Expression and characterization of a multivalent human respiratory syncytial virus protein.
Respiratory syncytial virus (RSV) has been recognized as one of the most common causes of severe respiratory tract infection in infants worldwide. As yet, a safe and effective vaccine has not been developed to protect humans from RSV. The F and G surface proteins have been widely investigated due to their potential to induce protective immunity. In addition, the M2 protein has been shown to be important in inducing a T-cell response. Our project involved the cloning of the immunodominant regions of the RSV F, M2 and G proteins into a bacterial vector, pET-32a (+). The recombinant RFM2G protein was expressed in Escherichia coli and purified using His Bind columns. The purified rRFM2G protein was analyzed by polyacrylamide gel electrophoresis and Western blotting. The predicted structure of the recombinant protein built by the Swiss PDB Viewer program suggested a rod shape with a distinct swollen head and neck which was confirmed by transmission electron microscopy and atomic force microscopy. BALB/c female mice were immunized with either RSV, rRFM2G alone, or rRFM2G in combination with flagellin as a mucosal adjuvant. Serum was collected on days 0, 14, 28 and 49 to assess the immune response by Enzyme-linked immunosorbent assay. Intranasal immunization of mice with the rRFM2G protein yielded significantly high serum IgG titers. Co-administration of the rRFM2G protein with flagellin did not augment the serum antibody response.
Subbarayan
P
P
Center for NanoBiotechnology Research, Alabama State University, 915 South Jackson Street, Montgomery, AL 36101, USA.
Qin
H
H
Pillai
S
S
Lee
J J
JJ
Pfendt
A P
AP
Willing
G
G
Miller
M E
ME
Dennis
V A
VA
Singh
S R
SR
D000276:Adjuvants, Immunologic; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood / Q000276:immunology; D000917:Antibody Formation / Q000187:drug effects / Q000276:immunology; D000939:Epitopes / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000494:pharmacology; D004926:Escherichia coli; D005260:Female; D005408:Flagellin / Q000276:immunology / Q000494:pharmacology; D015870:Gene Expression; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology / Q000494:pharmacology; D012136:Respiratory Syncytial Viruses; D014764:Viral Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000494:pharmacology
D000276:Adjuvants, Immunologic; D000914:Antibodies, Viral; D000939:Epitopes; D007074:Immunoglobulin G; C507355:M2-2 protein, respiratory syncytial virus; D011994:Recombinant Proteins; D014764:Viral Proteins; D005408:Flagellin