Protective vaccination against experimental canine visceral leishmaniasis using a combination of DNA and protein immunization with cysteine proteinases type I and II of .

Rafati, Sima; Nakhaee, Alireza; Taheri, Tahereh; Taslimi, Yasaman; Darabi, F.; Eravani, Davood; Sanos, Stéphanie L.; Kaye, Paul M.; Taghikhani, Mohammad; Jamshidi, Shahram; Rad, Mohammad Ali

doi:10.1016/j.vaccine.2005.02.009

Cited by 131 publications

(108 citation statements)

References 34 publications

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“…The most important step in developing an effective vaccine against leishmaniasis is to identify appropriate antigens, delivery systems (adjuvants) and to have the knowledge of the type of immune response generated in protected hosts. Previously, we have reported constructing eukaryotic plasmids harboring cpa and cpb and demonstrated that a cocktail vaccine containing these genes is possible and effective for leishmaniasis (8). However, recombinant naked pDNA is not proven as a powerful tool for DNA vaccination because of its rapid elimination from the circulation.…”

Section: Discussionmentioning

confidence: 99%

“…Both pGEM-cpa and pGEM-cpb were available from previous studies (8). Therefore, both cpa and cpb fragments from these constructs were cloned into pEGFP-N1.…”

Section: Pegfp-cpa and Pegfp-cpbmentioning

confidence: 99%

“…The entire full factorial design of experiment is listed in Table 1. 4 2.5 HSH S 5 2.5 ME S 6 2.5 M+H S 7 4 HSH S 8 4 ME S 9 4 M+H…”

Section: Preparation Of Csln Formulationsmentioning

confidence: 99%

“…Reproducibility of this assay was confirmed at 3, 7 and 14 days after loading, on the formulations stored at 4° C. The incubation time necessary for degrading 5 µg of naked pDNA was determined to be 60 min. The storage stability analysis of the fresh SD [1][2][3][4][5][6][7][8][9] formulations and those stored in dark for up to 4 weeks at different temperatures (4 ± 1°C and 25 ± 1°C) revealed different sizes and zeta potentials due to the DOTAP percents. DNA-loaded SLNs, when stored at refrigerated temperature (4 ± 1°C), were more stable in comparison with room temperature storage (Fig.…”

Section: Csln Formulation and Characterizationmentioning

confidence: 99%

“…Previously, our research team has shown that the native form of L. major CPs are recognized by lymphocytes taken from human and mice infected with Leishmania. Immunization experiments using CPs type I (CPB) and type II (CPA) have been carried out in mouse model as well as outbreed dogs with different strategies including DNA and recombinant protein vaccination (7,8). Recently, prime-boost vaccination using C-terminal extension (CTE) of L. infantum CPs type I have been performed in BALB/c mice and exhibited both type 1 and 2 immune responses.…”

Section: Introductionmentioning

confidence: 99%

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Cationic Solid Lipid Nanoparticles Loaded by Cystein Proteinase Genes as a Novel anti-Leishmaniasis DNA Vaccine Delivery System: Characterization and in vitro Evaluations

et al. 2010

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-Purpose. Leishmaniasis is a major health problem in many tropical and sub-tropical countries and development of a safe and easily-available vaccine has high priority. Although several antigens potentially capable of inducing protective immunity have been studied, in the absence of pharmaceutical industry interest they have remained as fine publications only. Amongst them, Cathepsin L-like cysteine proteinases (CPs) have received considerable attention and type I and II CPs have been used in a heterologous prime-boost vaccination regime for experimental visceral leishmaniasis in dogs. Due to the promising results of the mentioned vaccination regime, we aimed to evaluate cationic solid lipid nanoparticles (cSLNs) for in vitro delivery of cpa, cpb and cpb CTE intended to be used as a cocktail DNA vaccine in our forthcoming studies. Methods: cSLNs were formulated of cetyl palmitate, cholesterol, DOTAP and Tween 80 via melt emulsification method followed by high shear homogenization. Different formulations were prepared by anchoring pDNAs on the surface of cSLNs via charge interaction. The formulations were characterized according to their size and zeta potential as well as pDNA integrity and stability against DNase I treatment. Lipoplexes' cytotoxicity was investigated on COS-7 cells by MTT test. The effect of the DOTAP:pDNA ratio on protection ability and cytotoxicity was also studied. In vitro transfection efficiency was qualified by florescent microscopy and quantified using flow cytometry technique. Results: cSLN-pDNA complexes were formulated with suitable size and zeta potential. Efficiency/cytotoxicity ratio of cSLN-pDNAs formulations was comparable to linear PEI-25kDa-pDNAs polyplexes while exhibiting significantly lower cytotoxicity. Conclusion: Tested formulations were able to deliver immunogenic CP genes efficiently. This data proves the ability of this system as a promising DNA vaccine carrier for leishmaniasis to cover the main drawback of naked pDNA delivery that is rapidly elimination from the circulation.

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Section: Discussionmentioning

confidence: 99%

“…Both pGEM-cpa and pGEM-cpb were available from previous studies (8). Therefore, both cpa and cpb fragments from these constructs were cloned into pEGFP-N1.…”

Section: Pegfp-cpa and Pegfp-cpbmentioning

confidence: 99%

“…The entire full factorial design of experiment is listed in Table 1. 4 2.5 HSH S 5 2.5 ME S 6 2.5 M+H S 7 4 HSH S 8 4 ME S 9 4 M+H…”

Section: Preparation Of Csln Formulationsmentioning

confidence: 99%