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CBD-cellulose : a novel adjuvant system Tekant, Bahar

Abstract

Glycoprotein D gene of Herpes Simplex Virus Type 1 (gD.1) was cloned in Escherichia coli as a gene fusion to the endoglucanase A cellulose binding domain (CBD) gene of Cellulomonas fimi. The initial construct produced a form of CBDCenA-gD.1 fusion protein which severely inhibited the growth of E. coli. The inhibitory effect was attributed to the presence of the transmembrane region of the gD.1 protein. A cassette plasmid was constructed to express CBDCenA fused in frame to any other gene. A truncated gD.1 gene lacking the coding regions for the leader peptide and the transmembrane/cytoplasmic domains was inserted into this cassette, and the synthesis of 45.9 kDa fusion protein was shown. The fusion protein was severely degraded, and also, associated with the cell membranes. CBDCenA-PhoA, a chimeric protein containing alkaline phosphatase, was used to investigate the potential adjuvant effect of immunization with a CBD-fusion protein bound to cellulose. Immunogens tested included alkaline phosphatase alone, CBDCenA-PhoA alone, alkaline phosphatase in Avicel (micro crystalline cellulose) solution, CBDCenA-Pho A-Alum (aluminum hydroxide) and CBDCenA-PhoA-Avicel. Mice immunized with CBDCenA-PhoA-Avicel showed the highest anti-alkaline phosphatase and anti-CenA antibody titers. Animals immunized with alkaline phosphatase-Avicel, CBDCenA-PhoA-Alum, CBDCenA-PhoA or alkaline phosphatase alone showed mean secondary anti-alkaline phosphatase antibody titers that were 29%, 26%, 17% and 17% of the CBDCenAPhoA-Avicel titers, respectively. Animals immunized with CBDCenA-PhoA-Alum or CBDCenA-PhoA alone showed mean secondary anti-CenA antibody titers that were 57% and 42% of the CBDCenA-PhoA-Avicel titers, respectively. The results indicate that the use of CBD fusions to present an antigen bound to cellulose provides an effective adjuvant for antibody responses.

Item Metadata

Title
CBD-cellulose : a novel adjuvant system
Creator
Tekant, Bahar
Publisher
University of British Columbia
Date Issued
1991
Description
Glycoprotein D gene of Herpes Simplex Virus Type 1 (gD.1) was cloned in Escherichia coli as a gene fusion to the endoglucanase A cellulose binding domain (CBD) gene of Cellulomonas fimi. The initial construct produced a form of CBDCenA-gD.1 fusion protein which severely inhibited the growth of E. coli. The inhibitory effect was attributed to the presence of the transmembrane region of the gD.1 protein. A cassette plasmid was constructed to express CBDCenA fused in frame to any other gene. A truncated gD.1 gene lacking the coding regions for the leader peptide and the transmembrane/cytoplasmic domains was inserted into this cassette, and the synthesis of 45.9 kDa fusion protein was shown. The fusion protein was severely degraded, and also, associated with the cell membranes. CBDCenA-PhoA, a chimeric protein containing alkaline phosphatase, was used to investigate the potential adjuvant effect of immunization with a CBD-fusion protein bound to cellulose. Immunogens tested included alkaline phosphatase alone, CBDCenA-PhoA alone, alkaline phosphatase in Avicel (micro crystalline cellulose) solution, CBDCenA-Pho A-Alum (aluminum hydroxide) and CBDCenA-PhoA-Avicel. Mice immunized with CBDCenA-PhoA-Avicel showed the highest anti-alkaline phosphatase and anti-CenA antibody titers. Animals immunized with alkaline phosphatase-Avicel, CBDCenA-PhoA-Alum, CBDCenA-PhoA or alkaline phosphatase alone showed mean secondary anti-alkaline phosphatase antibody titers that were 29%, 26%, 17% and 17% of the CBDCenAPhoA-Avicel titers, respectively. Animals immunized with CBDCenA-PhoA-Alum or CBDCenA-PhoA alone showed mean secondary anti-CenA antibody titers that were 57% and 42% of the CBDCenA-PhoA-Avicel titers, respectively. The results indicate that the use of CBD fusions to present an antigen bound to cellulose provides an effective adjuvant for antibody responses.
Extent
5795520 bytes
Genre
Thesis/Dissertation
Type
Text
File Format
application/pdf
Language
eng
Date Available
2008-09-09
Provider
Vancouver : University of British Columbia Library
Rights
For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use.
DOI
10.14288/1.0086481
URI
http://hdl.handle.net/2429/1732
Degree
Master of Science - MSc
Program
Microbiology and Immunology
Affiliation
Science, Faculty of; Microbiology and Immunology, Department of
Degree Grantor
University of British Columbia
Graduation Date
1992-05
Campus
UBCV
Scholarly Level
Graduate
Aggregated Source Repository
DSpace

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For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use.