Vacunas contra el virus chikungunya: avances y perspectivas

El virus chikungunya (CHIKV) es un alfavirus cuya infección provoca una enfermedad caracterizada principalmente por fiebre y dolores articulares/musculares. Entre 25-50% de las infecciones se presentan con enfermedad crónica que puede durar de meses a años. El primer brote de CHIKV en Paraguay corresponde al año 2015, siendo el último en el año 2022/2023. Diversos candidatos vacunales contra CHIKV se encuentran en diferentes etapas de desarrollo, e incluso recientemente (noviembre/2023) fue aprobada la primera vacuna contra CHIKV llamada VLA1553 (Ixchiq). Adicionalmente, al menos 30 candidatos vacunales se encuentran en ensayos preclínicos/clínicos. Con la aprobación de la primera vacuna contra CHIKV y la posibilidad de otras que lleguen al mercado prontamente, debido al estado avanzado de otros candidatos vacunales, se abrirá un nuevo escenario en esta enfermedad. Se espera que la introducción de vacunas efectivas genere un avance importante para la prevención de esta enfermedad, disminuyendo los casos agudos y los efectos crónicos de la infección por el virus. En este trabajo de revisión se analiza el avance de las vacunas contra CHIKV, además de examinar los desafíos de vigilancia epidemiológica que plantean la introducción de estas vacunas.

Chikungunya virus (CHIKV) is an alphavirus that causes an illness characterized mainly by fever and joint/muscle pain. Between 25-50% of infections present with chronic diseases that can last from months to years. The first outbreak of CHIKV in Paraguay occurred in 2015, with the last outbreak occurring in 2022/2023. Several vaccine candidates against CHIKV are in different stages of development, and even recently (November/2023), the first vaccine against CHIKV, called VLA1553 (Ixchiq), was approved. In addition, at least 30 vaccine candidates are available for preclinical and clinical trials. With the approval of the first vaccine against CHIKV and the possibility of others coming to the market soon, due to the advanced status of other vaccine candidates, a new scenario will open for this disease. The introduction of effective vaccines is expected to generate an important advance in the prevention of this disease, reducing acute cases and the chronic effects of viral infection. This review analyzes the progress of CHIKV vaccines and examines the epidemiological surveillance challenges posed by the introduction of these vaccines.

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.

Extractos vegetales de tres especies del género Baccharis inducen la proliferación de células mononucleares humanas / Plant extracts of three species of the genus Baccharis induce human mononuclear cells proliferation

Las plantas de uso en medicina tradicional constituyen una fuente importante de compuestos con actividad inmunomoduladora; entre ellas las especies del género Baccharis, conocidas popularmente como "Jaguareteka´a" en nuestro país, son ampliamente empleadas. En este estudio se evaluó la actividad inmunomoduladora de extractos metanólicos de tres especies del género Baccharis (B. trimera, B. notosergilay B. punctulata) sobre la proliferación de células mononucleares humanas de sangre periférica. Los extractos de las tres especies estudiadas estimularon la proliferación de las células mononucleares. Específicamente, el extracto de B. notosergila estimuló la proliferación celular a todas las concentraciones probadas (5, 10, 25 y 50 µg/mL), mientras que los extractos de B. trimera y B. punctulata mostraron este efecto a 5 y 10 µg/mL. Además, por presentar mayor inducción de la proliferación, se realizó un fraccionamiento con diferentes solventes del extracto metanólico de B. notosergila y B. punctulata. La fracción de acetato de etilo de ambos extractos vegetales aumentó la proliferación celular, sugiriendo que compuestos de polaridad media son los responsables de esta actividad. Estos resultados demuestran que los extractos de B. trimera, B. notosergila y B. punctulata poseen actividad inmunomoduladora sobre células mononucleares humanas y servirán de base a otros estudios para determinar el o los componentes activos de los extractos sobre el sistema inmune(AU)

Plants used in traditional medicine are an important source of compounds with immunomodulatory activity. Species of the genus Baccharis, popularly known as "Jaguareteka'a" in our country, are used in folk medicine for the treatment of liver, gastrointestinal, inflammatory and infectious diseases. In this study, we evaluated the immunomodulatory activity of methanolic extracts of three species of the genus Baccharis (B. trimera, B. notosergila and B. punctulata) on the proliferation of human peripheral blood mononuclear cells. Extracts of the three species studied stimulated the proliferation of mononuclear cells. The extract of B. notosergila stimulated cell proliferation at all concentrations tested, while extracts of B. trimera and B. punctulata stimulated at 5 and 10 µg/mL. In addition, we carried out a separation with different solvents of the methanolic extract of B. notosergila and B. punctulata. The ethyl acetate fraction of both plant extracts induced the proliferation of immune cells. These results show that the extracts of B. trimera, B. notosergila and B. punctulata had immunomodulatory activity on human mononuclear cells. Future work will be required to identify the components responsible for the activity on the immune system(AU)

Rational evolution of the unusual Y-type oxyanion hole of Rhodococcus sp. CR53 lipase LipR.

Rhodococcus sp CR-53 lipase LipR was the first characterized member of bacterial lipase family X. Interestingly, LipR displays some similarity with α/ß-hydrolases of the C. antartica lipase A (CAL-A)-like superfamily (abH38), bearing a Y-type oxyanion hole, never found before among bacterial lipases. In order to explore this unusual Y-type oxyanion hole, and to improve LipR performance, two modification strategies based on site directed or saturation mutagenesis were addressed. Initially, a small library of mutants was designed to convert LipR Y-type oxyanion hole (YDS) into one closer to those most frequently found in bacteria (GGG(X)). However, activity was completely lost in all mutants obtained, indicating that the Y-type oxyanion hole of LipR is required for activity. A second approach was addressed to modify the two main oxyanion hole residues Tyr110 and Asp111, previously described for CAL-A as the most relevant amino acids involved in stabilization of the enzyme-substrate complex. A saturation mutagenesis library was prepared for each residue (Tyr110 and Asp111), and activity of the resulting variants was assayed on different chain length substrates. No functional LipR variants could be obtained when Tyr110 was replaced by any other amino acids, indicating that this is a crucial residue for catalysis. However, among the Asp111 variants obtained, LipR D111G produced a functional enzyme. Interestingly, this LipR-YGS variant showed less activity than wild type LipR on short- or mid- chain substrates but displayed a 5.6-fold increased activity on long chain length substrates. Analysis of the 3D model and in silico docking studies of this enzyme variant suggest that substitution of Asp by Gly produces a wider entrance tunnel that would allow for a better and tight accommodation of larger substrates, thus justifying the experimental results obtained.

Analysis of rotavirus non-structural protein NSP5 by mass spectrometry reveals a complex phosphorylation pattern.

Genomic replication and partial assembly of Rotavirus takes place in cytoplasmic viral structures called viroplasms. NSP5 is a viral phosphoprotein localized in viroplasms and its expression is imperative for viral cycle progress. During infection three isoforms of NSP5 can be observed by SDS-PAGE (26, 28 and 33-35kDa) and previous reports suggested that they differ in their phosphorylation patterns. In this study we obtained NSP5 from infected cells and by mass spectrometry we were able to identify nine phosphorylation sites. We detected that in all the isoforms the same residues can be found either phosphorylated or unmodified. Quantitative analysis showed that the 28kDa isoform has a higher phosphorylation level than the 26kDa isoform suggesting that migration properties depend on the total number of phosphorylated residues. Moreover, we identified two not previously described modifications for this protein: an N-acetylation in Serine-2 and an intramolecular disulfide bond in a highly conserved motif, CXXC which is located between two charged alpha-helix motifs.