Protection against filarial infection by 45-49 kDa molecules of Brugia malayi via IFN-γ-mediated iNOS induction

Shiv K Verma; Sujith K Joseph; Richa Verma; Vikas Kushwaha; Naveen Parmar; Pawan K Yadav; Jagadeshwar Reddy Thota; Susanta Kar; P Kalpana Murthy

doi:10.1016/j.vaccine.2014.11.041

Protection against filarial infection by 45-49 kDa molecules of Brugia malayi via IFN-γ-mediated iNOS induction

Vaccine. 2015 Jan 15;33(4):527-34. doi: 10.1016/j.vaccine.2014.11.041. Epub 2014 Nov 30.

Authors

Shiv K Verma¹, Sujith K Joseph¹, Richa Verma¹, Vikas Kushwaha¹, Naveen Parmar¹, Pawan K Yadav¹, Jagadeshwar Reddy Thota², Susanta Kar¹, P Kalpana Murthy³

Affiliations

¹ Division of Parasitology, CSIR-Central Drug Research Institute, New Campus, BS 10/1, Sector 10, Jankipuram Extension, Lucknow 226031, India.
² Mass Spectrometry Laboratory, Sophisticated Analytical Instrument Facility, CSIR-Central Drug Research Institute, New Campus, BS 10/1, Sector 10, Jankipuram Extension, Lucknow 226031, India.
³ Division of Parasitology, CSIR-Central Drug Research Institute, New Campus, BS 10/1, Sector 10, Jankipuram Extension, Lucknow 226031, India. Electronic address: [email protected].

PMID: 25454090
DOI: 10.1016/j.vaccine.2014.11.041

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.

Keywords: 2-DE; Brugia malayi; Cytokines; IgG and its subclasses; LTT assay; MALDI-MS; NO stimulating molecules; iNOS.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Antibodies, Helminth / blood
Antigens, Helminth / administration & dosage*
Antigens, Helminth / chemistry
Antigens, Helminth / isolation & purification
Brugia malayi / chemistry
Brugia malayi / immunology*
Cell Proliferation
Electrophoresis, Gel, Two-Dimensional
Electrophoresis, Polyacrylamide Gel
Enzyme Activators / administration & dosage*
Enzyme Activators / chemistry
Enzyme Activators / isolation & purification
Immunoglobulin G / blood
Interferon-gamma / metabolism*
Interleukins / metabolism
Leukocytes, Mononuclear / immunology
Male
Molecular Weight
Murinae
Nitric Oxide / metabolism
Nitric Oxide Synthase Type II / metabolism*
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Transforming Growth Factor beta / metabolism
Tumor Necrosis Factor-alpha / metabolism
Vaccination / methods

Substances

Antibodies, Helminth
Antigens, Helminth
Enzyme Activators
Immunoglobulin G
Interleukins
Transforming Growth Factor beta
Tumor Necrosis Factor-alpha
Nitric Oxide
Interferon-gamma
Nitric Oxide Synthase Type II