Drug – bio-affecting and body treating compositions – Designated organic active ingredient containing – Carbohydrate doai
Reexamination Certificate
1999-09-28
2001-09-04
Hauda, Karen M. (Department: 1632)
Drug, bio-affecting and body treating compositions
Designated organic active ingredient containing
Carbohydrate doai
C530S387300, C530S388150, C435S320100, C435S328000, C435S069100, C536S023500, C424S093600
Reexamination Certificate
active
06284742
ABSTRACT:
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates generally to immunology and adenoviral gene therapy. More specifically, the present invention relates to immunomodulation by genetic modification of dendritic cells and B-cells.
2. Description of the Related Art
An expanding body of evidence suggests that dendritic cells (DC) play a pivotal role in the immune system [Bancheareau, J. and R. M. Steinman. 1998, Dendritic cells and the control of immunity.
Nature.
392:2459]. Foremost, dendritic cells are recognized to serve as a key mediator of T-cell based immunity. Stemming from their important function, dendritic cells have been proposed for utility in a number of clinical strategies, especially vaccinations. It has become clear that genetic modification of these cells can promote immunity against pathogenic entities, both infectious and tumorigenic [Reeves, M. E., et al. 1996. Retroviral transduction of human dendritic cells with a tumor-associated antigen gene.
Cancer Res.
56:5672-7]. Importantly, all of these strategies are predicated upon efficient vectors for gene delivery to dendritic cells. To this end, a number of approaches have been utilized, albeit generally with poor efficiency of gene transfer [Arthur, J. F., et al. 1997. A comparison of gene transfer methods in human dendritic cells.
Cancer Gene Ther.
4:17-25; Van Tendeloo, V. F. I., et al. 1998. Nonviral transfection of distinct types of human dendritic cells: high-efficiency gene transfer by electroporation into hematopoetic progenitor- but not monocyte-derived dendritic cells.
Gene Ther.
5:700-7]. One candidate has been replication defective adenoviral vector. This vector has been suggested to be well suited for clinical applications by virtue of its high titer, efficiency gene delivery and exhuberant gene expression.
In spite of these theoretical advantages, the relative resistance of dendritic cells to adenoviral vector infection has confounded obtaining the full benefit of gene based immunotherapy strategies. [Arthur, J. F., et al. 1997. A comparison of gene transfer methods in human dendritic cells.
Cancer Gene Ther.
4:17-25; Dietz, A. B. and S. Vuk-Pavlovic. 1998. High efficiency adenovirus-mediated gene transfer to human dendritic cells.
Blood.
91:392-8]. The phenomenon of dendritic cell resistance to adenoviral mediated gene transfer may be based upon the paucity of adenoviral entry receptors. In permissive cells, the projecting adenoviral fiber-knob protein mediates binding to the cell surface coxsackie-adenovirus receptor (CAR) followed by interaction with and internalization of the virion by either of the av integrins avb3 or avb5 [Wickham, T. J., et al. 1993. Integrins &agr;v&bgr;3 and &agr;v&bgr;5 promote adenovirus internalization but not virus attachment. 73:309-19; Bergelson, J. M., et al. 1997. Isolation of a common receptor for Coxsackie B viruses and adenoviruses 2 and 5
, Science.
275:1320-3]. The present analysis has revealed an absence of CAR but adequate expression of the av integrin, &agr;v&bgr;5. High efficiency gene transfer independent of CAR expression by means of adenovirus targeted by bispecific entities to alternate cellular receptors has previously been shown [Douglas, J. T., et al. 1996. Targeted gene delivery by tropism modified adenoviral vectors.
Nature Biotech.
14:1574-8]. It was postulated that a similar strategy targeting the marker CD40, expressed on dendritic cells, might enhance gene transfer to dendritic cells.
A bispecific antibody was generated through chemical conjugation of a neutralizing anti-fiber-knob monoclonal antibody to a monoclonal antibody with affinity for the dendritic cell receptor, CD40. The present invention demonstrates that adenovirus complexed with this bispecific entity mediates dramatic enhancements in gene transfer to monocyte derived dendritic cells. More importantly, an upregulation of several dendritic cell maturational markers and enhanced allo-MLR performance after infection with CD40-targeted vector was observed, indicating the vector itself possesses maturational properties.
Thus, the prior art is deficient in methods of transducing dendritic cells and B-cells for immunomodulatory purposes. The present invention fulfills this long-standing need and desire in the art.
SUMMARY OF THE INVENTION
A bispecific antibody was generated through chemical conjugation of antibodies with affinities for the adenovirus fiber-knob and a dendritic cell receptor, CD40. The present invention shows that CD40 targeted adenovirus mediates dramatic enhancements in gene transfer to monocyte derived dendritic cells and that these enhancements can be attributed to a quantitative increase in the number of cells transduced. Additionally, the present invention shows that this enhancement is specific to the epitope recognized by the G28.5 antibody through successful blockade with the parent monoclonal, G28.5, and failure of the conjugate to mediate gene transfer on CD40 negative lines. Furthermore, an upregulation of several well documented dendritic cell maturational markers and enhanced allo-MLR by these cells was observed after infection with a retargeted vector. The dual role of CD40 in this scenario as both a surrogate adenovirus receptor and a powerful trigger of dendritic cell maturation may prove fortuitous as a retargeting strategy to this critical cell type of the immune system.
One object of the present invention is to provide a n adenovirus vector capable of targeting and transducing immune system cells, such as dendritic cells and B-cells, wherein transduction of B-cells results in maturation of the B-cells.
In an embodiment of the present invention, there is provided an immunomodulatory adenovirus, comprising: an adenoviral vector, and a bispecific antibody, comprising an antibody, or fragment thereof, recognizing a fiber-knob protein of said adenovirus conjugated to an antibody, or fragment thereof, recognizing a CD40 antigen, wherein said adenovirus is targeted to and transduces immune system cells resulting in modulation of said cells. Additionally, the bispecific antibody may be the product of a gene fusion.
In yet another embodiment, there is provided a n immunomodulatory adenovirus, comprising: a recombinant adenoviral vector, wherein the adenoviral gene encoding a fiber-knob protein has been replaced with a gene encoding an antibody, or fragment thereof, recognizing a CD40 antigen, or encoding the natural ligand of CD40, the trimeric CD40 ligand. When the adenovirus is targeted to and transduces immune system cells, the transduction results in modulation of the cells.
In yet another embodiment of the present invention, the adenoviral vector may express a therapeutic gene, selected from the group consisting of a gene encoding a tumor antigen, a gene encoding an antigen for an infectious agent, a gene encoding a cytotoxic agent and a gene encoding an immunomodulatory agent; the antibody recognizing the CD40 antigen is G28.5; the immune system cells are selected from the group consisting of dendritic cells and B-cells, as well as non-immune cells selected from the group consisting of vascular endothelium cells, epithelium cells, cells exhibiting chronic inflammation and cells and vessels of Karposi's sarcoma tumors; and maturation of the immune cells is indicative of modulation of the immune cells.
In yet another embodiment, there is provided a method of immunomodulation in an individual in need of such treatment, comprising the step of: administering to the individual a n immunomodulatory adenovirus, wherein the adenovirus modulates an immune response in the individual. This modulation is due to expression of a therapeutic gene by said adenovirus, and/or maturation of immune cells. The immune system cells are selected from the group consisting of dendritic cells and B-cells, as well as non-immune cells selected from the group consisting of vascular endothelium cells, epithelium cells, cells exhibiting chronic inflammation and cells and ves
Curiel David T.
Tillman Bryan Walter
Adler Benjamin Aaron
Beckerleg Anne Marie S.
Hauda Karen M.
UAB Research Foundation
LandOfFree
Immunomodulation by genetic modification of dendritic cells... does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Immunomodulation by genetic modification of dendritic cells..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Immunomodulation by genetic modification of dendritic cells... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2463221