An IgY-based immunoassay to evaluate the biomarker potential of the Tannerella forsythia virulence factor karilysin in human saliva.
Skottrup. Peter Durand PD; López. Rodrigo R; Ksiazek. Miroslaw M; Højrup. Peter P; Baelum. Vibeke V; Potempa. Jan J; Kaczmarek. Jakub Zbigniew JZ
Key Findings
- A high‑affinity avian IgY antibody against karilysin was created and characterized.
- An IgY‑based immunoassay can detect karilysin in saliva with low day‑to‑day variability.
- Karilysin levels were significantly higher in saliva of adolescents with periodontitis compared to healthy controls.
Practical Outcomes
- The new saliva test could let biohackers track gum‑related inflammation and possibly catch early signs of periodontal disease. While it doesn’t change how to use LL‑37, it highlights the role of bacterial enzymes that degrade LL‑37, suggesting oral health may influence overall immune balance.
Summary
Scientists made a new test that can spot a bacterial protein called karilysin in spit. People with gum disease had higher levels of this protein than healthy folks. The test is reliable and could be used to monitor oral health, but it doesn’t give direct tips on using the LL‑37 peptide or changing diet or supplements.
Abstract
Tannerella forsythia is a gram-negative anaerobic bacterium that is associated with the development of destructive periodontal disease. T. forsythia secretes the metalloprotease-like enzyme karilysin. Using in vitro systems karilysin has been shown to modulate the host immune response by degradation of complement system proteins and by inactivation of the antimicrobial peptide LL-37 by proteolytic cleavage. This makes karilysin a highly interesting virulence factor to study in the framework of drug development and diagnostics. However, to date the presence of karilysin in clinical samples has not been demonstrated due to the lack of specific probes. In the present work, a high titer and stable affinity-purified avian IgY antibody against karilysin was developed. By surface plasmon resonance imaging the IgY affinity was found to be in the low nanomolar range. The antibody could be used to detect karilysin in saliva samples by immuno-blotting and was specific when tested towards human MMP-3. Furthermore, an avian IgY-based immunoassay was developed, which demonstrated low intra- and interday assay variability (CV's below 10%). Application of the immunoassay on a well-characterized set of saliva samples from adolescents with or without signs of periodontitis showed that it was possible to detect karilysin in saliva. A significant difference in karilysin concentration was found between saliva from participants with signs of periodontitis and saliva from healthy controls (p = .0024). The median of karilysin levels among periodontitis cases was 957 pg/ml (IQR, 499-2132 pg/ml) and the median for controls was 569 pg/ml (IQR, 210-1343 pg/ml). Collectively our data confirm the presence of karilysin in clinical samples. The described IgY-based immunoassay may prove useful as part of protein-based biomarker screenings in the clinic or in point-of care settings.
Study Information
pubmed
2019
2019-03-14T00:00:00.000Z
10.1016/j.jim.2019.03.003
9
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