Secretion of IL-4 from human basophils. The relationship between IL-4 mRNA and protein in resting and stimulated basophils.
Secretion of IL-4 from human basophils. The relationship between IL-4 mRNA and protein in resting and stimulated basophils.
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DOI：
10.4049/jimmunol.152.6.3006
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发表时间：
1994-03
影响因子：
4.4
通讯作者：
D. MacGlashan;J. White;S. K. Huang;Santa Jeremy Ono;J. Schroeder;L. Lichtenstein
中科院分区：
文献类型：
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作者：
D. MacGlashan;J. White;S. K. Huang;Santa Jeremy Ono;J. Schroeder;L. Lichtenstein
用几种促分泌素刺激纯化的人嗜碱性粒细胞，检测其IL-4蛋白的分泌和IL-4 mRNA的表达。一般而言，这些研究在用促分泌素激发前使用IL-3预孵育15分钟。在这些条件下，用抗IgE Ab激发后，IL-4释放平均为30 pg/10（6）嗜碱性粒细胞（范围4-70）。FMLP和C5 a导致分泌水平略低。将纯度为88%至99%的嗜碱性粒细胞与来自相同制剂但纯度较低的嗜碱性粒细胞直接比较，表明IL-4分泌量与嗜碱性粒细胞的纯度成比例。IL-4 mRNA的存在（通过逆转录酶-PCR、竞争性逆转录酶-PCR或北方印迹法测定）也与嗜碱性粒细胞的纯度严格相关。IL-4 mRNA也被发现是组成性存在，并增加刺激后。最佳IL-4释放所需的多克隆抗IgE Ab的浓度略低于最佳组胺释放所需的浓度。IL-4分泌慢（t1/2的1.5小时）比组胺释放和放线菌酮抑制。在最后一系列研究中，我们发现IL-4分泌不需要IL-3;与IL-3预孵育15分钟导致与未用IL-3处理相比相同或略少的IL-4释放。与此相反，18小时的IL-3预处理导致IL-4分泌增加近10倍。我们的结论是，人类嗜碱性粒细胞分泌IL-4在几个促分泌素和IL-3引发是没有必要观察IL-4的分泌。
Purified human basophils have been examined for secretion of IL-4 protein and expression of IL-4 mRNA after stimulation with several secretagogues. In general, these studies used a 15-min preincubation with IL-3, before challenge with secretagogues. Under these conditions, IL-4 release averaged 30 pg/10(6) basophils (range 4-70) after challenge with anti-IgE Ab. FMLP and C5a led to somewhat lower levels of secretion. A direct comparison of basophils at 88 to 99% purity with basophils from the same preparations, but at lower purities, showed that the amount of IL-4 secretion was proportional to the purity of the basophils. The presence of mRNA for IL-4 (as determined by reverse transcriptase-PCR, competitive reverse transcriptase-PCR, or Northern blots) was also strictly related to the purity of the basophils. IL-4 mRNA was also found to be constitutively present and was increased after stimulation. The concentration of polyclonal anti-IgE Ab required for optimal IL-4 release was somewhat less than that required for optimal histamine release. IL-4 secretion was slower (t1/2 of 1.5 h) than histamine release and was inhibited by cycloheximide. In a final series of studies, we found that IL-3 was not required for IL-4 secretion; a short 15-min preincubation with IL-3 resulted in the same or slightly less IL-4 release than no treatment with IL-3. In contrast, an 18-h pretreatment with IL-3 resulted in a nearly tenfold increase in IL-4 secretion. We conclude that human basophils secrete IL-4 in response to several secretagogues and that IL-3 priming is not necessary to observe IL-4 secretion.