Preclinical characterization of amubarvimab and romlusevimab, a pair of non-competing neutralizing monoclonal antibody cocktail, against SARS-CoV-2

Monoclonal antibodies (mAbs) targeting the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein have demonstrated clinical efficacy in preventing or treating coronavirus disease 2019 (COVID-19), resulting in the emergency use authorization (EUA) for several SARS-CoV-2 targeting mAb by regulatory authority. However, the continuous virus evolution requires diverse mAb options to combat variants. Here we describe two fully human mAbs, amubarvimab (BRII-196) and romlusevimab (BRII-198) that bind to non-competing epitopes on the receptor binding domain (RBD) of spike protein and effectively neutralize SARS-CoV-2 variants. A YTE modification was introduced to the fragment crystallizable (Fc) region of both mAbs to prolong serum half-life and reduce effector function. The amubarvimab and romlusevimab combination retained activity against most mutations associated with reduced susceptibility to previously authorized mAbs and against variants containing amino acid substitutions in their epitope regions. Consistently, the combination of amubarvimab and romlusevimab effectively neutralized a wide range of viruses including most variants of concern and interest in vitro. In a Syrian golden hamster model of SARS-CoV-2 infection, animals receiving combination of amubarvimab and romlusevimab either pre- or post-infection demonstrated less weight loss, significantly decreased viral load in the lungs, and reduced lung pathology compared to controls. These preclinical findings support their development as an antibody cocktail therapeutic option against COVID-19 in the clinic.

Magnetically Enhanced Liquid SERS for Ultrasensitive Analysis of Bacterial and SARS-CoV-2 Biomarkers.

In this work, it is shown that surface-enhanced Raman scattering (SERS) measurements can be performed using liquid platforms to perform bioanalysis at sub-pM concentrations. Using magnetic enrichment with gold-coated magnetic nanoparticles, the high sensitivity was verified with nucleic acid and protein targets. The former was performed with a DNA fragment associated with the bacteria Staphylococcus aureus, and the latter using IgG antibody, a biomarker for COVID-19 screening. It is anticipated that this work will inspire studies on ultrasensitive SERS analyzers suitable for large-scale applications, which is particularly important for in vitro diagnostics and environmental studies.

Preliminary Evaluation of a New Initiative to Centralize Colorectal Cancer Care During the COVID-19 Epidemic in Shanghai, China: A Retrospective Study (preprint)

Background: A novel colorectal cancer center (CCC) was developed in the Tenth Hospital of Shanghai during the coronavirus disease 2019 (COVID-19) epidemic. We aimed to evaluate the CCC for the centralized management of colorectal cancer (CRC) on three dimensions during this distinctive period. Methods: This retrospective study used data from the Tenth Hospital’s patient databases. The research hypothesis was that the CCC reduces preoperative waiting time (PWT), length of stay (LOS) during hospitalization, and costs of hospitalization, without reducing the quality of surgery. Thus, we analyzed three dimensions: time, cost, and quality. We compared the expected outcomes between March 1–December 31, 2019, and March 1–December 31, 2020. Descriptive and inferential analyses of patient demographic characteristics, time, postoperative outcomes, and inpatient costs were conducted. Findings: A total of 965 hospitalizations for CRC were identified. In the CCC, PWT declined by 26.2 hours (p<0.01). Patients who entered the CCC express group only needed to wait for 24.5 hours before undergoing surgery, with shorter LOS during hospitalization than the normal group (p<0.01). No patients had any symptoms of COVID-19 or high-risk COVID-19 contacts, and the immediate postoperative complication incidence was low. The mean total inpatient cost (TIC) for all patients with CRC was 78309.824 Chinese yuan in 2020, which was slightly lower than this cost in 2019. Interpretation: This study examined the efficiency of the centralized management model for CRC care during the COVID-19 epidemic in terms of time, cost, and quality. Funding Information: This study was sponsored by the Clinical Research Plan of SHDC (No. SHDC2020CR5006-002), the National Natural Science Foundation of China (No.71804128, 71904145), the Special Funds for Fundamental Research Expenses of Central Universities (No. 22120200407) and the Personnel Development Plan of Shanghai Tenth People's Hospital of Tongji University (No. 2021SYPDRC014).Declaration of Interests: None declared.Ethics Approval Statement: This study has been approved by the Ethics Committee of the Shanghai Tenth People's Hospital of Tongji University (SHSY-IEC-4.1/20-272/01).

Identification of bis-benzylisoquinoline alkaloids as SARS-CoV-2 entry inhibitors from a library of natural products in vitro (preprint)

Coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a major public health issue. To screen for antiviral drugs for COVID-19 treatment, we constructed a SARS-CoV-2 spike (S) pseudovirus system using an HIV-1-based lentiviral vector with a luciferase reporter gene to screen 188 small potential antiviral compounds. Using this system, we identified nine compounds, specifically, bis-benzylisoquinoline alkaloids, that potently inhibited SARS-CoV-2 pseudovirus entry, with EC50 values of 0.1-10 M. Mechanistic studies showed that these compounds, reported as calcium channel blockers (CCBs), inhibited Ca2+-mediated membrane fusion and consequently suppressed coronavirus entry. These candidate drugs showed broad-spectrum efficacy against the entry of several coronavirus pseudotypes (SARS-CoV, MERS-CoV, SARS-CoV-2 [S-D614 and S-G614]) in different cell lines (293T, Calu-3, and A549). Antiviral tests using native SARS-CoV-2 in Vero E6 cells confirmed that four of the drugs (SC9/cepharanthine, SC161/hernandezine, SC171, and SC185/neferine) reduced cytopathic effect and supernatant viral RNA load. Among them, cepharanthine showed the strongest anti-SARS-CoV-2 activity. Collectively, this study offers new lead compounds for coronavirus antiviral drug discovery.

A Survey on Contact Tracing: the Latest Advancements and Challenges (preprint)

Infectious diseases are caused by pathogenic microorganisms, such as bacteria, viruses, parasites or fungi, which can be spread, directly or indirectly, from one person to another. Infectious diseases pose a serious threat to human health, especially COVID-19 that has became a serious worldwide health concern since the end of 2019. Contact tracing is the process of identifying, assessing, and managing people who have been exposed to a disease to prevent its onward transmission. Contact tracing can help us better understand the transmission link of the virus, whereby better interrupting its transmission. Given the worldwide pandemic of COVID-19, contact tracing has become one of the most critical measures to effectively curb the spread of the virus. This paper presents a comprehensive survey on contact tracing, with a detailed coverage of the recent advancements the models, digital technologies, protocols and issues involved in contact tracing. The current challenges as well as future directions of contact tracing technologies are also presented.

Longitudinal dynamics of the neutralizing antibody response to SARS-CoV-2 infection (preprint)

Background Coronavirus disease 2019 (COVID-19) is a global pandemic with no licensed vaccine or specific antiviral agents for therapy. Little is known about the longitudinal dynamics of SARS-CoV-2-specific neutralizing antibodies (NAbs) in COVID-19 patients. Methods Blood samples (n=173) were collected from 30 COVID-19 patients over a 3-month period after symptom onset and analyzed for SARS-CoV-2-specific NAbs, using the lentiviral pseudotype assay, coincident with the levels of IgG and proinflammatory cytokines. Results SARS-CoV-2-specific NAb titers were low for the first 7-10 d after symtom onset and increased after 2-3 weeks. The median peak time for NAbs was 33 d (IQR 24-59 d) after symptom onset. NAb titers in 93.3% (28/30) of the patients declined gradually over the 3-month study period, with a median decrease of 34.8% (IQR 19.6-42.4%). NAb titers increased over time in parallel with the rise in IgG antibody levels, correlating well at week 3 (r = 0.41, p < 0.05). The NAb titers also demonstrated a significant positive correlation with levels of plasma proinflammatory cytokines, including SCF, TRAIL, and M-CSF. Conclusions These data provide useful information regarding dynamic changes in NAbs in COVID-19 patients during the acute and convalescent phases.

The D614G mutation of SARS-CoV-2 spike protein enhances viral infectivity and decreases neutralization sensitivity to individual convalescent sera (preprint)

Coronavirus disease 2019 (COVID-19) is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The spike (S) protein that mediates SARS-CoV-2 entry into host cells is a major target for vaccines and therapeutics. Thus, insights into its sequence variations are key to understanding the infection and antigenicity of SARS-CoV-2. A dominant mutational variant at position 614 of the S protein (aspartate to glycine, D614G mutation) was observed in the SARS-CoV-2 genome sequence obtained from the Nextstrain database. Using a pseudovirus-based assay, we identified that S-D614 and S-G614 protein pseudotyped viruses share a common receptor, human angiotensin-converting enzyme 2 (ACE2), which could be blocked by recombinant ACE2 with the fused Fc region of human IgG1. However, S-D614 and S-G614 protein demonstrated functional differences. First, S-G614 protein could be cleaved by serine protease elastase-2 more efficiently. Second, S-G614 pseudovirus infected 293T-ACE2 cells significantly more efficiently than did the S-D614 pseudovirus, especially in the presence of elastase-2. Third, an elastase inhibitor approved for clinical use blocked elastase-enhanced S-G614 pseudovirus infection. Moreover, 93% (65/70) convalescent sera from patients with COVID-19 could neutralize both S-D614 and S-G614 pseudoviruses with comparable efficiencies, but about 7% (5/70) convalescent sera showed reduced neutralizing activity against the S-G614 pseudovirus. These findings have important implications for SARS-CoV-2 transmission and immune interventions.Competing Interest StatementThe authors have declared no competing interest.View Full Text