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1.
J Virol ; 97(4): e0021023, 2023 04 27.
Article in English | MEDLINE | ID: covidwho-2254654

ABSTRACT

Porcine enteric alphacoronavirus (PEAV) is a new bat HKU2-like porcine coronavirus, and its endemic outbreak has caused severe economic losses to the pig industry. Its broad cellular tropism suggests a potential risk of cross-species transmission. A limited understanding of PEAV entry mechanisms may hinder a rapid response to potential outbreaks. This study analyzed PEAV entry events using chemical inhibitors, RNA interference, and dominant-negative mutants. PEAV entry into Vero cells depended on three endocytic pathways: caveolae, clathrin, and macropinocytosis. Endocytosis requires dynamin, cholesterol, and a low pH. Rab5, Rab7, and Rab9 GTPases (but not Rab11) regulate PEAV endocytosis. PEAV particles colocalize with EEA1, Rab5, Rab7, Rab9, and Lamp-1, suggesting that PEAV translocates into early endosomes after internalization, and Rab5, Rab7, and Rab9 regulate trafficking to lysosomes before viral genome release. PEAV enters porcine intestinal cells (IPI-2I) through the same endocytic pathway, suggesting that PEAV may enter various cells through multiple endocytic pathways. This study provides new insights into the PEAV life cycle. IMPORTANCE Emerging and reemerging coronaviruses cause severe human and animal epidemics worldwide. PEAV is the first bat-like coronavirus to cause infection in domestic animals. However, the PEAV entry mechanism into host cells remains unknown. This study demonstrates that PEAV enters into Vero or IPI-2I cells through caveola/clathrin-mediated endocytosis and macropinocytosis, which does not require a specific receptor. Subsequently, Rab5, Rab7, and Rab9 regulate PEAV trafficking from early endosomes to lysosomes, which is pH dependent. The results advance our understanding of the disease and help to develop potential new drug targets against PEAV.


Subject(s)
Alphacoronavirus , Caveolae , Clathrin , Pinocytosis , Virus Internalization , rab GTP-Binding Proteins , Alphacoronavirus/physiology , rab GTP-Binding Proteins/metabolism , Endosomes/metabolism , Coronavirus Infections/metabolism , Hydrogen-Ion Concentration , Dynamins/metabolism , Caveolae/metabolism , Cholesterol/metabolism , Clathrin/metabolism , Pinocytosis/physiology , Vero Cells , Chlorocebus aethiops , Animals
2.
Pediatr Res ; 2023 Jan 03.
Article in English | MEDLINE | ID: covidwho-2185765

ABSTRACT

BACKGROUND: The safety of coronavirus disease 2019 (COVID-19) vaccines during pregnancy is a particular concern. Here, we addressed the neonatal outcomes after maternal vaccination of COVID-19 during pregnancy. METHODS: We systematically searched PubMed, EMBASE, and the WHO COVID-19 Database for studies on neonatal outcomes after maternal COVID-19 vaccination from inception to 3 July 2022. Main neonatal outcomes were related to preterm, small for gestation (SGA), NICU admission, low Apgar score at 5 min (<7), and additional neonatal outcomes such as gestation <34 weeks, low birth weight and some neonatal morbidity were all also analyzed. RESULTS: A total of 15 studies were included. We found that maternal vaccination during pregnancy was related to the reduction rates of Preterm, SGA, Low Apgar score at 5 min (<7). In addition, there was no evidence of a higher risk of adverse neonatal outcomes after maternal vaccination of COVID-19 during pregnancy, including NICU admission, preterm birth with gestation <34 weeks, low birth weight, very low birth weight, congenital anomalies, and so on. CONCLUSIONS: COVID-19 vaccination in pregnant women does not raise significant adverse effects on neonatal outcomes and is related to a protective effect on some neonatal outcomes. IMPACT: Present study has addressed the neonatal outcomes after maternal vaccination of COVID-19 during pregnancy. COVID-19 vaccination in pregnant women does not raise significant adverse effects on neonatal outcomes and is related to a protective effect on some neonatal outcomes. The present study could encourage pregnant women to be vaccinated against COVID-19.

3.
PLoS One ; 17(12): e0279071, 2022.
Article in English | MEDLINE | ID: covidwho-2162597

ABSTRACT

Although many studies have examined the effects of perceived stress on some specific stress responses during the COVID-19, a comprehensive study is still lacking. And the co-mediating role of coping style and resilience as important mediators of stress processes is also unclear. This study aimed to explore the effects of perceived stress on emotional, physical, and behavioral stress responses and the mediating roles of coping style and resilience in Chinese population during the recurrent outbreak of COVID-19 from a comprehensive perspective. 1087 participants were recruited to complete the anonymous online survey including the Perceived Stress Scale, the Stress Response Questionnaire, the Simplified Coping Style Questionnaire and the Emotional Resilience Questionnaire. Pearson's correlation and Hayes PROCESS macro 3.5 model 6 were used in the mediating effect analysis. Results showed that positive coping style and resilience both buffered the negative effects of perceived stress on emotional, physical, and behavioral responses through direct or indirect pathways, and resilience had the strongest mediating effects. The findings urged relevant authorities and individuals to take measures to promote positive coping style and resilience to combat the ongoing pandemic stress and protect public physical and mental health.


Subject(s)
COVID-19 , Humans , COVID-19/epidemiology , Adaptation, Psychological , Pandemics , Mental Health , Stress, Psychological
4.
Front Psychiatry ; 13: 962285, 2022.
Article in English | MEDLINE | ID: covidwho-2142284

ABSTRACT

Background: Quarantine as one of the most effective epidemic prevention measures, significantly increased people's stress levels. Ongoing monitoring of the stress status of people under quarantine during the pandemic is an important part of assessing the long-term impact of COVID-19 on mental health. This study aimed to gain a more comprehensive understanding of the stress status of people under quarantine, including perceived stress and stress responses, during the third year of the COVID-19 pandemic in China. Methods: An anonymous online survey was conducted among 464 participants from 39 cities in China from March 31 to April 12, 2022. The survey included three questionnaires: a self-designed questionnaire collecting demographic information and quarantine characteristics, the Perceived Stress Scale (PSS-10) and the Stress Response Questionnaire (SRQ). The t-test or one-way ANOVA or the Welch F-test were used to examine the differences among demographic and quarantine variables of perceived stress and stress responses, then multiple linear regressions were performed to identify the predictors of perceived stress and stress responses. Results: 428 valid respondents were finally included. The average scores of perceived stress, total stress response, emotional response, physical response, and behavioral response were 14.70 ± 7.02, 50.24 ± 22.48, 20.35 ± 9.99, 15.23 ± 7.25, and 11.39 ± 5.27, respectively. The regression analysis showed that the degree of financial worries and days of continuous quarantine were the predictors of perceived stress. The degree of financial worries was a vital factor in predicting total stress response, emotional response, physical response and behavioral response, and in predicting emotional response, age was also a significant predictor. Conclusion: The stress status of individuals under quarantine was generally stable but still needs further attention during the third year of the COVID-19 pandemic. People who are young, have a high degree of financial worries and have been quarantined for a long time may be at a higher risk of perceived stress and stress responses. Relevant authorities should pay closer attention to the risk groups, and additional support and assistance might be required for those mostly worried about their financial situations under quarantine.

5.
Frontiers in psychiatry ; 13, 2022.
Article in English | EuropePMC | ID: covidwho-2045528

ABSTRACT

Background Quarantine as one of the most effective epidemic prevention measures, significantly increased people's stress levels. Ongoing monitoring of the stress status of people under quarantine during the pandemic is an important part of assessing the long-term impact of COVID-19 on mental health. This study aimed to gain a more comprehensive understanding of the stress status of people under quarantine, including perceived stress and stress responses, during the third year of the COVID-19 pandemic in China. Methods An anonymous online survey was conducted among 464 participants from 39 cities in China from March 31 to April 12, 2022. The survey included three questionnaires: a self-designed questionnaire collecting demographic information and quarantine characteristics, the Perceived Stress Scale (PSS-10) and the Stress Response Questionnaire (SRQ). The t-test or one-way ANOVA or the Welch F-test were used to examine the differences among demographic and quarantine variables of perceived stress and stress responses, then multiple linear regressions were performed to identify the predictors of perceived stress and stress responses. Results 428 valid respondents were finally included. The average scores of perceived stress, total stress response, emotional response, physical response, and behavioral response were 14.70 ± 7.02, 50.24 ± 22.48, 20.35 ± 9.99, 15.23 ± 7.25, and 11.39 ± 5.27, respectively. The regression analysis showed that the degree of financial worries and days of continuous quarantine were the predictors of perceived stress. The degree of financial worries was a vital factor in predicting total stress response, emotional response, physical response and behavioral response, and in predicting emotional response, age was also a significant predictor. Conclusion The stress status of individuals under quarantine was generally stable but still needs further attention during the third year of the COVID-19 pandemic. People who are young, have a high degree of financial worries and have been quarantined for a long time may be at a higher risk of perceived stress and stress responses. Relevant authorities should pay closer attention to the risk groups, and additional support and assistance might be required for those mostly worried about their financial situations under quarantine.

6.
J Dev Behav Pediatr ; 43(9): 521-528, 2022 Dec 01.
Article in English | MEDLINE | ID: covidwho-2018245

ABSTRACT

OBJECTIVE: While 1 in 6 US children have a developmental disability (DD), and such children are disproportionately affected by COVID-19, little is known about their vaccination status. We surveyed New York State parents of children with DDs to ascertain willingness and concerns regarding COVID-19 vaccines. METHODS: An online survey was distributed to statewide DD networks from June to September 2021 (vaccines were authorized for adolescents in May 2021). We report associations between vaccine willingness and concerns and race/ethnicity, child age, in-person schooling, routine/flu vaccinations, and DD diagnoses. Willingness was categorized as "got/will get ASAP" (high), "wait and see/only if required," or "definitely not." RESULTS: A total of 352 parents (49.1% White) responded. Willingness differed by age ( p < 0.001). High willingness was reported for 73.9%, 50.0%, and 36.0% of children aged 12 to 17, 6 to 11, and 0 to 5 years, respectively. Willingness differed by autism diagnosis ( p < 0.01) and routine and flu vaccination status ( p < 0.001). Predominant concerns included side effects (89%) and children with disabilities not being in trials (80%). Less common concerns were COVID not serious enough in children to warrant vaccine (23%) and misinformation (e.g., microchips, 5G, DNA changes) (24%). Concerns about vaccine safety differed by age ( p < 0.01) and were highest for older and then the youngest children. In age-stratified adjusted models, attention-deficit/hyperactivity disorder was negatively associated with high willingness for age 5 or younger (OR = 0.02, 95% confidence interval, <0.001-0.622). CONCLUSION: Parents of children with DD in New York seemed highly willing for them to receive COVID-19 vaccines. Although few factors predicted willingness to vaccinate, addressing safety and developmental concerns regarding young children is warranted. Given their increased vulnerability, improved COVID-19 surveillance for children with DD is warranted.


Subject(s)
COVID-19 Vaccines , COVID-19 , Adolescent , Child , Child, Preschool , Humans , COVID-19/epidemiology , COVID-19/prevention & control , COVID-19 Vaccines/adverse effects , Developmental Disabilities/epidemiology , New York/epidemiology , Parents , Vaccines/adverse effects
7.
Front Psychiatry ; 13: 848844, 2022.
Article in English | MEDLINE | ID: covidwho-1855443

ABSTRACT

Background: Under the threat of COVID-19, many universities offer online courses to avoid student gatherings, which prevent teachers from collecting responses and optimizing courses. This work collected eye movement data to analyze attention allocation and proposed instruction for improving the courses. Methods: Subjects were recruited to watch three online courses. Meanwhile, their eye movement data were collected through Dikablis Glasses. Mayer's multimedia cognitive theory was adopted to discriminate the pivotal components of online course, and the Mann-Whitney relevance analysis demonstrated that different representations of courses affected the viewers' attention differently. Results: Three subjects watched three different types of political courses. Course 1, which combined text and explanation, attracted the most attention. Course 2 was shown to be less attractive than course 1 and better than course 3, but the subjects were distracted by the animations in course 2. Course 3, which did not use any technique to present learning content, attracts the least attention from the subjects. A correlation analysis shows that course 1 and course 3 have similar results compared with course 2. Conclusion: Online courses have become a norm during the COVID-19 pandemic. Improving the quality of online courses can effectively reduce the impact of the epidemic on teaching. These experiment results suggest that text + commentary in the design of online courses can effectively attract the attention of the listeners and achieve better learning results. Attention gradually rises in the early stage and then falls after reaching a peak. At this time, the proper introduction of animation can effectively reverse the attention curve, while individual text or commentary results in quickly losing the listener's attention.

8.
Front Public Health ; 10: 858780, 2022.
Article in English | MEDLINE | ID: covidwho-1809625

ABSTRACT

Background: The COVID-19 pandemic interfered with normal campus life, resulting in the need for the course to be conducted in an ideal online format. The purpose of this study is to analyze the impact of different styles of online political course videos on students' attention during the COVID-19 pandemic. Methods: Four college students participated in this small sample study. They were required to conduct two sessions of the experiment, in which they were required to watch three different styles of course videos in each session. While watching the videos, their EEG signals were acquired. For the acquired EEG signals, the sample entropy (SampEn) features were extracted. On the other hand, Mayer's theories of multimedia technology provide guidance for teachers' online courses to enhance students' attention levels. The results of EEG signals analysis and Mayer's theories of multimedia technology were combined to compare and analyze the effects of three styles of instructional videos. Results: Based on comparisons of the SampEn and Mayer's theories of multimedia technology analysis, the results suggest that online instruction in a style where the instructor and content appear on the screen at the same time and the instructor points out the location of the content as it is explained is more likely to elicit higher levels of students' attention. Conclusions: During the COVID-19 pandemic, online instructional methods have an impact on students' classroom attention. It is essential for teachers to design online instructional methods based on students' classroom attention levels and some multimedia instructional techniques to improve students' learning efficiency.


Subject(s)
COVID-19 , Attention , COVID-19/epidemiology , Humans , Pandemics , Students
9.
Frontiers in psychiatry ; 13, 2022.
Article in English | EuropePMC | ID: covidwho-1787183

ABSTRACT

Background Under the threat of COVID-19, many universities offer online courses to avoid student gatherings, which prevent teachers from collecting responses and optimizing courses. This work collected eye movement data to analyze attention allocation and proposed instruction for improving the courses. Methods Subjects were recruited to watch three online courses. Meanwhile, their eye movement data were collected through Dikablis Glasses. Mayer’s multimedia cognitive theory was adopted to discriminate the pivotal components of online course, and the Mann–Whitney relevance analysis demonstrated that different representations of courses affected the viewers’ attention differently. Results Three subjects watched three different types of political courses. Course 1, which combined text and explanation, attracted the most attention. Course 2 was shown to be less attractive than course 1 and better than course 3, but the subjects were distracted by the animations in course 2. Course 3, which did not use any technique to present learning content, attracts the least attention from the subjects. A correlation analysis shows that course 1 and course 3 have similar results compared with course 2. Conclusion Online courses have become a norm during the COVID-19 pandemic. Improving the quality of online courses can effectively reduce the impact of the epidemic on teaching. These experiment results suggest that text + commentary in the design of online courses can effectively attract the attention of the listeners and achieve better learning results. Attention gradually rises in the early stage and then falls after reaching a peak. At this time, the proper introduction of animation can effectively reverse the attention curve, while individual text or commentary results in quickly losing the listener’s attention.

10.
Nat Metab ; 4(1): 29-43, 2022 01.
Article in English | MEDLINE | ID: covidwho-1612214

ABSTRACT

Severe cases of infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are associated with elevated blood glucose levels and metabolic complications. However, the molecular mechanisms for how SARS-CoV-2 infection alters glycometabolic control are incompletely understood. Here, we connect the circulating protein GP73 with enhanced hepatic gluconeogenesis during SARS-CoV-2 infection. We first demonstrate that GP73 secretion is induced in multiple tissues upon fasting and that GP73 stimulates hepatic gluconeogenesis through the cAMP/PKA signaling pathway. We further show that GP73 secretion is increased in cultured cells infected with SARS-CoV-2, after overexpression of SARS-CoV-2 nucleocapsid and spike proteins and in lungs and livers of mice infected with a mouse-adapted SARS-CoV-2 strain. GP73 blockade with an antibody inhibits excessive glucogenesis stimulated by SARS-CoV-2 in vitro and lowers elevated fasting blood glucose levels in infected mice. In patients with COVID-19, plasma GP73 levels are elevated and positively correlate with blood glucose levels. Our data suggest that GP73 is a glucogenic hormone that likely contributes to SARS-CoV-2-induced abnormalities in systemic glucose metabolism.


Subject(s)
COVID-19/complications , COVID-19/virology , Glucose/metabolism , Hyperglycemia/etiology , Hyperglycemia/metabolism , Membrane Proteins/metabolism , SARS-CoV-2 , Animals , Biomarkers , Cyclic AMP-Dependent Protein Kinases/metabolism , Diet, High-Fat , Disease Models, Animal , Fasting , Gene Expression , Gluconeogenesis/drug effects , Gluconeogenesis/genetics , Host-Pathogen Interactions , Humans , Hyperglycemia/blood , Liver/metabolism , Liver/pathology , Membrane Proteins/antagonists & inhibitors , Membrane Proteins/blood , Membrane Proteins/genetics , Mice , Mice, Knockout , Organ Specificity/genetics
11.
Asia & the Pacific Policy Studies ; 8(2):243-252, 2021.
Article in English | ProQuest Central | ID: covidwho-1460144

ABSTRACT

This article explains how making a disease notifiable by law is a core component of a robust and well‐functioning health system. Mechanisms to rapidly detect and report existing or emerging infectious diseases in a timely manner are key to disease control and elimination. Using malaria in Asia‐Pacific as a case in point, we explore different policy considerations involved in making malaria a notifiable disease. These include the timing of legislative changes at different stages of elimination, investing in adequate infrastructure for a robust surveillance system that can support targeted interventions, and the importance of involving all sectors in the delivery of malaria services to detect, report and respond to every case. The article explains how frameworks to report on notifiable diseases, in this case malaria, contribute to improved regional health security.

12.
Disabil Health J ; 15(1): 101178, 2022 01.
Article in English | MEDLINE | ID: covidwho-1330742

ABSTRACT

BACKGROUND: People with intellectual and developmental disabilities (IDD) are at disproportionate risk for severe COVID-19 outcomes, particularly those living in congregate care settings. Yet, there is limited data on vaccine perceptions in the disability community. OBJECTIVE: To explore COVID-19 vaccine perceptions in individuals with IDD, their family members, and those who work with them, to inform a statewide vaccine information and messaging project. METHODS: A national survey, adapted in five languages for the IDD community, was distributed to a convenience sample of IDD organizations throughout New York State. Constructs included vaccine intention, reasons for vaccine hesitancy, and trusted sources of vaccine information. Zip code data were used to map respondent location and vaccine preferences. RESULTS: Of n = 825 respondents, approximately 75% intended to or had received the vaccine across roles (i.e., people with developmental disabilities, family members, direct care workers) and racial/ethnic groups. Greater vaccine hesitancy was reported in younger individuals and those making decisions on behalf of a person with IDD. Concerns included side effects and the swiftness of vaccine development. Black and Hispanic participants had heightened concerns about being an "experiment" for the vaccine. Trusted sources of information included healthcare providers and family members. Respondents who intended to/received the vaccine were dispersed throughout the state. CONCLUSIONS: Vaccine preferences in this New York State disability community sample align with national data. Identified concerns suggest the need for community education that addresses misperceptions. Age and race differences in perspectives highlight the need for tailored education, delivered by trusted messengers.


Subject(s)
COVID-19 , Disabled Persons , COVID-19 Vaccines , Child , Developmental Disabilities , Humans , New York , SARS-CoV-2 , Vaccination Hesitancy , Vaccine Development
13.
Liver Int ; 41(9): 2068-2075, 2021 09.
Article in English | MEDLINE | ID: covidwho-1171115

ABSTRACT

BACKGROUND AND AIMS: Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) associated acute liver injury (ALI) has been linked to poor outcomes in adults. Here we compare characteristics in children with elevated ALT (E-ALT) in two distinct manifestations of the infection, multisystem inflammatory syndrome-children (MIS-C) and coronavirus disease 2019 (COVID-19). METHODS: This is a retrospective study of patients ≤21 years of age with positive for SARS-CoV-2 PCR. E-ALT was defined as alanine aminotransferase (ALT) > 40 U/L. Bivariate analysis and multivariable logistic regression were obtained to describe differences in children with and without E-ALT in COVID-19 and MIS-C. RESULTS: E-ALT was detected in 36% of the 291 patients; 31% with COVID-19, and 51% with MIS-C. E-ALT in COVID-19 was associated with obesity (P < .001), immunocompromised status (P = .04), and chronic liver disease (P = .01). In the regression models, E-ALT in COVID-19 was associated with higher c-reactive protein (OR 1.08, P = .01) after adjusting for common independent predictors. Children with E-ALT and MIS-C were more often boys (P = .001), Hispanic (P = .04), or Black (P < .001). In MIS-C, male gender (OR 5.3, P = .02) and Black race (OR 4.4, P = .04) were associated with increased odds of E-ALT. Children with E-ALT in both cohorts had significantly higher multiorgan dysfunction, longer hospitalization, and ICU stay. Children with MIS-C had 2.3-fold increased risk of E-ALT compared to COVID-19. No association was found between E-ALT and mortality. CONCLUSION: E-ALT with SARS-CoV-2 presents as elevated transaminases without hepatic synthetic dysfunction. Patients with either manifestation of SARS-CoV-2 infection and E-ALT experienced more severe disease.


Subject(s)
COVID-19 , SARS-CoV-2 , Humans , Liver , Male , Phenotype , Retrospective Studies , Systemic Inflammatory Response Syndrome
14.
Nat Metab ; 2(12): 1391-1400, 2020 12.
Article in English | MEDLINE | ID: covidwho-947555

ABSTRACT

Responsible for the ongoing coronavirus disease 19 (COVID-19) pandemic, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infects host cells through binding of the viral spike protein (SARS-2-S) to the cell-surface receptor angiotensin-converting enzyme 2 (ACE2). Here we show that the high-density lipoprotein (HDL) scavenger receptor B type 1 (SR-B1) facilitates ACE2-dependent entry of SARS-CoV-2. We find that the S1 subunit of SARS-2-S binds to cholesterol and possibly to HDL components to enhance viral uptake in vitro. SR-B1 expression facilitates SARS-CoV-2 entry into ACE2-expressing cells by augmenting virus attachment. Blockade of the cholesterol-binding site on SARS-2-S1 with a monoclonal antibody, or treatment of cultured cells with pharmacological SR-B1 antagonists, inhibits HDL-enhanced SARS-CoV-2 infection. We further show that SR-B1 is coexpressed with ACE2 in human pulmonary tissue and in several extrapulmonary tissues. Our findings reveal that SR-B1 acts as a host factor that promotes SARS-CoV-2 entry and may help explain viral tropism, identify a possible molecular connection between COVID-19 and lipoprotein metabolism, and highlight SR-B1 as a potential therapeutic target to interfere with SARS-CoV-2 infection.


Subject(s)
COVID-19/metabolism , COVID-19/virology , Host-Pathogen Interactions , Lipoproteins, HDL/metabolism , SARS-CoV-2/physiology , Scavenger Receptors, Class B/metabolism , Virus Internalization , Cell Line , Cholesterol/metabolism , Disease Susceptibility , Humans , Protein Binding , Receptors, Virus , Spike Glycoprotein, Coronavirus/metabolism , Viral Tropism , Virus Attachment
15.
Eur J Clin Microbiol Infect Dis ; 40(5): 921-928, 2021 May.
Article in English | MEDLINE | ID: covidwho-921757

ABSTRACT

Serological test is a valuable diagnostic tool for coronavirus disease 2019 (COVID-19). However, considerable improvements to these tests are needed, especially in the detection sensitivity. In this study, six recombinant nucleocapsid and spike proteins of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) were prepared and evaluated, including three prokaryotic expression nucleocapsid proteins (rN, rN1, rN2) and three eukaryotic expression spike proteins (rS1, rS-RBD, rS-RBD-mFc). The recombinant proteins with the highest ELISA titers (rS1 and rS-RBD-mFc) were selected to develop a double-antigen sandwich colloidal gold immunochromatography assay (GICA) to detect total antibodies against SARS-CoV-2. The clinical evaluation results showed that the sensitivity and specificity of GICA were 92.09% (419/455) and 99.44% (706/710), respectively. Moreover, a significant number (65.63%, 21/32) of COVID-19 patients with undetectable viral RNA were correctly diagnosed by the GICA method. In conclusion, the eukaryotic expression spike proteins (rS1 and rS-RBD-mFc) are more suitable than the prokaryotic expression nucleocapsid proteins for serological diagnosis of SARS-CoV-2. The proposed GICA for detection of total antibodies could be a powerful complement to the current RNA tests for COVID-19.


Subject(s)
COVID-19 Serological Testing/methods , COVID-19/diagnosis , SARS-CoV-2/isolation & purification , Spike Glycoprotein, Coronavirus/immunology , Antibodies, Viral/blood , COVID-19/blood , COVID-19 Nucleic Acid Testing , Coronavirus Nucleocapsid Proteins/genetics , Coronavirus Nucleocapsid Proteins/immunology , Humans , Immunoassay , Phosphoproteins/genetics , Phosphoproteins/immunology , RNA, Viral/genetics , Recombinant Proteins/genetics , Recombinant Proteins/immunology , SARS-CoV-2/genetics , SARS-CoV-2/immunology , Sensitivity and Specificity , Spike Glycoprotein, Coronavirus/genetics
16.
Immunol Res ; 68(6): 325-339, 2020 12.
Article in English | MEDLINE | ID: covidwho-915244

ABSTRACT

Antibodies are considered as an excellent foundation to neutralize pathogens and as highly specific therapeutic agents. Antibodies are generated in response to a vaccine but little use as immunotherapy to combat virus infections. A new generation of broadly cross-reactive and highly potent antibodies has led to a unique chance for them to be used as a medical intervention. Neutralizing antibodies (monoclonal and polyclonal antibodies) are desirable for pharmaceutical products because of their ability to target specific epitopes with their variable domains by precise neutralization mechanisms. The isolation of neutralizing antiviral antibodies has been achieved by Phage displayed antibody libraries, transgenic mice, B cell approaches, and hybridoma technology. Antibody engineering technologies have led to efficacy improvements, to further boost antibody in vivo activities. "Although neutralizing antiviral antibodies have some limitations that hinder their full development as therapeutic agents, the potential for prevention and treatment of infections, including a range of viruses (HIV, Ebola, MERS-COV, CHIKV, SARS-CoV, and SARS-CoV2), are being actively pursued in human clinical trials."


Subject(s)
Antibodies, Monoclonal/therapeutic use , Antibodies, Neutralizing/therapeutic use , Antibodies, Viral/therapeutic use , Betacoronavirus/immunology , Coronavirus Infections/therapy , Pneumonia, Viral/therapy , Animals , Antibodies, Monoclonal/immunology , Antibodies, Neutralizing/immunology , Antibodies, Viral/immunology , COVID-19 , Coronavirus Infections/prevention & control , Epitopes/immunology , Humans , Immunotherapy/methods , Mice , Pandemics/prevention & control , Pneumonia, Viral/prevention & control , SARS-CoV-2
19.
J Virol ; 94(15)2020 07 16.
Article in English | MEDLINE | ID: covidwho-661225

ABSTRACT

The emergence of a novel coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), resulted in a pandemic. Here, we used X-ray structures of human ACE2 bound to the receptor-binding domain (RBD) of the spike protein (S) from SARS-CoV-2 to predict its binding to ACE2 proteins from different animals, including pets, farm animals, and putative intermediate hosts of SARS-CoV-2. Comparing the interaction sites of ACE2 proteins known to serve or not serve as receptors allows the definition of residues important for binding. From the 20 amino acids in ACE2 that contact S, up to 7 can be replaced and ACE2 can still function as the SARS-CoV-2 receptor. These variable amino acids are clustered at certain positions, mostly at the periphery of the binding site, while changes of the invariable residues prevent S binding or infection of the respective animal. Some ACE2 proteins even tolerate the loss or acquisition of N-glycosylation sites located near the S interface. Of note, pigs and dogs, which are not infected or are not effectively infected and have only a few changes in the binding site, exhibit relatively low levels of ACE2 in the respiratory tract. Comparison of the RBD of S of SARS-CoV-2 with that from bat coronavirus strain RaTG13 (Bat-CoV-RaTG13) and pangolin coronavirus (Pangolin-CoV) strain hCoV-19/pangolin/Guangdong/1/2019 revealed that the latter contains only one substitution, whereas Bat-CoV-RaTG13 exhibits five. However, ACE2 of pangolin exhibits seven changes relative to human ACE2, and a similar number of substitutions is present in ACE2 of bats, raccoon dogs, and civets, suggesting that SARS-CoV-2 may not be especially adapted to ACE2 of any of its putative intermediate hosts. These analyses provide new insight into the receptor usage and animal source/origin of SARS-CoV-2.IMPORTANCE SARS-CoV-2 is threatening people worldwide, and there are no drugs or vaccines available to mitigate its spread. The origin of the virus is still unclear, and whether pets and livestock can be infected and transmit SARS-CoV-2 are important and unknown scientific questions. Effective binding to the host receptor ACE2 is the first prerequisite for infection of cells and determines the host range. Our analysis provides a framework for the prediction of potential hosts of SARS-CoV-2. We found that ACE2 from species known to support SARS-CoV-2 infection tolerate many amino acid changes, indicating that the species barrier might be low. Exceptions are dogs and especially pigs, which revealed relatively low ACE2 expression levels in the respiratory tract. Monitoring of animals is necessary to prevent the generation of a new coronavirus reservoir. Finally, our analysis also showed that SARS-CoV-2 may not be specifically adapted to any of its putative intermediate hosts.


Subject(s)
Betacoronavirus/physiology , Coronavirus Infections/virology , Peptidyl-Dipeptidase A/metabolism , Pneumonia, Viral/virology , Spike Glycoprotein, Coronavirus/metabolism , Virus Attachment , Angiotensin-Converting Enzyme 2 , Animals , Animals, Domestic , Betacoronavirus/metabolism , COVID-19 , Chiroptera/virology , Coronavirus Infections/metabolism , Dogs , Glycosylation , Host-Pathogen Interactions , Humans , Models, Animal , Pandemics , Pets , Pneumonia, Viral/metabolism , Protein Binding , Protein Conformation , Protein Interaction Domains and Motifs , Raccoons/virology , SARS-CoV-2 , Sequence Alignment , Sequence Analysis, Protein , Swine , Viverridae/virology
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