Vaccibodies targeted to cross-presenting dendritic cells

A technology of vaccines and targeting units, applied in the field of vaccine bodies targeting cross-presented dendritic cells, can solve the problems of no treatment, no DNA vaccine, lack of efficacy, etc.

Inactive Publication Date: 2014-11-05
UNIVERSITY OF OSLO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Due to lack of potency, no DNA vaccines have been approved for human use to date
Additionally, no effective vaccines exist for several infectious diseases
In particular, no therapeutic DNA cancer vaccines have been approved for human use

Method used

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  • Vaccibodies targeted to cross-presenting dendritic cells
  • Vaccibodies targeted to cross-presenting dendritic cells
  • Vaccibodies targeted to cross-presenting dendritic cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0103] Materials and Methods

[0104] Cell Lines, Viruses and Antibodies:

[0105] HEK293E cells were used to express HA-Vaccibody and to transfect Xcrl-eGFP. Antibodies against Xcl1 were obtained from Lifespan Biosciences (C-16241), while antibodies α-HA (H-36-4-52), α-human IgG3 (HP-6017) and α-mCherry were purified in the laboratory. For serum immunoglobulin ELISA, α-mouse IgG1-bio (BD Pharmingen, clone 10.9), α-mouse IgG2a-bio (BD Pharmingen, clone 8.3), α-mouse IgG2b-bio (BD Pharmingen, clone R12 -3). Influenza virus strain A / PR / 8 / 34 (H1N1) was obtained from the Norwegian Institute of Public Health.

[0106] Purification of Xcl1-mCherry vaccibody:

[0107] 2x10 cells in PBS by electroporation with 40 μg Xcl1-mCherry or Xcl1(C11A)-mcherry DNA 7 NSO cells generate stable transfectants. Cells were transferred to fresh RPMI medium and recovered for 24 hours at 37°C in T-25 flasks without selection. The next day, G418 was added to a final concentration of 800 μg / ml, an...

Embodiment 2

[0131] Vaccine bodies were generated as described above, except Xcl2 was used instead of Xcl1 as the targeting unit.

[0132] HEK293E cells were transiently transfected with plasmids encoding murine Xcl1-HA (mXcl1), human Xcl1-HA (hXcl1), or human Xcl2-HA (hXcl2) vaccibody. Supernatants were harvested after 48 hours and analyzed for vaccibody secretion by ELISA. All three vaccibodies were efficiently expressed and secreted, with hXcl1 and hXcl2 giving better expression than mXcl1. The results are presented in Figure 8 middle. Next, Balb / c mice were immunized with 25 μg of DNA encoding mXcl1, hXcl1 or hXcl2-HA vaccibody. Fourteen days after immunization, blood samples were collected and serum titers of IgGl and IgG2a were determined by ELISA. Both hXcl1 and hXcl2 induced higher IgG1 and IgG2a responses than mXcl1. The results are presented in Figures 9a and 9b. Balb / c mice were subsequently immunized with 25 μg of DNA encoding mXcl1, hXcl1 or hXcl2-HA vaccibody and chall...

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PUM

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Abstract

The present invention relates to recombinant fusion proteins targeted to dendritic cells and uses thereof. In particular, the present invention relates to fusion proteins comprising an antibody component and a targeting components, and uses of such fusion proteins to trigger immune responses.

Description

[0001] Cross References to Related Applications [0002] This application claims priority to pending US Provisional Patent Application No. 61 / 538,186, filed September 23, 2012, the contents of which are incorporated by reference in their entirety. field of invention [0003] The invention relates to a recombinant fusion protein targeting dendritic cells and its application. In particular, the invention relates to fusion proteins (vaccibodies) comprising a targeting module, an antigen, a linker region and an antibody module, and methods for eliciting an immune response to such homodimeric fusion proteins or DNA encoding such fusion proteins. use. Background of the invention [0004] DNA vaccination is a technically simple method of inducing an immune response. However, the success in small animals has not been replicated in clinical trials. Several strategies are currently being pursued to increase the efficacy of DNA vaccines. [0005] Targeting of protein antigens to an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00A61K39/145C07K19/00
CPCA61K39/00A61K39/145C07K2319/33C07K14/47C07K16/18C07K14/005C07K2319/42C07K2318/10C07K2319/01A61K2039/53A61K2039/57C12N2760/16134A61K2039/6056A61K39/12A61P31/12A61P35/00A61P37/04
Inventor B.博根E.富瑟姆G.格罗德兰
Owner UNIVERSITY OF OSLO
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