No abstract

A major focus in gene therapy has been the use of recomand lesions at a young age. After transplantation of the binant viruses to deliver genes in vivo. Although this apoE secreting Pro 175 endothelial cells into apoEapproach shows much promise, there are many safety deficient mice, serum cholesterol levels were measured at concerns associated with the use of viral materials in the 2 week intervals. During the 3 months after the initiation of treatment of human diseases. Our alternative cell-based these experiments, levels of cholesterol in the animals havgene therapy approach utilizes endothelial cells (Pro 175) ing received the apoE secreting endothelial cells were statisolated from the murine embryonic yolk sac. These endoistically lower compared with the levels of age-matched thelial cells were evaluated for their potential use in gene controls having received non-secreting endothelial cells. therapy as a gene delivery platform. As a test model, we Concomitant with cholesterol reduction, atherosclerotic used these cells to deliver apolipoprotein E (apoE) in the aortic plaques were noticeably reduced in the experimental murine apoE knockout atherosclerosis model. The lack of apoE+ animals. These results highlight the potential of apoE protein in these animals results in high levels of these unique endothelial cells as an efficient delivery serum cholesterol and formation of severe aortic plaques platform for somatic gene therapy.

8Efficient pathogen detection is essential for the successful treatment and prevention of infectious 9 disease; however, current methods are often too time intensive to be clinically relevant in cases 10 requiring immediate intervention. We have developed a Surface Programmable Activation 11 Receptor (SPAR) diagnostic platform comprised of universal biosensor cells engineered for use 12 in combination with custom or commercial antibodies to achieve rapid and sensitive pathogen 13 detection. SPAR cells are stably transfected Jurkat T cells designed to constitutively express a 14 modified T cell mouse FcγRI receptor on the cell surface and a high level of the luminescent 15 reporter protein aequorin in the cytoplasm. The modified mFcγRI-CD3ζ receptor protein binds 16 with high affinity to the Fc region of any full-length mouse IgG2a and some IgG2 antibodies: 17 this allows customized target detection via the selection of specific antibodies. T-cell receptor 18 aggregation in response to target antigen binding results in signal transduction which, when 19 amplified via the endogenous T cell signal cascade, triggers the rapid intracellular release of 20 calcium. Increased Ca 2+ concentrations activate the expressed reporter protein aequorin resulting 21 in the immediate emission of detectable light. Testing demonstrates the accurate and specific 22 2detection of numerous targets including P. aeruginosa, E. coli O111, and E. coli O157. We 23 report that the SPAR biosensor cell platform is a reliable pathogen detection method that enables 24 the rapid identification of bacterial causative agents using standard laboratory instrumentation. 25The technology lends itself to the development of efficient point-of-care testing and may aid in 26 the implementation of effective and pathogen-specific clinical therapies. 27 Introduction 28The rapid and accurate identification of causative agents is critical to the prompt application of 29 directed, pathogen-specific antibiotic therapies. Effective and timely clinical intervention is 30 essential for the control of infectious disease as well as in the successful treatment of bacterial 31 infections. The observed increases in the frequency and severity of nosocomial infections (1) 32 and the increasing prevalence of antibiotic resistance induced by non-specific antibiotic use (2) 33 further highlight the need for informed antibiotic selection based upon precise and efficient 34 pathogen detection. 35 Current bacterial identification methods include both classic procedures and novel molecular 36 techniques. Traditional culture-based methods, while sensitive and reliable, are also labor-37 intensive and time-consuming and therefore often cannot provide definitive diagnostics within a 38 clinically relevant timeframe (3). Molecular diagnostic methods include immunological assays 39 such as ELISA (4), microarray immunoblot (5), or serological assays (6); nucleic acid-based 40 techniques including PCR (7), DNA sequencing (8), hybridization techniques (9), or DNA/RNA 41 microarrays (10);...