• Vaccine Name: C. parvum DNA vaccine encoding CpP2
• Target Pathogen: Cryptosporidium parvum
• Target Disease: Cryptosporidiosis
• Type: DNA vaccine
• Status: Research
• Host Species for Licensed Use: Human
• Antigen: CpP2: acidic ribosomal proteins P2 of C. parvum (Benitez et al., 2011)
• CpP2 gene engineering:
  • Type: Recombinant vector construction
  • Description: CpP2 antigen coding sequence was amplified by PCR and was ligated into the EcoRI and NotI restriction enzyme sites of the pUMVC4b expression vector. The ligation mix was then transformed into UltraMAX™ DH5a-FT™ Competent Cells and were selected on LB agar containing kanamycin (50 μg/ml). (Benitez et al., 2011)
  • Detailed Gene Information: Click Here.
• Immunization Route: Intramuscular injection (i.m.)

Mouse Response

• Host Strain: C57BL/6 interleukin-12p40 (IL-12p40) knockout (KO) mice (Benitez et al., 2011)
• Host age: Six-to-eight week-old (Benitez et al., 2011)
• Host gender: female (Benitez et al., 2011)
• Vaccination Protocol: Mice (10 per group) were injected with 100 μg plasmid pUMVC4b-CpP2 in TBE on days 0, 14, and 29. After primary immunization, mice in prime-boost regime were immunized with 3 μg of rP2 in Hunter’s TiterMax:PBS on days 43 and 57. The control group mice were injected with the empty pUMVC4b vector. (Benitez et al., 2011)
• Immune Response: Humoral: Anti-P2 antibody response was detected four weeks after primary dose injection and two weeks after the first booster dose injection. The response was further enhanced after the injection of the third dose. The anti-CpP2 antibody response was not observed in the control. Poor responses to the recombinant plasmid in wildtype mice were observed. Only IgG1 response was induced. IgG2a and IgA responses were not observed. (Benitez et al., 2011)
  Cellular: Stimulation of splenocyte cultures from immunized mice with different concentrations of rCpP2 protein resulted in a dose-dependent proliferative response. Increased levels of IFN-γ were observed in splenocyte cultures from immunized mice compared with both groups of controls (P<0.05). (Benitez et al., 2011)
• Challenge Protocol: The different treatment groups were challenged with a dose of 1 × 10^3 C. parvum oocysts by oral gavage 2 weeks after the last immunization. (Benitez et al., 2011)