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Journal of Dental Research
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A DNA Vaccine Encoding a Cell-surface Protein Antigen of Streptococcus mutans Protects Gnotobiotic Rats from Caries

M.W. Fan1,*, Z. Bian1, Z.X. Peng2, Y. Zhong1, Z. Chen1, B. Peng1 and R. Jia1

1 School of Stomatology, Wuhan University, and
2 Stomatological Center, Huazhong University of Science and Technology, Wuhan, People’s Republic of China 430079;


Figure 1
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  		Figure 1. Construction and verification of plasmid pCIA-P. (A) The A- and P-regions of PAc protein of S. mutans was cloned into eukaryotic expression vector pCI to obtain the recombinant plasmid pCIA-P. (B) Restriction enzyme digestion of pCIA-P and pCI. The sizes of pCIA-P, A-P fragment, and pCI were 6.2 kb, 2.2 kb, and 4.0 kb, respectively. Lane 1: pCI with XhoI digestion. Lane 2: pCIA-P with Bgl I digestion. Lane 3: pCIA-P with XhoI digestion. Lane 4: pCIA-P with Xho I/Sal I digestion. Lane 5: {lambda}DNA/HindIII marker.

 

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  		Figure 2. Recombinant PAc protein could be expressed in eukaryotic cells. (A) Recombinant PAc protein in the cytoplasm of ECV-304 cells transfected by pCIA-P. (B) ECV-304 cells transfected by pCI vector. Magnification: 200X.

 

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  		Figure 3. Expression of PAc protein in rats’ tissues after being immunized with pCIA-P by different routes. (A) PAc protein expressed in the sarcoplasm of parallel muscular fibers (lateral section). (B) Transverse section of A. (C) No PAc protein expression in control animals immunized with pCI vector. (D,E) PAc protein expression in both sides of a submandibular gland immunized with pCIA-P by TSG (injection side and opposite side, respectively). (F) No PAc protein in control animals whose TSG were immunized with pCI vector. Magnifications: A,B,C = 100X; D,E,F = 200X.

 

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  		Figure 4. The antibody level and caries scores of gnotobiotic rats (6 per group) immunized with various treatments. Data are expressed as means and standard deviations of antibody level and caries scores experiments. (A) The salivary and serum-specific anti-PAc antibody levels of gnotobiotic rats. {square} sera anti-PAc IgG. {blacksquare} salivary anti-PAc IgA. (B) Keyes caries scores of gnotobiotic rats. {square} Enamel lesion. {blacksquare} Dentinal slight lesion. X Dentinal moderate lesion. * p < 0.05.

 

Journal of Dental Research, Vol. 81, No. 11, 784-787 (2002)
DOI: 10.1177/154405910208101112
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