Publications by authors named "Ali Khamesipour"

152 Publications

Leishmanicidal potentials of extract and its fractions on in a murine model: parasite burden, gene expression, and histopathological profile.

J Med Microbiol 2021 Jun;70(6)

Leishmaniasis Research Center, Kerman University of Medical Sciences, Kerman, Iran.

Leishmaniasis is a neglected tropical and subtropical disease caused by over 20 protozoan species. Treatment of this complex disease with traditional synthetic drugs is a major challenge worldwide. Natural constituents are unique candidates for future therapeutic development. This study aimed to assess the anti-leishmanial effect of the extract, and its fractions compared to the standard drug (Glucantime, MA) in a murine model and explore the mechanism of action. Footpads of BALB/c mice were infected with stationary phase promastigotes and treated topically and intraperitoneally with extract, its fractions, or Glucantime, 4 weeks post-infection. The extract and fractions were prepared using the Soxhlet apparatus with chloroform followed by the column procedure. The crude extract significantly decreased the footpad parasite load and lesion size compared to the untreated control group (<0.05), as revealed by dilution assay, quantitative real-time PCR, and histopathological analyses. The primary mode of action involved an immunomodulatory role towards the Th1 response in the up-regulation of IFN-γ and IL-12 and the suppression of IL-10 gene expression profiling against cutaneous leishmaniasis caused by . This finding suggests that the extract possesses multiple combinatory effects of diverse bioactive phytochemical compositions that exert its mechanisms of action through agonistic-synergistic interactions. The topical extract formulation could be a suitable and unique candidate for future investigation and pharmacological development. Further studies are crucial to evaluate the therapeutic potentials of the extract alone and in combination with conventional drugs using clinical settings.
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http://dx.doi.org/10.1099/jmm.0.001333DOI Listing
June 2021

Taxonomy, Population Structure and Genetic Diversity of Iranian Leishmania Strains of Cutaneous and Visceral Leishmaniasis.

Acta Parasitol 2021 May 3. Epub 2021 May 3.

Foodborne and Waterborne Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Background: Despite the broad distribution of leishmaniasis in Iran, there is a little genetic information about the causative agents and epidemiological status of the disease. Genetic diversity of the parasite is suggested to be one of the factors, which influences the clinical manifestations of the disease. In this study, we investigated the genetic variations, population structure, and evolutionary history of Leishmania species from endemic foci of Iran.

Methods: Fifty-two isolates from humans, canines, and rodents from different endemic foci of Iran were used to sequence the N-acetyl glucosamine-1-phosphate transferase (Nagt) gene. Phylogenetic and structure analyses were performed to investigate inter- and intra-species diversity of the Leishmania isolates.

Results: In total, 10 haplotypes including L. major (n = 6), L. tropica (n = 2), L. infantum (n = 1) and L. turanica (n = 1) were identified across 52 isolates. Haplotype diversity (Hd) ranged from zero for L. infantum and L. turanica to 0.78 ± 0.136 for L. major. This study identified population structure of Leishmania isolates from different geographical regions of Iran. The results of the phylogenetic tree showed 4 distinct clades for each species of Leishmania. In addition, the highest intraspecies diversity was observed among L. major isolates. No correlation was observed between species and geographic distribution of haplotypes.

Conclusions: Leishmania isolates were identified at the species level using the Nagt gene, low variation within species indicates conservation of this gene in Leishmania. The results provide knowledge into the evolutionary history of Iranian Leishmania isolates.
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http://dx.doi.org/10.1007/s11686-021-00377-5DOI Listing
May 2021

Potential biomarkers of immune protection in human leishmaniasis.

Med Microbiol Immunol 2021 Jun 2;210(2-3):81-100. Epub 2021 May 2.

Center for Research and Training in Skin Diseases and Leprosy, Tehran University of Medical Sciences, 14155-6383, Tehran, Iran.

Leishmaniasis is a vector-borne neglected tropical disease endemic in over 100 countries around the world. Available control measures are not always successful, therapeutic options are limited, and there is no vaccine available against human leishmaniasis, although several candidate antigens have been evaluated over the last decades. Plenty of studies have aimed to evaluate the immune response development and a diverse range of host immune factors have been described to be associated with protection or disease progression in leishmaniasis; however, to date, no comprehensive biomarker(s) have been identified as surrogate marker of protection or exacerbation, and lack of enough information remains a barrier for vaccine development. Most of the current understanding of the role of different markers of immune response in leishmaniasis has been collected from experimental animal models. Although the data generated from the animal models are crucial, it might not always be extrapolated to humans. Here, we briefly review the events during Leishmania invasion of host cells and the immune responses induced against Leishmania in animal models and humans and their potential role as a biomarker of protection against human leishmaniasis.
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http://dx.doi.org/10.1007/s00430-021-00703-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8088758PMC
June 2021

Correction to: The leishmanicidal efect of Lucilia sericata larval saliva and hemolymph on in vitro Leishmania tropica.

Parasit Vectors 2021 Mar 15;14(1):155. Epub 2021 Mar 15.

Department of Medical Entomology and Vector Control, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.

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http://dx.doi.org/10.1186/s13071-021-04649-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7962219PMC
March 2021

The leishmanicidal effect of Lucilia sericata larval saliva and hemolymph on in vitro Leishmania tropica.

Parasit Vectors 2021 Jan 11;14(1):40. Epub 2021 Jan 11.

Department of Medical Entomology and Vector Control, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.

Background: Leishmaniasis is a major parasitic disease worldwide, except in Australia and Antarctica, and it poses a significant public health problem. Due to the absence of safe and effective vaccines and drugs, researchers have begun an extensive search for new drugs. The aim of the current study was to investigate the in vitro leishmanicidal activity of larval saliva and hemolymph of Lucilia sericata on Leishmania tropica.

Methods: The effects of different concentrations of larval products on promastigotes and intracellular amastigotes of L. tropica were investigated using the mouse cell line J774A.1 and peritoneal macrophages as host cells. The 3-(4.5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and direct observation and counting method were used to assess the inhibitory effects and cell cytotoxicity of the larval products. The effects of larval products on the amastigote form of L. tropica were quantitatively estimated by calculating the rate of macrophage infection, number of amastigotes per infected macrophage cell, parasite load and survival index.

Results: The 50% cytotoxicity concentration (CC) value of both larval saliva and hemolymph was 750 µg/ml, and the 50% inhibitory concentration (IC) values were 134 µg/ml and 60 µg/ml for larval saliva and larval hemolymph, respectively. The IC for Glucantime, used a positive control, was (11.65 µg/ml). Statistically significant differences in viability percentages of promastigotes were observed for different doses of both larval saliva and hemolymph when compared with the negative control (p ≤ 0.0001). Microscopic evaluation of the amastigote forms revealed that treatment with 150 µg/ml larval hemolymph and 450 µg/ml larval saliva significantly decreased the rate of macrophage infection and the number of amastigotes per infected macrophage cell.

Conclusion: Larval saliva and hemolymph of L. sericata have acceptable leishmanicidal properties against L. tropica.
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http://dx.doi.org/10.1186/s13071-020-04543-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7798311PMC
January 2021

Evolution of antigen-specific immune responses in cutaneous leishmaniasis patients.

Parasite Immunol 2021 Apr 15;43(4):e12814. Epub 2021 Feb 15.

Center for Research & Training in Skin Diseases & Leprosy (CRTSDL), Tehran University of Medical Sciences (TUMS), Tehran, Iran.

Aims: Despite immunization appearing to be the most appropriate strategy for long-term control of the vector-borne leishmaniases, no sustainable vaccine is currently available against any form of leishmaniasis. We therefore evaluated, in the context of vaccine antigen candidates, antigen-specific immune response at various stages of cutaneous leishmaniasis (CL).

Methods And Results: Peripheral blood mononuclear cells (PBMC) isolated from healthy volunteers and CL patients (caused by either Leishmania major or L tropica) were incubated with crude Leishmania proteins (soluble Leishmania antigen; SLA), single recombinant proteins (TSA, LeIF, LmSTI1) or chimeric fusion proteins (LEISH-F2 and LEISH-F3). The concentrations of immune modulatory cytokines were then determined. While we did not detect appreciable antigen-specific IL-5 secretion, SLA induced secretion of interleukin (IL)-10 in cultures from early active lesion CL patients and even from healthy individuals. Conversely, interferon (IFN)-γ responses to SLA and recombinant proteins followed a similar pattern, developing only in the late active CL lesion phase. Once established, antigen-specific IFN-γ responses persisted in cured CL patients.

Conclusion: Together, our results provide further insight into the development of immune responses during CL and further validate the selection of LEISH-F2 and LEISH-F3 as vaccine antigen candidates.
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http://dx.doi.org/10.1111/pim.12814DOI Listing
April 2021

Evaluation of a New Topical Treatment for the Control of Cutaneous Leishmaniasis.

Microorganisms 2020 Nov 17;8(11). Epub 2020 Nov 17.

Department of Biochemistry, University of Lausanne, 1066 Epalinges, Switzerland.

() causes cutaneous leishmaniasis in the Old World. The infection mostly induces a localized lesion restricted to the sand fly bite. The costs and the side effects of current treatments render imperative the development of new therapies that are affordable and easy to administrate. Topical treatment would be the ideal option for the treatment of cutaneous leishmaniasis. MF29 is a 3-haloacetamidobenzoate that was shown in vitro to inhibit tubulin assembly in . Here, we tested a topical cream formulated with MF29. BALB/c mice were infected in the ear dermis with metacyclic promastigotes and once the lesion appeared, mice were treated with different concentrations of MF29 and compared to the control group treated with the cream used as the vehicle. We observed that topical application of MF29 reduced the progression of the infection while control groups developed an unhealing lesion that became necrotic. The treatment decreased the type 2 immune response. Comparison with SinaAmphoLeish, another topical treatment, revealed that MF29 treatment once a day was sufficient to control lesion development, while application SinaAmphoLeish needed applications twice daily. Collectively, our data suggest that MF-29 topical application could be a promising topical treatment for cutaneous leishmaniasis.
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http://dx.doi.org/10.3390/microorganisms8111803DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7696824PMC
November 2020

Multilocus sequence typing analysis of Leishmania clinical isolates from cutaneous leishmaniasis patients of Iran.

Infect Genet Evol 2020 11 10;85:104533. Epub 2020 Sep 10.

Center for Research and Training in Skin Diseases and Leprosy, Tehran University of Medical Sciences, Tehran, Iran.

Cutaneous leishmaniasis (CL) is mainly caused by L. major and L. tropica in Old World and might be represented as typical skin lesion(s) or sometimes as a spectrum of atypical manifestations. We applied multilocus sequence typing (MLST) to explore genetic variations of Leishmania strains isolated from atypical vs. typical CL patients from Iran. A PCR-sequencing was performed for seven housekeeping genes (g6pd, mpi, asat, icd, 6pgd, fh, and trys) and genetic diversity indices and phylogenetic relationships were analyzed. A total of 41 isolates of L. major (28/41) and L. tropica (13/41) from 21 (51.2%) atypical CL and 20 (48.8%) typical CL cases were included. A set of additional sequences of 41 strains of 17 species of Leishmania were retrieved from databases. Different SNP variations were detected and the highest rate of heterozygous sites was found in g6pd and 6pgd genes (6 sites) for L. tropica and in asat and 6pgd genes (7 sites) for L. major strains. All strains were clustered into 58 unique sequence types (STs) including 17 STs related to 41 strains of Leishmania of this study. Concatenated tree clustered all strains in 6 main clades (A to F) including L. major (clade D) and L. tropica (clade B) strains. Two strains of L. major (codes 28 and 42) with highest nucleotide variations were more close to L. tropica and were grouped in Clade B. All of the STs were related in clonal complexes by using eBURST with the prediction of founder genotypes. A high rate of genetic variations and heterozygocity was evident in L. tropica and L. major strains; nevertheless, there was no significant difference in the diversity of Leishmania strains between typical CL and atypical CL groups. This study represents the first successful application of MLST approach to L. tropica and L. major strains in Iran.
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http://dx.doi.org/10.1016/j.meegid.2020.104533DOI Listing
November 2020

A Phospholipase-A Activity in Soluble Leishmania Antigens Causes Instability of Liposomes.

Curr Drug Deliv 2020 ;17(9):806-814

Nanotechnology Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran

Aim: This study aimed to investigate the existence of phospholipase-A (PLA) activity in Soluble L. major Antigens (SLA) because of no reports for it so far. Liposomes were used as sensors to evaluate PLA activity.

Objectives: Liposomal SLA consisting of Egg Phosphatidylcholine (EPC) or Sphingomyelin (SM) were prepared by two different methods in different pH or temperatures and characterized by Dynamic Light Scattering (DLS) and Thin Layer Chromatography (TLC).

Methods: Lipid hydrolysis led to the disruption of EPC liposomal SLA in both methods but the Film Method (FM) produced more stable liposomes than the Detergent Removal Method (DRM).

Result: The preparation of EPC liposomal SLA at pH 6 via FM protected liposomes from hydrolysis to some extent for a short time. EPC liposomes but not SM liposomes were disrupted in the presence of SLA.

Conclusion: Therefore, a phospholipid without ester bond such as SM should be utilized in liposome formulations containing PLA as an encapsulating protein.
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http://dx.doi.org/10.2174/1567201817666200731164002DOI Listing
January 2020

Immune response in cutaneous leishmaniasis patients with healing . non-healing lesions.

Iran J Microbiol 2020 Jun;12(3):249-255

Center for Research and Training in Skin Diseases and Leprosy, Tehran University of Medical Sciences, Tehran, Iran.

Background And Objectives: The outcome of infection mainly depends upon the species which causes the disease and the generation of the type of host immune response, the healing process and protection in leishmaniasis depends upon induction of Th1 response. In this study, the Th1/Th2 cytokine profile in cutaneous leishmaniasis (CL) is evaluated.

Materials And Methods: This study was carried out in leishmaniasis clinic of CRTSDL, TUMS, during March 2018 to March 2019. Peripheral blood mononuclear cells (PBMC) of volunteers with active healing and non-healing lesion (s) of cutaneous leishmaniasis (CL), volunteers with and without history of CL were cultured and stimulated with Soluble antigen (SLA). The supernatants were collected and the levels of IFN-γ, IL-5 and IL-10 were titrated using ELISA method.

Results: The results showed a significantly higher levels of IFN-γ in volunteers with active CL healing form (<0.005), history of CL (<0.005) than healthy volunteers. A significantly (<0.005) higher level of IFN-γ was seen in volunteers with active healing form of lesion than non-healing form. There was a significantly (<0.005) higher level of IL-10 in volunteers with a history of non-healing form and active non-healing form of CL. There was no significant difference in IL-5 production in PBMC of different groups.

Conclusion: IFN-γ production starts at early stage of cutaneous leishmaniasis and enhance during course of lesion healing, IFN-γ level is significantly higher in all patients compared to healthy volunteers, IFN-γ is significantly higher in patients with healing form than non-healing form of lesion.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7340611PMC
June 2020

Human immune response to salivary gland antigens in a leishmaniasis-endemic focus in Iran.

Pathog Glob Health 2020 09 9;114(6):323-332. Epub 2020 Jul 9.

Department of Medical Entomology and Vector Control, School of Public Health, Tehran University of Medical Sciences , Tehran, Iran.

Salivary proteins specific antibodies have been shown to be useful biomarkers of exposure to sand fly bites. This study aimed to investigate the level, duration, and dynamics of the human immune response against the SGL of  Parrot, 1917 (Diptera: Psychodidae), and to assess the immunoreactivity of human sera with SGL components in an endemic area of anthroponotic cutaneous leishmaniasis (ACL) in Iran. The study was carried out in 2-phase; longitudinal and cross-sectional. Sand flies were collected monthly from indoors and outdoors. In the longitudinal study, sera from healthy volunteers were collected monthly, and in the cross-sectional study, sera from healthy volunteers and patients with ACL lesion/s, were collected for immunoassay studies. The level of anti- saliva IgG was detected using the ELISA. Immunoreactivity of individual human sera with saliva components was also assessed by western blotting. was the predominant sand fly species in the study area. The maximum and minimum percentages of IgG responses were seen in October (66%) and March (29%), respectively. Additionally, the cross-sectional study showed that 59.3% of the healthy volunteers and 80% of the patients were IgG positive. The antibody response against  salivary gland was high during the sand fly active season and declined by the end of the activity of the vectors.  Antibody response against the SGL components of  was transient and individual-specific. Some individuals shared a strong reaction against certain individual antigens, which could be considered as vector exposure markers for further investigation.

List Of Abbreviations: ELISA: Enzyme-Linked Immunosorbent Assay; SDS PAGE: Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis; SGL: Salivary Gland Lysate; ACL: Anthroponotic Cutaneous Leishmaniasis; PBS: Phosphate Buffered Saline; BCA: Bicinchoninic Acid; PBS-T: Phosphate Buffered Saline Tween; FBS: Fetal Bovine Serum; HRP: Horseradish Peroxidase; TMB: 3,3',5,5'-Tetramethylbenzidine; PVDF: Polyvinylidene Difluoride; SGA: Salivary Gland Antigens; OD: Optical Density; KDa: Kilodalton; VL: Visceral Leishmaniasis; CL: Cutaneous Leishmaniasis; SGs: Salivary glands.
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http://dx.doi.org/10.1080/20477724.2020.1789399DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7480592PMC
September 2020

Cutaneous Leishmaniasis Situation and Predicting the Distribution of Phlebotomus papatasi and P. sergenti as Vectors of Leishmaniasis in Ardabil Province, Iran.

Korean J Parasitol 2020 Jun 26;58(3):229-236. Epub 2020 Jun 26.

Department of Public Health, School of Public Health, Ardabil University of Medical Sciences, Ardabil, Iran.

Cutaneous leishmaniosis (CL) is the most common form of leishmaniasis.CL caused by L. major and L. tropica is endemic in 17 provinces of Iran. This study was carried out to elucidate situation of CL in Ardabil province and to predict distribution of Phlebotomus papatasi and Phlebotomus sergenti (Diptera: Psychodidae) as vectors of CL in the region. In this cross-sectional study, data on CL patients were collected from local health centers of Ardabil province, Iran during 2006-2018 to establish a geodatabase using ArcGIS10.3. A total of 20 CL cases were selected randomly and skin samples were collected and analyzed by PCR method. MaxEnt 3.3.3 model was used to determine ecologically suitable niches for the main vectors. A total, 309 CL human cases were reported and the highest incidence rate of disease was occurred in Bilasavar (37/100,000) and Germi (35/100,000). A total of 2,794 sand flies were collected during May to October 2018. The environmentally suitable habitats for P. papatasi and P. sergenti were predicted to be present in northern and central areas of Ardabil province. The most variable that contributed ratio in the modeling were Isothermality and slope factors. Ardabil province is possibly an endemic are for CL. The presence of P. papatasi and P. sergenti justifies local transmission while the vectors of CL are existing in the northern and central areas of the province.
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http://dx.doi.org/10.3347/kjp.2020.58.3.229DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7338894PMC
June 2020

Computational Modeling and Analysis to Predict Intracellular Parasite Epitope Characteristics Using Random Forest Technique.

Iran J Public Health 2020 Jan;49(1):125-133

Department of Health Information Management, School of Allied Medical Sciences, Tehran University of Medical Sciences, Tehran, Iran.

Background: In a new approach, computational methods are used to design and evaluate the vaccine. The aim of the current study was to develop a computational tool to predict epitope candidate vaccines to be tested in experimental models.

Methods: This study was conducted in the School of Allied Medical Sciences, and Center for Research and Training in Skin Diseases and Leprosy, Tehran University of Medical Sciences, Tehran, Iran in 2018. The random forest which is a classifier method was used to design computer-based tool to predict immunogenic peptides. Data was used to check the collected information from the IEDB, UniProt, and AAindex database. Overall, 1,264 collected data were used and divided into three parts; 70% of the data was used to train, 15% to validate and 15% to test the model. Five-fold cross-validation was used to find optimal hyper parameters of the model. Common performance metrics were used to evaluate the developed model.

Results: Twenty seven features were identified as more important using RF predictor model and were used to predict the class of peptides. The RF model improves the performance of predictor model in comparison with the other predictor models (AUC±SE: 0.925±0.029). Using the developed RF model helps to identify the most likely epitopes for further experimental studies.

Conclusion: The current developed random forest model is able to more accurately predict the immunogenic peptides of intracellular parasites.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7152625PMC
January 2020

Relationship of Leishmania RNA Virus (LRV) and treatment failure in clinical isolates of Leishmania major.

BMC Res Notes 2020 Mar 4;13(1):126. Epub 2020 Mar 4.

Center for Research and Training in Skin Diseases and Leprosy, Tehran University of Medical Sciences, Tehran, Iran.

Objective: Leishmaniasis is caused by different Leishmania spp. Treatment failure (TF) of cutaneous leishmaniasis (CL) is a serious issue that may be due to various reasons, previous studies suggested Leishmania RNA virus (LRV) as a potential cause of TF. Two variant groups of LRV1 and LRV2 are reported. In this study, the presence of LRV1/LRV2 was compared in TF with treatment response (TR) isolates of L. major. Clinical isolates of 15 TF and 15 TR were collected from CL patients referred to the Health Centers of Isfahan. Genomic DNA was extracted to identify Leishmania spp. using ITS1-PCR-RFLP. Identification of LRV1/LRV2 was performed using SYBR Green Real-Time PCR. The statistical analysis to test relationship between the treatment response with Glucantime and the presence of LRV were performed using SPSS 16.0 with Fisher's Exact test. P value of less than 0.05 was considered significant.

Results: ITS1-PCR-RFLP results showed that every isolate was identified as L. major. The results showed no LRV1 in any of the samples but 7 TR isolates and 2 TF isolates showed positive for LRV2. Statistical analysis showed no significant difference between the presence of LRV2 and response to Glucantime (p-value = 0.1086). Therefore, other mechanisms might be responsible for TF.
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http://dx.doi.org/10.1186/s13104-020-04973-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7074996PMC
March 2020

Comparison of cytokine profile of IFN-γ, IL-5 and IL-10 in cutaneous leishmaniasis using PBMC whole blood.

Iran J Microbiol 2019 Oct;11(5):431-439

Center for Research and Training in Skin Diseases and Leprosy, Tehran University of Medical Sciences, Tehran, Iran.

Background And Objectives: The surrogate marker (s) of cure and protection in intracellular pathogens is not yet well defined. The aim of this study was to compare the cytokine profile using whole blood cells (WBC) vs. peripheral blood mononuclear cells (PBMC) in healthy and cutaneous leishmaniasis (CL) volunteers.

Materials And Methods: In this study, WBC and PBMC of the volunteers with history of CL (HCL), Active lesion (ACL) and healthy volunteers were collected. The WBC and PBMC were cultured and stimulated with either PHA or soluble antigens (SLA), after 72 hours, the supernatants were collected and the levels of IFN-γ, IL-5 and IL-10 were titrated using ELISA method.

Results: The mean ± SD of cytokines using WBC and PBMC in cutaneous leishmaniasis volunteers stimulated with phytohemagglutin (PHA) or SLA are as follow, PHA, IFN-γ=2295±995 vs. 2339±1115, IL-10=853±309 vs. 1330±966, and IL-5=299±136 vs. 352+156, SLA, IFN-γ, 931±824 vs. 825±532, IL-10, 233±78 vs. 408±381, and IL-5, 185±59 vs. 217±76, respectively. There was no significant difference between the IFN-γ, IL-5 and IL-10 levels using WBC vs. PBMC. There was a strong correlation between the cytokine profiles using WBC and PBMC in cutaneous leishmaniasis volunteers.

Conclusion: There was no significant difference between IFN-γ, IL-10, IL-5 levels in whole blood and PBMC of volunteers with active lesion or history of CL. Whole-blood culture which is easier, cheaper and more convenient could be used instead of PBMC to evaluate the cytokine profile in field conditions.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7049321PMC
October 2019

Immunization against infection in BALB/c mice using a subunit-based DNA vaccine derived from TSA, LmSTI1, KMP11, and LACK predominant antigens.

Iran J Basic Med Sci 2019 Dec;22(12):1493-1501

Department of Medical Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Objectives: To design a multivalent DNA vaccine encoding the most immunogenic regions of the antigens including TSA (Thiol-specific antioxidant protein), LmSTI1 ( stress-inducible protein1), LACK ( homologue of receptors for activated C Kinase), and KMP11 (kinetoplastid membrane protein-11) on BALB/c mice.

Materials And Methods: The chimeric construct was generated including the most immunogenic epitopes containing a combination of domains and oligopeptides of the aforementioned genes. The construct was cloned into pcDNA 3.1 plasmid and named "pleish-dom." Following intramuscular injection of mice, the capability of the vector pleish-dom alone and with pIL-12 (expressing murine IL-12) to raise protective cytokines and parasite burden was evaluated in the BALB/c mice as a susceptible animal model against .

Results: The immunized mice with pleish-dom/pIL-12 showed the highest and the lowest levels of interferon-gamma (IFN-γ) and interleukin-10 (IL-10), as well as the lowest leishmanin skin test (LST) reactions, were found through 8 weeks post-infection.

Conclusion: Although the obtained DNA vaccine from the immunogenic chimeric protein of antigens was able to induce a high level of IFN-γ, it partially protected mice against L. major. However co-administration with pIL-12 led to shift immune response to Th1 phenotype, granuloma formation, and lowered parasite burden, and consequently distinct protection was found.
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http://dx.doi.org/10.22038/IJBMS.2019.14051DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7043880PMC
December 2019

Development of nano-carriers for vaccine delivery.

Expert Opin Drug Deliv 2020 02 14;17(2):167-187. Epub 2020 Jan 14.

Center for Research and Training in Skin Diseases and Leprosy, Tehran University of Medical Sciences, Tehran, Iran.

: Leishmaniasis is a neglected tropical infection caused by several species of intracellular protozoan parasites of the genus Leishmania. It is strongly believed that the development of vaccines is the most appropriate approach to control leishmaniasis. However, there is no vaccine available yet and the lack of an appropriate adjuvant delivery system is the main reason.: Adjuvants are the utmost important part of a vaccine, to induce the immune response in the right direction. Limitations and drawbacks of conventional adjuvants have been necessitated the development of novel particulate delivery systems as adjuvants to obtain desirable protection against infectious diseases such as leishmaniasis. This review focused on particulate adjuvants especially nanoparticles that are in use to develop vaccines against leishmaniasis. The list of adjuvants includes generally lipids-, polymers-, or mineral-based delivery systems that target antigens specifically to the site of action within the host's body and enhance immune responses.: Over the past few years, there has been an increasing interest in developing particulate adjuvants as alternatives to immunostimulatory types. The composition of nano-carriers and particularly the physicochemical properties of nanoparticles have great potential to overcome challenges posed to leishmaniasis vaccine developments.
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http://dx.doi.org/10.1080/17425247.2020.1713746DOI Listing
February 2020

Conducting International Diploma Course on Leishmaniasis and Its Control in the Islamic Republic of Iran.

J Arthropod Borne Dis 2019 Sep 30;13(3):234-242. Epub 2019 Sep 30.

Department of Medical Entomology and Vector Control, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.

Background: Leishmaniasis represents the important public health problem in the world. One of the main objectives of World Health organization is capacity building of managers and authorities who are involved with diseases control activities.

Methods: The course was conducted in Esfahan Health Research and Training Center (E.H.R.T.C) in summer 2005 and 2009. The course carried out jointly by the Ministry of Health and Medical Education (MOH) of Iran, World Health Organization-Eastern Mediterranean Regional Office (WHO-EMRO) and School of Public health, Tehran University of Medical Sciences (SPH-TUMS) and designed for medical officers, senior technicians and managers involved in leishmaniasis control. Prior to initiate the course, pre-test evaluations including different subjects on leishmaniasis and its control were carried out. The examinations include multiple choice questions. The duration of the course was 3 weeks. A total of 206 contact hours were taught. The main subjects were Basic epidemiology, Leishmaniasis parasitology, Leishmaniasis entomology, control of vectors and reservoirs, principles of integrated pest management, Field work and Planning. Different methods of teaching including lecture, laboratory, workshop, team work, field exercise and presentation were used. Requirement for achievement of the course was to have at least 60% of the total mark for awarding the diploma certificate.

Results: A total of 45 participants from Iraq, Afghanistan and Iran graduated from this course.

Conclusion: The course is providing the skill for managers, how to combat against disease in their country and is parallel to the policy of the leishmaniasis control for capacity building in endemic areas of their countries.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6928383PMC
September 2019

Old world cutaneous leishmaniasis in Iran: clinical variants and treatments.

J Dermatolog Treat 2020 Jan 8:1-11. Epub 2020 Jan 8.

Center for Research and Training in Skin Diseases and Leprosy, Tehran University of Medical Sciences, Tehran, Iran.

Old World cutaneous leishmaniasis (OWCL) is endemic in Iran and most cases of cutaneous leishmaniasis (CL) are caused by , and then and rarely by . We aimed to describe clinical variants of OWCL and their treatments. Through literature search in PubMed, Scopus and Embase and google scholar, we have found articles about variant clinical pictures of OWCL and their treatments. The following clinical variants of OWCL namely; localized forms, zosteriform, erysipeloid, eczematoid, warty, localized lymphadenitis, sporotrichoid, hyperkeratotic, impetiginized, mucosal involvement in CL, lupoid leishmaniasis, chronic lesions due to leishmanization, disseminated cutaneous leishmaniasis, reactivation of CL after transplantation and coexistence of CL with other diseases, are reported from Iran. The mainstay of therapy remains pentavalent antimonial compounds and cryotherapy is an adjuvant to therapy. Treatment with antifungal agents, miltefosine, amphotericin B and herbal extract such as ZH-E have also been used. Treatment of CL in chronic cases and in immunosuppressed patients is difficult and relapse may occur. In clinical variants of CL with long duration and multiple lesions, systemic pentavalent antimonial compounds are first step of therapy. In case of incomplete response or resistant to classic treatment, combination therapy is indicated.
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http://dx.doi.org/10.1080/09546634.2019.1704214DOI Listing
January 2020

Comparative analysis between four model nanoformulations of amphotericin B-chitosan, amphotericin B-dendrimer, betulinic acid-chitosan and betulinic acid-dendrimer for treatment of : real-time PCR assay plus.

Int J Nanomedicine 2019 24;14:7593-7607. Epub 2019 Sep 24.

Department of Clinical Research, Pasteur Institute of Iran, Tehran, Iran.

Background: Amphotericin B (Amp) and Betulinic acid (BA) as antileishmanial agents have negligible water solubility and high toxicity. To solve these problems, for the first time, chitosan nanoparticles and Anionic Linear Globular Dendrimer (D) were synthesized for the treatment of (.

Method: Chitosan and dendrimer nanoparticles were synthesized, and Amp and BA were loaded into the nanoparticles. The particles were then characterized using various methods and their efficacy was evaluated in vitro and in vivo environments (parasite burden was confirmed using pathological studies and real-time PCR methods).

Result: The results of docking showed that Amp and BA can be loaded into chitosan and dendrimer nanoparticles. The results of physically drug loading efficiency for AK (Amphotericin B-chitosan), BK (Betulinic acid-chitosan), AD (Amphotericin B-Dendrimer) and BD (Betulinic acid- Dendrimer) were 90, 93, 84 and 96 percent, respectively. The characterization results indicated that the drugs were loaded into nanoparticles physically. Moreover, the increased solubility rate for AD=478, BD=790, AK=80 and BK=300 folds. Furthermore, the results of the drug delivery system showed the slow controlled drug release pattern with cellular uptake of more than 90%. The treatment results showed a 100 percent decrease of toxicity for the all nanodrugs was observed in vivo and in vitro environments. Moreover, AK10 and BK20 mg/kg reduced parasite burden by 83 percent (<0.001), while AD50 and BD40 mg/kg reduced it to a lesser extent compared to glucantime.

Conclusion: All the synthesized nanodrugs were completely succeeded by 100% to recovery the induced pathological effects in the infected footpad. Also, the results of present study were confirmed with real-time PCR and the results showed that AK and BK were succeeded in a large extent to the treatment of infection (<0.001), therefore AK and BK could be considered as proper alternatives of choices drugs.
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http://dx.doi.org/10.2147/IJN.S220410DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6831986PMC
February 2020

Salvage therapy with Sodium chlorosum (formerly DAC N-055) for cases of refractory lupoid cutaneous leishmaniasis: results from a compassionate use study with 0.09% Sodium chlorosum in amphiphilic basic cream.

BMC Infect Dis 2019 Nov 28;19(1):1005. Epub 2019 Nov 28.

Cutaneous Leishmaniasis Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.

Background: Lupoid cutaneous leishmaniasis (LCL) is known as a rare but serious complication of anthroponotic cutaneous leishmaniasis (ACL) resistant to conventional treatments. Sodium chlorosum, a pro-oxidative preparation of pharmaceutical sodium chlorite (NaClO), has been successfully used for the treatment of Old World cutaneous leishmaniasis lesions (OWCL) and of some LCL cases in Afghanistan. This clinical trial study aimed to evaluate the effect of a last resort therapy with topical 0.09% sodium chlorosum on LCL in Iran.

Methods: Twenty Iranian patients (12 women and 8 men) with LCL refractory to treatment were included in this salvage study. A magistral preparation of sodium chlorosum (10 mM NaClO in amphiphilic basic cream) was applied twice daily to the lesions for 6 weeks and continued up to 12 weeks in patients who showed a clinical response within the first 6 weeks. Responders were followed up for a maximum of 1 year. Lesions were photographed during weekly visits. Disappearance of erythema and indurated lesions were rated as complete clinical response.

Results: Patients with a mean age of 28.6 (±24.3) and with an ACL proven lesion history of 3.8 (±1.4) years were treated for an average of 7.9 (±1.8) weeks. At the end of the treatment period (12th week), a complete response was observed in 9 of 20 patients (45%). During the one-year follow-up period, LCL lesions recurred in 4 of these 9 patients (with one patient showing only a tiny lesion) and one case lost to follow up whereas the other four remained completely lesion-free. Mild temporary side-effects such as erythema and itching were seen in 4 of 20 patients (20%).

Conclusions: Topical sodium chlorosum showed promising therapeutic results and can be considered as safe, painless, and relatively effective treatment for LCL, an ethical prerequisite for a two-armed controlled trial.

Trial Registration: This study was registered in Iranian registry of clinical trials on 2019-02-02 with registration number IRCT20190114042356N1.
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http://dx.doi.org/10.1186/s12879-019-4518-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6883658PMC
November 2019

Development of a topical liposomal formulation of Amphotericin B for the treatment of cutaneous leishmaniasis.

Int J Parasitol Drugs Drug Resist 2019 12 23;11:156-165. Epub 2019 Sep 23.

Center for Research and Training in Skin Diseases and Leprosy, Tehran University of Medical Sciences, Tehran, Iran. Electronic address:

Background: Currently, there is no topical treatment available for any form of cutaneous leishmaniasis (CL) in most of the endemic areas. The aim of the current study was to develop a topical nano-liposomal Amphotericin B (AmB) for the treatment of CL.

Methodology/principal Findings: Liposomes containing 0.1, 0.2 and 0.4% AmB (Lip-AmB) were formulated and characterized for the size, entrapment efficiency, long term stability, and skin penetration properties using Franz diffusion cells. Liposomes diameters were around 100 nm with no change during more than 20 months' storage either at 4 °C or at room temperature. Franz diffusion cells studies showed that almost 4% of the applied formulations penetrated across the skin and the highest skin retention (73.92%) observed with Lip-AmB 0.4%. The median effective doses (ED), the doses of AmB required to kill 50% of L. major amastigotes were 0.151, 0.151, and 0.0856 (μg/mL) in Lip-AmB 0.1, 0.2, 0.4%, respectively. Lip-AmB 0.4% caused 80% reduction in fluorescence intensity of GFP+ L. tropica infected macrophages at 5 μg/mL of AmB concentration. Topical Lip-AmB was applied twice a day for 4 weeks to the skin of BALB/c mice to treat lesions caused by L. major. Results showed the superiority of Lip-AmB 0.4% compared to Lip-AmB 0.2 and 0.1%. The parasite was completely cleared from the skin site of infection and spleens at week 8 and 12 post-infection in mice treated with Lip-AmB 0.4%. The results suggest that topical Lip-AmB 0.4% may be a useful tool in the treatment of CL and merits further investigation.
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http://dx.doi.org/10.1016/j.ijpddr.2019.09.004DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6904837PMC
December 2019

Safety Evaluation of Topical Application of Nano-Liposomal Form of Amphotericin B (SinaAmpholeish) on Healthy Volunteers: Phase I Clinical Trial.

Iran J Parasitol 2019 Apr-Jun;14(2):197-203

Center for Research and Training in Skin Diseases and Leprosy, Tehran University of Medical Sciences, Tehran, Iran.

Background: We aimed to evaluate the safety of SinaAmpholeish in a double-blind, randomized, phase 1 clinical trial in healthy human volunteers.

Methods: The study was carried out in DermaLab of Center for Research and Training in Skin Diseases and Leprosy, Tehran University of Medical Sciences, Tehran, Iran in 2012. A topical Nano-liposomal formulation of 0.4% Amphotericin B was developed against under trade name of SinaAmpholeish. In this randomized, double-blind, right-left, comparative, phase I clinical trial, in 2 steps; 7 and 20 healthy volunteers were recruited and applied SinaAmpholeish on the right and its vehicle on the left volar side of forearm, twice a day for one week or 3 times a day for two weeks. Seven biophysical skin parameters were measured in standard conditions before and 2 wk after application.

Results: There was no adverse effect when SinaAmpholeish and its vehicle were used twice a day for seven days. However, when were used 3 times a day for two weeks, both SinaAmpholeish and its vehicle induced severe local skin reactions in 2 volunteers leading to discontinuation of application. Mild and temporary local reactions were observed in about half of the application sides and there was no significant difference between SinaAmpholeish and its vehicle.

Conclusion: The new formulation is safe and worth to be tested in further phase 2 clinical trial and since there was no adverse effect with twice a day application it was decided to use SinaAmpholeish twice a day in phase 2 clinical trial.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6737359PMC
September 2019

An overview of leishmanization experience: A successful control measure and a tool to evaluate candidate vaccines.

Acta Trop 2019 Dec 13;200:105173. Epub 2019 Sep 13.

Center for Research and Training in Skin Diseases and Leprosy, Tehran University of Medical Sciences, Tehran 14166-13675, Iran. Electronic address:

Leishmanization (LZ) is an intradermal inoculation of live Leishmania to induce an artificial cutaneous leishmaniasis (CL) lesion in a covered part of the body to protect against further natural CL lesion development. Leishmanization has been used from ancient times and when NNN medium was developed continued with using Leishmania from culture media. The objective of this study was to review LZ published experiences. This article is a review of LZ experiences and historical studies initiated since 1910 when Leishmania promastigotes were harvested from culture media and used for LZ. This review includes LZ experiences in Israel, some countries of Former Soviet Union and Iran. The results of LZ in Israel, some countries of Former Soviet Union and Iran showed that despite limitations, using this method significantly reduced the incidence rate of CL among leishmanized individuals in endemic areas. In conclusion, leishmanization using Leishmania major produced under GMP guideline is a valuable tool to protect against CL, there are limitations which need further study.
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http://dx.doi.org/10.1016/j.actatropica.2019.105173DOI Listing
December 2019

Associated-risk determinants for anthroponotic cutaneous leishmaniasis treated with meglumine antimoniate: A cohort study in Iran.

PLoS Negl Trop Dis 2019 06 12;13(6):e0007423. Epub 2019 Jun 12.

Regional Centre of Advanced Technologies and Materials, Faculty of Science, Palacký University in Olomouc, Šlechtitelů 27, Czech Republic.

Background: The control of cutaneous leishmaniasis (CL) is facilitated by knowledge of factors associated with the treatment failures in endemic countries. The aim of this evaluation was to identify the potential risk determinants which might affect the significance of demographic and clinical characteristics for the patients with anthroponotic CL (ACL) and the outcome of meglumine antimoniate (MA) (Glucantime) treatment.

Methodology/principal Findings: This current was executed as a cohort spanning over a period of 5 years which centered in southeastern part of Iran. Altogether, 2,422 participants were evaluated and 1,391 eligible volunteer patients with ACL caused by Leishmania tropica were included. Overall, 1,116 (80.2%) patients received MA intraleisionally (IL), once a week for 12 weeks along with biweekly cryotherapy, while 275 (19.8%) patients received MA alone (20 mg/kg/day for 3 weeks) (intramuscular, IM). The treatment failure rate in ACL patients was 11% using IL combined with cryotherapy plus IM alone, whilst 9% and 18.5% by IL along with cryotherapy or IM alone, respectively. Multivariate logistic regression model predicted 5 major associated-risk determinants including male (odds ratio (OR) = 1.54, confidence interval (CI) = 1.079-2.22, p = 0.018), lesion on face (OR = 1.574, CI = 1.075-2.303, p = 0.02), multiple lesions (OR = 1.446, CI = 1.008-2.075, p = 0.045), poor treatment adherence (OR = 2.041, CI = 1.204-3.46, p = 0.008) and disease duration > 4 months (OR = 2.739, CI = 1.906-3.936, p≤0.001).

Conclusions/significance: The present study is the original and largest cohort of ACL patients who treated with MA. A comprehensive intervention and coordinated action by the health authorities and policy-makers are crucial to make sure that patients strictly follow medical instructions. Early detection and effective therapy < 4 months following the onset of the lesion is critical for successful treatment of the patients. Since a significant number of patients are still refractory to MA, reducing man-vector exposure and development of new effective alternative drugs are essential measures against ACL due to L. tropica.
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http://dx.doi.org/10.1371/journal.pntd.0007423DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6590833PMC
June 2019

Niosomal formulation of amphotericin B alone and in combination with glucantime: In vitro and in vivo leishmanicidal effects.

Biomed Pharmacother 2019 Aug 30;116:108942. Epub 2019 May 30.

Center for Research and Training in Skin Diseases and Leprosy, Tehran University of Medical Sciences, Tehran, Iran.

This study aimed to evaluate the efficacy of glucantime and amphotericin B (AmB) encapsulated in niosome against cutaneous leishmaniasis (CL) using in vitro and in vivo models. The niosomal formulations of the drugs alone and in combination were prepared and characterized. Subsequent to the examination of their cytotoxicity, their efficacy was evaluated using an in vitro MTT assay, macrophage model, flow cytometry, and gene expression profiling. For evaluation of therapeutic effect of niosomal combination on the lesion induced by Leishmania major in inbred BALB/c mice, the size of lesions and number of parasites in spleen was assessed. The niosomal formulations demonstrated significantly greater inhibitory effects compared with the non-niosomal forms when the IC was considered. The niosomal combination showed an increase in the apoptotic values and gene expression levels of IL-12 and metacaspase and a decrease in the levels of IL-10 with a dose-response effect. The niosomal combination was also effective in reducing the lesion size and splenic parasite burden in mice. Our findings indicated that there is a synergistic effect between AmB and glucantime in niosomal form in the inhibition of intracellular and extracellular forms of L. tropica. Additionally, the in vivo results on L. major suggest that topical niosomal formulation could be useful in the treatment of CL.
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http://dx.doi.org/10.1016/j.biopha.2019.108942DOI Listing
August 2019

Generation of a CRISPR/Cas9-Based Vector Specific for Gene Manipulation in .

Iran J Parasitol 2019 Jan-Mar;14(1):78-88

Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.

Background: Gene manipulation strategies including gene knockout and editing are becoming more sophisticated in terms of mechanism of action, efficacy and ease of use. In classical molecular methods of gene knockout, homologous arms are designed for induction of crossing over event in double strand DNA. Recently, CRISPR/Cas9 system has been emerged as a precise and powerful tool for gene targeting. In this effort, we aimed to generate a CRISPR/Cas9-based vector specific for targeting genes in parasites.

Methods: U6 and DHFR promoters and neomycin-resistance gene were amplified from genome of (MHRO/IR/75/ER) and pEGFP-N1, respectively. U6 promoter was cloned in pX330 vector which is named as pX330-U6. DHFR promoter and neo resistance gene sequence fragments were fused using a combination of SOE (Splicing by overlap extension)-PCR and T/A cloning techniques. To generate pX-leish, fused fragments su-bcloned into the pX330-U6. Two sgRNAs were designed to target the gene and cloned in pX-leish.

Results: The pX-leish vector was designed for simultaneous expression of cas9 and G418 resistance proteins along with a self-cleaving 2A peptide for efficient separation of the two proteins. In this study pX-leish was designed with 3 features: 1) Compatible promoters with parasites. 2) Insertion of antibiotic selection marker 3) Designing an all-in-one vector containing all components required for CRISPR/Cas9 system.

Conclusion: This modified system would be valuable in genome manipulation studies in for vaccine research in future.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6511583PMC
May 2019

Evaluation of Immune Response against Leishmaniasis in BALB/c Mice Immunized with Cationic DOTAP/DOPE/CHOL Liposomes Containing Soluble Antigens.

Iran J Parasitol 2019 Jan-Mar;14(1):68-77

Biotechnology Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran.

Background: Whole killed vaccine reached phase III clinical trials but failed to display significant efficacy in human mainly due to limited Th1 inducer adjuvant. Liposomes consisting of 1, 2-dioleoyl-3trimethylammonium-propane (DOTAP) bearing an inherent adjuvanticity and 1, 2-dioleoyl-L-α-glycero-3-phosphatidylethanolamine (DOPE) is well known to intensify the efficacy of positively charged liposomes.

Methods: Soluble antigens (SLA) encapsulated in cationic liposomes using lipid film method in 2016). BALB/c mice were immunized subcutaneously (SC), three times in a 2-wk interval, with Lip (DOTAP)-SLA+, Lip (DOTAP/DOPE)-SLA+, Lip (DOTAP/DOPE/CHO)-SLA+, Lip (DOTAP/DOPE/CHO), Lip (DOPE/CHO), SLA or HEPES buffer. At week 2 after the last booster injection, immunized mice have challenged SC in the footpad with parasites. To investigate the rate of protection and the type of immune response generated in mice, lesions development was assessed, IL-4 and IFN-γ levels with the ratio of IgG2a/IgG1 isotype were studied to describe the type of generated immune response.

Results: Mice immunized with all liposomal form of SLA showed smaller footpad swelling and lower parasite burden in the spleen and footpad compared to the group of mice received buffer. However, these formulations did not show protection against leishmaniosis because of a generated mixed Th1/Th2 response in mice characterized by high production of IFN-γ and IL4 and a high titer of IgG1 and IgG2a antibody.

Conclusion: Immunization with Lip (DOTAP/DOPE/CHO)-SLA+ was not an appropriate strategy to protect mice against leishmaniosis.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6511587PMC
May 2019

Development of topical liposomes containing miltefosine for the treatment of Leishmania major infection in susceptible BALB/c mice.

Acta Trop 2019 Aug 16;196:142-149. Epub 2019 May 16.

Nanotechnology Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran; Department of Pharmaceutical Nanotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran; Biotechnology Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran. Electronic address:

Herein, we investigated the efficacy of liposomes for the topical delivery of miltefosine (ML) to treat cutaneous leishmaniasis (CL). Liposomes containing varying concentrations of ML (0.5, 1, 2 and 4%) were prepared and characterized by their size and entrapment efficiency. The liposome diameters were between 100-150 nm. The penetration of ML from liposomal formulations through and in the skin was assessed using ex-vivo Franz diffusion cells fitted with mouse skin at 37 °C for 24 h. Data indicated that Lip-ML-4% showed the highest percent of retention across mouse skin (82%). in vitro promastigote and amastigote assays showed that ML and Lip-ML inhibit the growth of parasites either in the culture medium or intracellularly. Lip-ML formulations were topically applied twice a day for 4 weeks to the skin of BALB/c mice infected with L. major. Results showed a significantly (p < 0.001) smaller lesion size in Lip-ML-2 and 4% when compared to controls. At week 8 post-infection, the number of parasites was higher in Lip-ML-0.5% compared to Lip-ML-2 and 4%, however, the difference was not significant. At week 12, the splenic parasite burden was significantly (p < 0.001) lower in mice treated with different Lip-ML formulations when compared to controls. The lesion parasite burden was significantly (p < 0.001) lower in mice treated with either Lip-ML-2 and 4% compared to Lip-ML-0.5% at week 12 post-infection. The results suggested that topical Lip-ML-4% showed optimal ex-vivo penetration and in vivo anti-leishmanial activity against CL caused by L. major when compared to ML cream and other liposomes and thus, merits further investigation.
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http://dx.doi.org/10.1016/j.actatropica.2019.05.018DOI Listing
August 2019

Leishmania major p27 gene knockout as a novel live attenuated vaccine candidate: Protective immunity and efficacy evaluation against cutaneous and visceral leishmaniasis in BALB/c mice.

Vaccine 2019 05 29;37(24):3221-3228. Epub 2019 Apr 29.

Department of Epidemiology and Biostatistics, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.

Leishmaniasis is a growing health problem in many parts of the world and efforts to find vaccine against the disease are a public health priority. Live attenuated vaccines are the gold standard for protection against intracellular pathogens such as Leishmania spp. Defined genetic alteration of the Leishmania genome can be achieved using a gene-targeted disruption strategy that allows for the selection of parasites lacking genes essential for long-term survival and virulence. Previously, we demonstrated that genetically modified live attenuated Leishmania major, lacking the p27gene (Lmp27) is safe and induces cellular immunity in BALB/c mice. p27 is a component of the COX complex that is responsible for ATP synthesis. In the current study, the Lmp27 strain was assessed as a live attenuated vaccine. Overall protective immunity and efficacy were evaluated at various time periods following Leishmania major (L. major) and Leishmania infantum (L. infantum) challenges separately in BALB/c mice. Cytokine and anti-Leishmania antibody levels, splenocyte proliferation, delayed type hypersensitivity (DTH), skin lesion development, and parasite burden in the liver and spleen were the measured variables. The results demonstrated that immunized mice had a significant T-helper type 1 (Th1) response, smaller skin lesions and lower parasite burdens in their liver and spleens following a L. major challenge. Furthermore, the Lmp27 mutant also granted cross-protection against L. infantum infection. These results suggest that immunization with Lmp27 parasites provide significant protective immunity and efficacy against infection with homologous as well as heterologous species of Leishmania parasites.
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http://dx.doi.org/10.1016/j.vaccine.2019.04.068DOI Listing
May 2019