Journal: Infection and Immunity

Article Title: Entamoeba gingivalis induces gingival cell death, collagen breakdown, and host immune response via VAMP8/-3-driven exocytosis pathways

doi: 10.1128/iai.00005-25

Figure Lengend Snippet: Protein co-localization in gingival cells. Cell cultures of gECs (A, B) and gFBs (C). After fixation, gingival epithelial cells were gECs, which were incubated and stained with an anti-VAMP8 antibody and (A) with an anti-MUCIN1 antibody conjugated with Alexa Fluor 546 or (B) with an anti-MUC21 antibody. In (C), gFBs were incubated and stained with an anti-MMP13 antibody and with an anti-VAMP3 antibody. Cell nuclei were stained with DAPI (blue). Co-localization was observed via fluorescent microscopy.

Article Snippet: ELISAs were performed according to the manufacturer’s protocols using ELISA Kits for human MUC1- (EHMUC1, ThermoFisher), MUC21 (HUFI00559, AssayGenie) and IL-8/CXCL8 (Sigma-Aldrich; RAB0319) on a microplate spectrophotometer (Multiskan GO, Thermo Fisher).

Techniques: Incubation, Staining, Microscopy

Journal: Infection and Immunity

Article Title: Entamoeba gingivalis induces gingival cell death, collagen breakdown, and host immune response via VAMP8/-3-driven exocytosis pathways

doi: 10.1128/iai.00005-25

Figure Lengend Snippet: Mucin and interleukin secretion after infection of gECs and gFBs with E. gingivalis and P. gingivalis . (A) Two hours after infection of parental gECs with E.g. , MUC1 and MUC21 secretion was increased 3.67-fold (mU/mL; P = 0.001) and 14.37-fold (ng/mL; P = 0.0031), respectively, compared with mucin secretion in mock-infected gECs. In gEC cells that were infected with P. gingivalis for 2 h, no significantly increased mucin secretion was observed. (B) In gEC-VAMP8(−/−) cells, secretion of MUC1 and MUC21 was not detectable. (C) IL-8 secretion was significantly increased in parental gECs after infection with E. gingivalis , but could not be detected in gECs-VAMP8(−/−) cells or in gECs infected with P. gingivalis . IL-8 secretion was similar in both parental gFB cultures and gFB-VAMP3(−/−), indicating that VAMP8, but not VAMP3, is involved in IL-8 secretion. An infection with E. gingivalis caused a stronger IL-8 secretion than an infection with P. gingivalis . (D) IL-1B secretion was significantly increased in parental gECs cultures after infection with E. gingivalis , but could not be detected in gEC-VAMP8(−/−) cells. In contrast, gFB-VAMP3(−/−) cells secreted IL1B, indicating that VAMP8, but not VAMP3, is involved in IL-1B secretion. The results of A-C were obtained by spectrometry, while the results of D were obtained by chemiluminescence detection. The dark turbidity of the P. gingivalis growth medium prevented an experiment with mock infections using chemiluminescence detection.

Article Snippet: ELISAs were performed according to the manufacturer’s protocols using ELISA Kits for human MUC1- (EHMUC1, ThermoFisher), MUC21 (HUFI00559, AssayGenie) and IL-8/CXCL8 (Sigma-Aldrich; RAB0319) on a microplate spectrophotometer (Multiskan GO, Thermo Fisher).

Techniques: Infection