Alam R, Lewis D M, Olenchock S A
Division of Respiratory Disease Studies, National Institute for Occupational Safety and Health, Morgantown, WV 26505.
J Immunol. 1988 Mar 15;140(6):1946-52.
The production of histamine-releasing factor (HRF) by human mononuclear cells has previously been reported. In this paper we describe the production of HRF by guinea pig spleen cells, thymocytes, and PBMC. Guinea pig lymphoid cells were cultured either alone or in the presence of mitogens (PHA and Con A) or specific Ag(OVA and keyhole limpet hemocyanin) and the dialyzed cell-free supernatant was tested for histamine-releasing activity on guinea pig lung mast cells and blood basophils. Lung mast cells were isolated by enzymatic digestion and partially purified by countercurrent elutriation and discontinuous Percoll gradient centrifugation. Guinea pig spleen cells, thymocytes, and PBMC spontaneously produced significant amounts of HRF. The production was enhanced upon stimulation with PHA or specific Ag in animals immunized with Ag in CFA. Two distinct species of HRF were identified with m.w. of 50,000 to 70,000 and 5000 to 8000 by gel chromatography. HRF is a trypsin- and chymotrypsin-sensitive heat-stable protein. It does not bind to Con A-Sepharose and its production is not inhibited by tunicamycin. HRF-induced histamine release from lung mast cells is a temperature-dependent process and is complete in 10 min at 37 degrees C. Intradermal injection of HRF caused an immediate ear-swelling reaction in guinea pigs. The most severe ear-swelling reactions did not resolve within 1 h, but instead evolved over a period of 12 to 24 h.
此前已有关于人单核细胞产生组胺释放因子（HRF）的报道。在本文中，我们描述了豚鼠脾细胞、胸腺细胞和外周血单核细胞（PBMC）产生HRF的情况。将豚鼠淋巴细胞单独培养，或在有丝分裂原（PHA和Con A）或特异性抗原（OVA和血蓝蛋白）存在的情况下培养，然后对透析后的无细胞上清液进行检测，观察其对豚鼠肺肥大细胞和血液嗜碱性粒细胞的组胺释放活性。通过酶消化分离肺肥大细胞，并通过逆流淘析和不连续Percoll梯度离心进行部分纯化。豚鼠脾细胞、胸腺细胞和PBMC可自发产生大量HRF。在用CFA免疫抗原的动物中，PHA或特异性抗原刺激后，HRF的产生会增强。通过凝胶色谱法鉴定出两种不同的HRF，分子量分别为50,000至70,000和5000至8000。HRF是一种对胰蛋白酶和糜蛋白酶敏感的热稳定蛋白。它不与Con A - Sepharose结合，其产生不受衣霉素抑制。HRF诱导肺肥大细胞释放组胺是一个温度依赖性过程，在37℃下10分钟内完成。皮内注射HRF会使豚鼠立即出现耳部肿胀反应。最严重的耳部肿胀反应在1小时内不会消退，而是在12至24小时内逐渐发展。
