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DOI: 10.1055/s-0044-1801302
Internalization and Colocalization of a Polyclonal Antibody Against Porphyromonas gingivalis FimA type I in Infected Cells
Funding This work was supported by Naresuan University (NU), and National Science, Research and Innovation Fund (NSRF) grant No. FRB660001/0179.
Abstract
Objective The aim of this work was to investigate the effect of a rabbit polyclonal antibody specific to Porphyromonas gingivalis FimA type I (FimI) protein internalized into P. gingivalis infected cells.
Materials and Methods Rabbits were immunized with P. gingivalis FimI protein and the serum was collected for immunoglobulin (Ig) purification. For visualization of the antibody inside the cells, it was labeled with Cy3 dye. Live P. gingivalis was labeled with PKH67 dye. Rabbit anti-FimI Ig-Cy3 was internalized into H357 cells infected with P. gingivalis-PKH67 by electroporation or coincubation. Location of the Ig or P. gingivalis was observed under fluorescence microscope or confocal microscope. Percentage of P. gingivalis-PKH67 infected cells was analyzed by flow cytometry.
Statistical Analysis Normality of data distribution was tested by Shapiro–Wilk test. The data failed normality test and were further analyzed by Kolmogorov–Smirnov test.
Results Rabbit anti-P. gingivalis FimI Ig-Cy3 and P. gingivalis-PKH67 were both located next to the nucleus. The rabbit anti-FimI Ig-Cy3 was able to enter H357 cells after the cells were cultured in the medium containing the labeled Ig for 16 hours. The location of the Ig was near the nucleus as found in cells electroporated with the Ig-Cy3. The percentage of P. gingivalis-PKH67 infected cells seemed to be decreased after the infected cells internalized anti-FimI Ig by electroporation. However, it was not statistically significance.
Conclusion Rabbit anti-P. gingivalis FimI Ig and P. gingivalis was colocalized near the nucleus. And the rabbit anti-FimI Ig was able to enter H357 cells by coincubation method.
Publication History
Article published online:
12 March 2025
© 2025. The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution License, permitting unrestricted use, distribution, and reproduction so long as the original work is properly cited. (https://creativecommons.org/licenses/by/4.0/)
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