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1.
Sci Rep ; 14(1): 25031, 2024 Oct 23.
Artigo em Inglês | MEDLINE | ID: mdl-39443608

RESUMO

Aiming at the difficulty of identification and localization in the detection of contraband targets in X-ray images, as well as the demand for upgrading and deployment of security equipment during the peak flow stage, a lightweight X-ray security contraband detection algorithm is proposed and optimized for YOLOv8. Firstly, the number of channels in the original network is reduced according to a predetermined ratio, which effectively reduces model parameters and computational complexity and achieves better results than the current mainstream lightweight methods, such as model pruning and detection backbone optimization. Secondly, due to the presence of objects of various scales and sizes in X-ray security inspection images, we introduced the BiFPN feature extraction strategy in the neck. We designed a simplified version of BiFPN based on the YOLOv8 network structure to effectively integrate feature information from different levels and scales. Finally, we incorporated the idea of Focal Loss to address the issue of imbalance between positive and negative samples and proposed a new regression loss function, Focal-GIoU Loss, which combines GIoU Loss. This increases the loss weight for important but difficult-to-detect samples, such as small targets and overlapping targets, making the model more focused on these critical samples. Results show that our proposed YOLO-CBF improves mAP by 1.5%, 0.9%, and 0.5% on three datasets with different levels of occlusion in PIDray, while reducing the parameter count and computational cost from the original 3.0M and 8.1G to 2.1M and 6.3G.Moreover, it performs well in comparative experiments between different models and in generalization experiments across different datasets.

2.
J Infect Dis ; 2024 Jul 12.
Artigo em Inglês | MEDLINE | ID: mdl-38996045

RESUMO

BACKGROUND: Thrombocytopenia is the major clinical feature associated with the severity of SFTS, but the mechanism by which it occurs remains unclear. METHODS: RNA transcriptome analyses were performed on platelets purified from SFTS patients and SFTSV-infected mice. The functions of differentially expressed genes (DEGs) in the platelets were characterized. ELISA, flow cytometry, and qRT-PCR were used to measure the levels of platelet activation, SFTSV infection in platelets, formation of neutrophil extracellular traps (NETs), transcription of DEGs and percent of platelets undergoing cell death. RESULTS: Enhanced neutrophil activation and interferon (IFN) signaling involved in the viral life cycle were common platelet responses in SFTS, which may consume increasing numbers of platelets. Other functional changes may be associated with different outcomes of SFTS. SFTSV infection led to platelet destruction by pyroptosis, apoptosis, necroptosis, and autophagy. In contrast to SFTS patients, platelets in SFTSV-infected mice mainly play a role in adaptive immunity, and platelet death was not as severe as in humans. CONCLUSIONS: The altered functions of platelets, such as mediating leukocyte activation and undergoing cell death, contribute to thrombocytopenia in SFTS patients. The different mechanisms of thrombocytopenia in mice, suggest that platelet functions should be considered in experimental animal models.

3.
Virol Sin ; 39(4): 537-545, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38679334

RESUMO

Ticks are a major parasite on the Qinghai-Tibet Plateau, western China, and represent an economic burden to agriculture and animal husbandry. Despite research on tick-borne pathogens that threaten humans and animals, the viromes of dominant tick species in this area remain unknown. In this study, we collected Dermacentor nuttalli ticks near Qinghai Lake and identified 13 viruses belonging to at least six families through metagenomic sequencing. Four viruses were of high abundance in pools, including Xinjiang tick-associated virus 1 (XJTAV1), and three novel viruses: Qinghai Lake virus 1, Qinghai Lake virus 2 (QHLV1, and QHLV2, unclassified), and Qinghai Lake virus 3 (QHLV3, genus Uukuvirus of family Phenuiviridae in order Bunyavirales), which lacks the M segment. The minimum infection rates of the four viruses in the tick groups were 8.2%, 49.5%, 6.2%, and 24.7%, respectively, suggesting the prevalence of these viruses in D. â€‹nuttalli ticks. A putative M segment of QHLV3 was identified from the next-generation sequencing data and further characterized for its signal peptide cleavage site, N-glycosylation, and transmembrane region. Furthermore, we probed the L, M, and S segments of other viruses from sequencing data of other tick pools by â€‹using the putative M segment sequence of QHLV3. By revealing the viromes of D. nuttalli ticks, this study enhances our understanding of tick-borne viral communities in highland regions. The putative M segment identified in a novel uukuvirus suggests that previously identified uukuviruses without M segments should have had the same genome organization as typical bunyaviruses. These findings will facilitate virus discovery and our understanding of the phylogeny of tick-borne uukuviruses.


Assuntos
Dermacentor , Genoma Viral , Filogenia , Viroma , Animais , Dermacentor/virologia , China , Viroma/genética , Genoma Viral/genética , RNA Viral/genética , Sequenciamento de Nucleotídeos em Larga Escala , Metagenômica , Vírus de RNA/genética , Vírus de RNA/classificação , Vírus de RNA/isolamento & purificação , Bunyaviridae/genética , Bunyaviridae/classificação , Bunyaviridae/isolamento & purificação
4.
Virol Sin ; 39(2): 194-204, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38360150

RESUMO

Haemaphysalis longicornis ticks, commonly found in East Asia, can transmit various pathogenic viruses, including the severe fever with thrombocytopenia syndrome virus (SFTSV) that has caused febrile diseases among humans in Hubei Province. However, understanding of the viromes of H. longicornis was limited, and the prevalence of viruses among H. longicornis ticks in Hubei was not well clarified. This study investigates the viromes of both engorged (fed) and free (unfed) H. longicornis ticks across three mountainous regions in Hubei Province from 2019 to 2020. RNA-sequencing analysis identified viral sequences that were related to 39 reference viruses belonging to unclassified viruses and seven RNA viral families, namely Chuviridae, Nairoviridae, Orthomyxoviridae, Parvoviridae, Phenuiviridae, Rhabdoviridae, and Totiviridae. Viral abundance and diversity in these ticks were analysed, and phylogenetic characteristics of the Henan tick virus (HNTV), Dabieshan tick virus (DBSTV), Okutama tick virus (OKTV), and Jingmen tick virus (JMTV) were elucidated based on their full genomic sequences. Prevalence analysis demonstrated that DBSTV was the most common virus found in individual H. longicornis ticks (12.59%), followed by HNTV (0.35%), whereas JMTV and OKTV were not detected. These results improve our understanding of H. longicornis tick viromes in central China and highlight the role of tick feeding status and geography in shaping the viral community. The findings of new viral strains and their potential impact on public health raise the need to strengthen surveillance efforts for comprehensively assessing their spillover potentials.


Assuntos
Haemaphysalis longicornis , Filogenia , Viroma , Animais , China , Genoma Viral , Haemaphysalis longicornis/virologia , Vírus de RNA/genética , Vírus de RNA/isolamento & purificação , Vírus de RNA/classificação , RNA Viral/genética , Análise de Sequência de RNA , Viroma/genética
5.
Nat Commun ; 14(1): 7365, 2023 11 14.
Artigo em Inglês | MEDLINE | ID: mdl-37963884

RESUMO

Crimean-Congo hemorrhagic fever virus (CCHFV) is a biosafety level-4 pathogen requiring urgent research and development efforts. The glycoproteins of CCHFV, Gn and Gc, are considered to play multiple roles in the viral life cycle by interactions with host cells; however, these interactions remain largely unclear to date. Here, we analyzed the cellular interactomes of CCHFV glycoproteins and identified 45 host proteins as high-confidence Gn/Gc interactors. These host molecules are involved in multiple cellular biological processes potentially associated with the physiological actions of the viral glycoproteins. Then, we elucidated the role of a representative cellular protein, HAX1. HAX1 interacts with Gn by its C-terminus, while its N-terminal region leads to mitochondrial localization. By the strong interaction, HAX1 sequestrates Gn to mitochondria, thus depriving Gn of its normal Golgi localization that is required for functional glycoprotein-mediated progeny virion packaging. Consistently, the inhibitory activity of HAX1 against viral packaging and hence propagation was further elucidated in the contexts of pseudotyped and authentic CCHFV infections in cellular and animal models. Together, the findings provide a systematic CCHFV Gn/Gc-cell protein-protein interaction map, but also unravel a HAX1/mitochondrion-associated host antiviral mechanism, which may facilitate further studies on CCHFV biology and therapeutic approaches.


Assuntos
Vírus da Febre Hemorrágica da Crimeia-Congo , Febre Hemorrágica da Crimeia , Animais , Vírus da Febre Hemorrágica da Crimeia-Congo/genética , Vírus da Febre Hemorrágica da Crimeia-Congo/metabolismo , Febre Hemorrágica da Crimeia/metabolismo , Glicoproteínas/genética , Glicoproteínas/metabolismo
6.
Risk Manag Healthc Policy ; 16: 1653-1665, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37641779

RESUMO

Background: Effective healthcare utilization throughout pregnancy is essential in protecting mother and child health, but the maternal healthcare utilization and its associated factors among migrant women are still underexplored. Methods: The data came from the 2018 China Migrants Dynamic Survey. Our analysis included 6337 pregnant migrant women. Prenatal healthcare utilization comprises receiving at least 5 antenatal care (ANC) times and establishing the maternal health record within the first 12 weeks of pregnancy. Postnatal healthcare utilization refers to whether an individual received a postpartum visit and a physical health examination within 28 days and 42 days following delivery. A multivariate binary logit model was employed to investigate the factors related to maternal healthcare utilization. Results: 67.15% of the 6337 participants established health records within the first 12 weeks of pregnancy, and 88.35% received at least five ANC visits. 76.88% and 84.20% of migrant pregnant women received a postpartum visit and a health examination respectively. Age was positively correlated with receiving at least five ANC visits (OR:1.245, 95% CI: 1.038-1.493), a postpartum visit within 28 days (OR: 1.272, 95% CI: 1.107-1.460) and a physical examination within 42 days after delivery (OR=1.174, 95% CI: 1.002-1.376). Education, household income, health insurance and maternal health education were positively associated with prenatal and postnatal healthcare utilization (P<0.05). Number of Children negatively correlated with ANC times (OR: 0.742, 95% CI: 0.613-0.898) and receiving health examination after delivery (OR: 0.720, 95% CI: 0.610-0.849). Conclusion: There is still potential for improvement in the maternal healthcare utilization, particularly in postnatal healthcare. Strengthening the follow-up, focusing on those who are younger, have lower socioeconomic status, and are members of ethnic minorities, and continuing to strengthen maternal health education for them can promote the maternal healthcare utilization before and after delivery.

7.
Electrophoresis ; 44(9-10): 775-783, 2023 05.
Artigo em Inglês | MEDLINE | ID: mdl-36891932

RESUMO

Conventional cancer diagnosis needs to excise diseased tissue from the patient's body for biopsy, causing severe injury to patients. Liquid biopsy (LB), with the superior advantage of minimal invasiveness, has shown its ability to cancer diagnosis in real-time and has been developing promising diagnostic instruments. However, until today, the developed instrument still cannot be an alternative to tissue biopsy in the majority of research and clinical settings. In this paper, we first summarize the challenges and limitations suffered by the existing LB instrument. Then, the opportunities and future progression of the next-generation instrument are discussed in detail. In all, we hope that the future LB instrument can be eventually integrated into the clinical workflow and serve as a validated and reliable tool for cancer diagnosis.


Assuntos
Neoplasias , Humanos , Biópsia Líquida , Neoplasias/diagnóstico
8.
BMC Med ; 20(1): 359, 2022 10 21.
Artigo em Inglês | MEDLINE | ID: mdl-36266654

RESUMO

BACKGROUND: The severe fever with thrombocytopenia syndrome disease (SFTS), caused by the novel tick-borne SFTS virus (SFTSV), was listed among the top 10 priority infectious disease by World Health Organization due to the high fatality rate of 5-30% and the lack of effective antiviral drugs and vaccines and therefore raised the urgent need to develop effective anti-SFTSV drugs to improve disease treatment. METHODS: The antiviral drugs to inhibit SFTSV infection were identified by screening the library containing 1340 FDA-approved drugs using the SFTSV infection assays in vitro. The inhibitory effect on virus entry and the process of clathrin-mediated endocytosis under different drug doses was evaluated based on infection assays by qRT-PCR to determine intracellular viral copies, by Western blot to characterize viral protein expression in cells, and by immunofluorescence assays (IFAs) to determine virus infection efficiencies. The therapeutic effect was investigated in type I interferon receptor defective A129 mice in vivo with SFTSV infection, from which lesions and infection in tissues caused by SFTSV infection were assessed by H&E staining and immunohistochemical analysis. RESULTS: Six drugs were identified as exerting inhibitory effects against SFTSV infection, of which anidulafungin, an antifungal drug of the echinocandin family, has a strong inhibitory effect on SFTSV entry. It suppresses SFTSV internalization by impairing the late endosome maturation and decreasing virus fusion with the membrane. SFTSV-infected A129 mice had relieving symptoms, reduced tissue lesions, and improved disease outcomes following anidulafungin treatment. Moreover, anidulafungin exerts an antiviral effect in inhibiting the entry of other viruses including SARS-CoV-2, SFTSV-related Guertu virus and Heartland virus, Crimean-Congo hemorrhagic fever virus, Zika virus, and Herpes simplex virus 1. CONCLUSIONS: The results demonstrated that the antifungal drug, anidulafungin, could effectively inhibit virus infection by interfering with virus entry, suggesting it may be utilized for the clinical treatment of infectious viral diseases, in addition to its FDA-approved use as an antifungal. The findings also suggested to further evaluate the anti-viral effects of echinocandins and their clinical importance for patients with infection of viruses, which may promote therapeutic strategies as well as treatments and improve outcomes pertaining to various viral and fungal diseases.


Assuntos
Anidulafungina , Infecções por Bunyaviridae , Viroses , Animais , Camundongos , Anidulafungina/farmacologia , Anidulafungina/uso terapêutico , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Antivirais/farmacologia , Antivirais/uso terapêutico , Infecções por Bunyaviridae/tratamento farmacológico , Clatrina , Receptor de Interferon alfa e beta , SARS-CoV-2 , Proteínas Virais , Viroses/tratamento farmacológico
9.
Lab Chip ; 22(18): 3545-3554, 2022 09 13.
Artigo em Inglês | MEDLINE | ID: mdl-35989675

RESUMO

Conventional bioparticle extraction requires labor-intensive operation, and expensive and bulky centrifuges. Herein, we report a miniaturized centrifuge by cascading four paralleled inertial spiral channels with a two-stage serpentine channel, allowing for the efficient washing and acquisition of concentrated bioparticles from background fluids. First, the effects of channel size and flow rate on particle focusing dynamics and solution exchange performances are explored to enable the optimization and wide application of our device. Then, the integrated device is fabricated and tested experimentally. The results indicate that 10-20 µm particles can be washed from the original samples with increased concentrations and with recovery efficiencies of >93%. Finally, to verify its versatility, we use our miniaturized centrifuge to successfully change the culture medium for cultured MCF-7 breast cancer cells, extract A549 lung cancer cells from a calcein-AM staining solution, purify white blood cells (WBCs) from lysed whole blood, and extract target cells from an unbonded magnetic microbead background. Compared with conventional centrifuges, our device has the advantages of simple fabrication, easy operation, and small footprint. More importantly, it offers outstanding capability for extracting bioparticles from various background fluids, and avoids bioparticle damage that may be caused by high-speed centrifugation. Therefore, we envision that our miniaturized centrifuge could be a promising alternative to traditional centrifuges in many applications.


Assuntos
Técnicas Analíticas Microfluídicas , Microfluídica , Centrifugação , Humanos , Leucócitos , Células MCF-7 , Microesferas
10.
Innovation (Camb) ; 3(1): 100181, 2022 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-34746904

RESUMO

Most COVID-19 convalescents can build effective anti-SARS-CoV-2 humoral immunity, but it remains unclear how long it can maintain and how efficiently it can prevent the reinfection of the emerging SARS-CoV-2 variants. Here, we tested the sera from 248 COVID-19 convalescents around 1 year post-infection in Wuhan, the earliest known epicenter. SARS-CoV-2 immunoglobulin G (IgG) was well maintained in most patients and potently neutralizes the infection of the original strain and the B.1.1.7 variant. However, varying degrees of immune escape was observed on the other tested variants in a patient-specific manner, with individuals showing remarkably broad neutralization potency. The immune escape can be largely attributed to several critical spike mutations. These results suggest that SARS-CoV-2 can elicit long-lasting immunity but this is escaped by the emerging variants.

11.
Emerg Infect Dis ; 27(12): 3166-3170, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34808086

RESUMO

We report the isolation of Tamdy virus from Hyalomma asiaticum ticks in northwest China and serologic evidence of human Tamdy virus infection in the same region. These findings highlight the need to further investigate a potential causal relationship between Tamdy virus and febrile illnesses of unknown etiology in that region.


Assuntos
Ixodidae , Carrapatos , Vírus , Animais , China/epidemiologia , Humanos
12.
Front Immunol ; 12: 722027, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34489971

RESUMO

Approximately half of the SARS-CoV-2 infections occur without apparent symptoms, raising questions regarding long-term humoral immunity in asymptomatic individuals. Plasma levels of immunoglobulin G (IgG) and M (IgM) against the viral spike or nucleoprotein were determined for 25,091 individuals enrolled in a surveillance program in Wuhan, China. We compared 405 asymptomatic individuals who mounted a detectable antibody response with 459 symptomatic COVID-19 patients. The well-defined duration of the SARS-CoV-2 endemic in Wuhan allowed a side-by-side comparison of antibody responses following symptomatic and asymptomatic infections without subsequent antigen re-exposure. IgM responses rapidly declined in both groups. However, both the prevalence and durability of IgG responses and neutralizing capacities correlated positively with symptoms. Regardless of sex, age, and body weight, asymptomatic individuals lost their SARS-CoV-2-specific IgG antibodies more often and rapidly than symptomatic patients did. These findings have important implications for immunity and favour immunization programs including individuals after asymptomatic infections.


Assuntos
Anticorpos Antivirais/sangue , Infecções Assintomáticas/epidemiologia , COVID-19/imunologia , Imunidade Humoral , SARS-CoV-2/imunologia , Adulto , Anticorpos Neutralizantes/imunologia , Formação de Anticorpos , COVID-19/epidemiologia , China , Monitoramento Epidemiológico , Feminino , Humanos , Imunoglobulina G/imunologia , Imunoglobulina M/imunologia , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , SARS-CoV-2/patogenicidade , Adulto Jovem
13.
Emerg Microbes Infect ; 10(1): 1975-1987, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34570681

RESUMO

ABSTRACTTick-borne viruses (TBVs) capable of transmitting between ticks and hosts have been increasingly recognized as a global public health concern. In this study, Hyalomma ticks and serum samples from camels were collected using recorded sampling correlations in eastern Kenya. Viromes of pooled ticks were profiled by metagenomic sequencing, revealing a diverse community of viruses related to at least 11 families. Five highly abundant viruses, including three novel viruses (Iftin tick virus, Mbalambala tick virus [MATV], and Bangali torovirus [BanToV]) and new strains of previously identified viruses (Bole tick virus 4 [BLTV4] and Liman tick virus [LMTV]), were characterized in terms of genome sequences, organizations, and phylogeny, and their molecular prevalence was investigated in individual ticks. Moreover, viremia and antibody responses to these viruses have been investigated in camels. MATV, BLTV4, LMTV, and BanToV were identified as viral pathogens that can potentially cause zoonotic diseases. The transmission patterns of these viruses were summarized, suggesting three different types according to the sampling relationships between viral RNA-positive ticks and camels positive for viral RNA and/or antibodies. They also revealed the frequent transmission of BanToV and limited but effective transmission of other viruses between ticks and camels. Furthermore, follow-up surveys on TBVs from tick, animal, and human samples with definite sampling relationships are suggested. The findings revealed substantial threats from the emerging TBVs and may guide the prevention and control of TBV-related zoonotic diseases in Kenya and in other African countries.


Assuntos
Camelus/virologia , Infecções por Vírus de RNA/transmissão , Infecções por Vírus de RNA/veterinária , Vírus de RNA/genética , Doenças Transmitidas por Carrapatos/virologia , Carrapatos/virologia , Animais , Genoma Viral/genética , Humanos , Quênia/epidemiologia , RNA Viral/genética , Infestações por Carrapato/epidemiologia , Doenças Transmitidas por Carrapatos/epidemiologia , Carrapatos/classificação , Viroma/genética
14.
Front Immunol ; 12: 708523, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34220870

RESUMO

Major advances have been made in understanding the dynamics of humoral immunity briefly after the acute coronavirus disease 2019 (COVID-19). However, knowledge concerning long-term kinetics of antibody responses in convalescent patients is limited. During a one-year period post symptom onset, we longitudinally collected 162 samples from 76 patients and quantified IgM and IgG antibodies recognizing the nucleocapsid (N) protein or the receptor binding domain (RBD) of the spike protein (S). After one year, approximately 90% of recovered patients still had detectable SARS-CoV-2-specific IgG antibodies recognizing N and RBD-S. Intriguingly, neutralizing activity was only detectable in ~43% of patients. When neutralization tests against the E484K-mutated variant of concern (VOC) B.1.351 (initially identified in South Africa) were performed among patients who neutralize the original virus, the capacity to neutralize was even further diminished to 22.6% of donors. Despite declining N- and S-specific IgG titers, a considerable fraction of recovered patients had detectable neutralizing activity one year after infection. However, neutralizing capacities, in particular against an E484K-mutated VOC were only detectable in a minority of patients one year after symptomatic COVID-19. Our findings shed light on the kinetics of long-term immune responses after natural SARS-CoV-2 infection and argue for vaccinations of individuals who experienced a natural infection to protect against emerging VOC.


Assuntos
Anticorpos Neutralizantes/sangue , Anticorpos Antivirais/sangue , COVID-19/imunologia , Imunoglobulina G/sangue , Imunoglobulina M/sangue , SARS-CoV-2/imunologia , Idoso , Formação de Anticorpos/imunologia , COVID-19/terapia , Convalescença , Proteínas do Nucleocapsídeo de Coronavírus/imunologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fosfoproteínas/imunologia , Glicoproteína da Espícula de Coronavírus/imunologia , Fatores de Tempo
15.
Nat Commun ; 12(1): 4351, 2021 07 16.
Artigo em Inglês | MEDLINE | ID: mdl-34272380

RESUMO

Small-molecule modulators of TLR8 have drawn much interests as it plays pivotal roles in the innate immune response to single-stranded RNAs (ssRNAs) derived from viruses. However, their clinical uses are limited because they can invoke an uncontrolled, global inflammatory response. The efforts described herein culminate in the fortuitous discovery of a tetrasubstituted imidazole CU-CPD107 which inhibits R848-induced TLR8 signaling. In stark contrast, CU-CPD107 shows unexpected synergistic agonist activities in the presence of ssRNA, while CU-CPD107 alone is unable to influence TLR8 signaling. CU-CPD107's unique, dichotomous behavior sheds light on a way to approach TLR agonists. CU-CPD107 offers the opportunity to avoid the undesired, global inflammation side effects that have rendered imidazoquinolines clinically irrelevant, providing an insight for the development of antiviral drugs.


Assuntos
Imidazóis/farmacologia , Transdução de Sinais/efeitos dos fármacos , Receptor 8 Toll-Like/agonistas , Receptor 8 Toll-Like/antagonistas & inibidores , Calorimetria , Células HEK293 , Humanos , Imidazóis/síntese química , Imidazóis/química , Inflamação , Simulação de Acoplamento Molecular , Quinolinas/química , Quinolinas/farmacologia , RNA/química , RNA/farmacologia , Proteínas Recombinantes , Transdução de Sinais/imunologia , Relação Estrutura-Atividade , Receptor 8 Toll-Like/química , Receptor 8 Toll-Like/metabolismo , Difração de Raios X
16.
PLoS Biol ; 19(5): e3001209, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33961621

RESUMO

The ongoing Coronavirus Disease 2019 (COVID-19) pandemic caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) threatens global public health and economy unprecedentedly, requiring accelerating development of prophylactic and therapeutic interventions. Molecular understanding of neutralizing antibodies (NAbs) would greatly help advance the development of monoclonal antibody (mAb) therapy, as well as the design of next generation recombinant vaccines. Here, we applied H2L2 transgenic mice encoding the human immunoglobulin variable regions, together with a state-of-the-art antibody discovery platform to immunize and isolate NAbs. From a large panel of isolated antibodies, 25 antibodies showed potent neutralizing activities at sub-nanomolar levels by engaging the spike receptor-binding domain (RBD). Importantly, one human NAb, termed PR1077, from the H2L2 platform and 2 humanized NAb, including PR953 and PR961, were further characterized and subjected for subsequent structural analysis. High-resolution X-ray crystallography structures unveiled novel epitopes on the receptor-binding motif (RBM) for PR1077 and PR953, which directly compete with human angiotensin-converting enzyme 2 (hACE2) for binding, and a novel non-blocking epitope on the neighboring site near RBM for PR961. Moreover, we further tested the antiviral efficiency of PR1077 in the Ad5-hACE2 transduction mouse model of COVID-19. A single injection provided potent protection against SARS-CoV-2 infection in either prophylactic or treatment groups. Taken together, these results shed light on the development of mAb-related therapeutic interventions for COVID-19.


Assuntos
Anticorpos Neutralizantes/imunologia , COVID-19/virologia , SARS-CoV-2/imunologia , Animais , Anticorpos Monoclonais/imunologia , Anticorpos Monoclonais/metabolismo , Anticorpos Neutralizantes/metabolismo , Anticorpos Neutralizantes/ultraestrutura , Anticorpos Antivirais/imunologia , COVID-19/epidemiologia , COVID-19/imunologia , COVID-19/metabolismo , Epitopos/imunologia , Humanos , Camundongos , Camundongos Transgênicos , Testes de Neutralização , Pandemias , Ligação Proteica , Domínios Proteicos , Receptores Virais/imunologia , Glicoproteína da Espícula de Coronavírus/imunologia
17.
Nat Commun ; 12(1): 2623, 2021 05 11.
Artigo em Inglês | MEDLINE | ID: mdl-33976198

RESUMO

COVID-19 pandemic caused by SARS-CoV-2 constitutes a global public health crisis with enormous economic consequences. Monoclonal antibodies against SARS-CoV-2 can provide an important treatment option to fight COVID-19, especially for the most vulnerable populations. In this work, potent antibodies binding to SARS-CoV-2 Spike protein were identified from COVID-19 convalescent patients. Among them, P4A1 interacts directly with and covers majority of the Receptor Binding Motif of the Spike Receptor-Binding Domain, shown by high-resolution complex structure analysis. We further demonstrate the binding and neutralizing activities of P4A1 against wild type and mutant Spike proteins or pseudoviruses. P4A1 was subsequently engineered to reduce the potential risk for Antibody-Dependent Enhancement of infection and to extend its half-life. The engineered antibody exhibits an optimized pharmacokinetic and safety profile, and it results in complete viral clearance in a rhesus monkey model of COVID-19 following a single injection. These data suggest its potential against SARS-CoV-2 related diseases.


Assuntos
Anticorpos Monoclonais/imunologia , Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/imunologia , COVID-19/imunologia , SARS-CoV-2/imunologia , Glicoproteína da Espícula de Coronavírus/imunologia , Animais , Anticorpos Monoclonais/metabolismo , Anticorpos Monoclonais/uso terapêutico , Anticorpos Neutralizantes/química , Anticorpos Neutralizantes/metabolismo , Anticorpos Antivirais/metabolismo , Especificidade de Anticorpos/imunologia , COVID-19/epidemiologia , Linhagem Celular Tumoral , Células Cultivadas , Chlorocebus aethiops , Feminino , Humanos , Macaca mulatta , Masculino , Mutação , Pandemias , Ligação Proteica , Domínios Proteicos , SARS-CoV-2/efeitos dos fármacos , SARS-CoV-2/metabolismo , Glicoproteína da Espícula de Coronavírus/genética , Glicoproteína da Espícula de Coronavírus/metabolismo , Resultado do Tratamento , Células Vero , Tratamento Farmacológico da COVID-19
18.
Chin Chem Lett ; 32(10): 3019-3022, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-33840982

RESUMO

The wide-spreading SARS-CoV-2 virus has put the world into boiling water for more than a year, however pharmacological therapies to act effectively against coronavirus disease 2019 (COVID-19) remain elusive. Chloroquine (CQ), an antimalarial drug, was found to exhibit promising antiviral activity in vitro and in vivo at a high dosage, thus CQ was approved by the FDA for the emergency use authorization (EUA) in the fight against COVID-19 in the US, but later was revoked the EUA status due to the severe clinical toxicity. Herein, we show that supramolecular formulation of CQ by a macrocyclic host, curcurbit[7]uril (CB[7]), reduced its non-specific toxicity and improved its antiviral activity against coronavirus, working in synergy with CB[7]. CB[7] was found to form 1:1 host-guest complexes with CQ, with a binding constant of ∼104 L/mol. The CQ-CB[7] formulation decreased the cytotoxicity of CQ against Vero E6 and L-02 cell lines. In particular, the cytotoxicity of CQ (60 µmol/L) against both Vero E6 cell line and L-02 cell lines was completely inhibited in the presence of 300 µmol/L and 600 µmol/L CB[7], respectively. Furthermore, the CB[7] alone showed astonishing antiviral activity in SARS-CoV-2 infected Vero E6 cells and mouse hepatitis virus strain A59 (MHV-A59) infected N2A cells, and synergistically improved the antiviral activity of CQ-CB[7], suggesting that CB[7]-based CQ formulation has a great potential as a safe and effective antiviral agent against SARS-CoV-2 and other coronavirus.

19.
J Hematol Oncol ; 14(1): 72, 2021 04 29.
Artigo em Inglês | MEDLINE | ID: mdl-33926500

RESUMO

Evidence suggests that platelets may directly interact with SARS-CoV-2, raising the concern whether ACE2 receptor plays a role in this interaction. The current study showed that SARS-CoV-2 interacts with both platelets and megakaryocytes despite the limited efficiency. Abundance of the conventional receptor ACE2 and alternative receptors or co-factors for SARS-CoV-2 entry was characterized in platelets from COVID-19 patients and healthy persons as well as human megakaryocytes based on laboratory tests or previously reported RNA-seq data. The results suggest that SARS-CoV-2 interacts with platelets and megakaryocytes via ACE2-independent mechanism and may regulate alternative receptor expression associated with COVID-19 coagulation dysfunction.


Assuntos
Plaquetas/virologia , COVID-19/sangue , Megacariócitos/virologia , Receptores Virais/fisiologia , SARS-CoV-2/fisiologia , Ligação Viral , Enzima de Conversão de Angiotensina 2/análise , Plaquetas/metabolismo , COVID-19/complicações , COVID-19/genética , Linhagem Celular , Humanos , Megacariócitos/metabolismo , Especificidade de Órgãos , Ativação Plaquetária , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , RNA Viral/isolamento & purificação , SARS-CoV-2/isolamento & purificação , Trombofilia/etiologia , Trombofilia/virologia , Transcrição Gênica , Internalização do Vírus
20.
Nat Commun ; 12(1): 1813, 2021 03 22.
Artigo em Inglês | MEDLINE | ID: mdl-33753738

RESUMO

Long-term antibody responses and neutralizing activities in response to SARS-CoV-2 infection are not yet clear. Here we quantify immunoglobulin M (IgM) and G (IgG) antibodies recognizing the SARS-CoV-2 receptor-binding domain (RBD) of the spike (S) or the nucleocapsid (N) protein, and neutralizing antibodies during a period of 6 months from COVID-19 disease onset in 349 symptomatic COVID-19 patients who were among the first be infected world-wide. The positivity rate and magnitude of IgM-S and IgG-N responses increase rapidly. High levels of IgM-S/N and IgG-S/N at 2-3 weeks after disease onset are associated with virus control and IgG-S titers correlate closely with the capacity to neutralize SARS-CoV-2. Although specific IgM-S/N become undetectable 12 weeks after disease onset in most patients, IgG-S/N titers have an intermediate contraction phase, but stabilize at relatively high levels over the 6 month observation period. At late time points, the positivity rates for binding and neutralizing SARS-CoV-2-specific antibodies are still >70%. These data indicate sustained humoral immunity in recovered patients who had symptomatic COVID-19, suggesting prolonged immunity.


Assuntos
COVID-19/imunologia , SARS-CoV-2/imunologia , Adulto , Idoso , Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/imunologia , Feminino , Humanos , Imunidade Humoral/imunologia , Imunoglobulina G/imunologia , Imunoglobulina M/imunologia , Masculino , Pessoa de Meia-Idade , Índice de Gravidade de Doença , Glicoproteína da Espícula de Coronavírus
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