Methods:
Bacterial cDNA was amplified using random DNA primers coupled with RNA polymerase promoter region. Fluorescence label cRNAs were amplified by IVT, and hybridized with custom-made DNA microarray of virulent factor genes, and monitored therapeutic targets. To develop passive immunotherapy, hybridoma against P. gingivalis virulent factors were constructed. Then, ScFv genes were molecular cloned and functionally expressed in Bacillus brevis hosts. Human type mAb was also constructed using transgenic mice carrying human immunoglobulin gene loci. To develop the mucosal immunization, we used P. gingivalis outer membrane protein and hemagglutinin as vaccine to mice models.
Results: DNA microarray analysis can determined the virulent factor gene expression in periodontal pocket. ScFv functionally expressed in Bacillus brevis hosts and capable to inhibited heamagglutinin and aggregation activity. Human type mAbs treatment reduced the alveolar bone in loss caused by oral infection with P. gingivalis in mice experimental model.
Conclusion: Identification of gene expression of virulent factors using microarray and recombinant passive antibodies might be useful for molecular target therapy against periodontal diseases.