Designing of multiepitope-based vaccine against Leptospirosis using Immuno-Informatics approaches (original) (raw)
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Discovery of Novel Leptospirosis Vaccine Candidates Using Reverse and Structural Vaccinology
Frontiers in immunology, 2017
Leptospira spp. are diderm (two membranes) bacteria that infect mammals causing leptospirosis, a public health problem with global implications. Thousands of people die every year due to leptospirosis, especially in developing countries with tropical climates. Prophylaxis is difficult due to multiple factors, including the large number of asymptomatic hosts that transmit the bacteria, poor sanitation, increasing numbers of slum dwellers, and the lack of an effective vaccine. Several leptospiral recombinant antigens were evaluated as a replacement for the inactivated (bacterin) vaccine; however, success has been limited. A prospective vaccine candidate is likely to be a surface-related protein that can stimulate the host immune response to clear leptospires from blood and organs. In this study, a comprehensive bioinformatics approach based on reverse and structural vaccinology was applied toward the discovery of novel leptospiral vaccine candidates. The Leptospira interrogans serovar...
Scientific reports, 2018
Leptospirosis is the most widespread zoonotic disease, estimated to cause severe infection in more than one million people each year, particularly in developing countries of tropical areas. Several factors such as variable and nonspecific clinical manifestation, existence of large number of serovars and asymptomatic hosts spreading infection, poor sanitation and lack of an effective vaccine make prophylaxis difficult. Consequently, there is an urgent need to develop an effective vaccine to halt its spread all over the world. In this study, an immunoinformatics approach was employed to identify the most vital and effective immunogenic protein from the proteome of Leptospira interrogans serovar Copenhageni strain L1-130 that may be suitable to stimulate a significant immune response aiding in the development of peptide vaccine against leptospirosis. Both B-cell and T-cell (Helper T-lymphocyte (HTL) and cytotoxic T lymphocyte (CTL)) epitopes were predicted for the conserved and most im...
Fems Microbiology Letters, 2005
Leptospirosis is an important global human and veterinary health problem. Humans can be infected by exposure to chronically infected animals and their environment. An important focus of the current leptospiral research is the identification of outer membrane proteins (OMPs). Due to their location, leptospiral OMPs are likely to be relevant in host-pathogen interactions, hence their potential ability to stimulate heterologous immunity. The existing whole-genome sequence of Leptospira interrogans serovar Copenhageni offers a unique opportunity to search for cell surface proteins. Predicted genes encoding potential surface proteins were amplified from genomic DNA by PCR methodology and cloned into an Escherichia coli expression system. The partially purified recombinant proteins were probed by Western blotting with sera from human patients diagnosed with leptospirosis. Sixteen proteins, out of a hundred tested, were recognized by antibodies present in human sera. Four of these proteins were conserved among eight serovars of L. interrogans and absent in the non-pathogenic Leptospira biflexa. These proteins might be useful for the diagnosis of the disease as well as potential vaccine candidates.
Computer aided subunit vaccine design against pathogenic Leptospira serovars
Interdisciplinary Sciences: Computational Life Sciences, 2012
Epitopes of Leptospira inducing CD4 + T-cell responses by binding to human MHC molecules could critically contribute to the development of subunit vaccines for leptospirosis. Herein, we have identified unique vaccine peptides from outer membrane proteins (OMPs) common to four sequenced pathogenic Leptospira serovars through in silico reverse vaccinology technique. The OMPs were explored for probable antigens using jemboss and screened in ProPred subsequently to predict thirty HLA-DRB epitopes. The HLA-DRB epitopes were validated through published positive control (HA307-PKYVKQNTLKLAT-319), SYFPEITHI and immune epitope database (IEDB) to list twelve epitopes as putative subunit vaccine peptides from nine OMPs. Cation efflux system membrane protein (czcA) having four subunit vaccine peptides, was modeled in Modeller9v7 and evaluated through Procheck, ProSA and ProQ. The HLA-DRB alleles and czcA 3D interactions were studied using Hex 5.1. Further, the T-cell epitopes present in czcA were docked individually with HLA-DRB alleles. The docking result revealed that czcA and its epitopes were interacting well with HLA-DRB alleles, hence would certainly produce cell mediated immune response in host. Thus, czcA and its four subunit vaccine peptides would be ideal T-cell driven efficacious vaccine against leptospirosis.
Frontiers in Cellular and Infection Microbiology
Leptospirosis is a neglected disease of man and animals that affects nearly half a million people annually and causes considerable economic losses. Current human vaccines are inactivated whole-cell preparations (bacterins) of Leptospira spp. that provide strong homologous protection yet fail to induce a cross-protective immune response. Yearly boosters are required, and serious side-effects are frequently reported so the vaccine is licensed for use in humans in only a handful of countries. Novel universal vaccines require identification of conserved surface-exposed epitopes of leptospiral antigens. Outer membrane β-barrel proteins (βb-OMPs) meet these requirements and have been successfully used as vaccines for other diseases. We report the evaluation of 22 constructs containing protein fragments from 33 leptospiral βb-OMPs, previously identified by reverse and structural vaccinology and cell-surface immunoprecipitation. Three-dimensional structures for each leptospiral βb-OMP were ...
Reverse Vaccinology: An Approach for Identifying Leptospiral Vaccine Candidates
International journal of molecular sciences, 2017
Leptospirosis is a major public health problem with an incidence of over one million human cases each year. It is a globally distributed, zoonotic disease and is associated with significant economic losses in farm animals. Leptospirosis is caused by pathogenic Leptospira spp. that can infect a wide range of domestic and wild animals. Given the inability to control the cycle of transmission among animals and humans, there is an urgent demand for a new vaccine. Inactivated whole-cell vaccines (bacterins) are routinely used in livestock and domestic animals, however, protection is serovar-restricted and short-term only. To overcome these limitations, efforts have focused on the development of recombinant vaccines, with partial success. Reverse vaccinology (RV) has been successfully applied to many infectious diseases. A growing number of leptospiral genome sequences are now available in public databases, providing an opportunity to search for prospective vaccine antigens using RV. Seve...
Molecular Immunology, 2018
Leptospirosis is known as a zoonotic disease of global importance originated from infection with the spirochete bacterium Leptospira. Although several leptospirosis vaccines have been tested, the vaccination is relatively unsuccessful in clinical application despite decades of research. Therefore, this study was conducted to construct a novel multi-epitope based vaccine against leptospirosis by using Hap1, LigA, LAg42, SphH and HSP58 antigens. T cell and IFN gamma epitopes were predicted from these antigens. In addition, to induce strong CD4 + helper T lymphocytes (HTLs) responses, Pan HLA DR-binding epitope (PADRE) and helper epitopes selected from Tetanus toxin fragment C (TTFrC) were applied. Moreover, for boosting immune response, Heparin-Binding Hemagglutinin (HBHA), a novel TLR4 agonist was added to the construct as an adjuvant. Finally, selected epitopes were linked together using EAAAK, GPGPG, AAY and HEYGAEALERAG linkers. Based on the predicted epitopes, a multi-epitope vaccine was construct with 490 amino acids. Physico-chemical properties, secondary and tertiary structures, stability, intrinsic protein disorder, solubility, and allergenicity of this vaccine construct were assessed by applying immunoinformatics analyses. A soluble, and non-allergic protein with a molecular weight of 53.476 kDa was obtained. Further analyses validated the stability of the chimeric protein and the predicted epitopes in the chimeric vaccine indicated strong potential to induce B-cell and T-cell mediated immune response. Therefore, immunoinformatics analysis showed that the modeled multi-epitope vaccine can properly stimulate the both T and B cells immune responses and could potentially be used for prophylactic or therapeutic usages.
International Journal of Infection
Leptospirosis is a widespread zoonotic disease caused by Leptospira interrogans. The conventional vaccines have some major problems. Therefore, recombinant vaccines such as multiple-epitope vaccine are suggested. OmpL1 and lipL32 are the most important proteins of Leptospira interrogans bacteria that can be used in epitope prediction process to design a multiple-epitope vaccine. Hence, in this study, the most reliable and accurate online servers were applied to predict B cell and T cell epitopes, the secondary and tertiary structures, enzyme digestion, and antigenicity score of ompL1 and lipL32. The results showed that epitopes located at 103-122, 210-232, and 272-291 amino acid residues are the common epitopes between T cell (MHCI) and B cell. 288-308 amino acid residues were introduced as common epitopes to stimulate both T cell (MHCI and MHCII) and B cell of ompL1 protein. In the case of LipL32 protein, 80-96 amino acid residues are recommended for T cell epitopes and 63-81 amino acid residues for stimulation of both B and T cells. All the mentioned epitopes can be considered as linear epitopes in designing a recombinant vaccine based on chimeric epitopes. It appears that these epitopes can be applied to design recombinant multiple-epitope vaccines against leptospirosis.
Leptospirosis Vaccine: Search for Subunit Candidates
Procedia in Vaccinology, 2009
Subunit vaccines are a potential intervention strategy against leptospirosis, which is a major public health problem in developing countries. Thus far, several proteins have been evaluated as potential vaccine candidates, but all those tested in aluminum hydroxide adjuvant have failed to protect animals against lethal challenge. Seven new leptospiral lipoproteins were evaluated as vaccine candidates. The coding sequences of these lipoproteins were amplified by PCR from Leptospira interrogans serovar Copenhageni, strain Fiocruz L1-130, cloned and expressed in Escherichia coli. The purified proteins were adsorbed in aluminum adjuvant and used in the immunization of four to six weeks old hamsters. After two doses of 60 g of recombinant protein, hamsters were challenged with a lethal dose of L. interrogans. All seven tested proteins failed to fully protect the animals from disease or death. Further study must be undertaken toward developing an efficient subunit vaccine against leptospirosis for humans, livestock and pets.
World’s Veterinary Journal, 2022
Leptospirosis can cause severe disease and probable death in humans. Antigenic epitopes from pathogenic strains of the bacteria have shown potential for serving as vaccine candidates and play a key role in the sensitivity and specificity of immunodiagnostic tests. This in-vitro analysis was undertaken to develop a prototype recombinant DNA vaccine using in-silico epitope prediction method. Epitope prediction software programs predicted the most antigenic linear B-cell epitopes of OmpL1, LipL32, LipL41, Loa22, and LigA. Thirteen epitopes were predicted, connected by the Gly-Ser linker, and synthesized. The purity of the concentrated recombinant multi-epitope protein was assessed by restriction enzyme digestion and gel electrophoresis. In-vitro expression on mammalian Chinese Hamster Ovary cell line indicated strong cytoplasmic fluorescence produced based on an indirect immunofluorescence antibody test. The green color of the cytoplasm indicates successful transcribed and translated DNA as against the blue-stained nucleus observed in the un-transfected control group based on the indirect immunofluorescence antibody test. The findings of the current study showed high antibody binding potentials of the vaccine constructs, which could be used for diagnostic applications or as polyvalent vaccine candidates.