Uptake and killing of Listeria monocytogenes by normal human peripheral blood granulocytes and monocytes as measured by flow cytometry and cell sorting.
Cellular components of innate immunity (NK cells, monocytes and granulocytes) play an important role in early resistance to Listeria monocytogenes in the mouse model. Minimally invasive methods of measuring the bacteriocidal capacity of these cells may be useful as a biomarker of susceptibility in humans. A technique was developed whereby the uptake and survival of L. monocytogenes could be measured in human granulocytes and monocytes using small volumes of peripheral blood. This method used flow cytometry to detect the presence of PKH-2-labeled bacteria within these cells. Survival of bacteria was determined by sorting of infected cells based on a combination of fluorescence and light scattering properties. Considerable variation in bacterial recovery was seen between normal volunteers. There was consistently greater survival of a fully virulent strain of L. monocytogenes within monocytes and granulocytes compared with an isogenic strain lacking the hemolysin, listeriolysin O, when measured at baseline. There was no evidence of longer-term bacterial survival or growth at 2 or 24 h. This technique may be useful for assessment of both host resistance and pathogen virulence.
Raybourne
R B
RB
Immunobiology Branch, Food and Drug Administration, MOD 1, 8301 Muirkirk Road, Laurel, MD 20708, USA. rraybour@cfsan.fda.gov
Roth
G
G
Deuster
P A
PA
Sternberg
E M
EM
Singh
A
A
D000328:Adult; D002469:Cell Separation; D003470:Culture Media; D005260:Female; D005434:Flow Cytometry; D005456:Fluorescent Dyes / Q000378:metabolism; D006098:Granulocytes / Q000276:immunology* / Q000382:microbiology; D006460:Hemolysin Proteins / Q000235:genetics; D006801:Humans; D008089:Listeria monocytogenes / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification / Q000472:pathogenicity; D008088:Listeriosis / Q000382:microbiology; D008297:Male; D009000:Monocytes / Q000276:immunology* / Q000382:microbiology; D009930:Organic Chemicals; D010587:Phagocytosis; D014774:Virulence
D003470:Culture Media; D005456:Fluorescent Dyes; D006460:Hemolysin Proteins; D009930:Organic Chemicals; C068241:PKH-2