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Nurulfiza Mat Isa, Nur Elina Abdul Mutalib, Noorjahan Banu Alitheen, Adelene Ai-Lian Song, Raha Abdul Rahim
This study demonstrates that cell wall treatment of Lactococcus lactis harbouring the internal ribosome entry site-incorporated lactococcal bicistronic vector pNZ:VIG mediated the delivery of genes into an eukaryotic cell line, DF1 cells, through bactofection. Bactofection analysis showed that the pNZ:VIG plasmid in L. lactis can be transferred into DF1 cells and that both the VP2 and gfp genes cloned in the plasmid can be transcribed and translated. The protein band relative to the Mr of VP2 protein (49 kDa) was successfully detected via Western blot analysis, while green fluorescence was successfully detected using a fluorescence microscope...
2017: Journal of Molecular Microbiology and Biotechnology
Nuriqmaliza M Kamal, M Zamri-Saad, Mas Jaffri Masarudin, Sarah Othman
BACKGROUND: Pasteurella multocida B:2 causes bovine haemorrhagic septicaemia (HS), leading to rapid fatalities in cattle and buffaloes. An attenuated derivative of P. multocida B:2 GDH7, was previously constructed through mutation of the gdhA gene and proved to be an effective live attenuated vaccine for HS. Currently, only two potential live attenuated vaccine candidates for HS are being reported; P. multocida B:2 GDH7 and P. multocida B:2 JRMT12. This study primarily aims to investigate the potential of P...
June 19, 2017: BMC Veterinary Research
Peter Celec, Roman Gardlik
Bacteria can be used for gene therapy via two strategies - either by transfection of eukaryotic host cells using bacteria (bactofection) or by alternative gene therapy that does not alter the host genome, but uses the prokaryotic expression system, which can be controlled or stopped from outside. While bactofection is optimal for gene substitution and DNA vaccination, alternative gene therapy is suitable for in situ delivery of proteins and treatment with intracellular bactochondria. A specific form of bacteria-mediated gene therapy is transkingdom RNA interference...
January 1, 2017: Frontiers in Bioscience (Landmark Edition)
Petra Kucerova, Monika Cervinkova
This review deals with the role of microorganisms in spontaneous regression of a tumour. Spontaneous cancer regression is a phenomenon that has been described for many centuries. One of the most well known methods of inducing spontaneous regression of cancer is the application of Coley's toxin (heat-killed Streptococcus pyogenes and Serratia marcescens), which has been used for the successful treatment of sarcomas, carcinomas, lymphomas, myelomas and melanomas. In clinical practice, the use of Bacillus Calmette-Guérin vaccine for the treatment of superficial urinary bladder cancer is the most common instance of the application of microorganisms for the treatment of cancer...
April 2016: Anti-cancer Drugs
Tai-Chun Chung, Charles H Jones, Akhila Gollakota, Mahmoud Kamal Ahmadi, Snehal Rane, Guojian Zhang, Blaine A Pfeifer
Bactofection offers a gene delivery option particularly useful in the context of immune modulation. The bacterial host naturally attracts recognition and cellular uptake by antigen presenting cells (APCs) as the initial step in triggering an immune response. Moreover, depending on the bacterial vector, molecular biology tools are available to influence and/or overcome additional steps and barriers to effective antigen presentation. In this work, molecular engineering was applied using Escherichia coli as a bactofection vector...
May 4, 2015: Molecular Pharmaceutics
X Yu, J Hua, G Hu, J Zhang, L Ma
The baculovirus-insect cell expression system (BES), one of the most popular systems for expression of eukaryotic proteins, was known to have drawbacks such as laborious manipulation of large-size baculovirus bacmids and the transfection procedure. These problems could be eliminated by direct infection of eukaryotic cells with nonpathogenic bacteria harbouring the respective gene - bactofection. However, it was unknown whether this system could be applied to insect cells. Therefore, in this study, the possibility of delivery of enhanced green fluorescent protein (EGFP) gene as a marker into the insect cell lines Sf9 and BmN-SWU1 using the above-mentioned approach with the Bac-to-Bac system was investigated...
2014: Acta Virologica
Joanne Cummins, Michelle Cronin, Jan Peter van Pijkeren, Cormac G M Gahan, Mark Tangney
Certain bacteria have emerged as biological gene vectors with natural tumor specificity, capable of specifically delivering genes or gene products to the tumor environment when intravenously (i.v.) administered to rodent models. Here, we describe procedures for studying this phenomenon in vitro and in vivo for both invasive and noninvasive bacteria suitable for exploitation as tumor-specific therapeutic delivery vehicles, due to their ability to replicate specifically within tumors and/or mediate bacterial-mediated transfer of plasmid DNA to mammalian cells (bactofection)...
2014: Methods in Molecular Biology
Kazuaki Ninomiya, Ryuji Yamada, Hitomi Meisaku, Nobuaki Shimizu
The present study demonstrates that ultrasound irradiation can facilitate bacteria-mediated gene delivery (bactofection). Escherichia coli modified with avidin were employed as a vehicle for delivery of the green fluorescent protein (GFP) gene, a model heterologous gene, into the breast cancer cell line MCF-7. Avidin-mediated binding of E. coli to MCF-7 cells enhanced the internalization of E. coli by approximately 17%, irrespective of the use of ultrasound irradiation. Furthermore, the use of ultrasound irradiation increased the internalization by approximately 5%, irrespective of the presence of avidin on the E...
May 2014: Ultrasonics Sonochemistry
Charles H Jones, Snehal Rane, Emily Patt, Anitha Ravikrishnan, Chih-Kuang Chen, Chong Cheng, Blaine A Pfeifer
Improvements to bacterial vectors have resulted in nonviral gene therapy vehicles that are easily prepared and can achieve high levels of transfection efficacy. However, these vectors are plagued by potential cytotoxicity and immunogenicity, prompting means of attenuation to reduce unwanted biological outcomes while maintaining transfection efficiency. In this study, listeriolysin O (LLO) producing Escherichia coli BL21(DE3) strains were pretreated with polymyxin B (PLB), a pore-forming antibiotic, and tested as a delivery vector for gene transfer to a murine RAW264...
November 4, 2013: Molecular Pharmaceutics
Sarah Othman, Andrew J Roe, Roger Parton, John G Coote
A reporter plasmid pSRG has been developed which expresses red fluorescent protein (RFP) from a constitutive prokaryotic promoter within Pasteurella multocida B:2 and green fluorescent protein (GFP) from a constitutive eukaryotic promoter within mammalian cells. This construct has been used to determine the location and viability of the bacteria when moving from the extracellular environment into the intracellular compartment of mammalian cells. Invasion assays with embryonic bovine lung (EBL) cells and an attenuated AroA(-) derivative of Pasteurella multocida B:2 (strain JRMT12), harbouring the plasmid pSRG, showed that RFP-expressing bacteria could be detected intracellularly at 3 h post-invasion...
2013: PloS One
Kumaran Narayanan, Choon Weng Lee, Aurelian Radu, Edmund Ui Hang Sim
Successful gene delivery into mammalian cells using bactofection requires entry of the bacterial vector via cell surface integrin receptors followed by release of plasmid DNA into the cellular environment. We show, for the first time, that addition of the DNA transfection reagent Lipofectamine improves entry of invasive Escherichia coli into HeLa cells and enhances up to 2.8-fold green fluorescent protein (GFP) expression from a reporter plasmid. The addition of Lipofectamine may be applicable to other bacterial vectors to increase their DNA delivery efficiency into mammalian cells...
August 15, 2013: Analytical Biochemistry
Li Qiu, Xinglong Wang, Huafang Hao, Guohui Mu, Ruyi Dang, Jia Wang, Shuxia Zhang, Enqi Du, Zengqi Yang
The use of attenuated Salmonella typhimurium as a bactofection vehicle for the oral delivery of a DNA vaccine against rabbit haemorrhagic disease virus (RHDV) was investigated. The DNA vaccine plasmid pcDNA3.1-VP60, which encodes the viral capsid protein VP60, was transformed into the attenuated S. typhimurium strain SL7207. The resulting recombinant bacteria, named as SL/pcDNA3.1-VP60, were orally used to immunise rabbits. The successful delivery of the DNA plasmid was confirmed by the detected VP60 transcription in the rabbit intestines through the reverse transcription polymerase chain reaction...
March 2013: Journal of Virological Methods
Cormac G M Gahan
Bacterial production of visible light is a natural phenomenon occurring in marine (Vibrio and Photobacterium) and terrestrial (Photorhabdus) species. The mechanism underpinning light production in these organisms is similar and involves the oxidation of an aldehyde substrate in a reaction catalysed by the bacterial luciferase enzyme. The genes encoding the luciferase and a fatty acid reductase complex which synthesizes the substrate are contained in a single operon (the lux operon). This provides a useful reporter system as cloning the operon into a recipient host bacterium will generate visible light without the requirement to add exogenous substrate...
February 1, 2012: Current Gene Therapy
Roman Gardlik
A number of genetically modified bacteria able to deliver a therapeutic gene into target cells has already been tested. Apart from the expected effects of bacterial therapy, the therapeutic bacterial strain also mediates a non-specific effect independent of the gene to be delivered. In this regard, we have recently shown that oral administration of the bacterial strain Escherichia coli XL1-Blue via gastric gavage to rats leads to a non-specific decrease in expression of vascular endothelial growth factor (VEGF) in intestinal wall without corresponding changes in other parameters...
September 2011: Bioengineered Bugs
Lubomíra Tóthová, Július Hodosy, Natália Kamodyová, Pavol Janega, Lívia Slobodníková, Adriana Liptáková, Peter Boor, Peter Celec
The role of innate immunity in the prevention of urinary tract infection is well-documented. Toll-like receptor 4 (TLR4) is a major determinant of innate immune response. In an animal model of urinary tract infection, bactofection-mediated gene transfer of TLR4 was tested in a preventive approach. Bactofection with TLR4 reduced the colonization with uropathogenic Escherichia coli by 91% in the kidney and by 41% in the bladder. Reduced colonization was associated with lower oxidative stress and expression of monocyte chemoattractant protein-1 and myeloperoxidase in the kidney...
June 2011: Current Microbiology
Roland Palffy, Roman Gardlik, Michal Behuliak, Peter Jani, Denisa Balakova, Ludevit Kadasi, Jan Turna, Peter Celec
Bacterial gene therapy - bactofection is a simple and effective method to deliver plasmid DNA into target tissue. We hypothesize that oral in vivo bactofection can be an interesting approach to influence the course of inflammatory bowel diseases. The aim of this study was to prove the effects of antioxidative and anti-inflammatory bactofection in dextran sulfate sodium (DSS)-treated mice. Attenuated bacteria Salmonella Typhimurium SL7207 carrying plasmids with genes encoding Cu-Zn superoxide dismutase and an N-terminal deletion mutant of monocyte chemoattractant protein-1 were prepared...
February 2011: Experimental Biology and Medicine
R Gardlik, M Behuliak, R Palffy, P Celec, C J Li
Several bacterial species have inherent ability to colonize solid tumors in vivo. However, their natural anti-tumor activity can be enhanced by genetic engineering that enables these bacteria express or transfer therapeutic molecules into target cells. In this review, we summarize latest research on cancer therapy using genetically modified bacteria with particular emphasis on blocking tumor angiogenesis. Despite recent progress, only a few recent studies on bacterial tumor therapy have focused on anti-angiogenesis...
May 2011: Gene Therapy
Roland Pálffy, Michal Behuliak, Roman Gardlík, Peter Jáni, L'udevít Kádasi, Ján Turna, Peter Celec
Salmonella typhimurium SL7207 carrying Cu-Zn superoxide dismutase and an N-terminal deletion mutant of monocyte chemoattractant protein-1 genes was applied to dextran sodium sulfate treated female Wistar rats. Stool quality, food and water intake were monitored. Markers of oxidative stress, interleukin 1, interleukin 6 and tumor necrosis factor alpha were quantified. No differences were found in body weights, markers of oxidative stress in plasma and inflammatory markers in colon homogenates. Plasma concentrations of I11, I16 were lower in the treatment groups than in the dextran sodium sulfate group...
2010: Folia Biologica (Krakow)
Roman Gardlik, Johannes H Fruehauf
Live bacterial vectors may be useful tools for the development of novel cancer therapies that can be added to the repertoire of existing drugs. Several bacterial strains effectively colonize solid tumors and act as antitumor therapeutics. The naturally occurring tumor-colonizing characteristics of bacterial species such as Salmonella sp, Clostridium sp and Escherichia coli can be further modified by genetic manipulations, making these bacterial systems excellent vehicles for the production and targeted delivery of therapeutic molecules into cancer cells...
October 2010: IDrugs: the Investigational Drugs Journal
Roman Gardlik, Julius Hodosy, Roland Palffy, Michal Behuliak, Pavol Janega, Peter Celec
BACKGROUND AND AIMS: Bactofection is delivery of therapeutic genes into target cells using bacteria penetrating the target cell membrane and releasing the gene into the cell. Hypoxia-inducible factor 1α (HIF-1α) represents a potential therapeutic gene to be used for gene delivery in ischemic diseases. The aim of this study was to prove the effects of bacteria-mediated transfer of hypoxia-inducible factor 1α (HIF-1α) in an experimental model of intestinal ischemia in rats. METHODS: Male Wistar rats with a surgically induced ischemia of colon (cecum) or sham-operated rats were treated by per os application of E...
July 2010: Archives of Medical Research
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