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
- Amino Acids, Diamino MeSH
- Angiotensin-Converting Enzyme 2 metabolism MeSH
- COVID-19 * prevention & control MeSH
- Cytokines metabolism MeSH
- Epithelial Cells * drug effects virology MeSH
- Fullerenes * pharmacology chemistry MeSH
- Spike Glycoprotein, Coronavirus * metabolism MeSH
- Cells, Cultured MeSH
- Humans MeSH
- Nanoparticles * chemistry MeSH
- Nasal Mucosa drug effects cytology MeSH
- Reactive Oxygen Species metabolism MeSH
- SARS-CoV-2 * drug effects MeSH
- Cytokine Release Syndrome * prevention & control MeSH
- Cell Survival * drug effects MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article 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
- Epitopes MeSH
- Spike Glycoprotein, Coronavirus MeSH
- Humans MeSH
- Mice MeSH
- Neutralization Tests MeSH
- Antibodies, Neutralizing * MeSH
- Antibodies, Viral MeSH
- SARS-CoV-2 MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Research Support, N.I.H., Extramural MeSH
Seven types of Coronaviruses (CoVs) have been identified that can cause infection in humans, including HCoV-229E, HCoV-NL63, HCoV-OC43, HCoV-HKU1, SARS-CoV, HCoV-MERS, and SARS-CoV-2. In this study, we investigated the genetic structure, the homology of the structural protein sequences, as well as the investigation of the active site of structural proteins. The active site of structural proteins was determined based on the previous studies, and the homology of their amino acid sequences and structure was compared. Multiple sequence alignment of Spike protein of HCoVs showed that the receptor-binding domain of SARS-CoV-2, SARS-CoV, and MERS-CoV was located at a similar site to the S1 subunit. The binding motif of PDZ (postsynaptic density-95/disks large/zona occludens-1) of the envelope protein, was conserved in SARS-CoV and SARS-CoV-2 according to multiple sequence alignment but showed different changes in the other HCoVs. Overall, spike protein showed the most variation in its active sites, but the other structural proteins were highly conserved. In this study, for the first time, the active site of all structural proteins of HCoVs as a drug target was investigated. The binding site of these proteins can be suitable targets for drugs or vaccines among HCoVs.
- MeSH
- Coronavirus * chemistry MeSH
- Spike Glycoprotein, Coronavirus * chemistry MeSH
- Catalytic Domain MeSH
- Humans MeSH
- SARS-CoV-2 * chemistry MeSH
- Severe acute respiratory syndrome-related coronavirus * chemistry MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Review MeSH
Virus SARS-CoV-2, který je příčinou pandemie, vstupuje do buněk po interakci mezi glykoproteinem S a buněčným membránovým enzymem ACE2. Všechny buněčné typy, které ACE2 exprimují, jsou virem SARS-CoV-2 infikovatelné. Onemocnění covid-19, které je následkem infekce SARS-CoV-2, probíhá s významnými odlišnostmi. Ty jsou způsobeny především individuální imunitní reaktivitou každého jedince. U nemocných, kteří nejsou schopni eliminovat dostatečně rychle infekci SARS-CoV-2, může dojít k neregulovanému poškozujícímu zánětu. Ten postihuje především dýchací systém. Virus SARS-CoV-2 lze považovat za neurotropní. Buněčné struktury CNS exprimují ACE2. Virus SARS-CoV-2 může do CNS vstupovat různými způsoby. Nemocní s roztroušenou sklerózou nejsou ve zvýšeném riziku infekce SARS-CoV-2. Profitují z léčby a aktivní imunizace u většiny z nich povede k navození protektivní imunity proti SARS-CoV-2 s minimálními riziky nežádoucích účinků očkování.
SARS-CoV-2 virus which is pandemic is entering host cells via interaction between viral spike glycoprotein S with membrane bound ACE2 enzyme. All cells expressing ACE2 are infectable by SARS-CoV-2 virus. The natural course of COVID-19 diseases which is caused by SARS-CoV-2 infection is very individual. It is caused especially by individual immune reactivities. Those infected people who are unable to control and eliminate SARS-CoV-2 infection effectively can progress into severe harm inflammatory response affecting especially respiratory tract. However, SARS-CoV-2 virus could be recognized as neurotropic. Cellular structure of CNS are expressing ACE2 viral receptor. There are several putative ways for SARS-CoV-2 to enter CNS. Patients with multiple sclerosis are not at increased risk of SARS-CoV-2 infection. These patients will profit from active immunisation against SARS-CoV-2. This immunisation will induced at least partial protection against SARS-CoV-2 with very limited risk of harm effects of immunisation.
- MeSH
- Angiotensin-Converting Enzyme 2 immunology MeSH
- B-Lymphocytes immunology MeSH
- Central Nervous System immunology physiopathology MeSH
- COVID-19 immunology physiopathology therapy MeSH
- Immunity MeSH
- Immunization MeSH
- Comorbidity MeSH
- Humans MeSH
- Risk Factors MeSH
- Multiple Sclerosis complications MeSH
- SARS-CoV-2 * immunology pathogenicity MeSH
- T-Lymphocytes immunology MeSH
- Check Tag
- Humans MeSH
- Publication type
- Research Support, Non-U.S. Gov't MeSH
INTRODUCTION: The COVID-19 vaccine was designed to provide protection against infection by the severe respiratory coronavirus 2 (SARS-CoV-2) and coronavirus disease 2019 (COVID-19). However, the vaccine's efficacy can be compromised in patients with immunodeficiencies or the vaccine-induced immunoprotection suppressed by other comorbidity treatments, such as chemotherapy or immunotherapy. To enhance the protective role of the COVID-19 vaccine, we have investigated a combination of the COVID-19 vaccination with ex vivo enrichment and large-scale expansion of SARS-CoV-2 spike glycoprotein-reactive CD4+ and CD8+ T cells. METHODS: SARS-CoV-2-unexposed donors were vaccinated with two doses of the BNT162b2 SARS-CoV-2 vaccine. The peripheral blood mononuclear cells of the vaccinated donors were cell culture-enriched with T cells reactive to peptides derived from SARS-CoV-2 spike glycoprotein. The enriched cell cultures were large-scale expanded using the rapid expansion protocol (REP) and the peptide-reactive T cells were evaluated. RESULTS: We show that vaccination with the SARS-CoV-2 spike glycoprotein-based mRNA COVID-19 vaccine-induced humoral response against SARS-CoV-2 spike glycoprotein in all tested healthy SARS-CoV-2-unexposed donors. This humoral response was found to correlate with the ability of the donors' PBMCs to become enriched with SARS-CoV-2 spike glycoprotein-reactive CD4+ and CD8+ T cells. Using an 11-day REP, the enriched cell cultures were expanded nearly 1000-fold, and the proportions of the SARS-CoV-2 spike glycoprotein-reactive T cells increased. CONCLUSION: These findings show for the first time that the combination of the COVID-19 vaccination and ex vivo T cell large-scale expansion of SARS-CoV-2-reactive T cells could be a powerful tool for developing T cell-based adoptive cellular immunotherapy of COVID-19.
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
- Adaptive Immunity MeSH
- CD4-Positive T-Lymphocytes immunology MeSH
- CD8-Positive T-Lymphocytes immunology MeSH
- COVID-19 immunology virology MeSH
- Cytokines immunology MeSH
- Spike Glycoprotein, Coronavirus immunology MeSH
- Middle Aged MeSH
- Humans MeSH
- Coronavirus 229E, Human immunology MeSH
- Prostatic Neoplasms immunology pathology MeSH
- SARS-CoV-2 immunology MeSH
- Aged MeSH
- Cross Reactions MeSH
- Check Tag
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Aged MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
