Early degranulation of human neutrophils: Immunocytochemical studies of surface and intracellular phagocytic events

Pryzwansky, Katherine B.; MacRae, Edith K.; Spitznagel, John K.; Cooney, M H

doi:10.1016/0092-8674(79)90215-0

Cited by 70 publications

(40 citation statements)

References 29 publications

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“…Some LF is observed on the cell surface not associated with zymosan. Similar staining is observed after phagocytosis of E. coli (14). Staining of this type for MPO was rarely observed.…”

Section: Resultssupporting

confidence: 58%

“…(Bar is 1 pm.) with 1% paraformaldehyde to detect only antigens associated with the cell surface (14,15). Zymosan particles associated with a neutrophil 30 sec after incubation are shown by phase microscopy and immunofluorescence staining ( Figure 5).…”

Section: Resultsmentioning

confidence: 99%

“…The methods used for immunofluorescent staining, specificity of the antisera and procedures used to label the antisera have been described (12,14,15). The cells were stained with labelled antiserum for 30 min at 4 ~ washed in phosphate buffered saline, mounted and examined with a Leitz Orthoplan fluorescent microscope using incident light excitation with filter pack H for fluorescein excitation and filter pack N for rhodamine excitation, both with edge filter K480 to minimize quenching.…”

Section: Immunofluorescence Microscopymentioning

confidence: 99%

“…Human venous blood was allowed to clot onto 8 mm glass coverslips as previously described (14). The cells were layered with Gey's tissue culture medium in Tris buffer pH 7.2 (19) containing 10% human AB serum and kept at 37 ~ for 10 min.…”

Section: Preparation Of Cell Monolayersmentioning

confidence: 99%

“…Due to the distribution of MPO and LF, they have been designated as specific markers for the azurophilic and specific granules, respectively (13,16). Using immunocytochemistry with antisera against MPO and LF, it has been possible to investigate neutrophil degranulation and secretion during early phases ofphagocytosis of bacteria (12,14). Degranulation was shown to be rapid and both granule markers, MPO and LF, are observed on the neutrophil surface within 5 sec after phagocytosis of E. coli has been initiated.…”

Section: Introductionmentioning

confidence: 99%

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Phagocytosis of zymosan by human neutrophils

MacRae

Pryzwansky

1984

Carlsberg Res. Commun.

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A rapid cellular response occurs when human neutrophils are challenged with the zymosan particle. Monolayers of neutrophils adhering to glass were incubated with opsonized zymosan at 37 ~ for 15 see to 3 min, fixed and prepared for observation by SEM, immunofluorescence or TEM. At 15 sec zymosan particles are seen by SEM in contact with the cell surface and by 30 sec partial engulfment into open cavities was apparent. The cavity, into which a zymosan particle is engulfed, forms by extension of lamellipodia around and over the particle. The zymosan particle is sequestered in an intracellular vacuole or phagosome by 1 min. At 3 min neutrophils often show bulging profiles of the internalized zymosan and a loss of surface projections. During early phagocytosis, simultaneous staining with contrasting fluorescent antisera to granule constituents, myeloperoxidase and lactoferrin, reveals granule antigens within the cavity of the forming phagosome. Sections through the cells examined by TEM after 1-3 min incubation with zymosan show granule contents in the phagosome peripheral space which retains connection to the extracellular environment.

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“…Some LF is observed on the cell surface not associated with zymosan. Similar staining is observed after phagocytosis of E. coli (14). Staining of this type for MPO was rarely observed.…”

Section: Resultssupporting

confidence: 58%

Section: Resultsmentioning

confidence: 99%

Section: Immunofluorescence Microscopymentioning

confidence: 99%

Section: Preparation Of Cell Monolayersmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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