Eimeria spp. are obligate intracellular apicomplexan protozoan parasites and are the cause of intestinal coccidiosis. Infections of the vertebrate host with these parasites result in the induction of potent protective immune responses, and protection against challenge infections is confined absolutely to homologous species, even though both T- and B-cell responses exhibit cross-species reactivity. Eimeria maxima is the most immunogenic of the seven species of Eimeria that infect the domestic chicken (Smith et al., 2002).
II. Vaccine Related Pathogen Genes
1. gam56
Gene Name :
gam56
Sequence Strain (Species/Organism) : Eimeria maxima
Molecule Role Annotation :
Animals vaccinated with Gam82 and challenged with E. maxima showed lower oocyst shedding and reduced intestinal pathology compared with non-vaccinated and parasite-challenged animals. Gam82 vaccination also stimulated the production of antigen-specific serum antibodies and induced greater levels of IL-2 and IL-15 mRNAs compared with non-vaccinated controls. These results demonstrate that the Gam82 recombinant protein protects against E. maxima and augments humoral and cell-mediated immunity (Jang et al., 2010).
Protein Note :
Von Willebrand factor type A (vWA) domain was originally found in the blood coagulation protein von Willebrand factor (vWF). Typically, the vWA domain is made up of approximately 200 amino acid residues folded into a classic a/b para-rossmann type of...; cl00057
Vaccination Protocol:
The DNA vaccine was injected intramuscularly to yellow feathered broilers of 1-week old at 3 dosages (25, 50, and 100 µg/chick). Injection was repeated once 1 week later (Xu et al., 2013).
Vaccine Immune Response Type:
VO_0000286
Immune Response:
Compared with control groups, the administration of pcDNA-Gam56 vaccine markedly increased the lymphocyte proliferation activity (P<0.05) at day 7 and 14 after the first immunization. The level of lymphocyte proliferation started to decrease on day 21 after the first immunization. A similar trend was seen in specific antibody levels (Xu et al., 2013).
Challenge Protocol:
One week after the second injection, birds were challenged orally with 5×104 sporulated oocysts of E. maxima, then weighed and killed at day 8 post challenge (Xu et al., 2013).
Efficacy:
Among the 3 pcDNA-Gam56 immunized groups, the median dosage group displayed the highest lymphocyte proliferation and antibody levels (P<0.05). The median dosage group had the greatest relative body weight gain (89.7%), and the greatest oocyst shedding reduction (53.7%) (Xu et al., 2013).
Description:
The Gam82 coding sequence was amplified by PCR using Proof Start DNA polymerase (Qiagen, Valencia, CA). Amplicons were cloned into the pET28a (+) plasmid vector (Novagen/EMD Chemicals, Gibbstown, NJ) downstream from an NH2-terminal His6 epitope tag, plasmid clones were verified by sequence analysis, and used to transform competent Escherichia coli BL21 Star (Invitrogen). Eluted protein fractions were visualized on 10% SDS-acrylamide gels stained with Coomassie brilliant blue and on Western blots probed with horseradish peroxidase-conjugated anti-His monoclonal antibody (1:3000; Qiagen), and stored at −20 °C (Jang et al., 2010).
Vaccination Protocol:
At 1 wk of age, chickens (30/group) were intramuscularly immunized with 100 μl of 300 or 600 μg/ml of recombinant Gam82 in Freund's complete adjuvant (FCA). Control animals received PBS in FCA. At 1 wk post-immunization, animals were given a booster injection with 100 μl of 300 or 600 μg/ml of recombinant Gam82, 3-1E, or PBS in Freund's incomplete adjuvant Ref1077:Jang et al., 2010].
Challenge Protocol:
At 7 d post-secondary immunization, animals were given PBS or 2.0 × 10^4 E. maxima sporulated oocysts by oral gavage using an 18-gauge needle (Jang et al., 2010).
Efficacy:
Animals vaccinated with Gam82 and challenged with E. maxima showed lower oocyst shedding and reduced intestinal pathology compared with non-vaccinated and parasite-challenged animals. Gam82 vaccination also stimulated the production of antigen-specific serum antibodies and induced greater levels of IL-2 and IL-15 mRNAs compared with non-vaccinated controls. These results demonstrate that the Gam82 recombinant protein protects against E. maxima and augments humoral and cell-mediated immunity (Jang et al., 2010).
Host Gene Response of
IL15
Gene Response:
Three and six days after second immunization with 60 micrograms of Gam82, there was a significant difference in IL-15 mRNA levels between the immunized and non immunized (PBS vaccinated) groups of chickens. Levels were determined in intestinal tissue (Jang et al., 2010).
Gene Response:
Three days after second immunization with 60 micrograms of Gam82, there was a significant difference in IL-2 mRNA levels between the immunized and non immunized (PBS vaccinated) groups of chickens. Levels were determined in intestinal tissue (Jang et al., 2010).
Chickens were vaccinated with Arlacel A injections prior to vaccination with E. maxima oocysts (Smith et al., 1994).
g.
Chicken Response
Host Strain:
Vedette
Vaccination Protocol:
Hens were injected intramuscularly with 0.25 ml of PBS in 0.25 ml of Arlacel A in weeks 0, 2, and 6 of the study (Smith et al., 1994).
Challenge Protocol:
At week 16 hens were infected with 20,000 oocysts of E. maxima (Smith et al., 1994).
Efficacy:
None of the hens vaccinated had clinical symptoms of infection (Smith et al., 1994).
V. References
1. Huang et al., 2015: Huang J, Zhang Z, Li M, Song X, Yan R, Xu L, Li X. Immune protection of microneme 7 (EmMIC7) against Eimeria maxima challenge in chickens. Avian pathology : journal of the W.V.P.A. 2015; 44(5); 392-400. [PubMed: 26181095].
2. Huang et al., 2015: Huang J, Zhang Z, Li M, Song X, Yan R, Xu L, Li X. Eimeria maxima microneme protein 2 delivered as DNA vaccine and recombinant protein induces immunity against experimental homogenous challenge. Parasitology international. 2015; 64(5); 408-416. [PubMed: 26072304].
3. Jang et al., 2010: Jang SI, Lillehoj HS, Lee SH, Lee KW, Park MS, Cha SR, Lillehoj EP, Subramanian BM, Sriraman R, Srinivasan VA. Eimeria maxima recombinant Gam82 gametocyte antigen vaccine protects against coccidiosis and augments humoral and cell-mediated immunity. Vaccine. 2010; 28(17); 2980-2985. [PubMed: 20178868].
4. Jang et al., 2010: Jang SI, Lillehoj HS, Lee SH, Lee KW, Park MS, Cha SR, Lillehoj EP, Subramanian BM, Sriraman R, Srinivasan VA. Eimeria maxima recombinant Gam82 gametocyte antigen vaccine protects against coccidiosis and augments humoral and cell-mediated immunity. Vaccine. 2010; 28(17); 2980-2985. [PubMed: 20178868].
5. Smith et al., 1994: Smith NC, Wallach M, Miller CM, Morgenstern R, Braun R, Eckert J. Maternal transmission of immunity to Eimeria maxima: enzyme-linked immunosorbent assay analysis of protective antibodies induced by infection. Infection and immunity. 1994; 62(4); 1348-1357. [PubMed: 8132342].
6. Smith et al., 2002: Smith AL, Hesketh P, Archer A, Shirley MW. Antigenic diversity in Eimeria maxima and the influence of host genetics and immunization schedule on cross-protective immunity. Infection and immunity. 2002; 70(5); 2472-2479. [PubMed: 11953384].
7. Wallach, 1997: Wallach M. The importance of transmission-blocking immunity in the control of infections by apicomplexan parasites. International journal for parasitology. 1997; 27(10); 1159-1167. [PubMed: 9394186].
8. Witcombe et al., 2004: Witcombe DM, Ferguson DJ, Belli SI, Wallach MG, Smith NC. Eimeria maxima TRAP family protein EmTFP250: subcellular localisation and induction of immune responses by immunisation with a recombinant C-terminal derivative. International journal for parasitology. 2004; 34(7); 861-872. [PubMed: 15157769].
9. Xu et al., 2013: Xu J, Zhang Y, Tao J. Efficacy of a DNA vaccine carrying Eimeria maxima Gam56 antigen gene against coccidiosis in chickens. The Korean journal of parasitology. 2013; 51(2); 147-154. [PubMed: 23710081].
