Designing a recombinant multiepitope vaccine against Leishmania donovani based immunoinformatics approaches

Hashemzadeh, Pejman; Rouzbahani, Arian Karimi; Bandehpour, Mojgan; Kheirandish, Farnaz; Dariushnejad, Hassan; Mohamadi, Mohsen

doi:10.23736/s1120-4826.20.02610-5

Cited by 3 publications

(1 citation statement)

References 40 publications

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“…The studies showed that there is anti-histone antibody H1 in serum of patients with Leishmaniasis. Histone H1 is a highly immunogenic protein, which is highly expressed in both promastigote and amastigote stages of Leishmania species (Hashemzadeh, Karimi Rouzbahani et al 2020). Leishmania A2 protein is mainly expressed in amastigote.…”

Section: Discussionmentioning

confidence: 99%

Design and evaluation of a novel multi-epitope antigen for evaluate the diagnostic immunity responses against Leishmania infantum infection

Hashemzadeh,

Bandehpour,

Kheirandish

et al. 2024

Preprint

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Background Leishmania infantum is the causative agent of visceral leishmaniasis in the Mediterranean region. The diagnosis of complex visceral leishmaniasis and delays in the diagnosis of the infection are associated with the death of patients. Proper diagnosis of infection is an important measure in controlling and preventing the disease. However, studies have shown that the accuracy of antigens used in current diagnostic tests is insufficient, for this reason, researchers are trying to identify multi-epitope antigens as diagnostic markers to increase the specificity and sensitivity of diagnostic tests. In this study, the design and expression of Leishmania infantum multi-epitope antigens were carried out in two parts of the structure design using bioinformatics tools and the laboratory part for the production of the recombinant protein. Materials and Methods The aim of this study was to design and computationally analyze and express Leishmania infantum multi-epitope antigens. In this study, nine antigenic proteins (CPB, H1, KMP11, GP63, HASPB, A2, K39, LACK, and PSA) were selected. Bioinformatics analyzes such as prediction of immune cell epitopes, design of recombinant structure, antigenicity, allergenicity, evaluation of physicochemical properties, solubility, prediction of secondary structure and tertiary structure, refinement and validation of 3D model structure and finally in silico cloning optimization of protein construct were performed. After synthesis of the designed recombinant gene fusion sequence in pUC57 cloning vector, its subcloning was performed in pET26b prokaryotic expression vector using BamHI/ HindIII restriction enzymes. The expression of recombinant multi-epitope antigen was performed in E. coli B (BL21) strain using IPTG inducer and confirmed by SDS-PAGE and western blotting techniques. Results The results of computational analysis showed that the complete structure, which is suitable for immunogenicity and is non-allergenic, was successfully cloned into pET-26b and expressed as a complete protein. Conclusion Finally, the protein was approved. Based on the expression of recombinant proteins and bioinformatics analysis, this structure can be studied in mouse models and its safety can be evaluated.

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Section: Discussionmentioning

confidence: 99%

Design and evaluation of a novel multi-epitope antigen for evaluate the diagnostic immunity responses against Leishmania infantum infection

Hashemzadeh,

Bandehpour,

Kheirandish

et al. 2024

Preprint

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Immunoinformatics analysis of Brucella melitensis to approach a suitable vaccine against brucellosis

Hashemzadeh,

nezhad,

Khoshkhabar

2023

Journal of Genetic Engineering and Biotechnology

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Background Brucellosis caused by B. melitensis is one of the most important common diseases between humans and livestock. Currently, live attenuated vaccines are used for this disease, which causes many problems, and unfortunately, there is no effective vaccine for human brucellosis. The aim of our research was to design a recombinant vaccine containing potential immunogenic epitopes against B. melitensis. Methods In this study, using immunoinformatics approaches, 3 antigens Omp31, Omp25, and Omp28 were identified and the amino acid sequence of the selected antigens was determined in NCBI. Signal peptides were predicted by SignaIP-5.0 server. To predict B-cell epitopes from ABCpred and Bcepred servers, to predict MHC-I epitopes from RANKPEP and SYFPEITHI servers, to predict MHC-II epitopes from RANKPEP and MHCPred servers, and to predict CTL epitopes were used from the CTLPred server. Potentially immunogenic final epitopes were joined by flexible linkers. Finally, allergenicity (AllerTOP 2.0 server), antigenicity (Vaxijen server), physicochemical properties (ProtParam server), solubility (Protein-sol server), secondary (PSIPRED and GRO4 servers) and tertiary structure (I-TASSER server), refinement (GalaxyWEB server), validation (ProSA-web, Molprobity, and ERRAT servers), and optimization of the codon sequence (JCat server) of the structure of the multi-epitope vaccine were analyzed. Results The analysis of immunoinformatics tools showed that the designed vaccine has high quality, acceptable physicochemical properties, and can induce humoral and cellular immune responses against B. melitensis bacteria. In addition, the high expression level of recombinant antigens in the E. coli host was observed through in silico simulation. Conclusion According to the results in silico, the designed vaccine can be a suitable candidate to fight brucellosis and in vitro and in vivo studies are needed to evaluate the research of this study.

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Bioinformatics Analysis to Designing a Multi-epitope-based Peptide Vaccine Combat Leishmania major

Rouzbahani

Kheirandish

Hashemzadeh

2022

Iran J Med Microbiol

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Background and Aim: Cutaneous leishmaniasis is a significant public health issue worldwide. Cutaneous leishmaniasis is the most prevalent in the world among the different types of leishmaniasis. Currently, available medications have had no discernible influence on the disease's progression. Up to now, there has been no approved cutaneous leishmaniasis vaccine. New developments in vaccination might be a potential way to come up with a vaccination that is successful for the treatment of cutaneous leishmaniasis. Materials and Methods:This research was conducted to learn more about an effective vaccine for Leishmania major, the ailment's primary cause of CL, which was designed using computational methods. Thus, a multiepitope protein was designed by utilizing potential immune system epitopes, including predicted MHC class I, MHC class II, Cytotoxic T lymphocytes, Bcell, and Interferon-gamma epitopes of Cysteine protease b (CPB), Leishmania homologue of activated C kinase (LACK), and Kinetoplastid membrane protein-11 (KMP-11) antigenic proteins. In order to enhance vaccine immunogenicity, two resuscitation-promoting factors of Mycobacterium tuberculosis were used as adjuvants. Final epitopes were matched with suitable linkers to construct the recombinant structure. The physicochemical and immune-based characteristics of the designed vaccine have been forecasted by using different tools. Moreover, homogeneity modeling was performed to obtain a high-quality 3D structure, followed by refinement and validation. Finally, the codon optimization based on E. coli resulted in a higher CAI value and optimal GC content, followed by combining it in the pET-14b cloning vector.Results: Evaluation of the various characteristics of the designed vaccine showed that it is an immunogenic and nonallergenic antigen that can induce immune responses against Leishmania major infection, which could be promising for cutaneous leishmaniasis. Conclusion:Research shows that a recombinant vaccine can be an effective candidate against cutaneous leishmaniasis.

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Designing a recombinant multiepitope vaccine against Leishmania donovani based immunoinformatics approaches

Cited by 3 publications

References 40 publications

Design and evaluation of a novel multi-epitope antigen for evaluate the diagnostic immunity responses against Leishmania infantum infection

Design and evaluation of a novel multi-epitope antigen for evaluate the diagnostic immunity responses against Leishmania infantum infection

Immunoinformatics analysis of Brucella melitensis to approach a suitable vaccine against brucellosis

Bioinformatics Analysis to Designing a Multi-epitope-based Peptide Vaccine Combat Leishmania major

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