RBD-mFc (10 μg/mouse) or PBS was mixed with AddaVax (Th1/Th2 adjuvant, Invitrogen), and these mixtures were subcutaneously injected into CAG-hACE2 mice on days –35 and –7 (Figure 8A ). Blood samples were taken with heparin sodium 2 days before SARS-CoV-2 infection to evaluate the production of RBD-sIgG. Mice were randomly assigned to 2 groups, PBS/AddaVax–immunized mice and RBD-mFc protein/AddaVax–immunized mice before SARS-CoV-2 infection: PBS/AddaVax with 2 × 103 TCID50 (n = 6), RBD-mFc protein/AddaVax with 2 × 103 TCID50 (n = 4), PBS/AddaVax with 2 × 104 TCID50 (n = 6), and RBD-mFc protein/AddaVax with 2 × 103 TCID50 (n = 4), respectively. RBD-immunized CAG-hACE2 mice and PBS-treated CAG-hACE2 mice were infected with 2 × 103 TCID50/25 μL and 1 × 104 TCID50/25 μL SARS-CoV-2 using the same method as above. The body weight and survival of the infected mice were monitored for up to 14 days.
Addavax
Manufactured by Thermo Fisher Scientific
Sourced in United States
AddaVax is a vaccine adjuvant used to enhance the immune response to vaccines. It is a water-in-oil emulsion that can be combined with various vaccine antigens to increase their immunogenicity.
Automatically generated - may contain errors
6 protocols using addavax
1
Evaluating RBD-mFc Protein Vaccine Efficacy
2
CORAVAX™ Immunogenicity in BALB/c Mice
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Six- to -eight-week-old female BALB/c mice were purchased from Charles River Laboratories (Wilmington, MA). Two groups of five mice were inoculated intramuscularly with 10 µg of chemically inactivated CORAVAX™ particles, with and without adjuvant (5 µg MPLAs in 2.5% AddaVax, Invitrogen, vac-mpls and vac-adx-10, respectively) in 100 µl PBS and boosted once with the same amount of virus ± adjuvant on day 21, respectively. Another group of five mice was immunized once with 108 ffu of live CORAVAX™. Blood samples were collected by retro-orbital bleed before the first immunization until day 56 post-immunization.
3
CORAVAX Immunization in Mice
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Groups of five 6–8 week old C57BL/6 or BALB/c mice were purchased from Charles River Laboratories. As per the adjuvant study, 5 µg MPLAs in 2.5% AddaVax (Invitrogen), and 100 µg Adju-Phos (alum, Invivogen) were formulated with the immunization. In all subsequent experiments, mouse immunizations were adjuvanted with MPLA-AddaVax. Mice were inoculated intramuscularly with 10 µg of chemically inactivated CORAVAX particles in 100 µL of PBS and boosted with the same amount of virus 4 weeks later. For pre-existing immunity experiments, 4 weeks before CORAVAX immunization, mice were immunized with either 10 µg of chemically inactivated BNSP333 or FILORAB1 in 100 µL of PBS. Those mice and an additional group of naive mice were immunized 4 weeks later with CORAVAX as described above and boosted 4 weeks later. Blood samples were collected by retro-orbital bleed before the first immunization until the study endpoint.
4
Heterologous Influenza Vaccine Boosting
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Mice were divided into 3 groups: homologous, heterologous, and control groups. Homologous-boosted mice (n = 10) were primed with the inactivated pandemic 2009 influenza vaccine and boosted at week 4 and 8 with the same vaccine strain. Heterologous-boosted mice (n = 10) were primed with the inactivated pandemic 2009 influenza vaccine and boosted with the inactivated A/Puerto Rico/8/1934 influenza vaccine at week 4 and the inactivated A/USSR/92/1977 influenza vaccine at week 8. The control group (n = 5) received PBS at the initial prime and at week 4 and 8. Mice were anesthetized by isoflurane inhalation. Ten μg antigen in PBS mixed with AddaVax (Invitrogen) adjuvant at a 1 : 1 ratio in a volume of 50 μl was administered intramuscularly for each immunization. Sera were collected 2 weeks after the third immunization. Mice were euthanized by overdose isoflurane inhalation. Blood was collected by direct cardiac puncture and allowed to clot for 1 hour at room temperature before centrifugation at 10,000 g for 15 minutes. Sera were stored at −20°C unit tested.
5
MERS-CoV Inactivated Vaccine Evaluation
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The hDPP4 mice were divided into nine groups (n = 5 per group). Each group was immunized twice with FA-inactivated MERS-CoV at 1, 5, or 10 μg dosage, respectively, without adjuvant (G1, G2, and G3) or with alum (G4, G5, and G6) or AddaVax® (Invitrogen, San Diego, CA, USA) (G7, G8, G9). The control group (G10, n = 5) was immunized with only alum. After that, all mice were challenged and euthanized, as described in Section 2.3.3 .
6
Intraperitoneal and Subcutaneous Mouse Immunization
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Mice were immunized intraperitoneally (i.p.) with 50 μg NP(40)-Ficoll (Biosearch Technologies) or 2x106 IU rSFV-βGal in a total volume of 100 μl PBS. NP(49)-CGG (Biosearch Technologies) was diluted in PBS and mixed 1:1 in AddaVax (Invitrogen). Mice were immunized i.p. with 5 µg or subcutaneously (s.c.) with 25 or 2.5 µg NP(49)-CGG on both flank regions in a volume of 100 μl PBS.
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