• Vaccine Ontology ID: VO_0011471
• Type: DNA vaccine
• Status: Research
• Antigen: E. tenella microneme 2 (MIC2)
• MIC2 gene engineering:
  • Type: DNA vaccine construction
  • Description: The EtMIC2 coding sequence was subcloned from EtMIC2-pGEX into the BamHI/HindIII sites of the pMal4c vector with a NH2-terminal maltose-binding protein tag, expressed in E. coli in TY broth (20 g/l tryptone, 10 g/l yeast extract, 10 g/l NaCl) containing 100 μg/ml ampicillin, the bacteria grown to OD600 = 0.5, induced with 1.0 mM isopropyl-β-d-thiogalactopyranoside for 3 h at 37 °C, collected by centrifugation, and disrupted by sonication on ice (Misonix, Farmingdale, NY). The EtMIC2 protein was isolated on an amylose affinity column (New England Biolabs, Beverly, MA) according to the manufacturer's instructions, digested with Factor Xa to release EtMIC2, and re-passed through the amylose column to remove maltose-binding protein (Ding et al., 2005).
  • Detailed Gene Information: Click Here.
• Vector: pcDNA vector
• Immunization Route: in ovo

• Vaccine Ontology ID: VO_0011449
• Type: DNA vaccine
• Status: Research
• Antigen: E. tenella sporozoite TA4 surface antigen
• TA4 gene engineering:
  • Type: DNA vaccine construction
  • Description: PCR amplification of TA4 (without stop codon) was performed using the gene-specific primers with pET28a-TA4 as template. After purification, the amplified TA4 (without stop codon) was digested with BamHI and EcoRI and cloned into the BamHI/EcoRI sites of pcDNA3.1b. The recombinant plasmid pcDNA3.1b-TA4 (without stop codon) was confirmed by PCR amplification and endonuclease cleavage. Using the same method above, amplified chIL-2 product was ligated into pcDNA3.1b-TA4 (without stop codon) and pcDNA4.0c (Invitrogen) to construct pcDNA3.1b-TA4-IL2 and pcDNA4.0c-IL-2 (Song et al., 2009).
  • Detailed Gene Information: Click Here.
• Vector: pcDNA3.1 (Song et al., 2009)
• Immunization Route: Intramuscular injection (i.m.)

• Vaccine Ontology ID: VO_0011469
• Type: DNA vaccine
• Status: Research
• Antigen: Eimeria tenella antigen gene (3-1E) and chicken interferon gamma gene (ChIFN-gamma)
• 3-1E gene engineering:
  • Type: DNA vaccine construction
  • Description: The Eimeria tenella antigen gene (3-1E) and chicken interferon gamma gene (ChIFN-gamma) were subcloned into the mammalian expression vector proVAX forming the plasmids proE and prol, and then linked by splicing overlap extension by polymerase chain reaction to construct the chimeric plasmid prolE; the chimeric protein (rlE) was expressed in Escherichia coli harboring the constructed plasmid pGEX/IE (Xu et al., 2006).
  • Detailed Gene Information: Click Here.
• Vector: proVAX
• Immunization Route: Intramuscular injection (i.m.)

• Vaccine Ontology ID: VO_0011470
• Type: Subunit vaccine
• Status: Research
• Antigen: E. tenella sporozoite antigen GX3262
• GX3262 gene engineering:
  • Type: Recombinant protein preparation
  • Description: To produce the 3-galactosidase-GX3262 fusion protein (Pgal-GX3262), recombinant E. coli cells. grown overnight in Luria-Bertani broth with ampicillin and induced with IPTG, were lysed by sonication and centrifuged, and the insoluble cell pellet was washed extensively with PBS (Miller et al., 1989).
  • Detailed Gene Information: Click Here.
• Adjuvant:
  • VO ID: VO_0000139
• Immunization Route: Subcutaneous injection

• Vaccine Ontology ID: VO_0011472
• Type: Recombinant vector vaccine
• Status: Research
• Antigen: Eimeria tenella rhomboid gene (Rho)
• Rho gene engineering:
  • Type: Recombinant vector construction
  • Description: The rhomboid gene segment (GenBank accession no. DQ323509) was generated from the cDNA of E. tenella strains by PCR with primers QF (5’-CTGACTGCAGATGTCGGACATCGAATCCCAGAG-3′) containing PstI restriction site (underlined) and QR (5’-GACTATCGATTTATGCGCATCCCATGGGCAAAGG-3’) containing ClaI restriction site (underlined), and cloned into the pMD18-T vector. The 774-bp rhomboid gene fragment was subcloned into the PstI/ClaI sites of pMV261. The plasmid pMV261-Rho was electrotransfected into BCG and selected by kanamycin (Sigma) (Wang et al., 2009).
  • Detailed Gene Information: Click Here.
• Vector: BCG
• Immunization Route: Intravenous injection (i.v.)

• Vaccine Ontology ID: VO_0011448
• Type: Recombinant vector vaccine
• Status: Research
• Antigen: E. tenella rhomboid gene (Rho)
• Rho gene engineering:
  • Type: Recombinant vector construction
  • Description: The rhomboid gene segment (GenBank accession no. DQ323509) was generated from the cDNA of E. tenella strains by PCR with primers QF (5’-CTGACTGCAGATGTCGGACATCGAATCCCAGAG-3′) containing PstI restriction site (underlined) and QR (5’-GACTATCGATTTATGCGCATCCCATGGGCAAAGG-3’) containing ClaI restriction site (underlined), and cloned into the pMD18-T vector. The 774-bp rhomboid gene fragment was subcloned into the PvuII/ClaI site of pMV361. The plasmid pMV361-Rho was electrotransfected into BCG and selected by kanamycin (Sigma) (Wang et al., 2009).
  • Detailed Gene Information: Click Here.
• Vector: BCG
• Immunization Route: Intranasal

Chicken Response

• Vaccination Protocol: To assess anti-EtMIC2 antibody titers and protective immunity to coccidiosis following in ovo vaccination with the EtMIC2 gene, 75 fertile eggs were distributed into five groups (15/group) and non-injected or injected with 100 μl of sterile PBS, 50 μg/egg of the pcDNA empty vector, or 25 or 50 μg/egg of EtMIC2-pcDNA. To determine the effects of post-vaccination boosting with the EtMIC2 gene, 150 fertile eggs were distributed into 10 groups (15/group) and non-injected or injected with 100 μl of sterile PBS, 50 μg/egg of the pcDNA empty vector, or 25 or 50 μg/egg of EtMIC2-pcDNA (Ding et al., 2005).
• Challenge Protocol: At day 11 post-hatching, chickens were infected with 10,000 sporulated oocysts of E. tenella. Serum samples were collected at days 1, 10, and 17 post-infection (days 10, 21, and 28 post-hatching) and anti-EtMIC2 antibody titers determined by ELISA (Ding et al., 2005).
• Efficacy: Birds vaccinated with Eimeria tenella microneme recombinant gene (EtMIC2) and encoded protein developed protective immunity against infection by E. tenella as assessed by significantly increased body weight gain and decreased fecal oocyst shedding compared with non-vaccinated controls. Vaccination with the EtMIC2 gene also led to protective immunity against infection by E. acervulina, but not E. maxima (Ding et al., 2005).

Chicken Response

• Vaccination Protocol: At 14 days of age, chickens were randomly distributed into seven groups (30/group), group 1 negative control, group 2 positive control, groups 3, 4, 5 and 6 immunized with 25 μg, 50 μg, 100 μg and 200 μg of DNA vaccine, respectively and group 7 vector control. Experimental groups were immunized with corresponding doses of DNA vaccine pcDNA3.1b–TA4–IL-2 by leg intramuscular injection. Control group chickens were injected with sterile TE or 100 μg pcDNA3.1b at the same injection site to the experimental groups. A booster immunization was given by the same method as the first immunization 7 days later (Song et al., 2009).
• Challenge Protocol: At 28 days of age, the chickens were weighed one-by-one and challenged with 5 × 10^4 sporulated oocysts of E. tenella JS except the negative control group. The chickens were weighed one-by-one and slaughtered 7 days post challenge (Song et al., 2009).
• Efficacy: The immunization procedure of DNA vaccine pcDNA-TA4-IL-2 of Eimeria tenella, including route, dose, time of immunization and age of primary immunization of chicken, was optimized. results illustrated that 25 microg was the optimal dose and intramuscular injection was the most effective route to induce protective immunity in Chinese Yellow chickens (Song et al., 2009).

Chicken Response

• Host Strain: Broiler
• Vaccination Protocol: Broilers were administered two intramuscular injections with the constructed DNA vaccines (50 microg); in the protein booster groups 100 microg of the rlE were given following the proIE prime (Xu et al., 2006).
• Challenge Protocol: One week following the booster immunization, all chickens except for the unchallenged control group were orally challenged with 5 x 10^4 sporulated E. tenella GS oocysts (Xu et al., 2006).
• Efficacy: The Eimeria tenella antigen gene (3-1E) and chicken interferon gamma gene (ChIFN-gamma) were subcloned into the mammalian expression vector proVAX forming the plasmids proE and prol. After challenge the proIE-vaccinated chickens showed protective immunity as demonstrated by significantly reduced oocyst shedding (Xu et al., 2006).

Chicken Response

• Host Strain: Hubbard broiler
• Vaccination Protocol: The basic experimental design involved immunizing birds subcutaneously or orally with partially purified recombinant antigen preparations, heatkilled bacterins. live E. coli cells, or control materials (Miller et al., 1989).
• Challenge Protocol: Broilers were challenged with E. tendell/i oocysts; and then evaluated for cecal lesions (Miller et al., 1989).
• Efficacy: A cDNA clone derived from sporulated oocysts of Eimeria tenella and encoding the expression product GX3262 was identified using a monoclonal antibody raised against Eimeria acervulina sporozoites. After challenge with an experimental E. tenella infection, the greatest degree of protection in immunized broiler chickens was seen after only a single immunization of 2-day-old birds with a live recombinant E. coli preparation (Miller et al., 1989).

Chicken Response

• Vaccination Protocol: Chickens were randomly assigned to 4 groups (40 birds in each group) and immunized intranasal (i.n.) with 10^6 CFU/200 μl of BCG or rBCG strains expressing rhomboid protein, respectivelly. Negative control was immunized i.n. with 200 μl PBS only. 2 weeks after the first vaccination, a second immunization was carried out with the same amounts of each sample (Wang et al., 2009).
• Challenge Protocol: All groups were challenged with 3 × 10^4 sporulated oocysts of E. tenella per bird 2 weeks after the final immunization (Wang et al., 2009).
• Efficacy: All the recombinant BCG immunized chickens developed specific immune responses, and there was a significant increases of the percentages of CD4(+) and CD8(+) cells compared to the control (P<0.05). Challenge experiments demonstrated that the two rBCG strains could provide significant protection against E. tenella challenge (Wang et al., 2009).

Chicken Response

• Vaccination Protocol: Chickens were randomly assigned to 4 groups (40 birds in each group) and immunized intranasal (i.n.) with 10^6 CFU/200 μl of BCG or rBCG strains expressing rhomboid protein, respectivelly. Negative control was immunized i.n. with 200 μl PBS only. 2 weeks after the first vaccination, a second immunization was carried out with the same amounts of each sample (Wang et al., 2009).
• Challenge Protocol: All groups were challenged with 3 × 10^4 sporulated oocysts of E. tenella per bird 2 weeks after the final immunization (Wang et al., 2009).
• Efficacy: All the recombinant BCG immunized chickens developed specific immune responses, and there was a significant increases of the percentages of CD4(+) and CD8(+) cells compared to the control (P<0.05). Challenge experiments demonstrated that the two rBCG strains could provide significant protection against E. tenella challenge (Wang et al., 2009).