Alizadeh H, Urban J F, Katona I M, Finkelman F D
J Immunol. 1986 Oct 15;137(8):2555-60.
To determine whether IgE+ cells in the intestinal mucosa of nematode-infected mice were of a mast cell or a lymphocyte lineage, the intestinal mucosae of mast cell-deficient w/wv mice were examined for IgE+ cells after inoculation with Trichinella spiralis muscle-stage larvae. Immunofluorescence staining techniques were used to detect IgE associated with cells in the intestinal mucosa. Comparisons were made among four strains of mice, w/wv (mast cell-deficient), +/+ (normal congenic littermates of w/wv), BALB/c, and SJL, that were either uninfected controls or inoculated with T. spiralis. Tissue sections from the small intestine of T. spiralis-infected BALB/c, SJL, and +/+ mice were fixed in ethanol and were stained with an affinity-purified F(ab')2 rabbit anti-mouse IgE followed by FITC goat anti-rabbit IgG. Large numbers of cells in the intestinal mucosa exhibited bright fluorescence. When other sections of intestines from these mice were processed in Carnoy's fixative and were stained with alcian blue at low pH (a metachromatic stain for mast cells) or alcian blue followed by immunofluorescence staining for IgE, large numbers of mast cells were observed in the intestinal mucosa, and 70 to 90% stained positively for IgE. There was a considerable number of cells in the intestinal mucosa which were IgE+ but which did not stain with alcian blue. Few alcian blue-positive cells and no IgE+ staining cells were present in the intestinal mucosa of control, uninfected +/+, BALB/c, and SJL mice. To determine whether these IgE+ alcian blue-negative cells were of a lymphocyte or a mast cell lineage, the mast cell-deficient w/wv mouse strain was examined after infection with T. spiralis. In contrast to BALB/c, SJL, or +/+ mice, few cells in the intestinal mucosa of T. spiralis-infected w/wv mice stained with alcian blue or were positive for IgE. However, when the IgE response in the MLN of the w/wv mice was compared to the IgE response of BALB/c, SJL, and +/+ mice, numerous IgE+ cells, but no alcian blue-positive cells, were observed in the parenchyma of the MLN from all four strains of T. spiralis-infected mice. In addition, flow microfluorometric analysis of MLN cells stained for surface IgE in suspension showed a comparable proportion of IgE-bearing cells, which were mostly B lymphocytes, among all four strains of T. spiralis-infected mice.(ABSTRACT TRUNCATED AT 400 WORDS)
为了确定线虫感染小鼠肠道黏膜中的IgE⁺细胞是肥大细胞谱系还是淋巴细胞谱系,在用旋毛虫肌幼虫接种后,对肥大细胞缺陷的w/wv小鼠的肠道黏膜进行了IgE⁺细胞检测。采用免疫荧光染色技术检测肠道黏膜中与细胞相关的IgE。对四组小鼠进行了比较,分别是w/wv(肥大细胞缺陷)、+/+(w/wv的正常同基因 littermates)、BALB/c和SJL,它们要么是未感染的对照,要么接种了旋毛虫。将旋毛虫感染的BALB/c、SJL和+/+小鼠小肠的组织切片用乙醇固定,并用亲和纯化的F(ab')2兔抗小鼠IgE染色,随后用异硫氰酸荧光素(FITC)标记的山羊抗兔IgG染色。肠道黏膜中的大量细胞呈现明亮的荧光。当将这些小鼠肠道的其他切片用卡诺氏固定液处理,并用低pH值的阿尔辛蓝(一种肥大细胞的异染性染色剂)染色或先用阿尔辛蓝染色再进行IgE免疫荧光染色时,在肠道黏膜中观察到大量肥大细胞,70%至90%的肥大细胞IgE染色呈阳性。肠道黏膜中有相当数量的细胞IgE呈阳性,但阿尔辛蓝染色阴性。在未感染的对照、+/+、BALB/c和SJL小鼠的肠道黏膜中,几乎没有阿尔辛蓝阳性细胞,也没有IgE⁺染色细胞。为了确定这些IgE⁺阿尔辛蓝阴性细胞是淋巴细胞谱系还是肥大细胞谱系,在用旋毛虫感染后对肥大细胞缺陷的w/wv小鼠品系进行了检测。与BALB/c、SJL或+/+小鼠不同,旋毛虫感染的w/wv小鼠肠道黏膜中几乎没有细胞被阿尔辛蓝染色或IgE呈阳性。然而,当比较w/wv小鼠肠系膜淋巴结(MLN)中的IgE反应与BALB/c、SJL和+/+小鼠的IgE反应时,在所有四组旋毛虫感染小鼠的MLN实质中均观察到大量IgE⁺细胞,但没有阿尔辛蓝阳性细胞。此外,对悬浮液中表面IgE染色的MLN细胞进行流式微荧光分析显示,在所有四组旋毛虫感染小鼠中,携带IgE的细胞比例相当,这些细胞大多是B淋巴细胞。(摘要截断于400字)