contracting the virus and developing the more severe forms of COVID-19. Prostate cancer (PCa) may be associated with both of these risks. We show that CD4+ T cells of SARS-CoV-2-unexposed patients with hormone-refractory (HR) metastatic PCa had decreased CD4+ T cell immune responses to antigens from SARS-CoV-2 spike glycoprotein but not from the spiked glycoprotein of the 'common cold'-associated human coronavirus 229E (HCoV-229E) as compared with healthy male volunteers who responded comparably to both HCoV-229E- and SARS-CoV-2-derived antigens. Moreover, the HCoV-229E spike glycoprotein antigen-elicited CD4+ T cell immune responses cross-reacted with the SARS-CoV-2 spiked glycoprotein antigens. PCa patients may have impaired responses to the vaccination, and the cross-reactivity can mediate antibody-dependent enhancement (ADE) of COVID-19. These findings highlight the potential for increased vulnerability of PCa patients to COVID-19.

• MeSH
• adaptivní imunita MeSH
• CD4-pozitivní T-lymfocyty imunologie   MeSH
• CD8-pozitivní T-lymfocyty imunologie   MeSH
• COVID-19 imunologie   virologie   MeSH
• cytokiny imunologie   MeSH
• glykoprotein S, koronavirus imunologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• lidský koronavirus 229E imunologie   MeSH
• nádory prostaty imunologie   patologie   MeSH
• SARS-CoV-2 imunologie   MeSH
• senioři MeSH
• zkřížené reakce MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Most studies investigating the characteristics of emerging SARS-CoV-2 variants have been focusing on mutations in the spike proteins that affect viral infectivity, fusogenicity, and pathogenicity. However, few studies have addressed how naturally occurring mutations in the non-spike regions of the SARS-CoV-2 genome impact virological properties. In this study, we proved that multiple SARS-CoV-2 Omicron BA.2 mutations, one in the spike protein and another downstream of the spike gene, orchestrally characterize this variant, shedding light on the importance of Omicron BA.2 mutations out of the spike protein.

• MeSH
• COVID-19 * MeSH
• glykoprotein S, koronavirus genetika   MeSH
• lidé MeSH
• mutace MeSH
• SARS-CoV-2 genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH