Dibbern Donald A, Palmer G William, Williams P Brock, Bock S Allan, Dreskin Stephen C
Division of Allergy and Clinical Immunology, University of Colorado Health Sciences Center, Medical School Building, Room 4627, 4200 East 9th Avenue, Denver, CO 80262, USA.
J Immunol Methods. 2003 Mar 1;274(1-2):37-45. doi: 10.1016/s0022-1759(02)00369-1.
Rat basophilic leukemia cells (RBL SX-38) express the alpha, beta, and gamma chains of human Fc epsilon RI. Following sensitization with IgE from a subset of allergic human donors, these cells can be triggered by exposure to anti-IgE or to very low concentrations of specific allergens. We examined 18 sera from patients who were highly sensitive to peanuts by history and had anti-peanut IgE by in vitro testing. The ability of these sera to sensitize the RBL SX-38 cells for degranulation with peanut allergens correlates very well with the absolute amount of anti-peanut IgE (r=0.95; p<0.001). The most effective sera contained at least 50 kU/l of total IgE and at least 15 kU/l of peanut-specific IgE. RBL SX-38 cells sensitized with these sera degranulated optimally upon exposure to anti-IgE (net degranulation of 40+/-8%, means+/-S.D.; n=8) and to a 10(5)-10(6) dilution of crude peanut extract (CPE) (37+/-7% net degranulation; 93+/-13% of that seen with anti-IgE). This assay is quite sensitive. Cells sensitized with selected sera are activated by exposure to a 1:10(7) dilution of the CPE containing picogram amounts of peanut allergens. This assay is also quite specific. Cells sensitized with sera from patients with anti-peanut IgE and no detectable IgE against soybean, walnut or grass pollen did not degranulate following exposure to these latter antigens. The converse was also true; cells sensitized with sera from patients without anti-peanut IgE did not react to peanut. These data demonstrate that RBL cells expressing human Fc epsilon RI form the basis of a useful model system for the detection of allergens and for the study of IgE-allergen interactions.
大鼠嗜碱性白血病细胞(RBL SX - 38)表达人FcεRI的α、β和γ链。在用来自一部分过敏人类供体的IgE致敏后,这些细胞可通过暴露于抗IgE或极低浓度的特异性变应原而被触发。我们检测了18份来自有花生过敏史且体外检测有抗花生IgE的患者的血清。这些血清使RBL SX - 38细胞对花生变应原脱颗粒致敏的能力与抗花生IgE的绝对量密切相关(r = 0.95;p < 0.001)。最有效的血清含有至少50 kU/l的总IgE和至少15 kU/l的花生特异性IgE。用这些血清致敏的RBL SX - 38细胞在暴露于抗IgE(净脱颗粒率为40±8%,平均值±标准差;n = 8)和10⁵ - 10⁶稀释的粗花生提取物(CPE)时达到最佳脱颗粒效果(净脱颗粒率为37±7%;是抗IgE诱导脱颗粒率的93±13%)。该检测方法相当灵敏。用选定血清致敏的细胞在暴露于含有皮克量花生变应原的1:10⁷稀释的CPE时被激活。该检测方法也相当特异。用有抗花生IgE且未检测到针对大豆、核桃或草花粉的IgE的患者血清致敏的细胞,在暴露于这些后一种抗原时不会脱颗粒。反之亦然;用无抗花生IgE的患者血清致敏的细胞对花生无反应。这些数据表明,表达人FcεRI的RBL细胞构成了一个用于检测变应原和研究IgE - 变应原相互作用的有用模型系统的基础。