Ying S, Durham S R, Barkans J, Masuyama K, Jacobson M, Rak S, Löwhagen O, Moqbel R, Kay A B, Hamid Q A
Department of Allergy, Royal Brompton National Heart and Lung Institute, London, United Kingdom.
Am J Respir Cell Mol Biol. 1993 Oct;9(4):356-60. doi: 10.1165/ajrcmb/9.4.356.
We have investigated the phenotype of interleukin-5 (IL-5) mRNA+ cells in the nasal mucosa of subjects with allergic rhinitis. Serial cryostat sections were cut from paraformaldehyde-fixed snap-frozen nasal biopsies from six patients, before and 24 h after local allergen provocation with grass pollen. Immunocytochemistry (APAAP) was followed by in situ hybridization on the same sections. For immunocytochemistry, antibodies against CD3, tryptase, and major basic protein (MBP) were used to identify T cells, mast cells, and eosinophils, respectively. Hybridization studies were performed using a Digoxigenin-labeled IL-5 riboprobe. Nitroblue tetrazolium (NBT) and X-phosphate-5-bromo-4-chloro-3- indoly phosphate (BCIP) served as chromogens to detect hybridized IL-5 mRNA signals. The majority of IL-5 mRNA+ cells were CD3+ (83.2%), whereas the remainder were either tryptase+ (11.3%) or MBP+ (5.4%). In contrast, only a few IL-5 mRNA+ cells were observed in nasal biopsies before challenge, all of which were co-localized to CD3+ cells. These results indicate that CD3+ cells are the principal cellular source of IL-5 transcripts in the nasal mucosa 24 h after allergen-induced late-phase nasal responses.
我们研究了变应性鼻炎患者鼻黏膜中白细胞介素-5(IL-5)mRNA+细胞的表型。从6例患者经多聚甲醛固定后速冻的鼻活检组织中切取连续的冰冻切片,分别在以草花粉进行局部变应原激发前和激发后24小时取材。在同一批切片上,先进行免疫细胞化学(碱性磷酸酶抗碱性磷酸酶法),然后进行原位杂交。免疫细胞化学检测时,分别用抗CD3、抗类胰蛋白酶和抗主要碱性蛋白(MBP)抗体来鉴定T细胞、肥大细胞和嗜酸性粒细胞。杂交研究采用地高辛标记的IL-5核糖探针。用硝基蓝四氮唑(NBT)和5-溴-4-氯-3-吲哚磷酸对硝基蓝四氮唑(BCIP)作为显色剂来检测杂交的IL-5 mRNA信号。大多数IL-5 mRNA+细胞为CD3+(83.2%),其余细胞要么为类胰蛋白酶+(11.3%),要么为MBP+(5.4%)。相比之下,激发前的鼻活检组织中仅观察到少数IL-5 mRNA+细胞,且所有这些细胞均与CD3+细胞共定位。这些结果表明,在变应原诱导的鼻黏膜迟发相反应24小时后, CD3+细胞是IL-5转录本的主要细胞来源。