tretinoin and Periodontal-Diseases

We studied the interactions between viable and heat-killed, opsonized and unopsonized periodontopathic bacteria with both uninduced and induced HL-60 promyelocytic leukemia cells. The cells were induced to differentiate into granulocyte-like cells by incubation with retinoic acid (RA) or dimethyl sulfoxide (DMSO). When unopsonized, Wolinella recta ATCC 33228 significantly suppressed the net proliferation of uninduced HL-60 cells, Actinobacillus actinomycetemcomitans strain Y4 was markedly lethal to the cells, and Bacteroides gingivalis ATCC 33277 had no effect. Unopsonized and opsonized A. actinomycetemcomitans and W. recta had equally potent lethal effects on induced HL-60 cells. Unopsonized B. gingivalis was not lethal to the induced cells in the dose used (100 bacteria/HL-60 cell), but opsonized B. gingivalis was lethal, especially in the first 24 h. The killing effects of A. actinomycetemcomitans and W. recta were largely eliminated if they were heated (56 degrees C, 30 min) before being added to the induced HL-60 cells. RA-induced HL-60 cells were more sensitive to the lethal effects of A. actinomycetemcomitans and W. recta than were DMSO-induced cells. The results suggest that the HL-60 cell line may be a useful model for studying granulocyte-bacteria interactions.

Topics: Actinobacillus; Bacteroides; Cell Differentiation; Dimethyl Sulfoxide; Granulocytes; Humans; Leukemia, Promyelocytic, Acute; Opsonin Proteins; Periodontal Diseases; Tretinoin; Tumor Cells, Cultured

As a first step toward elucidating the reasons for differences among periodontal pathogens in their cytotoxic effects on HL-60 cells, we used transmission electron microscopy to examine morphological aspects of granulocyte-bacteria interactions. Unopsonized Actinobacillus actinomycetemcomitans strain Y4 and Bacteroides gingivalis ATCC 33277 adhered to, and were phagocytosed by, retinoic acid-induced and dimethyl sulfoxide-induced HL-60 cells. In contrast, there was only minimal interaction between Wolinella recta ATCC 33238 and these induced granulocyte-like cells. Only isolated examples of adherence of W. recta to HL-60 cells were seen. In specimens prepared for routine transmission electron microscopy, ingested W. recta were not observed. In immunogold experiments, phagocytosed W. recta were noted, but only rarely. Opsonization of A. actinomycetemcomitans, B. gingivalis and W. recta with specific antisera appeared to increase their level of interaction with the HL-60 cells. We suggest that the HL-60 cell line may be useful in elucidating structure-function relationships between human neutrophil-like cells and putative periodontopathogens.

Topics: Actinobacillus; Bacterial Adhesion; Bacteroides; Cell Differentiation; Dimethyl Sulfoxide; Granulocytes; Humans; Immunohistochemistry; Leukemia, Promyelocytic, Acute; Opsonin Proteins; Periodontal Diseases; Tretinoin; Tumor Cells, Cultured