Abstract
The study describes the development of a vaccine using microcrystalline cellulose (Avicel PH-101) as a delivery carrier of recombinant protein-based antigen against erysipelas. Recombinant SpaA, surface protective protein, from a gram-positive pathogen Erysipelothrix rhusiopathiae was fused to a cellulose-binding domain (CBD) from Trichoderma harzianum endoglucanase II through a S3N10 peptide. The fusion protein (CBD-SpaA) was expressed in Escherichia coli and was subsequently bound to Avicel PH-101. The antigenicity of CBD-SpaA bound to the Avicel was evaluated by enzyme-linked immunosorbent (ELISA) and confocal laser scanning microscope (CLSM) assays. For the examination of its immunogenicity, groups of mice were immunized with different constructs (soluble CBD-SpaA, Avicel coated with CBD-SpaA, whole bacterin of E. rhusiopathiae (positive control), and PBS (negative control)). In two weeks after immunization, mice were challenged with 1x107 CFU of E. rhusiopathiae and Avicel coated with CBD-SpaA induced protective immunity in mice. In conclusion, this study demonstrates the feasibility of microcrystalline cellulose as the delivery system of recombinant protein subunit vaccine against E. rhusiopathiae infection in mice.
Highlights
Vaccines are the therapeutic formulations given to patients to elicit immune responses entailing antibody production or cell-mediated responses that will eventually fight variety of malignancies [1]
Forty SPF mice (5 weeks old) were randomly assigned to 4 groups of eight each and injected subcutaneously with 4 μg of soluble cellulose-binding domain (CBD)-surface protective antigen A (SpaA), Avicel coated with CBD-SpaA, ERT2T-A containing whole bacterin of E. rhusiopathiae serovar 15, and phosphate buffered saline (PBS) emulsified with an oil-based adjuvant, respectively
Most of the expressed CBD-SpaA were in soluble form, compared to our previous work in which extreme reduction of the solubility of E. coli-derived glutamate decarboxylase occurred after fusion with CBD [21]
Summary
Vaccines are the therapeutic formulations given to patients to elicit immune responses entailing antibody production (humoral) or cell-mediated responses that will eventually fight variety of malignancies [1]. Recombinant protein antigens, or their fragments, have been used and considered as novel vaccine candidates. Development of such kinds of vaccines avoids the safety issues with attenuating virus or cell cultures when making conventional viral vaccines [6, 7]. Some recombinant protein antigens have fatal handicaps (low stability and immunogenicity) caused by lack of key elements that can stimulate immune response [8]. Along with recombinant protein vaccines, has been suggested for improved immunogenicity [9]. Different strategies have been extensively explored in order to improve the immunogenicity by protecting antigens through immobilization on inorganic or organic matrices [10]
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