Additional file 5 of Immunogenicity and protective efficacy of a Streptococcus suis vaccine composed of six conserved immunogens

Additional file 5: SDS-PAGE and Western blot analysis of antigens included in the multicomponent vaccine. The purified proteins were run on SDS–polyacrylamide gel (A), transferred to membranes and probed with antisera raised in rabbits against each antigen (B-G). Numbers on the left are molecular masses in kDa

Additional file 3 of Immunogenicity and protective efficacy of a Streptococcus suis vaccine composed of six conserved immunogens

Additional file 3: Origin and activity of hyperimmune sera in opsonophagocytosis assays

Additional file 6 of Immunogenicity and protective efficacy of a Streptococcus suis vaccine composed of six conserved immunogens

Additional file 6: Scoring of fibrinosuppurative lesions of piglets challenged with S. suis cps14. Fourteen days after prime-booster immunization with multicomponent vaccine or placebo, sixteen growing piglets (n = 8 per group) were infected intranasally with 5 × 109 CFU of S. suis cps14 V3117/2. Five vaccinated animals and four placebo animals demonstrated clinical signs of severe disease. In both groups two piglets had to be euthanized for animal welfare reasons because of signs of polyarthritis. One vaccinated animal showed additional signs of central nervous system dysfunction (opisthotonus, generalized tremor, ataxia) and had to be euthanized as well. Surviving piglets were sacrificed fourteen days post-infection. Necropsies and histopathological screenings of the indicated tissues were conducted with all 16 piglets as described previously [10]

Additional file 2 of Immunogenicity and protective efficacy of a Streptococcus suis vaccine composed of six conserved immunogens

Additional file 2: Origin and bactericidal activity of sera of susceptible piglets, sera pre-infection and sera post-infection. Information on the original experimental infection and results of bactericidal as well as opsonophagocytosis assays of sera defined as sera of susceptible pigs, sera pre-infection and sera post-infection

Additional file 1 of Immunogenicity and protective efficacy of a Streptococcus suis vaccine composed of six conserved immunogens

Additional file 1: Oligonucleotides used for recombinant expression of S. suis antigens in E. coli. Name and sequences (5’-3’) of oligonucleotide primers used for construction of expression vectors of recombinant proteins included in the multicomponent vaccine (SSU0934, SSU1869, SSU0757, SSU1950, SSU1664, SSU0187)

Additional file 4 of Immunogenicity and protective efficacy of a Streptococcus suis vaccine composed of six conserved immunogens

Additional file 4: Multiple sequence alignment (tblastn) of antigens in different Streptococcus suis strains. Multiple sequence alignments of each antigen were conducted using sequences retrieved from the genome sequences of S. suis strains 10, 13-00283-02 and 16085/3b. As query served the respective sequence of strain P1/7. Dots represent identities, whereas differences are highlighted in red

Trapping and Proteomic Identification of Cellular Substrates of the ClpP Protease in <i>Staphylococcus aureus</i>

In the important human pathogen Staphylococcus aureus the cytoplasmic ClpP protease is essential for mounting cellular stress responses and for virulence. To directly identify substrates of the ClpP protease, we expressed in vivo a proteolytic inactive form of ClpP (ClpPtrap) that will retain but not degrade substrates translocated into its proteolytic chamber. Substrates captured inside the proteolytic barrel were co-purified along with the His-tagged ClpP complex and identified by mass spectrometry. In total, approximately 70 proteins were trapped in both of the two S. aureus strains NCTC8325-4 and Newman. About one-third of the trapped proteins are previously shown to be unstable or to be substrates of ClpP in other bacteria, supporting the validity of the ClpP-TRAP. This group of proteins encompassed the transcriptional regulators CtsR and Spx, the ClpC adaptor proteins McsB and MecA, and the cell division protein FtsZ. Newly identified ClpP substrates include the global transcriptional regulators PerR and HrcA, proteins involved in DNA damage repair (RecA, UvrA, UvrB), and proteins essential for protein synthesis (RpoB and Tuf). Our study hence underscores the central role of Clp-proteolysis in a number of pathways that contribute to the success of S. aureus as a human pathogen

Trapping and Proteomic Identification of Cellular Substrates of the ClpP Protease in <i>Staphylococcus aureus</i>

In the important human pathogen <i>Staphylococcus aureus</i> the cytoplasmic ClpP protease is essential for mounting cellular stress responses and for virulence. To directly identify substrates of the ClpP protease, we expressed in vivo a proteolytic inactive form of ClpP (ClpP^trap) that will retain but not degrade substrates translocated into its proteolytic chamber. Substrates captured inside the proteolytic barrel were co-purified along with the His-tagged ClpP complex and identified by mass spectrometry. In total, approximately 70 proteins were trapped in both of the two <i>S. aureus</i> strains NCTC8325-4 and Newman. About one-third of the trapped proteins are previously shown to be unstable or to be substrates of ClpP in other bacteria, supporting the validity of the ClpP-TRAP. This group of proteins encompassed the transcriptional regulators CtsR and Spx, the ClpC adaptor proteins McsB and MecA, and the cell division protein FtsZ. Newly identified ClpP substrates include the global transcriptional regulators PerR and HrcA, proteins involved in DNA damage repair (RecA, UvrA, UvrB), and proteins essential for protein synthesis (RpoB and Tuf). Our study hence underscores the central role of Clp-proteolysis in a number of pathways that contribute to the success of <i>S. aureus</i> as a human pathogen

DataSheet_4_Toxin exposure and HLA alleles determine serum antibody binding to toxic shock syndrome toxin 1 (TSST-1) of Staphylococcus aureus.pdf

Life-threatening toxic shock syndrome is often caused by the superantigen toxic shock syndrome toxin-1 (TSST-1) produced by Staphylococcus aureus. A well-known risk factor is the lack of neutralizing antibodies. To identify determinants of the anti-TSST-1 antibody response, we examined 976 participants of the German population-based epidemiological Study of Health in Pomerania (SHIP-TREND-0). We measured anti-TSST-1 antibody levels, analyzed the colonization with TSST-1-encoding S. aureus strains, and performed a genome-wide association analysis of genetic risk factors. TSST-1-specific serum IgG levels varied over a range of 4.2 logs and were elevated by a factor of 12.3 upon nasal colonization with TSST-1-encoding S. aureus. Moreover, the anti-TSST-1 antibody levels were strongly associated with HLA class II gene loci. HLA-DRB1*03:01 and HLA-DQB1*02:01 were positively, and HLA-DRB1*01:01 as well as HLA-DQB1*05:01 negatively associated with the anti-TSST-1 antibody levels. Thus, both toxin exposure and HLA alleles affect the human antibody response to TSST-1.</p

DataSheet_1_Toxin exposure and HLA alleles determine serum antibody binding to toxic shock syndrome toxin 1 (TSST-1) of Staphylococcus aureus.pdf

Life-threatening toxic shock syndrome is often caused by the superantigen toxic shock syndrome toxin-1 (TSST-1) produced by Staphylococcus aureus. A well-known risk factor is the lack of neutralizing antibodies. To identify determinants of the anti-TSST-1 antibody response, we examined 976 participants of the German population-based epidemiological Study of Health in Pomerania (SHIP-TREND-0). We measured anti-TSST-1 antibody levels, analyzed the colonization with TSST-1-encoding S. aureus strains, and performed a genome-wide association analysis of genetic risk factors. TSST-1-specific serum IgG levels varied over a range of 4.2 logs and were elevated by a factor of 12.3 upon nasal colonization with TSST-1-encoding S. aureus. Moreover, the anti-TSST-1 antibody levels were strongly associated with HLA class II gene loci. HLA-DRB1*03:01 and HLA-DQB1*02:01 were positively, and HLA-DRB1*01:01 as well as HLA-DQB1*05:01 negatively associated with the anti-TSST-1 antibody levels. Thus, both toxin exposure and HLA alleles affect the human antibody response to TSST-1.</p