Neutralizing antibodies from naturally infected individuals against SARS-CoV-2 Gamma and Delta variants in the Paraguayan population.

INTRODUCTION: Severe Acute Respiratory Syndrome-Coronavirus-2 Virus (SARS-CoV-2) is responsible for Coronavirus Disease 2019 (COVID-19). A substantial number of SARS-CoV-2 infection cases have been reported during the pandemic, and vaccination coverage in some regions, particularly in developing countries, remains very low. SARS-CoV-2 variants of concern (VOCs) have also emerged as some of the most pressing public health issues. In this scenario, it is crucial to know whether COVID-19 convalescent antibodies have cross-neutralizing action against VOCs to contribute to the analysis of the future progress of the pandemic. METHODOLOGY: The plasma of individuals infected with SARS-CoV-2 from June to November 2020 in Paraguay (before the first recorded infections associated with VOCs in the country) was selected. Anti-spike antibodies were determined in plasma samples (n = 626) obtained from this convalescent and unvaccinated group. Using a pseudotyped virus neutralization assay, we then investigated the neutralizing response against D614G variant and Gamma, and Delta VOCs. RESULTS: IgG antibodies against spike were detected in 85.6% of convalescent individuals. Samples from individuals previously infected by a non-VOC showed a 6.6- and 8.1-fold reduction in neutralizing capacity to the Gamma and Delta variants, respectively, when compared to the D614G variant. CONCLUSIONS: Our findings show that antibodies generated by non-VOC infection have reduced neutralizing capabilities against Gamma and Delta variants that appeared subsequently and might have implications for immunity strategies.

Inhibitory Activity of Saussurea costus Extract against Bacteria, Candida, Herpes, and SARS-CoV-2.

Medicinal herbs have long been utilized to treat various diseases or to relieve the symptoms of some ailments for extended periods. The present investigation demonstrates the phytochemical profile, molecular docking, anti-Candida activity, and anti-viral activity of the Saussurea costus acetic acid extract. GC-MS analysis of the extract revealed the presence of 69 chemical compounds. The chemical compounds were alkaloids (4%), terpenoids (79%), phenolic compounds (4%), hydrocarbons (7%), and sterols (6%). Molecular docking was used to study the inhibitory activity of 69 identified compounds against SARS-CoV-2. In total, 12 out of 69 compounds were found to have active properties exhibiting SARS-CoV-2 inhibition. The binding scores of these molecules were significantly low, ranging from -7.8 to -5.6 kcal/mol. The interaction of oxatricyclo [20.8.0.0(7,16)] triaconta-1(22),7(16),9,13,23,29-hexaene with the active site is more efficient. Furthermore, the extract exhibited significant antimicrobial activity (in vitro) against Candida albicans, which was the most susceptible microorganism, followed by Bacillus cereus, Salmonella enterica, Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa, respectively. On the other hand, its antiviral activity was evaluated against HSV-1 and SARS-CoV-2, and the results showed a significant positive influence against HSV-1 (EC50 = 82.6 g/mL; CC50 = 162.9 g/mL; selectivity index = 1.9). In spite of this, no impact could be observed in terms of inhibiting the entry of SARS-CoV-2 in vitro.

Antiviral activity of two Acanthospermum species against herpes simplex virus 1.

ETHNOPHARMACOLOGICAL RELEVANCE: Acanthospermum species are used in traditional medicine for treating various pathologies, including bacterial and viral infections. In a screening study, we identified the activity of the ethanolic extracts of Acanthospermum australe and Acanthospermum hispidum against herpes simplex virus 1 (HSV-1). AIM OF THE STUDY: In this work, we analyzed the phytochemical profile and antiviral activity of the chemical fractionation products of Acanthospermum australe and Acanthospermum hispidum. Additionally, we identified the effect of these fractions on different steps of the viral cycle. MATERIALS AND METHODS: Acanthospermum samples were extracted with methanol and further partitioned with solvents of increasing polarities: hexane, chloroform, ethyl acetate, and butanol. Cytotoxicity and antiviral activity were analyzed for each fraction. The active fractions were tested to identify the virucidal effect and the inhibition of virus-cell binding. Further, the effect of these fractions on the replication and viral gene was quantitated by qPCR, and the expression of gD protein was evaluated by Western blot. RESULTS: The chloroform and hexane fractions of Acanthospermum hispidum and Acanthospermum australe showed dose-dependent antiviral activity. The chloroform fraction inhibited the virus-cell binding and virus cycle in a post-entry mechanism by decreasing replication and the expression of early and late viral genes. The hexane fraction did not inhibit virus binding; however, it showed antiviral activity in post-entry events by inhibiting the immediate-early, early, and late genes. We identified in both species the presence of 3.6-dimetoxiapigenin, axillarin, and penduletin in the chloroform fraction and methyl-(Z,Z)-9,12-octadecadienoate and phytol in the hexane fraction. CONCLUSIONS: Acanthospermum hispidum and Acanthospermum australe possess antiviral activity against HSV-1 and affect different steps of the viral cycle. These characteristics make them good candidates for developing phytotherapeutic products against HSV-1.

Screening of Natural Products Inhibitors of SARS-CoV-2 Entry.

The COVID-19 pandemic has led to the search for new molecules with antiviral activity against SARS-CoV-2. The entry of the virus into the cell is one of the main targets for inhibiting SARS-CoV-2 infection. Natural products are an important source of new therapeutic alternatives against diseases. Pseudotyped viruses allow the study of SARS-CoV-2 viral entry inhibitors, and due to their simplicity, they allow the screening of a large number of antiviral candidates in Biosafety Level 2 facilities. We used pseudotyped HIV-1 with the D614G SARS-CoV-2 spike glycoprotein to test its ability to infect ACE2-expressing HEK 293T cells in the presence of diverse natural products, including 21 plant extracts, 7 essential oils, and 13 compounds from plants and fungi. The 50% cytotoxic concentration (CC50) was evaluated using the resazurin method. From these analyses, we determined the inhibitory activity of the extract of Stachytarpheta cayennensis, which had a half-maximal inhibitory concentration (IC50) of 91.65 µg/mL, a CC50 of 693.5 µg/mL, and a selectivity index (SI) of 7.57, indicating its potential use as an inhibitor of SARS-CoV-2 entry. Moreover, our work indicates the usefulness of the pseudotyped-virus system in the screening of SARS-CoV-2 entry inhibitors.