Flow microfluorometric analysis of phagocyte degranulation in bacteria infected whole human blood cell cultures

Alexander L. Kravtsov; Elena V. Bobyleva; Tatyana P. Grebenyukova; Oleg S. Kuznetsov; Youri V. Kulyash

doi:10.1117/12.475607

16 July 2002 Flow microfluorometric analysis of phagocyte degranulation in bacteria-infected whole human blood cell cultures

Alexander L. Kravtsov, Elena V. Bobyleva, Tatyana P. Grebenyukova, Oleg S. Kuznetsov, Youri V. Kulyash

Author Affiliations +

Proceedings Volume 4707, Saratov Fall Meeting 2001: Optical Technologies in Biophysics and Medicine III; (2002) https://doi.org/10.1117/12.475607
Event: Saratov Fall Meeting 2001, 2001, Saratov, Russian Federation

Abstract

A quantitative flow microfluorometric method was used to study the intensity of human blood phagocyte degranulation in response to viable staphylococcus aureus or Yersinia pestis cells. Microorganisms were added directly to defibrinated whole blood. Uninfected and infected blood samples were incubated at 37 degrees C to 8 h. The results were recorded in dynamics after the staining of whole blood with acridine orange solution. Lymphocytes with a low azurophilic granule per cell content were discriminated from phagocytes by the measurement of single cell red cytoplasmic granule fluorescence. 30,000 cells in each sample were examined. S. aureus cells caused a dose-dependent decrease in the number of phagocytes having a high red cytoplasmic fluorescence intensity and a corresponding increase in the weakly fluorescence cell population. In the presence of an initial S. aureus-to-phagocyte ratio more than 1:1, degranulation was measured after 3 h of incubation and to 8 h the percentage of degranulated phagocytes was at least 100 percent Y. pestis cells grown for 48 h at 28 degrees C caused at same condition as the degranulation only about 50 percent of cells. Y.pestis EV cells preincubated in broth for 12 h at 37 degrees C did no stimulate the phahocyte degranulation. The results of these studies suggest that analysis of cell populations via flow microfluorimeter technology may be a powerful tool in analysis bacterial infection.

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Alexander L. Kravtsov, Elena V. Bobyleva, Tatyana P. Grebenyukova, Oleg S. Kuznetsov, and Youri V. Kulyash "Flow microfluorometric analysis of phagocyte degranulation in bacteria-infected whole human blood cell cultures", Proc. SPIE 4707, Saratov Fall Meeting 2001: Optical Technologies in Biophysics and Medicine III, (16 July 2002); https://doi.org/10.1117/12.475607

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