RESUMEN
The present study provides the first multiepitope vaccine construct using the 3CL hydrolase protein of SARS-CoV-2. The coronavirus 3CL hydrolase (Mpro) enzyme is essential for proteolytic maturation of the virus. This study was based on immunoinformatics and structural vaccinology strategies. The design of the multiepitope vaccine was built using helper T-cell and cytotoxic T-cell epitopes from the 3CL hydrolase protein along with an adjuvant to enhance immune response; these are joined to each other by short peptide linkers. The vaccine also carries potential B-cell linear epitope regions, B-cell discontinuous epitopes, and interferon-γ-inducing epitopes. Epitopes of the constructed multiepitope vaccine were found to be antigenic, nonallergic, nontoxic, and covering large human populations worldwide. The vaccine construct was modeled, validated, and refined by different programs to achieve a high-quality three-dimensional structure. The resulting high-quality model was applied for conformational B-cell epitope selection and docking analyses with toll-like receptor-3 for understanding the capability of the vaccine to elicit an immune response. In silico cloning and codon adaptation were also performed with the pET-19b plasmid vector. The designed multiepitope peptide vaccine may prompt the development of a vaccine to control SARS-CoV-2 infection.
Asunto(s)
Vacunas contra la COVID-19/inmunología , COVID-19/prevención & control , Proteasas 3C de Coronavirus/inmunología , Epítopos de Linfocito B/inmunología , Epítopos de Linfocito T/inmunología , SARS-CoV-2/inmunología , Receptor Toll-Like 3/inmunología , Secuencia de Aminoácidos , Sitios de Unión , COVID-19/inmunología , COVID-19/virología , Vacunas contra la COVID-19/genética , Clonación Molecular/métodos , Biología Computacional/métodos , Proteasas 3C de Coronavirus/química , Proteasas 3C de Coronavirus/genética , Epítopos de Linfocito B/química , Epítopos de Linfocito B/genética , Epítopos de Linfocito T/química , Epítopos de Linfocito T/genética , Vectores Genéticos/química , Vectores Genéticos/inmunología , Antígenos HLA/química , Antígenos HLA/genética , Antígenos HLA/inmunología , Humanos , Inmunidad Innata/efectos de los fármacos , Inmunogenicidad Vacunal , Interferón gamma/genética , Interferón gamma/inmunología , Simulación del Acoplamiento Molecular , Unión Proteica , Dominios y Motivos de Interacción de Proteínas , SARS-CoV-2/efectos de los fármacos , SARS-CoV-2/patogenicidad , Linfocitos T Citotóxicos/química , Linfocitos T Citotóxicos/inmunología , Linfocitos T Citotóxicos/virología , Linfocitos T Colaboradores-Inductores/química , Linfocitos T Colaboradores-Inductores/inmunología , Linfocitos T Colaboradores-Inductores/virología , Receptor Toll-Like 3/química , Receptor Toll-Like 3/genética , Interfaz Usuario-Computador , Vacunas de SubunidadRESUMEN
Anopheles culicifacies mosquitoes are able to transmit both falciparum and vivax malaria in India. More than 65% of malaria cases reported annually spread through this vector. Despite the fact that it poses major vectorial burden in India, the molecular basis of its immune role against Plasmodium development has not been explored intensively. Here, we characterized An. culicifacies SOCS (suppressor of cytokine signaling) gene, a regulator of STAT pathway and its expression analysis upon Plasmodium infection. Our analysis has demonstrated that An. culicifacies SOCS gene shares strikingly high level of sequence similarity in SH2 domain and SOCS box region with other mosquito species. However, its N-terminal identity is limited to Anophelines mosquito only, suggesting its genus specific role. SOCS mRNA is expressed in all developmental stages of mosquito and its expression is higher in male than female adults. SOCS mRNA is significantly induced after Plasmodium infection in midgut tissue indicating its involvement in the immune defense responses. This is the first evidence of involvement of SOCS as an immune gene in Indian malaria vector An. culicifacies.
Asunto(s)
Anopheles/parasitología , Plasmodium berghei , Proteínas Supresoras de la Señalización de Citocinas/genética , Secuencia de Aminoácidos , Animales , Anopheles/genética , Anopheles/inmunología , Secuencia de Bases , Femenino , Insectos Vectores , Masculino , Datos de Secuencia Molecular , Filogenia , Proteínas Supresoras de la Señalización de Citocinas/químicaRESUMEN
Global statistical data shed light on an alarming trend that every year thousands of people die due to adverse drug reactions as each individual responds in a different way to the same drug. Pharmacogenomics has come up as a promising field in drug development and clinical medication in the past few decades. It has emerged as a ray of hope in preventing patients from developing potentially fatal complications due to adverse drug reactions. Pharmacogenomics also minimizes the exposure to drugs that are less/non-effective and sometimes even found toxic for patients. It is well reported that drugs elicit different responses in different individuals due to variations in the nucleotide sequences of genes encoding for biologically important molecules (drug-metabolizing enzymes, drug targets and drug transporters). Single nucleotide polymorphisms (SNPs), the most common type of polymorphism found in the human genome is believed to be the main reason behind 90% of all types of genetic variations among the individuals. Therefore, pharmacogenomics may be helpful in answering the question as to how inherited differences in a single gene have a profound effect on the mobilization and biological action of a drug. In the present review, we have discussed clinically relevant examples of SNP in associated diseases that can be utilized as markers for "better management of complex diseases" and attempted to correlate the drug response with genetic variations. Attention is also given towards the therapeutic consequences of inherited differences at the chromosomal level and how associated drug disposition and/or drug targets differ in various diseases as well as among the individuals.
Asunto(s)
Farmacogenética , Variantes Farmacogenómicas , Polimorfismo de Nucleótido Simple , Marcadores Genéticos , Predisposición Genética a la Enfermedad , Humanos , Fenotipo , Factores de Riesgo , Resultado del TratamientoRESUMEN
Effect of anti-mosquito-fat body antibodies on the development of the malaria parasite, Plasmodium vivax has been studied by feeding Anopheles stephensi mosquitoes with infected blood supplemented with serum from immunized rabbits. Immunogenic polypeptides were identified by western blot. Mosquitoes that ingested anti-fat body antibodies along with infectious blood meal had significantly fewer oocysts than the mosquitoes in the control group. Effect of anti-mosquito fat body antibodies on fecundity, hatchability, mortality and engorgement of mosquitoes has also been reported. A significant reduction in fecundity and hatchability was observed, however, effect on mortality and engorgement was variable and statistically insignificant. Results indicated that fat body antibodies have the potential to disrupt reproductive physiology of malaria vector An. stephensi.
Asunto(s)
Anopheles/fisiología , Anticuerpos/farmacología , Cuerpo Adiposo/inmunología , Malaria/prevención & control , Plasmodium vivax/fisiología , Animales , Anopheles/inmunología , Anopheles/parasitología , Anticuerpos/inmunología , Western Blotting , Reacciones Cruzadas , Ensayo de Inmunoadsorción Enzimática , Fertilidad/efectos de los fármacos , Humanos , Sueros Inmunes , Insectos Vectores , Malaria/sangre , Malaria/transmisión , Plasmodium vivax/patogenicidad , ConejosRESUMEN
Rabbit antibodies to hemolymph antigens (102.5, 101, 100, 96, 88, 80, 64, 55, 43, 29, and 23 kDa) of Anopheles stephensi reduced fecundity as well as viability in An. stephensi. However, ingestion of these antibodies was not associated with a marked effect on the engorgement of mosquitoes but egg laying was significantly delayed. Antisera raised against hemolymph proteins were also used to identify cross reactive antigens/epitopes present in other tissues by Western blotting, as well as by in vivo ELISA. In addition, a significant reduction in oocyst development was also observed in An. stephensi mosquitoes that ingested anti-hemolymph antibodies along with Plasmodium vivax. The results confirmed the feasibility of targeting mosquito antigens as a novel anti-mosquito strategy, as well as confirmed the usefulness of such antigens for the development of a transmission-blocking vaccine.