Nizze H
Exp Pathol (Jena). 1975;11(5-6):219-32.
Repeated intraperitoneal injections of anti-mouse pancreas rabbit serum or of anti-mouse pancreas guinea pig serum produce a chronical sclerotizing pancreatitis. This study has the aim to contribute to the further elucidation of the changes which occur in the acinar cells, as well as to the etiology and pathogenesis of immune pancreatitis, by means of immunohistological, enzyme histochemical and electron microscopic studies.
Anti-mouse pancreas rabbit serum was obtained by sensitization of rabbits with an admixture of AB-mouse pancreas extract (100,000 g - supernatant) and complete Freund's adjuvant [details see NIZZE, Exp. Path. (1975a)]. The presence of precipitating mouse pancreas antibodies in the rabbit serum was ascertained by the agargel diffusion test according to Duchterlony (1958). The experiments were performed with 54 adult male white mice (AB colony strain) of 22 to 30 g.b.s. (averagely 26 g). The animals were divided into 4 groups which were treated as follows: 1. 24 mice with anti-mouse pancreas rabbit serum, 2. 12 mice with rabbit normal serum, 3. 12 mice with physiological saline, 4. 6 mice remained untreated (controls) Always 4 animals of the group 1 as well as each 2 of the groups 2 and 3 were administered in total 1, 3, 5, 9, 17 or 33 intraperitoneal injections of 0.3 ml of the correspondent serum or with physiological saline within 3 hours, 1, 2, 4, 8 or 16 days. The last injection was regularly applied 3 hours before sacrification by decapitation. The time of sacrification was always at 11.00 o'clock a.m. For immunohistological and enzyme histochemical investigations 10 mum thick cryostat sections were prepared consisting of pancreatic specimens piled up to a bloc. In each case the tissue samples were taken from the experimental animals and from one control animal sacrificed at the same day. The sections were incubated in FITC-labelled anti-rabbit globulin goat serum at room temperature for 30 min in a moist chamber. For control of specificity were employed: a) initial incubation of equal sections with unlabelled anti-rabbit globulin goat serum for 30 min (""blocking test''), b) pancreatic tissue specimens of each one untreated control animal present in the cryostat sections and thus incubated like the pancreatic tissue of the experimental animals, c) native nonincubated cryostat sections from the same bloc to exclude nonspecific autofluorescence. Evaluation of the sections was done in a Zeiss-Lg-microscope with HBO-50 high pressure mercury lamp. Exciter filters were UG 1/3.5 and 1/1.5, the eyepiece was screened with a GG 9/1 filter photographs were taken on ORWO X-ray film RS 2 (VEB Filmfabrik Wolfen). The enzyme histochemical studies were performed on cryostat sections of the same tissue bloc using the following methods: lead nitrate- or calcium-Co-method after GOMORI (1952) for demonstration of acid and alkaline phosphatase, naphthylacetate method (NACHLAS and SELIGMAN 1945) for nonspecific esterase, MTT-co-method (PEARSE et al...
反复腹腔注射抗小鼠胰腺兔血清或抗小鼠胰腺豚鼠血清会引发慢性硬化性胰腺炎。本研究旨在通过免疫组织学、酶组织化学和电子显微镜研究,进一步阐明腺泡细胞中发生的变化,以及免疫性胰腺炎的病因和发病机制。
用AB小鼠胰腺提取物(100,000g - 上清液)与完全弗氏佐剂的混合物致敏兔子,获得抗小鼠胰腺兔血清[详细方法见NIZZE,《实验病理学》(1975a)]。根据Duchterlony(1958)的琼脂凝胶扩散试验确定兔血清中沉淀性小鼠胰腺抗体的存在。实验用54只体重22至30克(平均26克)的成年雄性白色小鼠(AB菌落株)进行。动物分为4组,处理如下:1. 24只小鼠注射抗小鼠胰腺兔血清;2. 12只小鼠注射兔正常血清;3. 12只小鼠注射生理盐水;4. 6只小鼠不处理(对照组)。每组中始终有4只第1组的动物以及第2组和第3组各2只动物,在3小时、1天、2天、4天、8天或16天内,总共进行1、3、5、9、17或33次腹腔注射0.3毫升相应血清或生理盐水。最后一次注射在断头处死前3小时常规进行。处死时间总是在上午11点。为进行免疫组织学和酶组织化学研究,制备了由堆积成块的胰腺标本组成的10μm厚的低温恒温切片。每种情况下,组织样本均取自实验动物和同一天处死的一只对照动物。切片在室温下于湿润小室中用异硫氰酸荧光素标记的抗兔球蛋白山羊血清孵育30分钟。用于特异性对照的方法有:a)将相同切片与未标记的抗兔球蛋白山羊血清初始孵育30分钟(“阻断试验”);b)低温恒温切片中每只未处理对照动物的胰腺组织标本,其孵育方式与实验动物的胰腺组织相同;c)来自同一块的未孵育的天然低温恒温切片,以排除非特异性自发荧光。切片在配备HBO - 50高压汞灯的蔡司Lg显微镜下进行评估。激发滤光片为UG 1/3.5和1/1.5,目镜用GG 9/1滤光片筛选,照片拍摄于ORWO X射线胶片RS 2(VEB Filmfabrik Wolfen)。酶组织化学研究在同一组织块的低温恒温切片上采用以下方法进行:GOMORI(1952)的硝酸铅或钙 - 钴法用于显示酸性和碱性磷酸酶,萘乙酸法(NACHLAS和SELIGMAN 1945)用于显示非特异性酯酶,MTT - 钴法(PEARSE等人...