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Vaccine Comparison

C. parvum DNA vaccine encoding CP15/60 C. parvum DNA vaccine encoding CP23 C. parvum DNA vaccine encoding CpP2 Cp15-23 pVAX-15-23 pVAX1-C-Cp12-Cp21
Vaccine Information Vaccine Information Vaccine Information Vaccine Information Vaccine Information Vaccine Information
  • Vaccine Ontology ID: VO_0011502
  • Type: DNA vaccine
  • Status: Research
  • Antigen: C. parvum sporozoite surface antigen CP15/60
  • CP15/60 gene engineering:
    • Type: DNA vaccine construction
    • Description: Recombinant pCP15/60 plasmid DNA was prepared using standard procedures and resuspended to 1 mg/ml in 0.15 M NaCl. Sera was collected from multiparous Holstein–Fresian cows at 8–12 weeks prior to parturition and tested by IFA for presence of Ab to C. parvum oocysts (Jenkins et al., 1999).
    • Detailed Gene Information: Click Here.
  • Vector: pCP15/60 (Jenkins et al., 1999)
  • Immunization Route: Parenchymal injection
  • Vaccine Ontology ID: VO_0011362
  • Type: DNA vaccine
  • Status: Research
  • Antigen: Cp23 antigen
  • CP23 gene engineering:
    • Type: DNA vaccine construction
    • Description: The C. parvum 27-kDa antigen coding sequence (GenBank accession number U34390) was amplified from a Cp23- pGEX 4T-2 clone. The fragment was cloned into the EcoRI and NotI restriction enzyme sites of the pUMVC4b expression vector (Adevron, Fargo, ND, USA). This vector has a cytomegalovirus promoter and immunoadjuvant site that enhances immune responses. Plasmids containing inserts (pUMVC4b-Cp23) and plasmids without inserts (pUMVC4b) were transformed into UltraMAX™ DH5a-FT™ Competent Cells (Invitrogen, Carlsbad, CA, USA), purified, and dissolved in sterile Tris–borate–ethylenediamine tetraacetic acid (TBE). After cloning, the sequence of the resulting clone was confirmed by automated DNA sequencing (Ehigiator et al., 2007).
    • Detailed Gene Information: Click Here.
  • Vector: pUMVC4b expression vector
  • Immunization Route: Subcutaneous injection
  • 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.
  • Vector: pUMVC4b (Benitez et al., 2011)
  • Immunization Route: Intramuscular injection (i.m.)
  • Type: Subunit vaccine
  • Status: Research
  • Host Species for Licensed Use: Human
  • Antigen: Cp15: 15 kDa sporozoite surface antigen (Liu et al., 2010); Cp23: a glycoprotein, geographically conserved among C. parvum isolates, present in both the sporozoite and merozoite stages (Liu et al., 2010)
  • CP15 gene engineering:
    • Type: Recombinant protein preparation
    • Detailed Gene Information: Click Here.
  • CP23 gene engineering:
    • Type: Recombinant protein preparation
    • Detailed Gene Information: Click Here.
  • Immunization Route: Intramuscular injection (i.m.)
  • Description: C. parvum recombinant protein vaccine that uses Cp15-23 fused protein as antigen and uses Freund’s adjuvant. (Liu et al., 2010)
  • Type: DNA vaccine
  • Status: Research
  • Host Species for Licensed Use: Human
  • Antigen: Cp15: 15 kDa sporozoite surface antigen (Wang et al., 2010); Cp23(Wang et al., 2010): a glycoprotein, geographically conserved among C. parvum isolates, present in both the sporozoite and merozoite stages (Liu et al., 2010)
  • CP15 gene engineering:
    • Type: DNA vaccine construction
    • Description: Amplified by PCR using a reverse primer containing a synthetic linker sequence. The amplified Cp15 was linked to Cp23 and was inserted into the pVAX1 expression vector. (Wang et al., 2010)
    • Detailed Gene Information: Click Here.
  • CP23 gene engineering:
    • Type: DNA vaccine construction
    • Description: Amplified by PCR. The amplified Cp23 was linked to Cp15 and was inserted into the pVAX1 expression vector. (Wang et al., 2010)
    • Detailed Gene Information: Click Here.
  • Vector: pVAX1: plasmid vector encoding antigen DNA (Wang et al., 2010); pMEM12R: plasmid vector encoding adjuvant sequence (Wang et al., 2010).
  • Immunization Route: Intramuscular injection (i.m.)
  • Description: C. parvum DNA vaccine that express Cp15 and Cp23 in pVAX1 as antigen and express IL-12 cytokine in pMEM12R as adjuvant. (Wang et al., 2010)
  • Type: DNA vaccine
  • Status: Research
  • Host Species for Licensed Use: Human
  • Antigen: Cp12 (Yu et al., 2010): a 12kDA surface adherence protein of C.parvum (Yao et al., 2007);Cp21: a 21kDA surface adherence protein of C.parvum (Yu et al., 2010):
  • Cp12 gene engineering:
    • Type: DNA vaccine construction
    • Description: Amplified by PCR using a forward primer containing the booster sequence encoding for CpG-ODN. The amplified C-Cp12 was linked to Cp21 and formed C-Cp12-Cp21, and was inserted into the pVAX1 expression vector. (Yu et al., 2010)
    • Detailed Gene Information: Click Here.
  • Cp21 gene engineering:
    • Type: DNA vaccine construction
    • Description: Amplified by PCR using a forward primer containing a synthetic linker sequence. The amplified Cp21 with the linker sequence was inked to C-Cp12 and formed C-Cp12-Cp21, and was inserted into the pVAX1 expression vector. (Yu et al., 2010)
    • Detailed Gene Information: Click Here.
  • Vector: pVAX1 expression vector (Yu et al., 2010)
  • Immunization Route: Intramuscular injection (i.m.) or Nasal spray
  • Description: C. parvum DNA vaccine that express Cp12 and Cp21 in pVAX1 as antigen and use CpG-ODN as adjuvant. (Yu et al., 2010)
Host Response Host Response Host Response Host Response Host Response Host Response

Mouse Response

  • Host Strain: C57BL/6NCr
  • Vaccination Protocol: Cows (n=2) with negligible titers (<1:50) to C. parvum oocysts were immunized at 6, 4 and 2 weeks prior to parturition with 1 mg of plasmid DNA by needle-syringe injection into the parenchymal tissue of the mammary gland. Colostrum from the cows was also tested for conferring passive immunity against C. parvum infection by oral administration to immunosuppressed adult inbred mice. Immune colostrum and control colostrum were administered to separate groups of dexamethasone (DEX)-treated adult C57BL/6NCr mice beginning 12 h before and at 12 h intervals for 3 days after oral C. parvum oocyst infection (Jenkins et al., 1999).
  • Challenge Protocol: In three separate experiments, DEX-treated mice (n=4 per group, six groups per treatment) received 250 μl of HBC or NBC by gastric intubation using a 26 ga. gavage needle 12 h prior to C. parvum infection. At 0 h, the mice were infected with either 103 or 104 C. parvum oocysts and given 250 μl of either HBC or NBC. Control mice were given H2O and either infected or not infected with C. parvum oocysts (Jenkins et al., 1999).
  • Efficacy: Mice receiving immune colostrum showed partial protection (about 50% reduction) against intestinal C. parvum development compared to mice receiving control colostrum. This protection was evident at a challenge dose of 10^3 C. parvum oocysts per mouse (Jenkins et al., 1999).

Mouse Response

  • Host Strain: C57BL/6 KO
  • Vaccination Protocol: Mice (ten per group) were injected subcutaneously in the ear with 100-μg plasmid pUMVC4b-Cp23 in TBE on days 0, 14, and 29. Mice in the control group were injected with either the vector pUMVC4b without the insert or with PBS (Ehigiator et al., 2007).
  • Challenge Protocol: The different treatment groups of IL-12p40KO mice were challenged with a dose of 1,000 C. parvum oocysts by oral gavage 2 weeks after the last immunization (Ehigiator et al., 2007).
  • Efficacy: Cp23-DNA vaccination induced a 50-60% reduction in oocysts shedding, indicating a partial protection against C. parvum infection in IL-12 KO mice (Ehigiator et al., 2007).
  • Host Ighg1 response
    • Description: Six weeks post immunization, mice were bled and antibody responses were measured by ELISA. IgG1 responses were significantly higher in Cp23 vaccinated mice than vector (control) immunized mice (Ehigiator et al., 2007).
    • Detailed Gene Information: Click Here.
  • Host Ighv1-9 response
    • Description: Six weeks post immunization, mice were bled and antibody responses were measured by ELISA. IgG2a responses were significantly higher in Cp23 vaccinated mice than vector (control) immunized mice (Ehigiator et al., 2007).
    • Detailed Gene Information: Click Here.

Mouse Response

  • Host Strain: C57BL/6 interleukin-12p40 (IL-12p40) knockout (KO) mice (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)

Mouse Response

  • Host Strain: BALB/c mice (Liu et al., 2010)
  • Vaccination Protocol: The mice were randomly divided into different groups and were immunized subcutaneously with 10 μg of rCp15–23, rCp23, or crude extract of C. parvum in complete Freund’s adjuvant. Subsequent immunizations on days 14 and 28 were performed with 10 μg proteins in incomplete Freund’s adjuvant. The control group were given adjuvant alone. (Liu et al., 2010)
  • Immune Response: Humoral: All immunized groups have specific antibody responses. The concentrations of IgG remained at low levels until days 14 after the first vaccination, whereas the second dose of vaccine rapidly and significantly boosted the responses. A peak concentration was observed after third boost. Both rCp15–23and rCp23 induced stronger antibody response than crude extract (P<0.05). (Liu et al., 2010)
    Cellular: The number of CD4+ and CD8+ T cells was increased in all immunized groups compared with the control (P<0.01), whereas the number of CD4+ T cells was much more than that of CD8+ T cells. The stimulation of cells from rCp15–23 immunized mice generated higher CD4+, CD8+ T cells and the ratio of CD4+/CD8+ than the other two immunized groups (P<0.01) (Liu et al., 2010)
    Cytokine: Significantly higher concentrations of IFN-γ or IL-12 were found in the spleen cells of all immunized groups. The IFN-γ and IL-12 levels were significantly higher in rCp15–23 immunized mice compared with the crude extract immunized mice (P<0·05). No significant difference was observed in crude extract group compared with the control group. Very low level of IL-4 was found in all the groups and no difference was found between different groups. (Liu et al., 2010)
  • Challenge Protocol: 10 mice were randomly selected from each group and were challenged with 1×10^6 oocysts orally 2 weeks after the last immunization. (Liu et al., 2010)

Mouse Response

  • Host Strain: C57BL/6 interleukin-12p40 (IL-12p40) knockout (KO) mice (Wang et al., 2010)
  • Vaccination Protocol: Each mouse was vaccinated with with 100 μg of multivalent (pVAX-15–23) or single gene (pVAX-15 or pVAX-23) DNA vaccine alone or plus pMEM12R or in TBE on days 0, 14, and 28. Non-recombinant pVAX-1 plasmid was used as negative control. (Wang et al., 2010)
  • Immune Response: Cellular: splenocytes from mice vaccinated with pVAX15–23 expressed significant INF-γ levels in response of both antigens. IFN-γ expression was significantly boosted by co-administration of pMEM12R, inducting Th1 type immune response. (Wang et al., 2010)
    Humoral: Splenocytes from vaccinated mice exhibited a strong lymphoproliferation responses 2 and 10 weeks after the final immunization, revealing sustained response of to stimulating proteins on the term of time. Stimulation of splenocytes derived from mice immunized 10 weeks before cell harvest showed significant (p < 0.01) pVAX15–23-specific proliferative response in vitro culture. pVAX15–23-DNA vaccine could induce a more sustained cellular immune response than pVAX15 and pVAX23. (Wang et al., 2010)
  • Challenge Protocol: Mice were orally challenged with a single dose of 1 × 10^5 C. parvum oocysts in 200 μL of 0.15 M phosphate-buffered saline (PBS, pH 7.2) 2 weeks after the last immunization. (Wang et al., 2010)
  • Efficacy: The pattern of oocysts shedding was similar in all experimental groups: shedding peak on day 10-12 post challenge and steady decline with the time course. DNA vaccination mitigated the intensity of the infection with significant decrease for mice in pVAX-15-23 group. Mice injected pVAX-15-23 and pMEM12R resolved the infection earlier than other groups. (Wang et al., 2010)

Mouse Response

  • Host Strain: BALB/c mice (Yu et al., 2010)
  • Vaccination Protocol: Two hundred mice were randomly divided into 10 groups. Mice in different groups were immunized at weeks 0, 3, and 5 with different schemes: 1) 100 μg of pVAX1-CP12 musculature, 2) 100 μg of pVAX1-CP21 musculature, 3) 100 μg of pVAX1-CP12 nasal, 4) 100 μg of pVAX1-CP21 nasal, 5) 100 μg of pVAX1-CP12-CP21 musculature, 6) 100 μg of pVAX1-C-CP12-CP21 musculature, 7) 100 μg of pVAX1-CP12-CP21 nasal, 8) 100 μg of pVAX1-C-CP12-CP21 nasal, 9) 100 μg of pVAX1, 10) 0.1mL of PBS (Yu et al., 2010)
  • Immune Response: Humoral: pVAX1-C-Cp12-Cp21 nasal spray group had significantly higher level of IgG compared to pVAX1 vector group and PBS group (P < 0.01). The IgG level in the pVAX1-C-Cp12-Cp21 nasal spray group was also higher than that of pVAX1-Cp12, pVAX1-Cp21, and pVAX1-Cp12-Cp21 group but was not significant.
    Cellular: The percentages of CD4+ T cells in pVAX1-C-Cp12-Cp21 nasal group were significantly higher than that in the groups treated with pVAX1 vector, PBS, pVAX1-Cp12, and pVAX1-Cp21 (P < 0.01). The percentages of CD8+ T cells in pVAX1-C-Cp12-Cp21 nasal spray group was significantly higher than that in the PBS group (P < 0.01). (Yu et al., 2010)
  • Challenge Protocol: Each mouse was orally inoculated with 1 × 10^6 C. parvum oocysts in 0.5 ml of water 2 weeks after the last immunization. (Yu et al., 2010)
  • Efficacy: Mice in pVAX1-Cp12-Cp21 and pVAX1-C-Cp12-Cp21 groups excreted less numbers of oocysts than that in other groups (P < 0.05). Mice in the pVAX1-C-Cp12-Cp21 nasal spray group have a 77.5% reduction in the level of oocysts shedding. (Yu et al., 2010)
References References References References References References
Jenkins et al., 1999: Jenkins MC, O'Brien C, Trout J, Guidry A, Fayer R. Hyperimmune bovine colostrum specific for recombinant Cryptosporidium parvum antigen confers partial protection against cryptosporidiosis in immunosuppressed adult mice. Vaccine. 1999; 17(19); 2453-2460. [PubMed: 10392628].
Ehigiator et al., 2007: Ehigiator HN, Romagnoli P, Priest JW, Secor WE, Mead JR. Induction of murine immune responses by DNA encoding a 23-kDa antigen of Cryptosporidium parvum. Parasitology research. 2007; 101(4); 943-950. [PubMed: 17487508].