303 research outputs found
Ariel - Volume 2 Number 2
Editors
Delvyn C. Case, Jr.
Paul M. Fernhoff
News Editors
Richard Bonanno
Daniel B. Gould
Ronald A. Hoffman
Lay-Out Editor
Carol Dolinskas
Sports Editor
James J. Nocon
Contributing Editors
MichaeI J. Blecker
Lin Sey Edwards
Jack Guralnik
W. Cherry Light
Features Editor
Donald A. Bergman
Stephen P. Flynn
Business Manager
Nick Grego
Public Relations
Robin A. Edward
Efficient delivery of small interfering RNA for inhibition of IL-12p40 expression in vivo
Background: RNA interference is an evolutionary conserved immune response mechanism that
can be used as a tool to provide novel insights into gene function and structure. The ability to
efficiently deliver small interfering RNA to modulate gene expression in vivo may provide new
therapeutic approaches to currently intractable diseases.
Methods: In vitro, siRNA targeting IL-12p40 was delivered to the murine macrophage cell line
(J774A.1) encapsulated in a liposome with an IL-12 inducing agent (LPS/IFN-γ) over a number of
time points. Controls included a variety of non-target specific siRNA reagents. Supernatants were
analyzed for cytokine production while the cells were removed for mRNA profiling.
In vivo, siRNA-targeting IL-12p40 was delivered to the murine peritoneal cavity in a therapeutic
fashion, after endotoxin (LPS) challenge. Cells from the peritoneal cavity were removed by lavage
and analyzed by flow cytometry. Levels of IL-12 present in lavage and in serum were also examined
by ELISA.
Results: In this report, we show that IL-12p40 siRNA can specifically silence macrophage
expression of IL-12p40 mRNA and IL-12p70 protein in vitro. We extend this finding to demonstrate
that delivery of liposome encapsulated siRNA targeting IL-12p40 to the murine peritoneal cavity
can modulate an inflammatory stimulus in vivo. Furthermore, specific siRNA can be used
therapeutically after endotoxin challenge to reduce both the local and systemic inflammatory
response. Thus, the delivery of siRNA can be used to elicit specific non-permanent inhibition of
endogenous protein expression.
Conclusion: In vitro silencing of IL-12p40 using siRNA at selected doses leads to specific
knockdown of IL-12p70 protein production without inducing type I interferons. Furthermore,
siRNA targeting murine IL-12p40 can be used therapeutically to counter an inflammatory response
in vivo
Efficient delivery of small interfering RNA for inhibition of IL-12p40 expression in vivo
Background: RNA interference is an evolutionary conserved immune response mechanism that
can be used as a tool to provide novel insights into gene function and structure. The ability to
efficiently deliver small interfering RNA to modulate gene expression in vivo may provide new
therapeutic approaches to currently intractable diseases.
Methods: In vitro, siRNA targeting IL-12p40 was delivered to the murine macrophage cell line
(J774A.1) encapsulated in a liposome with an IL-12 inducing agent (LPS/IFN-γ) over a number of
time points. Controls included a variety of non-target specific siRNA reagents. Supernatants were
analyzed for cytokine production while the cells were removed for mRNA profiling.
In vivo, siRNA-targeting IL-12p40 was delivered to the murine peritoneal cavity in a therapeutic
fashion, after endotoxin (LPS) challenge. Cells from the peritoneal cavity were removed by lavage
and analyzed by flow cytometry. Levels of IL-12 present in lavage and in serum were also examined
by ELISA.
Results: In this report, we show that IL-12p40 siRNA can specifically silence macrophage
expression of IL-12p40 mRNA and IL-12p70 protein in vitro. We extend this finding to demonstrate
that delivery of liposome encapsulated siRNA targeting IL-12p40 to the murine peritoneal cavity
can modulate an inflammatory stimulus in vivo. Furthermore, specific siRNA can be used
therapeutically after endotoxin challenge to reduce both the local and systemic inflammatory
response. Thus, the delivery of siRNA can be used to elicit specific non-permanent inhibition of endogenous protein expression.
Conclusion: In vitro silencing of IL-12p40 using siRNA at selected doses leads to specific knockdown of IL-12p70 protein production without inducing type I interferons. Furthermore, siRNA targeting murine IL-12p40 can be used therapeutically to counter an inflammatory response
in vivo
Ariel - Volume 3 Number 1
Editors
Richard J. Bonanno
Robin A. Edwards
Associate Editors
Steven Ager
Stephen Flynn
Tom Williams
Lay-out Editor
Eugenia Miller
Contributing Editors
Michael J. Blecker
Milton Parker
James J. Nocon
Lynne Porter
Editors Emeritus
Delvyn C. Case, Jr.
Paul M. Fernhof
Efficient delivery of small interfering RNA for inhibition of IL-12p40 expression in vivo
Background: RNA interference is an evolutionary conserved immune response mechanism that
can be used as a tool to provide novel insights into gene function and structure. The ability to
efficiently deliver small interfering RNA to modulate gene expression in vivo may provide new
therapeutic approaches to currently intractable diseases.
Methods: In vitro, siRNA targeting IL-12p40 was delivered to the murine macrophage cell line
(J774A.1) encapsulated in a liposome with an IL-12 inducing agent (LPS/IFN-γ) over a number of
time points. Controls included a variety of non-target specific siRNA reagents. Supernatants were
analyzed for cytokine production while the cells were removed for mRNA profiling.
In vivo, siRNA-targeting IL-12p40 was delivered to the murine peritoneal cavity in a therapeutic
fashion, after endotoxin (LPS) challenge. Cells from the peritoneal cavity were removed by lavage
and analyzed by flow cytometry. Levels of IL-12 present in lavage and in serum were also examined
by ELISA.
Results: In this report, we show that IL-12p40 siRNA can specifically silence macrophage
expression of IL-12p40 mRNA and IL-12p70 protein in vitro. We extend this finding to demonstrate
that delivery of liposome encapsulated siRNA targeting IL-12p40 to the murine peritoneal cavity
can modulate an inflammatory stimulus in vivo. Furthermore, specific siRNA can be used
therapeutically after endotoxin challenge to reduce both the local and systemic inflammatory
response. Thus, the delivery of siRNA can be used to elicit specific non-permanent inhibition of
endogenous protein expression.
Conclusion: In vitro silencing of IL-12p40 using siRNA at selected doses leads to specific
knockdown of IL-12p70 protein production without inducing type I interferons. Furthermore,
siRNA targeting murine IL-12p40 can be used therapeutically to counter an inflammatory response
in vivo
Ariel - Volume 2 Number 7
Editors
Richard J. Bonanno
Robin A. Edwards
Associate Editors
Steven Ager
Stephen Flynn
Shep Dickman
Tom Williams
Lay-out Editor
Eugenia Miller
Contributing Editors
Michael J. Blecker
W. Cherry Light
James J. Nocon
Lynne Porter
Editors Emeritus
Delvyn C. Case, Jr.
Paul M. Fernhof
White Light-Informed Optical Properties Improve Ultrasound-Guided Fluorescence Tomography of Photoactive Protoporphyrin IX
Subsurface fluorescence imaging is desirable for medical applications, including protoporphyrin-IX (PpIX)-based skin tumor diagnosis, surgical guidance, and dosimetry in photodynamic therapy. While tissue optical properties and heterogeneities make true subsurface fluorescence mapping an ill-posed problem, ultrasound-guided fluorescence-tomography (USFT) provides regional fluorescence mapping. Here USFT is implemented with spectroscopic decoupling of fluorescence signals (auto-fluorescence, PpIX, photoproducts), and white light spectroscopy-determined bulk optical properties. Segmented US images provide a priori spatial information for fluorescence reconstruction using region-based, diffuse FT. The method was tested in simulations, tissue homogeneous and inclusion phantoms, and an injected-inclusion animal model. Reconstructed fluorescence yield was linear with PpIX concentration, including the lowest concentration used, 0.025 μg/ml . White light spectroscopy informed optical properties, which improved fluorescence reconstruction accuracy compared to the use of fixed, literature-based optical properties, reduced reconstruction error and reconstructed fluorescence standard deviation by factors of 8.9 and 2.0, respectively. Recovered contrast-to-background error was 25% and 74% for inclusion phantoms without and with a 2-mm skin-like layer, respectively. Preliminary mouse-model imaging demonstrated system feasibility for subsurface fluorescence measurement in vivo. These data suggest that this implementation of USFT is capable of regional PpIX mapping in human skin tumors during photodynamic therapy, to be used in dosimetric evaluations
Ariel - Volume 2 Number 6
Editors
Richard J. Bonanno
Robin A. Edwards
Associate Editors
Steven Ager
Stephen Flynn
Shep Dickman
Tom Williams
Lay-out Editor
Eugenia Miller
Contributing Editors
Michael J. Blecker
W. Cherry Light
James J. Nocon
Lynne Porter
Editors Emeritus
Delvyn C. Case, Jr.
Paul M. Fernhof
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