Intracellular location of Bartonella henselae cocultivated with Vero cells and used for an indirect fluorescent-antibody

Bartonella henselae, the major causative agent of cat scratch disease, was cocultivated with Vero cells on chamber slides and visualized by indirect immunofluorescence by using a patient serum containing specific antibodies. Confocal microscopy localized the granular B. henselae-specific fluorescence mainly around the nuclei of Vero cells. By transmission electron microscopy, these granules were identified as clusters of multiple intracellular organisms. Fixed slides with the monolayers of Vero cells with intracellular B. henselae were used for an indirect fluorescent-antibody test to investigate the seroprevalence of specific immunoglobulin G in 100 serum samples from blood donors. Seventy-four serum samples were negative; 19, 3, and 4 were positive at dilutions of 1:64, 1:128, and 1:256, respectively. In our population, a serum titer of 1:256 or greater should stimulate further investigations. Moreover, elucidation of the mechanism by which B. henselae enters the cells may help to understand the pathogenesis of cat scratch disease. Bartonella (Rochalimea) henselae has been isolated from lymph nodes of patients with cat scratch disease (CSD) (3) and recently was also detected by PCR (1). The role of B. henselae as the major causative agent of CSD is largely based on sero-logical surveys with an indirect fluorescent-antibody (IFA) test