Measurement of intracellular calcium ions and oxygen radicals in polymorphonuclear leucocyte-erythrocyte 'ghost' hybrids

The Journal of Physiology
A K Campbell, M B Hallett

Abstract

Sendai virus induced fusion between rat polymorphonuclear leucocytes and human erythrocyte 'ghosts' containing the Ca-activated photoprotein obelin. This resulted in the production of more than 90% of the cell population as viable hybrid cells, containing active photoprotein. Substances entrapped originally within the 'ghosts' could be transferred to the hybrids as shown morphologically by fluorescence microscopy, and immunologically by the use of antibodies specific for each of the cell types. Resting free Ca2+ in the hybrids was estimated to be 0.1-0.3 microM. The relationship between intracellular Ca2+ within the hybrids and the production by the hybrids of oxygen radicals, as measured by luminol chemiluminescence, was investigated. The Ca ionophore A23187 stimulated both a rise in intracellular Ca2+ and oxygen radical production, the maximum rate of oxygen radical production being dependent upon the intracellular Ca2+ concentration. Complement activation at the cell surface, chemotactic peptide, and concanavilin A stimulated a rise in intracellular Ca2+, each with different characteristics: complement activation increased intracellular Ca2+ to 8 microM for at least 60 sec; chemotactic peptide raised it to approximately 0.6 ...Continue Reading

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