Toxoplasma gondii: proteomic analysis of antigenicity of soluble tachyzoite antigen.
The obligate intracellular parasite Toxoplasma gondii is an important pathogen of humans and animals. The tachyzoite of T. gondii is the main life-cycle stage that is responsible for toxoplasmosis. Study of the antigenicity of soluble tachyzoite antigen (STAg) is important for discovery of protective antigens which will aid in the detection and prevention of toxoplasmosis. At present, no complete proteome map of T. gondii STAg is established, although a large-scale whole proteomic analysis of tachyzoites is underway. In this study, 1227 protein spots of T. gondii soluble tachyzoite antigen (STAg) were fractionated by 2-dimensional electrophoresis (2-DE) at pH range 3-10. By mass spectrometry (MS) analysis, among the separated 1227 protein spots, 426 were identified by searching the Swissport and NCBI nr databases. Two hundred and thirty of these identified spots (230/426, 54%) were demonstrated to be T. gondii protein by MS. Of the 21 Toxoplasma protein spots identified by Western blot with rabbit anti-T. gondii serum, 16 had immunoregulatory functions and five had immune defense functions. Due to multiple spots for a single protein, these 16 spots represented 11 proteins: a putative protein disulfide isomerase (PDI), heat shock protein 60 (Hsp60), a pyruvate kinase (PK), a putative glutamate dehydrogenase (GDH), a coronin, a heat shock protein 70 (Hsp70), a protein kinase C receptor 1 (RACK1), a malate dehydrogenase (MDH), a major surface antigen 1 (SAG1), an uridine phosphorylase (UPase) and a peroxiredoxin (Prx). Among the identified 11 proteins, except that the antigenicity and immunogenicity of the SAG1 has been reported and antigenicity of Hsp70 has been disputed, the remaining antigenic proteins were first identified in this study. In conclusion, we obtained nine novel types of immunogenic proteins that might be potential candidates of vaccine development for toxoplasmosis, which we will confirm in later studies.
Ma
Guang-Yuan
GY
Department of Parasitology, Shanxi Medical University, No. 56 Xinjian Nan Road, Taiyuan, Shanxi 030001, PR China.
Zhang
Jian-Zhong
JZ
Yin
Guo-Rong
GR
Zhang
Jian-Hong
JH
Meng
Xiao-Li
XL
Zhao
Fei
F
D000818:Animals; D000953:Antigens, Protozoan / Q000032:analysis / Q000276:immunology*; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D008297:Male; D013058:Mass Spectrometry; D051379:Mice; D008807:Mice, Inbred BALB C; D040901:Proteomics; D015800:Protozoan Proteins / Q000032:analysis / Q000276:immunology; D016052:Protozoan Vaccines / Q000032:analysis / Q000276:immunology; D011817:Rabbits; D014122:Toxoplasma / Q000276:immunology*
D000953:Antigens, Protozoan; D015800:Protozoan Proteins; D016052:Protozoan Vaccines; C094052:soluble tachyzoite antigen, Toxoplasma gondii