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Treponema pallidum subsp. pallidum TP0136 protein is heterogeneous among isolates and binds cellular and plasma fibronectin via its NH2-terminal end.

Ke W, Molini BJ, Lukehart SA, Giacani L - PLoS Negl Trop Dis (2015)

Bottom Line: To deepen our understanding of the TP0136-Fn interaction dynamics, we used two naturally-occurring sequence variants of the TP0136 protein to investigate which region of the protein is responsible for Fn binding, and whether TP0136 would adhere to human cellular Fn in addition to plasma Fn and super Fn as previously reported.As a complementary approach, we tested inhibition of T. pallidum binding to Fn by recombinant full-length TP0136 proteins and fragments, as well as by anti-TP0136 immune sera.Our results show that TP0136 adheres more efficiently to cellular Fn than to plasma Fn, that the TP0136 NH2-terminal conserved region of the protein is primarily responsible for binding to plasma Fn but that binding sites for cellular Fn are also present in the protein's central and COOH-terminal regions.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Division of Allergy and Infectious Diseases, University of Washington, Harborview Medical Center, Seattle, Washington, United States of America; Graduate School, Southern Medical University, Guangzhou, PR China; Division of STD, Guangdong Provincial Center for STI & Skin Diseases Control and Prevention, Guangzhou, PR China.

ABSTRACT
Adherence-mediated colonization plays an important role in pathogenesis of microbial infections, particularly those caused by extracellular pathogens responsible for systemic diseases, such as Treponema pallidum subsp. pallidum (T. pallidum), the agent of syphilis. Among T. pallidum adhesins, TP0136 is known to bind fibronectin (Fn), an important constituent of the host extracellular matrix. To deepen our understanding of the TP0136-Fn interaction dynamics, we used two naturally-occurring sequence variants of the TP0136 protein to investigate which region of the protein is responsible for Fn binding, and whether TP0136 would adhere to human cellular Fn in addition to plasma Fn and super Fn as previously reported. Fn binding assays were performed with recombinant proteins representing the two full-length TP0136 variants and their discrete regions. As a complementary approach, we tested inhibition of T. pallidum binding to Fn by recombinant full-length TP0136 proteins and fragments, as well as by anti-TP0136 immune sera. Our results show that TP0136 adheres more efficiently to cellular Fn than to plasma Fn, that the TP0136 NH2-terminal conserved region of the protein is primarily responsible for binding to plasma Fn but that binding sites for cellular Fn are also present in the protein's central and COOH-terminal regions. Additionally, message quantification studies show that tp0136 is highly transcribed during experimental infection, and that its message level increases in parallel to the host immune pressure on the pathogen, which suggests a possible role for this protein in T. pallidum persistence. In a time where syphilis incidence is high, our data will help in the quest to identify suitable targets for development of a much needed vaccine against this important disease.

No MeSH data available.


Related in: MedlinePlus

TP0136 message quantification during experimental infection.For message quantification, a biopsy from the leading edge of a dermal lesion was obtained from each of the three infected rabbits every three days for 30 days. Each sample was amplified in triplicate. The data were reported as the mean values ± standard error (SE) for triplicate experiments. Left y axis shows real-time qPCR analysis of TP0136 message normalized to TP0547 mRNA (orange line) during progression of primary syphilitic lesions in the rabbit model. Although biopsies were obtained at day 0 and day 3 as well, no message quantification was possible from these samples. Newman-Keuls Multiple Comparison Test was used to assess significant differences in TP0136 message level between time points (*p<0.05) whenever a significant difference between sample means was found by ANOVA. Right y axis shows absolute quantification data for TP0574 message (black bars), reflecting absolute T. pallidum burden.
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pntd.0003662.g002: TP0136 message quantification during experimental infection.For message quantification, a biopsy from the leading edge of a dermal lesion was obtained from each of the three infected rabbits every three days for 30 days. Each sample was amplified in triplicate. The data were reported as the mean values ± standard error (SE) for triplicate experiments. Left y axis shows real-time qPCR analysis of TP0136 message normalized to TP0547 mRNA (orange line) during progression of primary syphilitic lesions in the rabbit model. Although biopsies were obtained at day 0 and day 3 as well, no message quantification was possible from these samples. Newman-Keuls Multiple Comparison Test was used to assess significant differences in TP0136 message level between time points (*p<0.05) whenever a significant difference between sample means was found by ANOVA. Right y axis shows absolute quantification data for TP0574 message (black bars), reflecting absolute T. pallidum burden.

Mentions: Quantitative analysis of tp0136 mRNA expression in primary dermal lesions of rabbits infected with the Nichols Seattle strain showed that transcription of tp0136 is approximately at the same level as tp0574 during early lesion development (day 6–18 post-infection), then subsequently increases (day 21–30 post-infection) (Fig. 2). tp0136 message levels calculated at day 27 and 30 were significantly higher with respect to earlier time points, but not with respect to each other. tp0574 absolute message level (black bars; Fig. 2) reflects the increase and decline of treponemal burden in experimental lesions due to initial growth of the organism followed by reduction in treponemal numbers due to the host’s adaptive immune response to T. pallidum.


Treponema pallidum subsp. pallidum TP0136 protein is heterogeneous among isolates and binds cellular and plasma fibronectin via its NH2-terminal end.

Ke W, Molini BJ, Lukehart SA, Giacani L - PLoS Negl Trop Dis (2015)

TP0136 message quantification during experimental infection.For message quantification, a biopsy from the leading edge of a dermal lesion was obtained from each of the three infected rabbits every three days for 30 days. Each sample was amplified in triplicate. The data were reported as the mean values ± standard error (SE) for triplicate experiments. Left y axis shows real-time qPCR analysis of TP0136 message normalized to TP0547 mRNA (orange line) during progression of primary syphilitic lesions in the rabbit model. Although biopsies were obtained at day 0 and day 3 as well, no message quantification was possible from these samples. Newman-Keuls Multiple Comparison Test was used to assess significant differences in TP0136 message level between time points (*p<0.05) whenever a significant difference between sample means was found by ANOVA. Right y axis shows absolute quantification data for TP0574 message (black bars), reflecting absolute T. pallidum burden.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4368718&req=5

pntd.0003662.g002: TP0136 message quantification during experimental infection.For message quantification, a biopsy from the leading edge of a dermal lesion was obtained from each of the three infected rabbits every three days for 30 days. Each sample was amplified in triplicate. The data were reported as the mean values ± standard error (SE) for triplicate experiments. Left y axis shows real-time qPCR analysis of TP0136 message normalized to TP0547 mRNA (orange line) during progression of primary syphilitic lesions in the rabbit model. Although biopsies were obtained at day 0 and day 3 as well, no message quantification was possible from these samples. Newman-Keuls Multiple Comparison Test was used to assess significant differences in TP0136 message level between time points (*p<0.05) whenever a significant difference between sample means was found by ANOVA. Right y axis shows absolute quantification data for TP0574 message (black bars), reflecting absolute T. pallidum burden.
Mentions: Quantitative analysis of tp0136 mRNA expression in primary dermal lesions of rabbits infected with the Nichols Seattle strain showed that transcription of tp0136 is approximately at the same level as tp0574 during early lesion development (day 6–18 post-infection), then subsequently increases (day 21–30 post-infection) (Fig. 2). tp0136 message levels calculated at day 27 and 30 were significantly higher with respect to earlier time points, but not with respect to each other. tp0574 absolute message level (black bars; Fig. 2) reflects the increase and decline of treponemal burden in experimental lesions due to initial growth of the organism followed by reduction in treponemal numbers due to the host’s adaptive immune response to T. pallidum.

Bottom Line: To deepen our understanding of the TP0136-Fn interaction dynamics, we used two naturally-occurring sequence variants of the TP0136 protein to investigate which region of the protein is responsible for Fn binding, and whether TP0136 would adhere to human cellular Fn in addition to plasma Fn and super Fn as previously reported.As a complementary approach, we tested inhibition of T. pallidum binding to Fn by recombinant full-length TP0136 proteins and fragments, as well as by anti-TP0136 immune sera.Our results show that TP0136 adheres more efficiently to cellular Fn than to plasma Fn, that the TP0136 NH2-terminal conserved region of the protein is primarily responsible for binding to plasma Fn but that binding sites for cellular Fn are also present in the protein's central and COOH-terminal regions.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Division of Allergy and Infectious Diseases, University of Washington, Harborview Medical Center, Seattle, Washington, United States of America; Graduate School, Southern Medical University, Guangzhou, PR China; Division of STD, Guangdong Provincial Center for STI & Skin Diseases Control and Prevention, Guangzhou, PR China.

ABSTRACT
Adherence-mediated colonization plays an important role in pathogenesis of microbial infections, particularly those caused by extracellular pathogens responsible for systemic diseases, such as Treponema pallidum subsp. pallidum (T. pallidum), the agent of syphilis. Among T. pallidum adhesins, TP0136 is known to bind fibronectin (Fn), an important constituent of the host extracellular matrix. To deepen our understanding of the TP0136-Fn interaction dynamics, we used two naturally-occurring sequence variants of the TP0136 protein to investigate which region of the protein is responsible for Fn binding, and whether TP0136 would adhere to human cellular Fn in addition to plasma Fn and super Fn as previously reported. Fn binding assays were performed with recombinant proteins representing the two full-length TP0136 variants and their discrete regions. As a complementary approach, we tested inhibition of T. pallidum binding to Fn by recombinant full-length TP0136 proteins and fragments, as well as by anti-TP0136 immune sera. Our results show that TP0136 adheres more efficiently to cellular Fn than to plasma Fn, that the TP0136 NH2-terminal conserved region of the protein is primarily responsible for binding to plasma Fn but that binding sites for cellular Fn are also present in the protein's central and COOH-terminal regions. Additionally, message quantification studies show that tp0136 is highly transcribed during experimental infection, and that its message level increases in parallel to the host immune pressure on the pathogen, which suggests a possible role for this protein in T. pallidum persistence. In a time where syphilis incidence is high, our data will help in the quest to identify suitable targets for development of a much needed vaccine against this important disease.

No MeSH data available.


Related in: MedlinePlus