Squiquera L, Galimberti R, Morelli L, Plotkin L, Milicich R, Kowalckzuk A, Leoni J
Immunodermatology Unit, Hospital Italiano de Buenos Aires, Argentina.
Clin Exp Dermatol. 1994 Jul;19(4):289-93. doi: 10.1111/j.1365-2230.1994.tb01197.x.
In order to analyse the humoral immune response to the commensal yeast Pityrosporum ovale, we developed a western immunoblot technique with a salt soluble extract of P. ovale cytoplasm. In the present study, we tested sera from patients with psoriasis (n = 15), seborrhoeic dermatitis (n = 10), pityriasis versicolor (n = 8), and normal controls (n = 10). Seventy-three per cent (11/15) of the patients with psoriasis showed specific reactivity with a protein derived from P. ovale of estimated molecular mass 120 kDa, and 46% (7/15) of the cases recognized a 100-kDa protein. Sera from pityriasis versicolor and normal donors showed nonspecific reactivity with several bands of lower molecular weight. To characterize the location of the 100 and 120-kDa proteins, we performed a lyticase digestion of the cell wall, and analysed the soluble digested products by western blotting. The sera from psoriasis patients detected several bands in the range 100-120 kDa. The finding of the immunoreactive 120-kDa protein in this fraction suggests its location at the space between cell wall and membrane (periplasmic space). As a control, we performed an extraction of the cytoplasmic proteins of the dimorphic yeast Candida albicans. C. albicans showed a different pattern of banding in SDS-PAGE. Immunoblots with C. albicans did not allow the detection of any related band. A smear was observed in the high molecular weight range consistent with the presence of lipopolysaccharides. The role of the immune response in infection by P. ovale has not yet been fully explored.(ABSTRACT TRUNCATED AT 250 WORDS)
为了分析对共生酵母卵形糠秕孢子菌的体液免疫反应,我们开发了一种用卵形糠秕孢子菌细胞质盐溶性提取物进行的蛋白质免疫印迹技术。在本研究中,我们检测了银屑病患者(n = 15)、脂溢性皮炎患者(n = 10)、花斑糠疹患者(n = 8)和正常对照者(n = 10)的血清。73%(11/15)的银屑病患者对一种源自卵形糠秕孢子菌、估计分子量为120 kDa的蛋白质表现出特异性反应,46%(7/15)的病例识别出一种100 kDa的蛋白质。花斑糠疹患者和正常供体的血清与几条较低分子量的条带表现出非特异性反应。为了确定100 kDa和120 kDa蛋白质的位置,我们对细胞壁进行了溶菌酶消化,并通过蛋白质免疫印迹分析了可溶性消化产物。银屑病患者的血清在100 - 120 kDa范围内检测到几条条带。在该部分中发现免疫反应性120 kDa蛋白质表明其位于细胞壁和细胞膜之间的空间(周质空间)。作为对照,我们对双态酵母白色念珠菌的细胞质蛋白进行了提取。白色念珠菌在SDS - PAGE中显示出不同的条带模式。用白色念珠菌进行的免疫印迹未检测到任何相关条带。在高分子量范围内观察到一条涂片,与脂多糖的存在一致。卵形糠秕孢子菌感染中免疫反应的作用尚未得到充分探索。(摘要截断于250字)