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Cutaneous leishmaniasis (CL), caused by differentAdvances in high-throughput sequencing technologies, particularly their application for evaluation of the global gene expression changes, have made transcriptomics as a powerful tool to understand the pathogen-host molecular interactions.In this review, we focus on the transcriptomics studies that have been performed so far on human blood or tissue-driven samples to investigate

Additional file 1: Table S1. Nucleic acid and protein sequences of all components needed for construct design. Figure S1. Complete sequence of Utr1 and Utr2 of LEXSYcon2 vector. Trans-splicing sites used in mRNA synthesis are highlighted in blue (longest polypyrimidine tract), red (splice acceptor site), and purple (polyA site). Utr1 and Utr2 sequences are obtained from the LEXSYcon2 Expression Kit’s manual (Jena Bioscience). Figure S2. Optimal and centroid secondary structure analysis by RNAfold for all mRNA combinations. The full-sized mRNA sequence of all combinations was submitted to the RNAfold web server for secondary structure prediction. This server predicts two secondary structures, including optimal and centroid structures. a-column: Optimal secondary structure. b-column: Centroid structure. Table S2. Characteristics of Galaxy refined models. Figure S3. Superimposition of all combinations with the original proteins. The validated 3D model of each combination was compared with the original proteins, PpSP15, PsSP9, and PdSP15, using UCSF Chimera v1.15 software. The Superimposition was applied to indicate similarity between two overlaid protein structures by calculating RMSD. For closely homologous proteins, the RMSD is as small as 3 Å. The 3D model color: copper for PpSp15-T2A-PsSp9, PpSp15–PsSp9, PpSP15-Histag-T2A, and PpSP15-Histag. Yellow for PsSP9-T2A-PpSP15, PsSP9–PpSP15, and PsSP9-His-tag-T2A. Red for PpSp15, PsSP9, and PdSP15. Table S3. High-scored HLA-II junctional epitope. Table S4. High scored helper and CTL epitopes of vaccine constituent proteins. Figure S4. Estimation of population coverage of PpSP15 + PsSP9 combined vaccine in the Old World. Population coverage of the predicted epitopes was evaluated with the IEDB population coverage tool. Table S5. Physicochemical characteristics of the vaccine construct compared with original PpSP15 and PsSP9 components. Table S6. Antigenicity and allergenicity of the candidate vaccine construct compared with the original PpSP15 and PsSP9 components. Figure S5. In silico cloning of the vaccine candidate into the pLEXSY expression vector using SnapGene software. The red part shows the PpSP15-T2A-PsSP9 target sequence between BglII and NheI restriction sites. Figure S6. In frame cloning of PpSP15-T2A-PsSP9 construct into the pLEXSY expression vector by SnapGene software.

Neda Moazzezy,1 Mohammad Reza Asadi Karam,1 Sima Rafati,2 Saeid Bouzari,1 Mana Oloomi1 1Molecular Biology Department, Pasteur Institute of Iran, Tehran, Iran; 2Immunotherapy and Leishmania Vaccine Research Department, Pasteur Institute of Iran, Tehran, IranCorrespondence: Saeid Bouzari; Mana Oloomi Email manaoloomi@yahoo.com; saeidbouzari@yahoo.comIntroduction: A urinary tract infection (UTI), which is often caused by uropathogenic E. coli (UPEC) strains, affects many people worldwide annually. UPEC causes the production of pro-inflammatory cytokines by the bladder epithelial cells; however, it has been proven that the UPEC can inhibit the early activation of the innate immune system.Methods: This study aimed to examine the antibacterial and immunomodulatory effects of different doses of truncated alpha-defensins (human neutrophil peptide (HNP)-1) analog 2Abz23S29 on the mouse UTI model. Experimentally uropathogenic E. coli CFT073-infected mice were treated with low-dose 2Abz23S29 (250μg/mL), high-dose 2Abz23S29 (750μg/mL), ciprofloxacin (cip) (800μg/mL), or high-dose 2Abz23S29plus cip once a day 24 h post-infection. The 2Abz23S29 and cip treatment were given for two consecutive days.Results: The in vivo results showed that fewer UPEC were recovered from the bladders of mice treated transurethrally with 2Abz23S29. Moreover, low-dose 2Abz23S29 significantly decreased the level of the interleukin-6 (IL-6), whereas high-dose 2Abz23S29 increased pro-inflammatory cytokines including IL-6, macrophage inflammatory protein/2 (MIP/2), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) in infected bladders of mice. Besides, the levels of cytokines IL-6 and MIP/2 in infected mice treated with a combination of high-dose 2Abz23S29 and cip were significantly higher than the untreated mice. In contrast, CFT073-infected mice treated with a combination of high-dose 2Abz23S29 and cip showed no changes in cytokines TNF-α and IL-1β levels, indicating that ciprofloxacin may play an anti-inflammatory role.Conclusion: Collectively, apart from the direct antibacterial role of 2Abz23S29, our data illustrated that 2Abz23S29 modulates pro-inflammatory cytokine production of bladder in a dose-dependent manner, which has implications for the development of new anti-infective agents.Keywords: urinary tract infection, pro-inflammatory cytokines, human neutrophil peptide (HNP)-1