Recombinant Calponin of human filariid Brugia malayi: Secondary structure and immunoprophylactic potential |
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| Institution: | 1. Division of Parasitology, CSIR-Central Drug Research Institute, New Campus, BS 10/1, Sector 10, Jankipuram Extension, Lucknow 226 031, India;2. Division of Molecular and Structural Biology, CSIR-Central Drug Research Institute, New Campus, BS 10/1, Sector 10, Jankipuram Extension, Lucknow 226 031, India;1. George Mason University, Department of Global and Community Health, 4400 University Drive, Fairfax, VA 22030, United States;2. Emory University, Rollins School of Public Health, Department of Behavioral Sciences and Health Education, 1518 Clifton Road, Atlanta, GA 30322, United States;3. Emory University, Emory Vaccine Center, Hope Clinic, 500 Irvin Court, Suite 200, Decatur, GA 30030, United States;4. Emory University, School of Medicine, Division of Infectious Diseases, 500 Irvin Court, Suite 200, Decatur, GA 30030, United States;1. Institut National de la Santé et de la Recherche Médicale, Unit 1018, Centre for Research in Epidemiology and Population Health, Villejuif, France;2. Internal Medicine and Infectious Diseases, Groupe Hospitalier Universitaire Paris Centre, APHP, Paris, France;3. Samusocial International, Ivry sur Seine, France;4. Samusocial Casablanca, Morocco;5. Samusocial Bamako, Mali;6. Samusocial Ouagadougou, Burkina Faso;7. Samusocial Bruxelles, Belgium;8. Samusocial Dakar, Senegal;9. Service d’Aide Médicale Urgente (SAMU), Hôpital Necker, APHP, Paris, France;1. Ehime University School of Medicine, Japan;2. Department of Clinical Oncology, Ehime University Graduate School of Medicine, Japan;1. School of Public Health and Community Medicine, Faculty of Medicine, University of New South Wales, Sydney, NSW 2052, Australia;2. The Kirby Institute, University of New South Wales, Sydney, NSW 2052, Australia;1. Avenue JP Rullens 9, 1200 Brussels, Belgium;2. Center for Global Health, Cincinnati Children''s Hospital, Cincinnati, OH, USA;1. Division of Infectious Diseases, Department of Pediatrics, Washington University School of Medicine, St. Louis, MO 63110, USA;2. Division of Laboratory and Genomic Medicine, Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA;3. Neisseria and Streptococcus Reference Laboratory, Department of Microbiology and Infection Control, Statens Serum Institut, Copenhagen, Denmark;4. Division of Immunobiology, Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA |
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| Abstract: | In the search for potential vaccine candidates for the control of human lymphatic filariasis, we recently identified calponin-like protein, that regulates actin/myosin interactions, in a proinflammatory fraction F8 (45.24–48.64 kDa) of Brugia malayi adult worms. In the present study, the gene was cloned, expressed, and the recombinant Calponin of B. malayi (r-ClpBm) was prepared and characterized. r-ClpBm bears homology with OV9M of Onchocerca volvulus, a non-lymphatic filariid that causes loss of vision and cutaneous pathology. r-ClpBm was found to be a ～45 kDa protein that folds into a predominantly α-helix conformation. The protective efficacy of r-ClpBm against B. malayi infection in Mastomys coucha was investigated by assessing the course of microfilaraemia and adult worm burden in the host immunized with r-ClpBm and subsequently infected with infective third stage larvae (L3). Expression of the Calponin was detected in all life stages (microfilariae, L3, L4, L5 and adults) of the parasite and immunization with r-ClpBm partially protected M. coucha against establishment of infection as inferred by ～42% inhibition in parasite burden. Upregulated cellular proliferation, TNF-α, IFN-γ, IL-1β, IL-4, nitric oxide (NO) release, expression of iNOS, and specific IgG, IgG1 and IgG2b in immunized animals correlated with parasitological findings. r-ClpBm immunization caused degranulation in majority of mast cells indicating possible involvement of mast cell products in reducing the parasite survival. It appears that complex mechanisms including Th1, Th2, NO and mast cells are involved in the clearance of infection. To the best of our knowledge this is the first report on cloning, expression of the gene and purification of r-ClpBm, determination of its secondary structure and its ability to partially prevent establishment of B. malayi infection. Thus, r-ClpBm may further be studied and developed in combination with other protective molecules of B. malayi as a component of potential filarial cocktail vaccine candidate. |
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| Keywords: | Calponin iNOS COX-2 Cytokines IgG and its subclasses LTT assay |
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