Table 1.
Mice immunization and bleeding strategies
| Immunization formulation | Antigen/adjuvant(s) | |||||
|---|---|---|---|---|---|---|
| rPfCelTOS (µg/mouse) | CpG (µg/mouse) | Poly I:C (µg/mouse) | CFA/IFA (µL) | |||
| Prime | Boosts | |||||
| Vaccine groups | ||||||
| 1 | rPfCelTOS (Ag) | 10 | 5 | – | – | – |
| 2 | Ag/CpG | 10 | 5 | 10 | – | – |
| 3 | Ag/Poly I:C | 10 | 5 | – | 10 | – |
| 4 | Ag/CpG + Poly I:C | 10 | 5 | 5 | 5 | – |
| 5 | Ag/CFA/IFA | 10 | 5 | – | – | 100 |
| Control groups | ||||||
| 6 | CpG | – | – | 10 | – | – |
| 7 | Poly I:C | – | – | – | 10 | – |
| 8 | CpG + Poly I:C | – | – | 5 | 5 | – |
| 9 | CFA/IFA | – | – | – | – | 100 |
| 10 | PBS 1× | – | – | – | – | – |
Mice received 200 μL rPfCelTOS (10 μg at prime and 5 μg at boost) alone (non-adjuvanted vaccine group) and in the presence of CpG (10 μg/mouse, vaccine grade type, InvivoGen, San Diego, CA, USA), Poly I:C (10 μg/mouse, vaccine grade type, InvivoGen, San Diego, CA, USA), and combination of both CpG (5 μg/mouse) and Poly I:C (5 μg/mouse) adjuvants (adjuvanted vaccine groups) subcutaneously. Also, the mice were immunized with rPfCelTOS emulsified in CFA (complete Freund’s adjuvant) as reference adjuvant (the first dose) or IFA (incomplete Freund’s adjuvant; subsequent doses), with 1:1 volume/volume ratio. Control mice immunized with sterile PBS 1× (pH 7.4), CFA/IFA (as the reference adjuvant), CpG, Poly I:C, and CpG + Poly I:C. Sera samples were collected from the tail vein 10 days after each immunization (days 10, 24, and 38 after the first immunization)
Ag rPfCelTOS antigen, CFA complete Freund’s adjuvant, IFA incomplete Freund’s adjuvant, CpG synthetic oligodeoxynucleotides (ODNs) containing unmethylated CpG motifs, Poly I:C polyinosinic:polycytidylic acid