Phagocytosis of S. pneumoniae by Human Granulocytes

Experiments investigating phagocytosis were performed using a flow cytometry assay including S. pneumoniae labeled with 5, 6-carboxyfluorescein succinimidyl ester (FAM-SE; Molecular Probes) and human HL-60 cells (CCL-240; ATCC) differentiated to granulocytes [14 (link),15 (link),27 (link),28 (link)]. Infection assays were performed with a ratio of 10 bacteria per cell. Results were expressed as a RFI defined as the proportion of positive cells for fluorescent bacteria multiplied by the geometric mean of fluorescence intensity, which correlates with the amount of bacteria phagocytosed per cell [26 (link),27 (link)].

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Aguinagalde L., Corsini B., Domenech A., Domenech M., Cámara J., Ardanuy C., García E., Liñares J., Fenoll A, & Yuste J. (2015). Emergence of Amoxicillin-Resistant Variants of Spain9V-ST156 Pneumococci Expressing Serotype 11A Correlates with Their Ability to Evade the Host Immune Response. PLoS ONE, 10(9), e0137565.

Publication 2015

FAM-SE CCL-240

6 carboxyfluorescein Assays Bacteria Cell Ester Flow cytometry Fluorescence Granulocytes Hl 60 cells Human Infection Molecular probes S pneumoniae

Corresponding Organization : Centro de Investigación Biomédica en Red de Enfermedades Respiratorias

Other organizations : Bellvitge University Hospital, Institut d'Investigació Biomédica de Bellvitge, Centro de Investigaciones Biológicas Margarita Salas

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Variable analysis

independent variables

Ratio of 10 bacteria per cell

dependent variables

Proportion of positive cells for fluorescent bacteria
Geometric mean of fluorescence intensity, which correlates with the amount of bacteria phagocytosed per cell

control variables

S. pneumoniae labeled with 5, 6-carboxyfluorescein succinimidyl ester (FAM-SE; Molecular Probes)
Human HL-60 cells (CCL-240; ATCC) differentiated to granulocytes

controls

Positive control: Not specified
Negative control: Not specified

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