Use of in vivo-induced antigen technology (IVIAT) for the identification of Streptococcus suis serotype 2 in vivo-induced bacterial protein antigens.
BACKGROUND
Streptococcus suis serotype 2 (SS2) is a zoonotic agent that causes death and disease in both humans and swine. A better understanding of SS2-host molecular interactions is crucial for understanding SS2 pathogenesis and immunology. Conventional genetic and biochemical approaches used to study SS2 virulence factors are unable to take into account the complex and dynamic environmental stimuli associated with the infection process. In this study, in vivo-induced antigen technology (IVIAT), an immunoscreening technique, was used to identify the immunogenic bacterial proteins that are induced or upregulated in vivo during SS2 infection.


RESULTS
Convalescent-phase sera from pigs infected with SS2 were pooled, adsorbed against in vitro antigens, and used to screen SS2 genomic expression libraries. Upon analysis of the identified proteins, we were able to assign a putative function to 40 of the 48 proteins. These included proteins implicated in cell envelope structure, regulation, molecule synthesis, substance and energy metabolism, transport, translation, and those with unknown functions. The in vivo-induced changes in the expression of 10 of these 40 genes were measured using real-time reverse transcription (RT)-PCR, revealing that the expression of 6 of the 10 genes was upregulated in the in vivo condition. The strain distribution of these 10 genes was analyzed by PCR, and they were found in the most virulent SS2 strains. In addition, protein sequence alignments of the newly identified proteins demonstrate that three are putative virulence-associated proteins.


CONCLUSIONS
Collectively, our results suggest that these in vivo-induced or upregulated genes may contribute to SS2 disease development. We hypothesize that the identification of factors specifically induced or upregulated during SS2 infection will aid in our understanding of SS2 pathogenesis and may contribute to the control SS2 outbreaks. In addition, the proteins identified using IVIAT may be useful potential vaccine candidates or virulence markers.
Gu
Hongwei
H
Key Lab Animal Disease Diagnostic & Immunology, Ministry of Agriculture, Nanjing Agricultural University, Nanjing, PR China. hongweigu999@yahoo.com.cn
Zhu
Haodan
H
Lu
Chengping
C
D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D000942:Antigens, Bacterial / Q000032:analysis*; D019295:Computational Biology; D004797:Enzyme-Linked Immunosorbent Assay; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D015698:Genomic Library; D012329:RNA, Bacterial / Q000235:genetics; D013290:Streptococcal Infections / Q000382:microbiology / Q000662:veterinary*; D017011:Streptococcus suis / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D013552:Swine / Q000382:microbiology; D013553:Swine Diseases / Q000382:microbiology*; D013556:Swine, Miniature / Q000382:microbiology
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D012329:RNA, Bacterial