Woo T Y, Hogan V A, Patel H, Anhalt G J, Labib R S, Voorhees J J, Diaz L A
J Invest Dermatol. 1983 Jul;81(1 Suppl):115s-21s. doi: 10.1111/1523-1747.ep12540871.
IgG isolated from sera of patients with pemphigus vulgaris (PV) has been shown to induce cell detachment when added to primary epidermal cell cultures (PECC). We studied the specificity of this phenomenon. IgG fractions were purified from the sera of five patients with PV and control IgG fractions from the sera of normal donors and patients with bullous pemphigoid (BP), systemic lupus erythematosus (SLE), and anti-AB blood group sera (anti-AB). IgG fractions were added to PECC either at initial plating (0 hours), at media change (48 hours), or sequentially at both times, and cell detachment was quantitated at 72 and 96 hours. Significant cell detachment occurred only when PV IgG was added to the growth media sequentially at 0 and 48 hours (p = 0.001), and this effect was dose-dependent for either dose. Substitution of an unrelated IgG (BP, SLE, or anti-AB) at either time points reduced cell detachment to near control values. Furthermore, cell detachment was inhibited by the addition of the proteinase inhibitors alpha 2 macroglobulin (70% inhibition of detachment), aprotinin (63% inhibition), soybean and lima bean trypsin inhibitor (62 and 64%, respectively), and pepstatin (49%), but not by the inhibitors chymostatin, leupeptin, or antipain. These data confirm that PV IgG induces increased cell detachment in PECC and shows that this effect is specific for PV IgG, is dose-dependent, and may be inhibited by certain proteinase inhibitors.
从寻常型天疱疮(PV)患者血清中分离出的IgG,已被证明添加到原代表皮细胞培养物(PECC)中时可诱导细胞脱离。我们研究了这一现象的特异性。从5例PV患者的血清中纯化IgG组分,并从正常供体、大疱性类天疱疮(BP)、系统性红斑狼疮(SLE)患者的血清以及抗AB血型血清(抗AB)中获取对照IgG组分。IgG组分在初始接种时(0小时)、换液时(48小时)或在这两个时间点依次添加到PECC中,并在72小时和96小时对细胞脱离进行定量。仅当在0小时和48小时依次将PV IgG添加到生长培养基中时才会发生显著的细胞脱离(p = 0.001),并且这种效应在任何一种剂量下均呈剂量依赖性。在任一时刻用无关的IgG(BP、SLE或抗AB)替代可将细胞脱离减少至接近对照值。此外,添加蛋白酶抑制剂α2巨球蛋白(抑制脱离70%)、抑肽酶(63%抑制)、大豆和利马豆胰蛋白酶抑制剂(分别为62%和64%)以及胃蛋白酶抑制剂(49%)可抑制细胞脱离,但糜蛋白酶抑制剂、亮抑酶肽或抗蛋白酶则无此作用。这些数据证实PV IgG可诱导PECC中细胞脱离增加,并表明这种效应对PV IgG具有特异性、呈剂量依赖性,且可能被某些蛋白酶抑制剂所抑制。