30 research outputs found
Bacterial outer-membrane vesicles promote VĪ³9VĪ“2 T cell oncolytic activity
BACKGROUND: Increasing evidence suggests the immune activation elicited by bacterial outer-membrane vesicles (OMVs) can initiate a potent anti-tumor immunity, facilitating the recognition and destruction of malignant cells. At present the pathways underlying this response remain poorly understood, though a role for innate-like cells such as Ī³Ī“ T cells has been suggested. METHODS: Peripheral blood mononuclear cells (PBMCs) from healthy donors were co-cultured with E. coli MG1655 Īpal ĪlpxM OMVs and corresponding immune activation studied by cell marker expression and cytokine production. OMV-activated Ī³Ī“ T cells were co-cultured with cancer cell lines to determine cytotoxicity. RESULTS: The vesicles induced a broad inflammatory response with Ī³Ī“ T cells observed as the predominant cell type to proliferate post-OMV challenge. Notably, the majority of Ī³Ī“ T cells were of the VĪ³9VĪ“2 type, known to respond to both bacterial metabolites and stress markers present on tumor cells. We observed robust cytolytic activity of VĪ³9VĪ“2 T cells against both breast and leukaemia cell lines (SkBr3 and Nalm6 respectively) after OMV-mediated expansion. CONCLUSIONS: Our findings identify for the first time, that OMV-challenge stimulates the expansion of VĪ³9VĪ“2 T cells which subsequently present anti-tumor capabilities. We propose that OMV-mediated immune activation leverages the anti-microbial/anti-tumor capacity of VĪ³9VĪ“2 T cells, an axis amenable for improved future therapeutics
Multifunctional, self-assembling, anionic peptide-lipid nanocomplexes for targeted siRNA delivery
Formulations of cationic liposomes and polymers readily self-assemble by electrostatic interactions with siRNA to form cationic nanoparticles which achieve efficient transfection and silencing inĀ vitro. However, the utility of cationic formulations inĀ vivo is limited due to rapid clearance from the circulation, due to their association with serum proteins, as well as systemic and cellular toxicity. These problems may be overcome with anionic formulations but they provide challenges of self-assembly and transfection efficiency. We have developed anionic, siRNA nanocomplexes utilizing anionic PEGylated liposomes and cationic targeting peptides that overcome these problems. Biophysical measurements indicated that at optimal ratios of components, anionic PEGylated nanocomplexes formed spherical particles and that, unlike cationic nanocomplexes, were resistant to aggregation in the presence of serum, and achieved significant gene silencing although their non-PEGylated anionic counterparts were less efficient. We have evaluated the utility of anionic nanoparticles for the treatment of neuronal diseases by administration to rat brains of siRNA to BACE1, a key enzyme involved in the formation of amyloid plaques. Silencing of BACE1 was achieved inĀ vivo following a single injection of anionic nanoparticles by convection enhanced delivery and specificity of RNA interference verified by 5' RACE-PCR and Western blot analysis of protein
Antigen-Specific Gene Therapy after Immunisation Reduces the Severity of Collagen-Induced Arthritis
Reestablishment of tolerance induction in rheumatoid arthritis (RA) would be an optimal treatment with few, if any, side effects. However, to develop such a treatment further insights in the immunological mechanisms governing tolerance are needed. We have developed a model of antigen-specific tolerance in collagen type II (CII) induced arthritis (CIA) using lentivirus-based gene therapy. The immunodominant epitope of CII was inserted into a lentivirus vector to achieve expression on the MHC class II molecule and the lentiviral particles were subsequently intravenously injected at different time points during CIA. Injection of lentiviral particles in early phases of CIA, that is, at day 7 or day 26 after CII immunisation, partially prevented development of arthritis, decreased the serum levels of CII-specific IgG antibodies, and enhanced the suppressive function of CII-specific T regulatory cells. When lentiviral particles were injected during manifest arthritis, that is, at day 31 after CII immunisation, the severity of arthritis progression was ameliorated, the levels of CII-specific IgG antibodies decreased and the proportion of T regulatory cells increased. Thus, antigen-specific gene therapy is effective when administered throughout the inflammatory course of arthritis and offers a good model for investigation of the basic mechanisms during tolerance in CIA
Image_2_Bacterial outer-membrane vesicles promote VĪ³9VĪ“2 T cell oncolytic activity.jpeg
BackgroundIncreasing evidence suggests the immune activation elicited by bacterial outer-membrane vesicles (OMVs) can initiate a potent anti-tumor immunity, facilitating the recognition and destruction of malignant cells. At present the pathways underlying this response remain poorly understood, though a role for innate-like cells such as Ī³Ī“ T cells has been suggested.MethodsPeripheral blood mononuclear cells (PBMCs) from healthy donors were co-cultured with E. coli MG1655 Īpal ĪlpxM OMVs and corresponding immune activation studied by cell marker expression and cytokine production. OMV-activated Ī³Ī“ T cells were co-cultured with cancer cell lines to determine cytotoxicity.ResultsThe vesicles induced a broad inflammatory response with Ī³Ī“ T cells observed as the predominant cell type to proliferate post-OMV challenge. Notably, the majority of Ī³Ī“ T cells were of the VĪ³9VĪ“2 type, known to respond to both bacterial metabolites and stress markers present on tumor cells. We observed robust cytolytic activity of VĪ³9VĪ“2 T cells against both breast and leukaemia cell lines (SkBr3 and Nalm6 respectively) after OMV-mediated expansion.ConclusionsOur findings identify for the first time, that OMV-challenge stimulates the expansion of VĪ³9VĪ“2 T cells which subsequently present anti-tumor capabilities. We propose that OMV-mediated immune activation leverages the anti-microbial/anti-tumor capacity of VĪ³9VĪ“2 T cells, an axis amenable for improved future therapeutics.</p
Table_1_Bacterial outer-membrane vesicles promote VĪ³9VĪ“2 T cell oncolytic activity.docx
BackgroundIncreasing evidence suggests the immune activation elicited by bacterial outer-membrane vesicles (OMVs) can initiate a potent anti-tumor immunity, facilitating the recognition and destruction of malignant cells. At present the pathways underlying this response remain poorly understood, though a role for innate-like cells such as Ī³Ī“ T cells has been suggested.MethodsPeripheral blood mononuclear cells (PBMCs) from healthy donors were co-cultured with E. coli MG1655 Īpal ĪlpxM OMVs and corresponding immune activation studied by cell marker expression and cytokine production. OMV-activated Ī³Ī“ T cells were co-cultured with cancer cell lines to determine cytotoxicity.ResultsThe vesicles induced a broad inflammatory response with Ī³Ī“ T cells observed as the predominant cell type to proliferate post-OMV challenge. Notably, the majority of Ī³Ī“ T cells were of the VĪ³9VĪ“2 type, known to respond to both bacterial metabolites and stress markers present on tumor cells. We observed robust cytolytic activity of VĪ³9VĪ“2 T cells against both breast and leukaemia cell lines (SkBr3 and Nalm6 respectively) after OMV-mediated expansion.ConclusionsOur findings identify for the first time, that OMV-challenge stimulates the expansion of VĪ³9VĪ“2 T cells which subsequently present anti-tumor capabilities. We propose that OMV-mediated immune activation leverages the anti-microbial/anti-tumor capacity of VĪ³9VĪ“2 T cells, an axis amenable for improved future therapeutics.</p
Image_1_Bacterial outer-membrane vesicles promote VĪ³9VĪ“2 T cell oncolytic activity.jpeg
BackgroundIncreasing evidence suggests the immune activation elicited by bacterial outer-membrane vesicles (OMVs) can initiate a potent anti-tumor immunity, facilitating the recognition and destruction of malignant cells. At present the pathways underlying this response remain poorly understood, though a role for innate-like cells such as Ī³Ī“ T cells has been suggested.MethodsPeripheral blood mononuclear cells (PBMCs) from healthy donors were co-cultured with E. coli MG1655 Īpal ĪlpxM OMVs and corresponding immune activation studied by cell marker expression and cytokine production. OMV-activated Ī³Ī“ T cells were co-cultured with cancer cell lines to determine cytotoxicity.ResultsThe vesicles induced a broad inflammatory response with Ī³Ī“ T cells observed as the predominant cell type to proliferate post-OMV challenge. Notably, the majority of Ī³Ī“ T cells were of the VĪ³9VĪ“2 type, known to respond to both bacterial metabolites and stress markers present on tumor cells. We observed robust cytolytic activity of VĪ³9VĪ“2 T cells against both breast and leukaemia cell lines (SkBr3 and Nalm6 respectively) after OMV-mediated expansion.ConclusionsOur findings identify for the first time, that OMV-challenge stimulates the expansion of VĪ³9VĪ“2 T cells which subsequently present anti-tumor capabilities. We propose that OMV-mediated immune activation leverages the anti-microbial/anti-tumor capacity of VĪ³9VĪ“2 T cells, an axis amenable for improved future therapeutics.</p
Image_7_Bacterial outer-membrane vesicles promote VĪ³9VĪ“2 T cell oncolytic activity.jpeg
BackgroundIncreasing evidence suggests the immune activation elicited by bacterial outer-membrane vesicles (OMVs) can initiate a potent anti-tumor immunity, facilitating the recognition and destruction of malignant cells. At present the pathways underlying this response remain poorly understood, though a role for innate-like cells such as Ī³Ī“ T cells has been suggested.MethodsPeripheral blood mononuclear cells (PBMCs) from healthy donors were co-cultured with E. coli MG1655 Īpal ĪlpxM OMVs and corresponding immune activation studied by cell marker expression and cytokine production. OMV-activated Ī³Ī“ T cells were co-cultured with cancer cell lines to determine cytotoxicity.ResultsThe vesicles induced a broad inflammatory response with Ī³Ī“ T cells observed as the predominant cell type to proliferate post-OMV challenge. Notably, the majority of Ī³Ī“ T cells were of the VĪ³9VĪ“2 type, known to respond to both bacterial metabolites and stress markers present on tumor cells. We observed robust cytolytic activity of VĪ³9VĪ“2 T cells against both breast and leukaemia cell lines (SkBr3 and Nalm6 respectively) after OMV-mediated expansion.ConclusionsOur findings identify for the first time, that OMV-challenge stimulates the expansion of VĪ³9VĪ“2 T cells which subsequently present anti-tumor capabilities. We propose that OMV-mediated immune activation leverages the anti-microbial/anti-tumor capacity of VĪ³9VĪ“2 T cells, an axis amenable for improved future therapeutics.</p
Image_6_Bacterial outer-membrane vesicles promote VĪ³9VĪ“2 T cell oncolytic activity.jpeg
BackgroundIncreasing evidence suggests the immune activation elicited by bacterial outer-membrane vesicles (OMVs) can initiate a potent anti-tumor immunity, facilitating the recognition and destruction of malignant cells. At present the pathways underlying this response remain poorly understood, though a role for innate-like cells such as Ī³Ī“ T cells has been suggested.MethodsPeripheral blood mononuclear cells (PBMCs) from healthy donors were co-cultured with E. coli MG1655 Īpal ĪlpxM OMVs and corresponding immune activation studied by cell marker expression and cytokine production. OMV-activated Ī³Ī“ T cells were co-cultured with cancer cell lines to determine cytotoxicity.ResultsThe vesicles induced a broad inflammatory response with Ī³Ī“ T cells observed as the predominant cell type to proliferate post-OMV challenge. Notably, the majority of Ī³Ī“ T cells were of the VĪ³9VĪ“2 type, known to respond to both bacterial metabolites and stress markers present on tumor cells. We observed robust cytolytic activity of VĪ³9VĪ“2 T cells against both breast and leukaemia cell lines (SkBr3 and Nalm6 respectively) after OMV-mediated expansion.ConclusionsOur findings identify for the first time, that OMV-challenge stimulates the expansion of VĪ³9VĪ“2 T cells which subsequently present anti-tumor capabilities. We propose that OMV-mediated immune activation leverages the anti-microbial/anti-tumor capacity of VĪ³9VĪ“2 T cells, an axis amenable for improved future therapeutics.</p