title: Immunological characterization of recombinant Wuchereria bancrofti cuticular collagen (COL-4) as putative vaccine candidate for human lymphatic filariasis.
abstract: OBJECTIVE
  To elucidate immunoprophylactic potential of recombinant Wuchereria bancrofti (W. bancrofti) cuticular collagen (COL-4) in BALB/c mice and filarial clinical samples.
  
  
  METHODS
  col-4 gene was PCR amplified from W. bancrofti L3 cDNA library and cloned in pRSET B vector. Recombinant COL-4 was over expressed in salt inducible system and was purified by nickel affinity chromatography. Humoral and cellular responses were measured by ELISA and peripheral blood mononuclear cells (PBMC) of various filarial clinical samples respectively using purified recombinant COL-4 antigen. Then the protective immune responses of COL-4 immunized BALB/c mice were characterized.
  
  
  RESULTS
  Sequence analysis of COL-4 with human host proteins reveals lack of homology. The recombinant COL-4 was found to be at 15 kDa fusion protein. The affinity purified COL-4 showed significant reactivity with putatively immune sera and in a similar fashion it demonstrated marked proliferation in PBMC samples. Immunization studies in experimental filarial host (mice) elicited significant titers with protective antibody isotype profile (IgM and IgG). Cellular immune responses were also significant in terms of splenocytes proliferation assay on mice samples.
  
  
  CONCLUSIONS
  Our immunological findings in experimental host suggest Th2 mediated immune response. Hence, we propose that W. bancrofti COL-4 could be an efficacious vaccine candidate against lymphatic filariasis.
authors: Centre for Biotechnology, Anna University, Chennai - 600025, India.
authors: Chakkaravarthy
authors: C
authors: Arunkumar
authors: Centre for Biotechnology, Anna University, Chennai - 600025, India.
authors: Pandurangan
authors: P
authors: Pandiaraja
authors: Centre for Biotechnology, Anna University, Chennai - 600025, India.
authors: P R
authors: PR
authors: Prince
authors: Centre for Biotechnology, Anna University, Chennai - 600025, India. Electronic address: pk.kaliraj@gmail.com.
authors: Perumal
authors: P
authors: Kaliraj
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003094:Collagen / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D004605:Elephantiasis, Filarial / Q000517:prevention & control*; D015801:Helminth Proteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D007963:Leukocytes, Mononuclear; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D014614:Vaccines, Synthetic / Q000276:immunology*; D014958:Wuchereria bancrofti / Q000235:genetics / Q000276:immunology*
chemical_list: D000909:Antibodies, Helminth; D015801:Helminth Proteins; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; D003094:Collagen
keywords: Cuticular collagen; Lymphatic filariasis; Vaccine; Wuchereria bancrofti