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Research Article Free access | 10.1172/JCI107667

Myeloperoxidase-Mediated Iodination by Granulocytes INTRACELLULAR SITE OF OPERATION AND SOME REGULATING FACTORS

Richard K. Root and Thomas P. Stossel

Infectious Disease Section, Department of Medicine, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19174

Department of Hematology, Children's Hospital Medical Center, Boston, Massachusetts 02115

Find articles by Root, R. in: PubMed | Google Scholar

Infectious Disease Section, Department of Medicine, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19174

Department of Hematology, Children's Hospital Medical Center, Boston, Massachusetts 02115

Find articles by Stossel, T. in: PubMed | Google Scholar

Published May 1, 1974 - More info

Published in Volume 53, Issue 5 on May 1, 1974
J Clin Invest. 1974;53(5):1207–1215. https://doi.org/10.1172/JCI107667.
© 1974 The American Society for Clinical Investigation
Published May 1, 1974 - Version history
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Abstract

The intracellular site of operation of the myeloperoxidase-H2O2-halide antibacterial system of granulocytes has been determined by utilizing measurements of the fixation of iodide to trichloracetic acid (TCA) precipitates of subcellular fractions, including intact phagocytic vesicles. Na125I was added to suspensions of guinea pig granulocytes in Krebs-Ringer phosphate buffer, and they were then permitted to phagocytize different particles. Phagocytic vesicles were formed by allowing cells to ingest a paraffin oil emulsion (POE) and collected by flotation on sucrose after homogenization. Measurement of 125I bound to TCA precipitates of the different fractions and the homogenates disclosed that the lysosome-rich fraction obtained by centrifugation from control (nonphagocytizing) cells accounted for a mean 93.1% of the total cellular activity. With phagocytosis of POE, TCA-precipitable iodination increased two- to sevenfold, and the lysosomal contribution fell to a mean 36.9% of the total. The appearance of activity within phagocytic vesicles accounted for almost the entire increase seen with phagocytosis (a mean 75.7%), and iodide was bound within these structures with high specific activity. More iodide was taken up by cells than fixed, regardless of iodide concentration, and was distributed widely throughout the cell rather than selectively trapped within the vesicles.

The amount of iodide taken up and fixed varied considerably with the phagocytic particle employed. Yeast particles were found to stimulate iodination to a far greater degree than the ingestion of POE or latex. Such observations are consistent with the concept that the ingested particle is a major recipient of the iodination process. Measurements of metabolic activities related to the formation and utilization of peroxide by cells phagocytizing different particles were made and correlated with iodination. The findings suggest that mechanisms must exist within granulocytes to collect or perhaps even synthesize H2O2 within phagocytic vesicles to serve as substrate for myeloperoxidase. The simultaneous stimulation of other metabolic pathways for peroxide disposal and its release into the medium by phagocytizing cells is consistent with the high diffusability of this important bactericidal substance.

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