title: Protection against filarial infection by 45-49 kDa molecules of Brugia malayi via IFN-γ-mediated iNOS induction.
abstract: Nitric oxide (NO) mediated mechanisms have been implicated in killing of some life-stages of Brugia malayi/Wuchereria bancrofti and protect the host through type 1 responses and IFN-γ stimulated toxic mediators' release. However, the identity of NO stimulating molecules of the parasites is not known. Three predominantly NO-stimulating SDS-PAGE resolved fractions F8 (45.24-48.64 kDa), F11 (33.44-38.44 kDa) and F12 (28.44-33.44 kDa) from B. malayi were identified and their proteins were analyzed by 2-DE and MALDI-TOF/TOF. Tropomyosin, calponin and de novo peptides were identified by 2-DE and MALDI-TOF/TOF in F8 and immunization with F8 conferred most significant protection against L3-initiated infection in Mastomys coucha. Immunized animals showed upregulated F8-induced NO, IFN-γ, TNF-α, IL-1β, IL-10, TGF-β release, cellular proliferative responses and specific IgG and IgG1. Anti-IFN-γ, anti-TNF-α, and anti-IL-1β significantly reduced F8-mediated NO generation and iNOS induction at protein levels. Anti-IFN-γ treated cells showed maximum reduction (>74%) in NO generation suggesting a predominant role of IFN-γ in iNOS induction. In conclusion, the findings suggest that F8 which contains tropomyosin, calponin and de novo peptides protects the host via IFN-γ mediated iNOS induction and may hold promise as vaccine candidate(s). This is also the first report of identification of tropomyosin and calponin in B. malayi.
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, New Campus, BS 10/1, Sector 10, Jankipuram Extension, Lucknow 226031, India.
authors.forename: Shiv K
authors.initials: SK
authors.lastname: Verma
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, New Campus, BS 10/1, Sector 10, Jankipuram Extension, Lucknow 226031, India.
authors.forename: Sujith K
authors.initials: SK
authors.lastname: Joseph
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, New Campus, BS 10/1, Sector 10, Jankipuram Extension, Lucknow 226031, India.
authors.forename: Richa
authors.initials: R
authors.lastname: Verma
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, New Campus, BS 10/1, Sector 10, Jankipuram Extension, Lucknow 226031, India.
authors.forename: Vikas
authors.initials: V
authors.lastname: Kushwaha
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, New Campus, BS 10/1, Sector 10, Jankipuram Extension, Lucknow 226031, India.
authors.forename: Naveen
authors.initials: N
authors.lastname: Parmar
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, New Campus, BS 10/1, Sector 10, Jankipuram Extension, Lucknow 226031, India.
authors.forename: Pawan K
authors.initials: PK
authors.lastname: Yadav
authors.affiliation: Mass Spectrometry Laboratory, Sophisticated Analytical Instrument Facility, CSIR-Central Drug Research Institute, New Campus, BS 10/1, Sector 10, Jankipuram Extension, Lucknow 226031, India.
authors.forename: Jagadeshwar Reddy
authors.initials: JR
authors.lastname: Thota
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, New Campus, BS 10/1, Sector 10, Jankipuram Extension, Lucknow 226031, India.
authors.forename: Susanta
authors.initials: S
authors.lastname: Kar
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, New Campus, BS 10/1, Sector 10, Jankipuram Extension, Lucknow 226031, India. Electronic address: drpkmurthy@yahoo.com.
authors.forename: P Kalpana
authors.initials: PK
authors.lastname: Murthy
mesh_terms: D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D000947:Antigens, Helminth / Q000008:administration & dosage* / Q000737:chemistry / Q000302:isolation & purification; D017178:Brugia malayi / Q000737:chemistry / Q000276:immunology*; D049109:Cell Proliferation; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D020536:Enzyme Activators / Q000008:administration & dosage* / Q000737:chemistry / Q000302:isolation & purification; D007074:Immunoglobulin G / Q000097:blood; D007371:Interferon-gamma / Q000378:metabolism*; D007378:Interleukins / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008297:Male; D008970:Molecular Weight; D051189:Murinae; D009569:Nitric Oxide / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D016212:Transforming Growth Factor beta / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D014611:Vaccination / Q000379:methods
chemical_list: D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D020536:Enzyme Activators; D007074:Immunoglobulin G; D007378:Interleukins; D016212:Transforming Growth Factor beta; D014409:Tumor Necrosis Factor-alpha; D009569:Nitric Oxide; D007371:Interferon-gamma; D052247:Nitric Oxide Synthase Type II
keywords: 2-DE; Brugia malayi; Cytokines; IgG and its subclasses; LTT assay; MALDI-MS; NO stimulating molecules; iNOS