Early events and implication of F-actin and annexin I associated structures in the phagocytic uptake of Brucella suis by the J-774A.1 murine cell line and human monocytes

Abstract

Brucella spp. are facultative, intracellular pathogenic bacteria that cause brucellosis, a zoonosis affecting mammalian species. Brucella entry into myelomonocytic cell lines is highly enhanced by opsonization. Few studies have been undertaken to unravel the first interactions between these bacteria and their host cells. This paper deals with early events following contact of Brucella suis with the J-774A.1 phagocytic cell line and differentiated monocytes. Phagocytic uptake of bacteria was documented under a fluorescence microscope using GFP-expressing B. suis. Unlike entry in the J-774A. 1 cell line, non-opsonized Brucella entered differentiated human monocytes as efficiently as opsonized bacteria. However, following 1 h infections, a mean of only three bacteria were phagocytized and the whole monocyte population was only infected after a 4 h infection. Contact of non-opsonized Brucella with phagocytes did not induce marked structural changes at the cell surface, as revealed by scanning electron microscopy. Contact of Brucella (opsonized or not) elicited transient local recruitment of F-actin, revealed by phalloidin labelling, and of annexin I-associated structures, revealed by immunofluorescence staining. Finally, bacteria appeared to be rapidly internalized in monocytes once they had adhered to the cell surface. A low percentage of infected cells and few adhered and/or internalized bacteria following short-term infections could have resulted either from the fact that there were few sites of entry or the weak bacterial initial interactions with the host-cell membrane or the bacterial receptor.