APC-targeted DNA vaccination to enhance specific immune responses against infectious diseases
1 Formål
The purpose of this project is to test the delivery of Nykode DNA vaccines containing tumor antigens, T cell epitopes, either alone or in combination with antigens from various infectious diseases and evaluate the immune response. With this, we may explore the immune response (both B and T cell) generated, especially focusing on the tailored T cell response that may be less sensitive to mutations. To mimic the clinical settings in our clinical trials NCT04405349 and NCT03548467, the animals will receive either weekly, biweekly, or triweekly immunizations up to week 12. To monitor antigen-specific T cell response, blood will be collected weekly for flow analysis.
2 Skadevirkninger
Moderate burden for the animals are expected from these experiments. In these experiments, DNA plasmids are injected intramuscularly in combination with electroporation to transfect nearby muscle cells. The animals are sedated (sevoflurane or ZRF) under the procedures. The animals may only experience potential fright/discomfort in conjunction with blood sampling from the saphenous vein. It is not expected that the animals will experience any significant weakening of health or their condition in general. With increased frequency of vaccination, the animals may experience possible scarring and/or inflammation. To minimise these side effects, we will alternate the administration of the pDNA between quadriceps and tibialis anterior.
3 Forventet nytteverdi
Nykode Therapeutics ASA is a Norwegian biotech company that has developed an unique DNA vaccine platform technology that may be used for both therapeutic and prophylactic vaccine purposes against both infectious diseases. Through pre-clinical experiments, Nykode will gain scientific data to aid the preparation of applications to implement clinical trials with vaccines against infectious diseases.
4 Antall dyr og art
The specific immune responses the various antigens may induce in mice will be monitored at different time points and with various methods including serum ELISA, flow, ELISpot, FluoroSpot and neutralizing antibody assays. For this work we will require 480 BALB/c mice, 480 C57BL/6 mice and 480 HLA.A2 mice, in total 1440 mice.
5 Hvordan etterleve 3R
To reduce the number of animals in experiments, several Nykode plasmids are tested and quality controlled in vitro to assess and confirm proper binding and folding of the vaccine proteins. Only the most promising constructs will proceed to animal experiments. The immune system consists of a complex network of cells and organs that cannot be imitated in vitro, and the specific immune response that the different antigens may elicit can only be tested in vivo. The animal welfare in these experiments is well cared for and all procedures (except blood samples from leg vain) will happen under anesthesia. Human endpoints include: Weight loss: Not more than 10%. General signs of disease or discomfort like brittle fur, strained respiration, humpback, reduced food intake, wounds, lack of grooming or reduced activity may be reasons for euthanasia, although the weight loss is not >10%.
The purpose of this project is to test the delivery of Nykode DNA vaccines containing tumor antigens, T cell epitopes, either alone or in combination with antigens from various infectious diseases and evaluate the immune response. With this, we may explore the immune response (both B and T cell) generated, especially focusing on the tailored T cell response that may be less sensitive to mutations. To mimic the clinical settings in our clinical trials NCT04405349 and NCT03548467, the animals will receive either weekly, biweekly, or triweekly immunizations up to week 12. To monitor antigen-specific T cell response, blood will be collected weekly for flow analysis.
2 Skadevirkninger
Moderate burden for the animals are expected from these experiments. In these experiments, DNA plasmids are injected intramuscularly in combination with electroporation to transfect nearby muscle cells. The animals are sedated (sevoflurane or ZRF) under the procedures. The animals may only experience potential fright/discomfort in conjunction with blood sampling from the saphenous vein. It is not expected that the animals will experience any significant weakening of health or their condition in general. With increased frequency of vaccination, the animals may experience possible scarring and/or inflammation. To minimise these side effects, we will alternate the administration of the pDNA between quadriceps and tibialis anterior.
3 Forventet nytteverdi
Nykode Therapeutics ASA is a Norwegian biotech company that has developed an unique DNA vaccine platform technology that may be used for both therapeutic and prophylactic vaccine purposes against both infectious diseases. Through pre-clinical experiments, Nykode will gain scientific data to aid the preparation of applications to implement clinical trials with vaccines against infectious diseases.
4 Antall dyr og art
The specific immune responses the various antigens may induce in mice will be monitored at different time points and with various methods including serum ELISA, flow, ELISpot, FluoroSpot and neutralizing antibody assays. For this work we will require 480 BALB/c mice, 480 C57BL/6 mice and 480 HLA.A2 mice, in total 1440 mice.
5 Hvordan etterleve 3R
To reduce the number of animals in experiments, several Nykode plasmids are tested and quality controlled in vitro to assess and confirm proper binding and folding of the vaccine proteins. Only the most promising constructs will proceed to animal experiments. The immune system consists of a complex network of cells and organs that cannot be imitated in vitro, and the specific immune response that the different antigens may elicit can only be tested in vivo. The animal welfare in these experiments is well cared for and all procedures (except blood samples from leg vain) will happen under anesthesia. Human endpoints include: Weight loss: Not more than 10%. General signs of disease or discomfort like brittle fur, strained respiration, humpback, reduced food intake, wounds, lack of grooming or reduced activity may be reasons for euthanasia, although the weight loss is not >10%.