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Presumable role of outer membrane proteins of Salmonella containing sialylated lipopolysaccharides serovar Ngozi, sv. Isaszeg and subspecies arizonae in determining susceptibility to human serum.

Futoma-Kołoch B, Godlewska U, Guz-Regner K, Dorotkiewicz-Jach A, Klausa E, Rybka J, Bugla-Płoskońska G - Gut Pathog (2015)

Bottom Line: Weaker C3 deposition ratio on the Salmonella sv.Immunoblotting revealed that OMP isolated from the tested strains also bound C3 protein fragments.We suggest that activation of C3 serum protein is dependent on the sialic acid contents in the LPS as well as on the presence of OMP in the range of molecular masses of 35-48 kDa.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Institute of Genetics and Microbiology, University of Wrocław, Przybyszewskiego 63-77, 51-148 Wrocław, Poland.

ABSTRACT

Background: The O48 group comprises Salmonella bacteria containing sialic acid in the lipopolysaccharide (LPS). Bacteria with sialylated surface structures are described as pathogens that avoid immunological response of the host by making similar their surface antigens to the host's tissues (molecular mimicry). It is known that the smooth-type LPS of Salmonella enterica and outer membrane proteins (OMP) PgtE, PagC and Rck mediate serum resistant phenotype by affecting complement system (C). The aim of this study was to investigate C3 component activation by Salmonella O48 LPS and OMP.

Findings: In the present study, we examined C3 component deposition on the three Salmonella O48 strains: S. enterica subspecies enterica serovar Ngozi, S. enterica subsp. enterica sv. Isaszeg, and S. enterica subsp. arizonae containing sialic acid in the O-specific part of LPS. The greatest C3 deposition occurred on Salmonella sv. Isaszeg cells (p < 0.005) as well as on their LPS (low content of sialic acid in LPS) (p < 0.05) after 45 min of incubation in 50% human serum. Weaker C3 deposition ratio on the Salmonella sv. Ngozi (high content of sialic acid in LPS) and Salmonella subsp. arizonae (high content of sialic acid in LPS) cells correlated with the lower C3 activation on their LPS. Immunoblotting revealed that OMP isolated from the tested strains also bound C3 protein fragments.

Conclusions: We suggest that activation of C3 serum protein is dependent on the sialic acid contents in the LPS as well as on the presence of OMP in the range of molecular masses of 35-48 kDa.

No MeSH data available.


Related in: MedlinePlus

C3 complement protein depositions on immobilized LPS. Direct Sandwich ELISA. Microtiter plate wells were coated for 2 h at 37°C with polyclonal rabbit anti-C3c diluted 1/500 in 0.1 M sodium carbonate buffer (pH 9.6). Mixtures of LPS (0.6; 6.0; 60.0 μg/ml) and 80% HS were incubated for 45 min at 37°C. Activation of C3 differs significantly if the p values are less than p < 0.05 (*).
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Fig4: C3 complement protein depositions on immobilized LPS. Direct Sandwich ELISA. Microtiter plate wells were coated for 2 h at 37°C with polyclonal rabbit anti-C3c diluted 1/500 in 0.1 M sodium carbonate buffer (pH 9.6). Mixtures of LPS (0.6; 6.0; 60.0 μg/ml) and 80% HS were incubated for 45 min at 37°C. Activation of C3 differs significantly if the p values are less than p < 0.05 (*).

Mentions: The results obtained for the surface antigens indicated that both LPS (direct sandwich ELISA) and OMP (immunoblots) antigens bound C3. It was observed C3 component activation occurred at a similar rate for LPS isolated from Salmonella at 15 min (p > 0.05) but different after 45 min of incubation (Figure 4) with the highest A492 for Salmonella sv. Isaszeg (p < 0.05). The low absorbance (A492 = 0.4) was noted for control sets containing HS or PBS. Considering OMP, C3 binding was obtained when OMP were electrophoresed in nonreducing conditions (BN-PAGE) (Figure 5). SDS–PAGE produced additional information that efficient C3 fixation occurred on the OMP bands of the molecular masses in the range of 35–48 kDa (Figure 6).Figure 4


Presumable role of outer membrane proteins of Salmonella containing sialylated lipopolysaccharides serovar Ngozi, sv. Isaszeg and subspecies arizonae in determining susceptibility to human serum.

Futoma-Kołoch B, Godlewska U, Guz-Regner K, Dorotkiewicz-Jach A, Klausa E, Rybka J, Bugla-Płoskońska G - Gut Pathog (2015)

C3 complement protein depositions on immobilized LPS. Direct Sandwich ELISA. Microtiter plate wells were coated for 2 h at 37°C with polyclonal rabbit anti-C3c diluted 1/500 in 0.1 M sodium carbonate buffer (pH 9.6). Mixtures of LPS (0.6; 6.0; 60.0 μg/ml) and 80% HS were incubated for 45 min at 37°C. Activation of C3 differs significantly if the p values are less than p < 0.05 (*).
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4504086&req=5

Fig4: C3 complement protein depositions on immobilized LPS. Direct Sandwich ELISA. Microtiter plate wells were coated for 2 h at 37°C with polyclonal rabbit anti-C3c diluted 1/500 in 0.1 M sodium carbonate buffer (pH 9.6). Mixtures of LPS (0.6; 6.0; 60.0 μg/ml) and 80% HS were incubated for 45 min at 37°C. Activation of C3 differs significantly if the p values are less than p < 0.05 (*).
Mentions: The results obtained for the surface antigens indicated that both LPS (direct sandwich ELISA) and OMP (immunoblots) antigens bound C3. It was observed C3 component activation occurred at a similar rate for LPS isolated from Salmonella at 15 min (p > 0.05) but different after 45 min of incubation (Figure 4) with the highest A492 for Salmonella sv. Isaszeg (p < 0.05). The low absorbance (A492 = 0.4) was noted for control sets containing HS or PBS. Considering OMP, C3 binding was obtained when OMP were electrophoresed in nonreducing conditions (BN-PAGE) (Figure 5). SDS–PAGE produced additional information that efficient C3 fixation occurred on the OMP bands of the molecular masses in the range of 35–48 kDa (Figure 6).Figure 4

Bottom Line: Weaker C3 deposition ratio on the Salmonella sv.Immunoblotting revealed that OMP isolated from the tested strains also bound C3 protein fragments.We suggest that activation of C3 serum protein is dependent on the sialic acid contents in the LPS as well as on the presence of OMP in the range of molecular masses of 35-48 kDa.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Institute of Genetics and Microbiology, University of Wrocław, Przybyszewskiego 63-77, 51-148 Wrocław, Poland.

ABSTRACT

Background: The O48 group comprises Salmonella bacteria containing sialic acid in the lipopolysaccharide (LPS). Bacteria with sialylated surface structures are described as pathogens that avoid immunological response of the host by making similar their surface antigens to the host's tissues (molecular mimicry). It is known that the smooth-type LPS of Salmonella enterica and outer membrane proteins (OMP) PgtE, PagC and Rck mediate serum resistant phenotype by affecting complement system (C). The aim of this study was to investigate C3 component activation by Salmonella O48 LPS and OMP.

Findings: In the present study, we examined C3 component deposition on the three Salmonella O48 strains: S. enterica subspecies enterica serovar Ngozi, S. enterica subsp. enterica sv. Isaszeg, and S. enterica subsp. arizonae containing sialic acid in the O-specific part of LPS. The greatest C3 deposition occurred on Salmonella sv. Isaszeg cells (p < 0.005) as well as on their LPS (low content of sialic acid in LPS) (p < 0.05) after 45 min of incubation in 50% human serum. Weaker C3 deposition ratio on the Salmonella sv. Ngozi (high content of sialic acid in LPS) and Salmonella subsp. arizonae (high content of sialic acid in LPS) cells correlated with the lower C3 activation on their LPS. Immunoblotting revealed that OMP isolated from the tested strains also bound C3 protein fragments.

Conclusions: We suggest that activation of C3 serum protein is dependent on the sialic acid contents in the LPS as well as on the presence of OMP in the range of molecular masses of 35-48 kDa.

No MeSH data available.


Related in: MedlinePlus