Canine lyme disease vaccine
A vaccine, alpha virus technology, applied in the field of canine Lyme disease vaccine, can solve the problem of no report
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Embodiment 1
[0077] Construction of OspA and OspC vaccines delivered by alphavirus RNA replicon particles
[0078] RNA viruses have been used as vector-vehicles for the introduction of vaccine antigens that have been genetically engineered into their genomes. However, to date, their use has been largely limited to the incorporation of viral antigens into RNA viruses, followed by introduction of the virus into recipient hosts. The result is the induction of protective antibodies against the incorporated viral antigens. For example, alphavirus replicon vectors have been used to express the C. botulinum neurotoxin H C or B. anthracis protective antigen to protect mice from botulinum neurotoxin and anthrax [Lee et al., Vaccine24(47-48) 6886-6892(2006)]. Alphavirus RNA replicon particles have been used to encode pathogenic antigens. Such alphavirus replicon platforms have been developed by several different alphaviruses, including Venezuelan equine encephalitis virus (VEE) [Pushko et al., Vi...
Embodiment 2
[0084] Vaccines with RP-OspA constructs
[0085] Materials and methods
[0086] Constructs: RP-OspA constructs were generated as described above using the nucleotide sequence encoding an antigen comprising an immunogenic epitope of outer surface protein A.
[0087] Animals: 5-month-old Beagle dogs (Marshall Bioresources) were housed together in a kennel, with free access to food and water.
[0088] Preparation of RP-OspA vaccine: OspA RNA was electroporated into Vero cells together with helper RNA. After the co-electroporation process, OspA was packaged into RP, resulting in RP-OspA. Then, RP-OspA was mixed with stabilizers (sucrose, N-Z amine, gelatin), 0.9% saline, amphotericin B, and gentamicin so that a 1.0 mL dose contained a target concentration of 1.0×10 8 replicon particles / mL. Afterwards, the vaccine is lyophilized.
[0089] Vaccination and serum collection: Dogs were vaccinated subcutaneously in the neck of the dog with a 1 mL dose of RP-OspA vaccine, and booste...
Embodiment 3
[0099] Vaccines with RP-OspC constructs
[0100] Materials and methods
[0101] Constructs: RP-OspC constructs were generated as described above using the nucleotide sequence encoding an antigen comprising an immunogenic epitope of outer surface protein C.
[0102] Animals: 5-month-old Beagle dogs (Marshall Bioresources) were housed together in a kennel, with free access to food and water.
[0103] Preparation of RP-OspC vaccine: OspC RNA was electroporated into Vero cells together with helper RNA. After the co-electroporation process, OspC was packaged into RP, resulting in RP-OspC. Then, RP-OspC was mixed with stabilizers (sucrose, N-Z amine, gelatin), 0.9% saline, amphotericin B, and gentamicin so that a 1.0 mL dose contained a target concentration of 1.0×10 8 replicon particles / mL. Afterwards, the vaccine is lyophilized.
[0104] Vaccination and serum collection: Dogs were vaccinated subcutaneously in the neck of the dog with a 1 mL dose of RP-OspC vaccine, and booste...
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