IMMUNOREACTIVITIES OF GASTRIN (G-) CELLS .2. NONSPECIFIC-BINDING OF IMMUNOGLOBULINS TO G-CELLS BY IONIC INTERACTIONS
IMMUNOREACTIVITIES OF GASTRIN (G-) CELLS .2. NONSPECIFIC-BINDING OF IMMUNOGLOBULINS TO G-CELLS BY IONIC INTERACTIONS
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DOI：
10.1007/bf00494642
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发表时间：
1980-01-01
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影响因子：
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通讯作者：
GRUBE, D
中科院分区：
文献类型：
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作者：
GRUBE, D
各种胃肠胰（GEP）内分泌细胞已被证明同时含有针对多种肽激素的免疫反应性。在本研究中，通过未标记抗体酶 (PAP) 技术，使用免疫细胞化学染色序列中单个步骤的修改，研究了大鼠胃胃泌素 (G-) 细胞针对 21 种特异性抗血清和 10 种对照血清的“免疫反应性”。结果表明，免疫球蛋白可以通过非特异性离子相互作用与胃泌素细胞颗粒明显结合。即使在 GEP 内分泌系统免疫组织化学研究中常用的血清稀释范围内，免疫球蛋白的这种非特异性结合也会发生。由于“吸附对照”（抗血清与其各自抗原的预吸附）不会区分免疫球蛋白与 GEP 内分泌细胞的特异性和非特异性结合，因此需要额外的特异性对照。与通过“建立的”抗血清对各种GEP内分泌细胞进行免疫染色以及通过胃泌素抗血清对G细胞进行免疫染色相反，来自非免疫动物的血清的免疫球蛋白以及抗促肾上腺皮质激素-促脂激素相关肽的抗体可以通过改变用作稀释剂或漂洗溶液的缓冲液的NaCl含量而从其在G细胞中的结合位点被取代。为了排除免疫球蛋白非特异性结合对 GEP 内分泌细胞的免疫染色，建议对这些细胞进行免疫细胞化学研究时采用以下工作程序： 1. 使用低浓度的高效价抗血清（稀释 1:1,500 或更多）。 2. 将用作稀释剂或冲洗溶液的缓冲液的盐 (NaCl) 含量升高至 0.5 M。 3. 只有考虑到第 1 点和第 2 点，吸附控制才会显示可靠的结果。
Various gastro-entero-pancreatic (GEP) endocrine cells have been shown to contain concomitantly immunoreactivities against several peptide hormones. In the present study the “immunoreactivities” of gastrin (G-) cells of the rat stomach against 21 specific antisera and 10 control sera were investigated by means of the unlabelled antibody enzyme (PAP) technique using modifications of single steps in the immunocytochemical staining sequence. The results indicate that immunoglobulins can bind to gastrin cell granules obviously by non-specific ionic interactions. This non-specific binding of immunoglobulins occurs even in dilution ranges of the sera commonly used in immunohistochemical investigations of the GEP endocrine system. Since “adsorption controls” (preadsorption of the antisera with their respective antigens) will not discriminate between specific and nonspecific binding of immunoglobulins to GEP endocrine cells additional specificity controls are necessary. In contrast to the immunostaining of various GEP endocrine cells by “established” antisera and of G-cells by gastrin antiserum immunoglobulins of sera from non-immunized animals as well as antibodies against corticotropin-lipotropin related peptides could be displaced from their binding sites in G-cells by alterations of the NaCl content of the buffers used as diluents or as rinsing solutions. To exclude immunostaining of GEP endocrine cells by nonspecific binding of immunoglobulins the following working procedures are recommended for immunocytochemical investigations of these cells: 1. Use of high titer antisera at low concentrations (diluted 1:1,500 or more). 2. Elevation of the salt (NaCl) content up to 0.5 M of the buffer used as diluent or as rinsing solution. 3. Adsorption controls will show reliable results only if point 1. and 2. have been taken into account.