title: Molecular and functional characterization of macrophage migration inhibitory factor (MIF) homolog of human from lymphatic filarial parasite Wuchereria bancrofti.
abstract: The ability of nematode parasites to survive in a highly complex immune system involves diverse strategies including production of a variety of host immune modulators. Various parasite-associated surface antigens or excretory and secretory products may possibly play a role in the host-parasite interactions and successful survival of parasite in their respective host. One among these molecules is a human cytokine homolog, macrophage migration inhibitory factor-1 (MIF-1) in various parasites. We identified a homolog of this cytokine from human lymphatic filarial parasite, Wuchereria bancrofti, expression cloned and investigated its molecular characteristics and catalytic properties. We also assessed the humoral reactivity of the recombinant MIF-1 of W. bancrofti (rWb-MIF-1) against sera belonging to different categories of individuals viz. microfilaremic, chronic patients, endemic normal, and non-endemic normal. Our results showed that the complete coding sequence of W. bancrofti is 1,078 bp, comprising two introns and three exons: first and second introns being 577 and 153 bp long, while the three exons I, II, and III being 108, 173, and 67 bp long, respectively. The rWb-MIF-1 was overexpressed in a salt-inducible host, Escherichia coli GJ 1158, and its functional activity was determined by dopachrome tautomerase and insulin reduction assays. The results of both the assays showed that the purified protein is functionally active and hence folded appropriately. The rWb-MIF-1 protein did not show elevation of specific IgG4 antibodies in microfilaremic cases, a hallmark in case of lymphatic filariasis, while it showed IgE reactivity in some of these cases (five out of ten).
authors.affiliation: Vector Control Research Centre (ICMR) Indira Nagar, Medical Complex, Pondicherry 605 006, India.
authors.forename: Rohit
authors.initials: R
authors.lastname: Sharma
authors.forename: S L
authors.initials: SL
authors.lastname: Hoti
authors.forename: R L
authors.initials: RL
authors.lastname: Meena
authors.forename: V
authors.initials: V
authors.lastname: Vasuki
authors.forename: T
authors.initials: T
authors.lastname: Sankari
authors.forename: P
authors.initials: P
authors.lastname: Kaliraj
mesh_terms: D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D003001:Cloning, Molecular; D004605:Elephantiasis, Filarial / Q000276:immunology / Q000469:parasitology; D004926:Escherichia coli / Q000235:genetics; D005091:Exons; D015870:Gene Expression; D015801:Helminth Proteins / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood; D007438:Introns; D008263:Macrophage Migration-Inhibitory Factors / Q000235:genetics* / Q000378:metabolism*; D016366:Open Reading Frames; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D014958:Wuchereria bancrofti / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D000909:Antibodies, Helminth; D015801:Helminth Proteins; D008263:Macrophage Migration-Inhibitory Factors; D011994:Recombinant Proteins; D007073:Immunoglobulin E