Irani A M, Huang C, Xia H Z, Kepley C, Nafie A, Fouda E D, Craig S, Zweiman B, Schwartz L B
Department of Pediatrics, Virginia Commonwealth University, Richmond 23298, USA.
J Allergy Clin Immunol. 1998 Mar;101(3):354-62. doi: 10.1016/S0091-6749(98)70248-9.
Human basophils are difficult to detect with classic histochemical stains at sites of allergic inflammation. The 2D7 anti-basophil monoclonal antibody was used to identify basophils in skin during the late-phase response to a cutaneous allergen challenge.
The 2D7 monoclonal antibody was used on protease-digested sections of skin biopsy specimens obtained 6 and 24 hours after an allergen or buffer challenge. The skin chamber technique was used to compare buffer- and allergen-challenged sites at 6 hours, and intradermal injection of allergen was used to compare allergen-challenged sites at 6 and 24 hours.
Dramatic increases in the numbers of 2D7+ cells and in tissue staining by 2D7 were observed 6 hours after allergen challenge compared with buffer challenge. Histamine levels in skin chamber fluid varied with 2D7+ cell concentrations. By 24 hours, 2D7+ cells and tissue staining appeared to diminish but were still detectable in the allergen-challenged sites. Basophils localized primarily in and around blood vessels, whereas mast cells remained mostly in the superficial dermis. Mast cells were 2D7- in both the allergen- and buffer-challenged skin. Metachromatic staining of 2D7+ basophils with toluidine blue was absent in these tissue sections.
The 2D7 monoclonal antibody provides a more sensitive and precise marker than histochemical staining for human basophil involvement during the late-phase response to an allergen challenge. Basophil infiltration was observed at 6 hours only after allergen challenge and persisted at similar levels by 24 hours.
在变应性炎症部位，经典组织化学染色很难检测到人类嗜碱性粒细胞。2D7抗嗜碱性粒细胞单克隆抗体被用于在皮肤变应原激发后的迟发反应期识别皮肤中的嗜碱性粒细胞。
在变应原或缓冲液激发后6小时和24小时获取的皮肤活检标本经蛋白酶消化的切片上使用2D7单克隆抗体。采用皮肤小室技术在6小时时比较缓冲液激发部位和变应原激发部位，并采用变应原皮内注射在6小时和24小时时比较变应原激发部位。
与缓冲液激发相比，变应原激发后6小时观察到2D7+细胞数量和2D7组织染色显著增加。皮肤小室液中的组胺水平随2D7+细胞浓度而变化。到24小时时，2D7+细胞和组织染色似乎减少，但在变应原激发部位仍可检测到。嗜碱性粒细胞主要定位于血管内及血管周围，而肥大细胞大多仍位于浅表真皮层。在变应原激发和缓冲液激发的皮肤中，肥大细胞均为2D7阴性。在这些组织切片中，2D7+嗜碱性粒细胞未出现甲苯胺蓝异染性染色。
对于变应原激发迟发反应期人类嗜碱性粒细胞的参与情况，2D7单克隆抗体提供了一种比组织化学染色更敏感和精确的标志物。仅在变应原激发后6小时观察到嗜碱性粒细胞浸润，到24小时时仍维持在相似水平。
