The focus of this proposal is to develop a vaccine to prevent respiratory infection with Pseudomonas, the major pathogen in cystic fibrosis into a hybrid of a DC and activated CD4+ T cell. The central hypothesis is that in vivo administration of DC that have been genetically modified to express CD40L and primed with heat killed Pseudomonas will induce sufficiently robust anti-Pseudomonas host defenses to prevent respiratory infection with Pseudomonas. The preliminary data demonstrates that the CD40L/DC/Pseudomonas strategy works to induce specific anti- Pseudomonas immunity in vitro and in vivo. Strikingly, using components from CD4-/- mice, the CD40L genetically modified, Pseudomonas primed DC induce naive B cell to produce Pseudomonas- specific immunoglobins in vitro and protect CD4-/- mice in vivo from pulmonary challenge with Pseudomonas. CD40L modified, Pseudomonas primed DC induce naive B cell to produce Pseudomonas specific immunoglobulins in vitro and protect CD4-/- mice in vivo from pulmonary challenge with Pseudomonas. CD40L modified, Pseudomonas primed DC by enhancing the efficiency of gene transfer to the DC using Ad vectors modified to optimally bind to, and enter, DC, or by using vectors that may permit longer duration of expression of the CD40L transgene.
Specific aim 2 evaluates the hypothesis that protection can be accomplished with reduced numbers of CD40L transgene.
Specific aim 2 evaluates the hypothesis that protection can be accomplished with reduced numbers of CD40L/DC/Pseudomonas by also modifying the DC to express genes coding for effector mediators produced by activated DC (e.g., IL12), molecules that will function to suppress the induction of apoptosis in DC (crmA, TRANCE), or CD4+ co-stimulatory molecules in addition to CD40L (CD28).
Specific aim 3 examines the hypothesis that the anti-Pseudomonas immunity induced with the CD40L/DC/Pseudomonas strategy using the laboratory Pseudomonas strain PA01 is sufficient to protect against a variety of isolates of Pseudomonas from individuals with CR, and that the protection afforded by this strategy in c57Bl/6 mice is universal for other strains of mice, and for CFTR-/- mice.
Specific aim 4 assesses the hypothesis that the anti- Pseudomonas immunity induced with the CD40L/DC/Pseudomonas strategy established in the murine system is applicable to human components.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project--Cooperative Agreements (U01)
Project #
5U01HL066952-02
Application #
6501585
Study Section
Special Emphasis Panel (ZHL1)
Project Start
2001-09-01
Project End
2002-08-31
Budget Start
Budget End
Support Year
2
Fiscal Year
2001
Total Cost
$199,380
Indirect Cost
Name
Weill Medical College of Cornell University
Department
Type
DUNS #
201373169
City
New York
State
NY
Country
United States
Zip Code
10065
Chiuchiolo, Maria J; Crystal, Ronald G (2016) Gene Therapy for Alpha-1 Antitrypsin Deficiency Lung Disease. Ann Am Thorac Soc 13 Suppl 4:S352-69
Nolan, Daniel J; Ginsberg, Michael; Israely, Edo et al. (2013) Molecular signatures of tissue-specific microvascular endothelial cell heterogeneity in organ maintenance and regeneration. Dev Cell 26:204-19
Wang, Lan; Rosenberg, Jonathan B; De, Bishnu P et al. (2012) In vivo gene transfer strategies to achieve partial correction of von Willebrand disease. Hum Gene Ther 23:576-88
Krause, Anja; Whu, Wen Zhu; Xu, Yaqin et al. (2011) Protective anti-Pseudomonas aeruginosa humoral and cellular mucosal immunity by AdC7-mediated expression of the P. aeruginosa protein OprF. Vaccine 29:2131-9
Kobayashi, Hideki; Butler, Jason M; O'Donnell, Rebekah et al. (2010) Angiocrine factors from Akt-activated endothelial cells balance self-renewal and differentiation of haematopoietic stem cells. Nat Cell Biol 12:1046-56
Shmelkov, Sergey V; Hormigo, Adília; Jing, Deqiang et al. (2010) Slitrk5 deficiency impairs corticostriatal circuitry and leads to obsessive-compulsive-like behaviors in mice. Nat Med 16:598-602, 1p following 602
Rabbany, Sina Y; James, Daylon; Rafii, Shahin (2010) New dimensions in vascular engineering: opportunities for cancer biology. Tissue Eng Part A 16:2157-9
Wang, G; Qiu, J; Wang, R et al. (2010) Persistent expression of biologically active anti-HER2 antibody by AAVrh.10-mediated gene transfer. Cancer Gene Ther 17:559-70
Yamamoto, Masaya; James, Daylon; Li, Hui et al. (2010) Generation of stable co-cultures of vascular cells in a honeycomb alginate scaffold. Tissue Eng Part A 16:299-308
Watanabe, M; Boyer, J L; Crystal, R G (2010) AAVrh.10-mediated genetic delivery of bevacizumab to the pleura to provide local anti-VEGF to suppress growth of metastatic lung tumors. Gene Ther 17:1042-51

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