Immunolocalization and serum antibody responses to Brugia malayi pepsin inhibitor homolog (Bm-33).
cDNA coding for Brugia malayi pepsin inhibitor homolog (Bm-33) from the human filarial parasite was cloned in pRSET for large-scale expression and functional characterization. The pRSET-B cloned gene did not yield recombinant protein expression and the reason was attributed to the presence of an N-terminal signal peptide. The gene was subcloned in pRSET-A without a signal peptide and the 33 kDa histidine-tagged recombinant protein was purified by IMAC. All individuals from an endemic area generated IgG responses against Bm-33 in the order MF>CP>EN. Isotype analysis indicated an elevated IgG4 reactivity in the order MF>EN>CP. Bm-33-specific IgE levels were elevated in MF, CP and EN compared to non-endemic normals with no significant differences among the groups. Paraffin-embedded sections of Setaria digitata (cattle filarial parasite) stained with mouse anti-Bm-33 antibodies exhibited the hypodermal nature of Bm-33. These findings suggest that Bm-33 is an immunodominant antigen and contributes to filarial pathogenesis.
Krushna
Nagampalli S A
NS
Centre for Biotechnology, Anna University, Chennai-600025, Tamil Nadu, India.
Shiny
Chandanpurath
C
Dharanya
Srinivasan
S
Sindhu
Arivazhagan
A
Aishwarya
Sridharan
S
Narayanan
Rangarajan B
RB
D000293:Adolescent; D000328:Adult; D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood*; D000947:Antigens, Helminth / Q000032:analysis / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification*; D001483:Base Sequence; D017178:Brugia malayi / Q000737:chemistry* / Q000276:immunology*; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000302:isolation & purification; D020405:Dermis / Q000737:chemistry; D005260:Female; D005370:Filarioidea / Q000737:chemistry; D015801:Helminth Proteins / Q000032:analysis* / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007073:Immunoglobulin E / Q000032:analysis; D007074:Immunoglobulin G / Q000097:blood; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification; D016415:Sequence Alignment; D055815:Young Adult
D000909:Antibodies, Helminth; D000947:Antigens, Helminth; C540446:Bm-33 protein, Brugia malayi; D018076:DNA, Complementary; D015801:Helminth Proteins; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D007073:Immunoglobulin E