Abstract
Bartonella henselae is a causative agent of anemia, cat scratch disease, bacillary angiomatosis, recurrent fever, hepatitis, endocarditis, chronic lymphadenopathy, joint and neurological disorders. B. henselae are intra-erythrocytic bacteria. The goal of this study was to visualize the B. henselae invasion into enucleated human red blood cells in real time using bacterium endogenous fluorescence. We took advantage of the unique fluorescence emission spectral profile of the bacteria. We used a linear unmixing approach to separate the fluorescence emission spectra of human erythrocytes from native B. henselae when excited at 488nm. Human blood samples were inoculated with B. henselae and incubated for 60 hours. 3-D live images were captured at select intervals using multi-photon laser scanning microscopy. Uninfected blood samples were also analyzed. This study revealed bacteria entering mature erythrocytes over a 60 hour time period.
Highlights
Bartonella spp. are Gram-negative, fastidious, and facultative intracellular bacteria [1] which can cause Peruvian bartonellosis (Carion’s disease), cat scratch disease, bacillary angiomatosis, trench fever, endocarditis, bacteremia [2], hepatitis, chronic lymphadenopathy, joint and neurological disorders
Unmixing is used with synthetic fluorescence reporters but we were able to take advantage of the relatively strong endogenous fluorescence of the erythrocytes and the bacteria
We unmixed all images with the same couple of spectra that we got from red blood cells (RBC) and B. henselae in the image
Summary
Bartonella spp. are Gram-negative, fastidious, and facultative intracellular bacteria [1] which can cause Peruvian bartonellosis (Carion’s disease), cat scratch disease, bacillary angiomatosis, trench fever, endocarditis, bacteremia [2], hepatitis, chronic lymphadenopathy, joint and neurological disorders. Recognition of the clinical spectrum of Bartonella spp. infection in humans has greatly increased in the last 20 years [3,4]. The infection may be fatal even to immunocompetent patients [5]. B. henselae is the most frequently documented species of the genus to cause human infection [6]. It can infect erythrocytes of many mammals including cats, dogs, mice, rats, horse, dolphins, etc., causing cyclic bacteremia. B. henselae can infect endothelial cells [7], macrophages [8], and pericytes [9]
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