Identification of a human immunodominant B-cell epitope within the GRA1 antigen of Toxoplasma gondii by phage display of cDNA libraries.
Excreted secreted antigens of the protozoan parasite Toxoplasma gondii play a key role in stimulating the host immune system during acute and chronic infection. With the aim of identifying the immunodominant epitopes of T. gondii antigens involved in the human B-cell response against the parasite, we employed a novel immunological approach. A library of cDNA fragments from T. gondii tachyzoites was displayed as fusion proteins to the amino-terminus of lambda bacteriophage capsid protein D. The lambda D-tachyzoite library was then affinity-selected by using a panel of sera of pregnant women, all infected with the parasite. Some of the clones identified through this procedure matched the sequence of the dense granule GRA1 protein (p24), allowing us to identify its antigenic regions. In particular, the analysis of human antibody response against the recombinant GRA1 antigen fragments revealed the existence of an immunodominant epitope (epi-24 peptide).
Beghetto
E
E
Kenton Laboratories, Rome, Italy.
Pucci
A
A
Minenkova
O
O
Spadoni
A
A
Bruno
L
L
Buffolano
W
W
Soldati
D
D
Felici
F
F
Gargano
N
N
D000595:Amino Acid Sequence; D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics / Q000276:immunology*; D010582:Bacteriophage lambda / Q000737:chemistry; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D018985:Epitopes, B-Lymphocyte / Q000235:genetics / Q000276:immunology*; D005260:Female; D015723:Gene Library; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D016056:Immunodominant Epitopes / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D019151:Peptide Library; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D017386:Sequence Homology, Amino Acid; D014122:Toxoplasma / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D014123:Toxoplasmosis / Q000276:immunology*
D000953:Antigens, Protozoan; D016054:DNA, Protozoan; D018985:Epitopes, B-Lymphocyte; C412913:GRA1 antigen; D016056:Immunodominant Epitopes; D019151:Peptide Library; D005982:Glutathione Transferase