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Blood Sample Collecting and Processing for Perio VAL Cohort

 Introduction

Chronic periodontitis (ChP) is a prevalent inflammatory disease affecting 46% of the US population. ChP produces a profound local inflammatory response to dysbiotic oral microbiota that leads to the destruction of alveolar bone and tooth loss. ChP is also associated with systemic illnesses, including cardiovascular diseases, malignancies, and adverse pregnancy outcomes. The mechanisms underlying oral and systemic diseases are poorly understood. Our previous cohort demarcates systemic and cell-specific immune dysfunctions in patients with ChP, which can be temporarily reversed by the local treatment of ChP . Here, we conducted a validation cohort (Perio VAL) to validate our previous results.

Method

We consent 16 patients and 12 healthy controls in the Bell Dental Center (San Leandro, CA). Whole-blood samples were collected at baseline, 3 weeks, and 3 months after standard nonsurgical treatment for ChP. Blood was left unstimulated or stimulated with Porphyromonas gingivalis lipopolysaccharide (PgLPS), tumor necrosis factor α (TNFα), interleukins 12 (IL-12), and a cytokine cocktail containing interleukins 2/4/6 (IL-2/4/6) and stored at -80 degree. Before CyTOF analysis on white blood cells, samples were lysed to remove red blood cells.

Result

A total of about 420 samples were lysed and stored for CyTOF analysis.

Conclusion

We prepared the samples in Perio VAL cohort for the CyTOF analysis. In the future, we will conduct the multiplex single-cell analysis to investigate the oral and systemic immunological link in ChP.





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