Grab D J, Baldwin C L, Brown W C, Innes E A, Lonsdale-Eccles J D, Verjee Y
International Laboratory for Research on Animal Diseases, Nairobi, Kenya.
Infect Immun. 1992 Sep;60(9):3892-6. doi: 10.1128/iai.60.9.3892-3896.1992.
Theileria parva is a protozoan parasite that infects and transforms bovine lymphocytes. Here we report the partial purification of a T. parva-specific protein from infected lymphocytes that is recognized by CD4+ parasite-specific T-cell clones derived from immune cattle. T. parva-infected lymphocytes were homogenized in Dulbecco's phosphate-buffered saline in the presence of protease inhibitors. The antigen was purified from a postmicrosomal supernatant by using a combination of DEAE-cellulose chromatography and hydroxylapatite column chromatography. After labelling with 125I, the antigen preparation was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and found to contain 8 to 10 proteins. This preparation was subjected to chromatography in phosphate-buffered saline on HPLC TSK-250/125 columns coupled in tandem. A radiolabelled protein of M(r) 24,000 correlated with antigenic activity.
小泰氏梨浆虫是一种感染并转化牛淋巴细胞的原生动物寄生虫。在此我们报告从感染的淋巴细胞中部分纯化出一种小泰氏梨浆虫特异性蛋白,该蛋白可被源自免疫牛的CD4 +寄生虫特异性T细胞克隆识别。在蛋白酶抑制剂存在的情况下,将感染小泰氏梨浆虫的淋巴细胞在杜尔贝科磷酸盐缓冲盐水中匀浆。通过结合使用DEAE - 纤维素色谱法和羟基磷灰石柱色谱法,从微粒体后上清液中纯化抗原。用125I标记后,将抗原制剂进行十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳,发现含有8至10种蛋白质。将该制剂在磷酸盐缓冲盐水中通过串联连接的HPLC TSK - 250/125柱进行色谱分析。一种相对分子质量为24,000的放射性标记蛋白与抗原活性相关。