Expression and purification of a Plasmodium vivax antigen - PvTARAg55 tryptophan- and alanine-rich antigen and its immunological responses in human subjects.
Despite the immense global efforts, the malaria vaccine is not yet available and requires the identification of newer target molecules. Since tryptophan-rich proteins of P. yoelii have been proposed as vaccine candidates, we describe here the expression, purification and immunological characterization of a 55kDa Plasmodium vivax tryptophan- and alanine-rich antigen (PvTARAg55). This protein consists of 480 aa residues with a calculated molecular mass of 55.0kDa. It shows 42% aa sequence identity (64% homology) with PyPAg1 of P. yoelii and shares positional conservation of tryptophan residues. Sequence analysis of PvTARAg55 from different P. vivax isolates revealed that typtophan-rich domain which contains most of the B-cell epitopes was highly conserved in the parasite population while the alanine-rich domain showed polymorphism. Exon-2 covering major part (420 aa) of the protein including both the domains was PCR amplified, cloned, expressed in Escherichia coli, and the recombinant protein purified to its homogeneity. Majority of P. vivax-infected individuals (82.5%, n=40) produced antibodies against this antigen. Proliferative responses to the recombinant PvTARAg55 were observed in 60% (n=20) of individuals who had recently been exposed to the P. vivax infection. Measurement of Th1- (IFN-gamma, TNF-alpha, and IL-12) and Th2-type (IL-4 and IL-10) cytokine production in response to this recombinant antigen revealed a mixed type T-cell response with a Th2 response being more pronounced. These results demonstrate that PvTARAg55 elicits high humoral and cellular immune responses thus establishes its immunogenecity in humans.
Department of Biotechnology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi 110029, India.
Asim A
AA
Siddiqui
Neeru
N
Singh
Yagya D
YD
Sharma
D000409:Alanine; D000595:Amino Acid Sequence; D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood*; D000953:Antigens, Protozoan / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D016207:Cytokines / Q000096:biosynthesis; D005091:Exons; D006801:Humans; D008213:Lymphocyte Activation; D017780:Malaria Vaccines / Q000276:immunology*; D008969:Molecular Sequence Data; D010966:Plasmodium vivax / Q000276:immunology*; D020449:Repetitive Sequences, Amino Acid; D014364:Tryptophan; D014614:Vaccines, Synthetic / Q000276:immunology*
D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D016207:Cytokines; D017780:Malaria Vaccines; D014614:Vaccines, Synthetic; D014364:Tryptophan; D000409:Alanine