In silico and in vivo analysis of Toxoplasma gondii epitopes by correlating survival data with peptide-MHC-I binding affinities.
BACKGROUND
Protein antigens comprising peptide motifs with high binding affinity to major histocompatibility complex class I (MHC-I) molecules are expected to induce a stronger cytotoxic T-lymphocyte response and thus provide better protection against infection with microorganisms where cytotoxic T-cells are the main effector arm of the immune system.


METHODS
Data on cyst formation and survival were extracted from past studies on the DNA immunization of mice with plasmids coding for Toxoplasma gondii antigens. From in silico analyses of the vaccine antigens, the correlation was tested between the predicted affinity for MHC-I molecules of the vaccine peptides and the survival of immunized mice after challenge with T. gondii. ELISPOT analysis was used for the experimental testing of peptide immunogenicity.


RESULTS
Predictions for the Db MHC-I molecule produced a strong, negative correlation between survival and the dissociation constant of vaccine-derived peptides. The in silico analyses of nine T. gondii antigens identified peptides with a predicted dissociation constant in the interval from 10nM to 40μM. ELISPOT assays with splenocytes from T. gondii-infected mice further supported the importance of the peptide affinity for MHC-I.


CONCLUSIONS
In silico analysis clearly helped the search for protective vaccine antigens. The ELISPOT analysis confirmed that the predicted T-cell epitopes were immunogenic by their ability to release interferon gamma in spleen cells.
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province, PR China; Biophysical Immunology Laboratory, Department of Biomedicine, Aarhus University, Aarhus, Denmark.
Si-Yang
SY
Huang
Biophysical Immunology Laboratory, Department of Biomedicine, Aarhus University, Aarhus, Denmark.
Maria Risager
MR
Jensen
Biophysical Immunology Laboratory, Department of Biomedicine, Aarhus University, Aarhus, Denmark.
Carina Agerbo
CA
Rosenberg
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province, PR China.
Xing-Quan
XQ
Zhu
Department of Infectious Diseases, Clinical Institute, Faculty of Health Sciences, Aarhus University Hospital, Skejby, Denmark.
Eskild
E
Petersen
Biophysical Immunology Laboratory, Department of Biomedicine, Aarhus University, Aarhus, Denmark. Electronic address: vorup-jensen@biomed.au.dk.
Thomas
T
Vorup-Jensen
D000818:Animals; D003198:Computer Simulation; D058501:Enzyme-Linked Immunospot Assay; D018984:Epitopes, T-Lymphocyte / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D005260:Female; D015395:Histocompatibility Antigens Class I / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007114:Immunization; D007371:Interferon-gamma / Q000276:immunology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D010455:Peptides / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D014122:Toxoplasma / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D014123:Toxoplasmosis / Q000276:immunology* / Q000469:parasitology; D019444:Vaccines, DNA / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics / Q000276:immunology
D018984:Epitopes, T-Lymphocyte; D015395:Histocompatibility Antigens Class I; D010455:Peptides; D019444:Vaccines, DNA; D007371:Interferon-gamma
In silico prediction; MHC class I; Toxoplasma gondii; Vaccine antigens