Immunoprotection of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from Lactococcus garvieae against Lactococcosis in tilapia

• Published in: Journal of general and applied microbiology Vol. 59; no. 6; pp. 437 - 449
• Main Authors: Tsai, Ming-An, Wang, Pei-Chi, Cao, Thanh-Trung, Liao, Pei-Chih, Liaw, Li-Ling, Chen, Shih-Chu
• Format: Journal Article
• Language: English
• Published: Japan Applied Microbiology, Molecular and Cellular Biosciences Research Foundation 01.01.2013; Japan Science and Technology Agency
• Subjects: Animals; Antibodies, Bacterial - blood; Bacterial Proteins - genetics; Bacterial Proteins - immunology; Bacterial Proteins - isolation & purification; Escherichia coli - enzymology; Escherichia coli - genetics; Fish Diseases - microbiology; Fish Diseases - prevention & control; Gene Expression; glyceraldehyde-3-phosphate dehydrogenase (GAPDH); Glyceraldehyde-3-Phosphate Dehydrogenases - genetics; Glyceraldehyde-3-Phosphate Dehydrogenases - immunology; Glyceraldehyde-3-Phosphate Dehydrogenases - isolation & purification; Immunization; immunoprotection; Lactococcus - enzymology; Lactococcus - genetics; Lactococcus - immunology; Lactococcus garvieae; Rabbits; Recombinant Proteins; Streptococcus; Tilapia - microbiology
• ISSN: 0022-1260; 1349-8037
• DOI: 10.2323/jgam.59.437

In this study, the gene encoding 40 kDa GAPDH of L. garvieae was determined and overexpressed by using the Escherichia coli expression system. Analysis results indicated that the sequences of GAPDH of L. garvieae nucleotide and its amino acid are highly homologous (80.4−100%) to several products of GAPDH from L. garvieae and other Streptococcus-related bacteria. According to Western blotting results, rabbit antiserum and tilapia infection serum reacted strongly to the recombinant GAPDH protein. In another experiment, tilapia were immunized intraperitoneally with formalin-killed L. garvieae whole cells, recombinant GAPDH (50 μg fish−1) from L. garvieae or both. ISA 763A was used as an adjuvant for vaccine and saline was used as a negative control. The fish challenged at 4 weeks after immunization with GAPDH+WC+ISA had the highest survival rate at 100%, followed by fish immunized with WC+ISA or GAPDH+ISA, which had RPS values of 87.5% and 50%, respectively. Additionally, specific antibody responses against L. garvieae whole cells and GAPDH were based on enzyme-linked immunosorbent assay. Following 4 weeks of immunization, the specific antibody level of all vaccine groups significantly increased, except for antibody responses against L. garvieae GAPDH of those immunized with formalin-killed L. garvieae whole cells. Our results further demonstrated that GAPDH from L. garvieae protected tilapia from experimental L. garvieae infection, implying the potential use of L. garvieae GAPDH as a vaccine against L. garvieae.