Immunity induced by DNA vaccine of plasmid encoding the rhoptry protein 1 gene combined with the genetic adjuvant of pcIFN-gamma against Toxoplasma gondii in mice.
OBJECTIVE
To construct the eukáryotic expression recombinant plasmid, pcIFN-gamma, as a genetic adjuvant and observe the immune responses elicited by pcDNA3-rhoptry protein 1 (pc-ROP1) combined with pcIFN-gamma against Toxoplasma gondii (T. gondii) infection in mice.


METHODS
A fragment of the IFN-gamma gene was directly inserted into the pcDNA3 plasmid and identified by two restriction endonucleases digestion. pcIFN and pcROP1 DNA was injected into the left leg muscle of mice at a dosage of 100 micrograms, and a booster vaccination was given at the same dosage after two weeks. Control groups were injected with pcDNA3 blank plasmid or normal saline. At 30, 50 and 70 days after booster injection, kinetic tests were carried out: MTT assay for the proliferation response of T lymphocyte cells and the activity of NK cells, sandwich ABC-ELISA for the determination of IFN-gamma, IL-2 and IL-10; a serum enzymetic aassay for nitric oxide (NO) in sera and ELISA for the titer of IgG antibody in sera.


RESULTS
The recombinant plasmid, pcIFN-gamma was constructed. The proliferation response of spleen T lymph cells, NK cell killing activity, and serum levels of IFN-gamma, IL-2 and NO in mice injected with pcROP1 and pcIFN-gamma were higher than in those injected with pcROP1 alone. There was no difference in IgG antibody levels between the two groups.


CONCLUSIONS
The genetic adjuvant, pcIFN-gamma, could enhance the cellular immune response induced by DNA vaccine of pcROP1 in mice against Toxoplasma gondii infection.
Guo
H
H
Institute of Parasitology, Sun Yat-sen University of Medical Sciences, Guangzhou 510089, China.
Chen
G
G
Lu
F
F
Chen
H
H
Zheng
H
H
D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood; D002455:Cell Division / Q000276:immunology; D004274:DNA, Recombinant / Q000235:genetics; D007110:Immunity, Active / Q000276:immunology*; D007074:Immunoglobulin G / Q000097:blood; D007371:Interferon-gamma / Q000097:blood / Q000235:genetics / Q000276:immunology*; D016753:Interleukin-10 / Q000097:blood; D007376:Interleukin-2 / Q000097:blood; D007694:Killer Cells, Natural / Q000276:immunology; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009569:Nitric Oxide / Q000097:blood; D010957:Plasmids / Q000235:genetics; D013154:Spleen / Q000166:cytology / Q000276:immunology; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology; D014122:Toxoplasma / Q000276:immunology*; D014124:Toxoplasmosis, Animal / Q000276:immunology / Q000469:parasitology / Q000517:prevention & control; D019444:Vaccines, DNA / Q000235:genetics / Q000276:immunology*
D000913:Antibodies, Protozoan; D004274:DNA, Recombinant; D007074:Immunoglobulin G; D007376:Interleukin-2; D008565:Membrane Proteins; C117001:ROP 1 protein, Toxoplasma gondii; D019444:Vaccines, DNA; D016753:Interleukin-10; D009569:Nitric Oxide; D007371:Interferon-gamma