Delivery of a cocktail DNA vaccine encoding cysteine proteinases type I, II and III with solid lipid nanoparticles potentiate protective immunity against Leishmania major infection

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 cpbCTE 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-25KD-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 rapid elimination from the circulation.

Download Full-text

Cationic solid lipid nanoparticles loaded by cystein proteinase genes as a novel anti-leishmaniasis DNA vaccine delivery system: Characterization and in vitro evaluations

Journal of Controlled Release ◽

10.1016/j.jconrel.2010.07.079 ◽

2010 ◽

Vol 148 (1) ◽

pp. e105-e106 ◽

Cited By ~ 5

Author(s):

Delaram Doroud ◽

Alireza Vatanara ◽

Farnaz Zahedifard ◽

Elham Gholami ◽

Rouhollah Vahabpour ◽

...

Keyword(s):

Dna Vaccine ◽

Delivery System ◽

Solid Lipid Nanoparticles ◽

Vaccine Delivery ◽

Lipid Nanoparticles ◽

Vaccine Delivery System ◽

Solid Lipid ◽

System Characterization

Download Full-text

C-Terminal Domain Deletion Enhances the Protective Activity of cpa/cpb Loaded Solid Lipid Nanoparticles against Leishmania major in BALB/c Mice

PLoS Neglected Tropical Diseases ◽

10.1371/journal.pntd.0001236 ◽

2011 ◽

Vol 5 (7) ◽

pp. e1236 ◽

Cited By ~ 15

Author(s):

Delaram Doroud ◽

Farnaz Zahedifard ◽

Alireza Vatanara ◽

Yasaman Taslimi ◽

Rouholah Vahabpour ◽

...

Keyword(s):

Solid Lipid Nanoparticles ◽

Leishmania Major ◽

Lipid Nanoparticles ◽

Protective Activity ◽

Solid Lipid ◽

Terminal Domain

Download Full-text

Arg-Gingipain A DNA Vaccine Induces Protective Immunity against Infection by Porphyromonas gingivalis in a Murine Model

Infection and Immunity ◽

10.1128/iai.69.5.2858-2864.2001 ◽

2001 ◽

Vol 69 (5) ◽

pp. 2858-2864 ◽

Cited By ~ 36

Author(s):

Hideo Yonezawa ◽

Kazuyuki Ishihara ◽

Katsuji Okuda

Keyword(s):

Porphyromonas Gingivalis ◽

Dna Vaccine ◽

Protective Immunity ◽

Defense Mechanisms ◽

Type I Collagen ◽

Periodontal Pathogen ◽

Enzyme Linked Immunosorbent Assay ◽

Type I ◽

Cysteine Proteinases ◽

Connective Tissues

ABSTRACT Arginine-specific cysteine proteinases (RgpA and RgpB) produced by the periodontal pathogen Porphyromonas gingivalis are suspected virulence factors and are involved in interrupting host defense mechanisms as well as in penetrating and destroying periodontal connective tissues. To induce a protective immune response againstP. gingivalis, we constructed an rgpA DNA vaccine. BALB/c mice were immunized intradermally by Gene Gun with plasmid DNA carrying rgpA. Antibody responses againstP. gingivalis were determined by an enzyme-linked immunosorbent assay. The rgpA DNA vaccine induced high levels of serum antibodies against P. gingivalis. Sera from the rgpA DNA vaccine-immunized mice diminished the proteolytic activity of RgpA and RgpB and inhibited the binding ofP. gingivalis to a type I collagen sponge. Moreover, the sera effectively reduced the hemagglutination of P. gingivalis, indicating that the hemagglutinin activity of the organism is associated with RgpA. We found with a murine abscess model that mice immunized with the rgpA DNA vaccine were resistant to an invasive P. gingivalis W50 challenge. These results suggest that the rgpA DNA vaccine induced specific antibodies against the enzyme and that this vaccine could confer protective immunity against P. gingivalis infection.

Download Full-text

Antileishmanial activity of conventional and solid lipid nanoparticles of Amphotericin B on Leishmania major

Infectious Disorders - Drug Targets ◽

10.2174/1871526519666191015170627 ◽

2019 ◽

Vol 19 ◽

Author(s):

Shahrzad Soltani ◽

Hoda Mojiri -Forushani ◽

Sheyda Soltani ◽

Mehdi Sagha Kahvaz ◽

Masoud Foroutan

Keyword(s):

Amphotericin B ◽

Solid Lipid Nanoparticles ◽

Leishmania Major ◽

Colorimetric Assay ◽

Lipid Nanoparticles ◽

Antileishmanial Activity ◽

Control Group ◽

Dependent Manner ◽

Solid Lipid

Background: Obligate intracellular parasites of Leishmania genus belong to family Trypanosomatidae and more than twenty species causes this neglected vector-borne infection throughout the globe. The current study was aimed to assess the antileishmanial activity of Amphotericin B (AmB) and AmB formulated into solid lipid nanoparticles (SLNs) in vitro and in vivo. Materials and methods: In the present research, microemulsification and high shear homogenization methods were used to prepare SLNs. Leishmania major (L. major) promastigotes were cultured in RPMI 1640 and incubated for three time points of 24, 48 and 72 h at 25±1°C. Then, the MTT colorimetric assay was employed for obtaining of 50% inhibitory concentration (IC50). Finally, we evaluated the efficacy of AmB and AmB-SLN for the treatment of experimental cutaneous leishmaniasis (CL) in BALB/c mice. Results: The average diameter sizes of prepared AmB-SLN were <180 nm and monodisperse preparations with polydispersity index 0.21±0.29. The antileishmanial activity of AmB and AmB-SLN revealed a dose and time-dependent manner in vitro. The IC50 values of AmB (38.18±1.33, 25.06±2.00, and 13.87±0.61 μg/ml) and AmB-SLN (0.40±0.02, 0.26±0.02, and 0.14±0.01 μg/ml) were estimated after 24, 48 and 72 hours, respectively. In all BALB/c treatment groups, the diameter of lesions were significantly smaller than the control group. Conclusion: Discovery of new effective drugs based on nanocarriers, such as SLN, is practical and opens a new window for the treatment of CL. More studies are needed in the future.

Download Full-text