Department of Microbiology, Immunology and Parasitology, Universidade Federal de São Paulo, São Paulo, Brazil.
Exp Parasitol. 2010 Feb;124(2):153-8. doi: 10.1016/j.exppara.2009.09.003. Epub 2009 Sep 6.
A 500 bp fragment encoding an isoform of cysteine proteinase from Leishmania (Leishmania) amazonensis was subcloned and expressed in the pHis vector, resulting in a recombinant protein of 24 kDa, rLacys24. In Western blots of L. (L.) amazonensis extracts, antibodies directed to rLacys24 recognized a cysteine proteinase isoform of 30 kDa. Analysis by fluorescence-activated cell sorter showed a significantly higher expression of CD8(+) lymphocytes in animals immunized with rLacys24 plus CFA, whereas a low expression of CD4(+) lymphocytes was observed in these animals. The cytotoxicity of lymphocytes isolated from mice immunized with rLacys24 plus CFA on L. (L.) amazonensis-infected macrophages was significantly higher than that observed in the presence of lymphocytes from control animals. Immunization of BALB/c mice with rLacys24 plus CFA resulted in a low but significant decrease of foot lesions after challenge with L. (L.) amazonensis compared to those exhibited by control mice.
一个 500bp 的编码利什曼原虫（Leishmania）亚马逊变体胱氨酸蛋白酶的片段被亚克隆并在 pHis 载体中表达，产生了一个 24kDa 的重组蛋白 rLacys24。在利什曼原虫（Leishmania）亚马逊提取物的 Western blot 中，针对 rLacys24 的抗体识别出一种 30kDa 的胱氨酸蛋白酶同工型。流式细胞术分析显示，用 rLacys24 加 CFA 免疫的动物中 CD8+淋巴细胞的表达显著增加，而这些动物中 CD4+淋巴细胞的表达则较低。从用 rLacys24 加 CFA 免疫的小鼠中分离的淋巴细胞对感染巨噬细胞的利什曼原虫（Leishmania）亚马逊的细胞毒性明显高于对照动物的淋巴细胞。用 rLacys24 加 CFA 免疫 BALB/c 小鼠可导致在受到利什曼原虫（Leishmania）亚马逊挑战后足部病变的低度但显著降低，与对照小鼠相比。
