title: Disorganized muscle protein-1 (DIM-1) of filarial parasite Brugia malayi: cDNA cloning, expression, purification, structural modeling and its potential as vaccine candidate for human filarial infection.
abstract: We have recently identified disorganized muscle protein-1 (DIM-1) in one of the proinflammatory fractions of the human filaria Brugia malayi adult worm. The present study was undertaken to characterize B. malayi DIM-1 (DIM-1bm) and explore its vaccine potential. In this study we cloned and expressed the DIM-1bm gene, investigated its sequence homology with other nematodes, constructed in silico structural model, purified the recombinant DIM-1bm (rDIM-1bm) protein, and studied the effect of immunization with rDIM-1bm on the establishment of B. malayi infection in Mastomys coucha. DIM-1bm showed similarity with DIM-1 of Caenorhabditis elegans, Ascaris suum and Loa loa. Structural modeling revealed three immunoglobulin domains in DIM-1bm indicating that it is a member of immunoglobulin superfamily (IgSF) and 'blastn' results showed that DIM-1bm coding sequence (CDS) have almost no homology with human and mouse nucleotide sequences. Immunization with rDIM-1bm partially protected M. coucha against establishment of infection as inferred by a low recovery of microfilariae (37-64%) and parasite burden (∼50%). The enhanced activity of macrophages, and IFN-γ and NO responses, and elevated levels of specific IgG, IgG1, IgG2a and IgG2b correlated with parasitological findings. This is the first report on cloning, expression, structural modeling and purification of rDIM-1bm and its ability to partially prevent establishment of B. malayi infection. DIM-1bm's almost complete lack of homology with the human counterpart makes it an attractive protein for exploring its vaccine potential.
authors: Division of Parasitology, CSIR-Central Drug Research Institute, New Campus, BS 10/1, Sector 10, Jankipuram Vistar, Lucknow 226031, India.
authors: Vikas
authors: V
authors: Kushwaha
authors: Molecular and Structural Biology, CSIR-Central Drug Research Institute, New Campus, BS 10/1, Sector 10, Jankipuram Vistar, Lucknow 226031, India.
authors: Vikash
authors: V
authors: Kumar
authors: Division of Parasitology, CSIR-Central Drug Research Institute, New Campus, BS 10/1, Sector 10, Jankipuram Vistar, Lucknow 226031, India.
authors: Shiv K
authors: SK
authors: Verma
authors: Department of Biosciences, Integral University, Kursi Road, Lucknow 226026, India.
authors: Rolee
authors: R
authors: Sharma
authors: Molecular and Structural Biology, CSIR-Central Drug Research Institute, New Campus, BS 10/1, Sector 10, Jankipuram Vistar, Lucknow 226031, India.
authors: M I
authors: MI
authors: Siddiqi
authors: Division of Parasitology, CSIR-Central Drug Research Institute, New Campus, BS 10/1, Sector 10, Jankipuram Vistar, Lucknow 226031, India. Electronic address: drpkmurthy@yahoo.com.
authors: P K
authors: PK
authors: Murthy
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000947:Antigens, Helminth / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D017178:Brugia malayi / Q000235:genetics*; D003001:Cloning, Molecular; D005260:Female; D005849:Gerbillinae; D007074:Immunoglobulin G / Q000097:blood; D007371:Interferon-gamma / Q000276:immunology; D008264:Macrophages / Q000276:immunology; D008297:Male; D008958:Models, Molecular; D008969:Molecular Sequence Data; D051189:Murinae; D009124:Muscle Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D009569:Nitric Oxide / Q000378:metabolism; D010587:Phagocytosis
chemical_list: D000947:Antigens, Helminth; D007074:Immunoglobulin G; D009124:Muscle Proteins; D009569:Nitric Oxide; D007371:Interferon-gamma
keywords: Brugia malayi; Disorganized muscle protein-1; IFN-γ; IgG and its subclasses; Macrophage activity; Nitric oxide