Campbell A K, Hallett M B
J Physiol. 1983 May;338:537-50. doi: 10.1113/jphysiol.1983.sp014688.
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 microM for a prolonged period (at least 10 min) and concanavilin A stimulated a transient (t1/2 = approximately 80 sec) rise to approximately 0.6 microM. Un-opsinized particles, latex beads (diam. approximately equal to 1 micron), which stimulate oxygen radical production, did not produce a detectable rise in intracellular Ca2+. Intracellular EGTA (approximately, 2.5 mM) inhibited both the oxygen radical production and the rises in intracellular Ca2+ induced by chemotactic peptide and concanavilin A, and delayed both the rise in intracellular Ca2+ and the onset of oxygen radical production induced by complement. Intracellular EGTA had no effect on oxygen radical production stimulated by latex particles. An increase in intracellular Ca2+ triggered oxygen radical production by the hybrids in response to the following stimuli: concanavilin A, chemotactic peptide, complement and the Ca ionophore A23187. However, the extent and duration of the increase in cytoplasmic Ca2+ was different for each stimulus, and the apparent relationship between cytoplasmic Ca2+ and oxygen radical production was different for each stimulus. Un-opsinized particles stimulated oxygen radical production without requiring a rise in intracellular Ca2+ concentration.
仙台病毒诱导大鼠多形核白细胞与含有钙激活光蛋白奥贝林的人红细胞“血影”发生融合。这导致超过90%的细胞群体成为含有活性光蛋白的存活杂交细胞。如通过荧光显微镜在形态学上以及通过使用针对每种细胞类型的特异性抗体在免疫学上所显示的，最初包裹在“血影”内的物质可转移至杂交细胞。杂交细胞中静息游离钙离子浓度估计为0.1 - 0.3微摩尔。研究了杂交细胞内钙离子与通过鲁米诺化学发光法测定的杂交细胞产生氧自由基之间的关系。钙离子载体A23187刺激细胞内钙离子升高以及氧自由基产生，氧自由基产生的最大速率取决于细胞内钙离子浓度。细胞表面补体激活、趋化肽和伴刀豆球蛋白A刺激细胞内钙离子升高，各自具有不同特点：补体激活使细胞内钙离子至少60秒内升高至8微摩尔；趋化肽使其在较长时间（至少10分钟）内升高至约0.6微摩尔，伴刀豆球蛋白A刺激短暂（半衰期约为80秒）升高至约0.6微摩尔。未致敏颗粒、直径约为1微米的乳胶珠可刺激氧自由基产生，但未引起细胞内钙离子可检测到的升高。细胞内乙二醇双四乙酸（约2.5毫摩尔）抑制趋化肽和伴刀豆球蛋白A诱导的氧自由基产生以及细胞内钙离子升高，并延迟补体诱导的细胞内钙离子升高和氧自由基产生起始。细胞内乙二醇双四乙酸对乳胶颗粒刺激的氧自由基产生无影响。细胞内钙离子升高会触发杂交细胞对以下刺激产生氧自由基：伴刀豆球蛋白A、趋化肽、补体和钙离子载体A23187。然而，每种刺激引起的细胞质钙离子升高的程度和持续时间不同，并且细胞质钙离子与氧自由基产生之间的明显关系因每种刺激而异。未致敏颗粒刺激氧自由基产生，而无需细胞内钙离子浓度升高。
