CFm40L-ADAPTATION IMPROVES ADENOVIRAL VECTOR DELIVERY TO ANTIGENPRESENTING CELLS
Document Type
Presentation
Start Date
22-10-2010 1:00 PM
End Date
22-10-2010 1:15 PM
Description
OBJECTIVE: Adenoviruses (Ad) are widely-used as transduction vectors in gene therapy and as vaccine vectors for immunization but are inefficient at transducing professional antigen-presenting cells (APC), such as dendritic cells, due to poor expression of the coxsackie-adenovirus receptor. CFm40L, a recombinant bispecific molecular adaptor, exploits APC expression of the immune signaling molecule CD40 in an attempt to re-target Ad vectors to these cells. Here, the capacity of CFm40L-Ad conjugates to target CD40-expressing cells is evaluated in vitro and in vivo.
METHODS: Transduction efficiency of CFm40L-Ad vectors encoding fluorescent proteins was measured in RAW264.7 cells, B lymphoma cell lines, and mouse splenic DCs by flow cytometry. In vivo transgene delivery and biodistribution following intranasal administration of CFm40L-Ad vectors was observed by biophotonic in vivo imaging and quantified using a luciferase reporter assay.
RESULTS: CFm40L-adaptation induced approximately ten-fold higher transduction efficiency of CD40-expressing cell lines by Ad vectors. Following intranasal administration, CFm40L-Ad vectors demonstrated altered patterns in biodistribution of the luc transgene. In vivo imaging demonstrated a greater localization of luminescence in the lungs of mice inoculated with CFm40L-Ad/Luc as compared to animals receiving nonadapted Ad/Luc. These results were confirmed using a luciferase reporter assay to measure luciferase activity in local tissues.
CONCLUSIONS: CFm40L adaptation represents a simple, efficient, and effective strategy for redirecting Ad vectors to CD40-expressing cells, particularly dendritic cells. Improved gene delivery to antigen-presenting cells represents a potential means for increasing the efficacy of Ad vaccine vectors and warrants further study of CFm40L as a component of novel immunization strategies in specific disease models.
Recommended Citation
Auten, Matthew; Huang, Weitao; Kutner, Robert; and Marrero, Luis, "CFm40L-ADAPTATION IMPROVES ADENOVIRAL VECTOR DELIVERY TO ANTIGENPRESENTING
CELLS" (2010). Dr. Joseph M. Moerschbaecher, III Graduate Research Day. 1.
https://digitalscholar.lsuhsc.edu/grad_rs/2010/presentation2/1
CFm40L-ADAPTATION IMPROVES ADENOVIRAL VECTOR DELIVERY TO ANTIGENPRESENTING CELLS
OBJECTIVE: Adenoviruses (Ad) are widely-used as transduction vectors in gene therapy and as vaccine vectors for immunization but are inefficient at transducing professional antigen-presenting cells (APC), such as dendritic cells, due to poor expression of the coxsackie-adenovirus receptor. CFm40L, a recombinant bispecific molecular adaptor, exploits APC expression of the immune signaling molecule CD40 in an attempt to re-target Ad vectors to these cells. Here, the capacity of CFm40L-Ad conjugates to target CD40-expressing cells is evaluated in vitro and in vivo.
METHODS: Transduction efficiency of CFm40L-Ad vectors encoding fluorescent proteins was measured in RAW264.7 cells, B lymphoma cell lines, and mouse splenic DCs by flow cytometry. In vivo transgene delivery and biodistribution following intranasal administration of CFm40L-Ad vectors was observed by biophotonic in vivo imaging and quantified using a luciferase reporter assay.
RESULTS: CFm40L-adaptation induced approximately ten-fold higher transduction efficiency of CD40-expressing cell lines by Ad vectors. Following intranasal administration, CFm40L-Ad vectors demonstrated altered patterns in biodistribution of the luc transgene. In vivo imaging demonstrated a greater localization of luminescence in the lungs of mice inoculated with CFm40L-Ad/Luc as compared to animals receiving nonadapted Ad/Luc. These results were confirmed using a luciferase reporter assay to measure luciferase activity in local tissues.
CONCLUSIONS: CFm40L adaptation represents a simple, efficient, and effective strategy for redirecting Ad vectors to CD40-expressing cells, particularly dendritic cells. Improved gene delivery to antigen-presenting cells represents a potential means for increasing the efficacy of Ad vaccine vectors and warrants further study of CFm40L as a component of novel immunization strategies in specific disease models.
Comments
See abstract book page 37