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.
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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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