| 抗斯氏按蚊中肠蛋白组分的抗体对约氏疟原虫卵囊的作用 |
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| 引用本文： | 魏秋芬,曾令娥,孙宝清,邵长玲,王凤芸,诸欣平.抗斯氏按蚊中肠蛋白组分的抗体对约氏疟原虫卵囊的作用[J].中国寄生虫学与寄生虫病杂志,2006,24(6):441-444. |
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| 作者姓名： | 魏秋芬 曾令娥 孙宝清 邵长玲 王凤芸 诸欣平 |
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| 作者单位： | 1. 首都医科大学燕京医学院免疫学与病原生物学教研室,北京,101300
2. 首都医科大学基础医学院病原生物学教研室,北京,100069 |
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| 基金项目： | 首都医科大学自然科学基金 |
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| 摘 要： | 目的 观察抗斯氏按蚊中肠蛋白组分的抗体对约氏疟原虫卵囊的抑制作用。 方法 解剖实验室饲养斯氏按蚊雌蚊，取中肠（胃）制备中肠蛋白抗原并免疫BALB/c小鼠（8只， 100 μg/只)， 共免疫4次， 每次间隔7～10 d，末次免疫后10 d，腋窝动脉取血，分离血清。用蛋白质印迹（Western blotting）分析中肠蛋白的免疫活性抗原。用葡聚糖凝胶过滤法获得相对分子质量（Mr）为38 000~50 000的蛋白。用该中肠蛋白免疫小鼠（12只， 100 μg/只)， 共免疫4次，每次间隔7~10 d。同时设PBS对照组。末次免疫后7 d，ELISA检测小鼠血清中的抗体，抗体效价≥1 : 2 560时，该免疫组小鼠和对照组小鼠经腹腔接种感染约氏疟原虫（约含2×107个感染疟原虫的红细胞），感染后3 d取小鼠尾血镜检，雌配子体数＞2/10个视野的小鼠作为供血鼠，斯氏按蚊成蚊吸血后9 d解剖，计数中肠的卵囊数量。 结果 Western blotting显示斯氏按蚊中肠蛋白抗原的显色区带有8条，其中Mr 38 000~50 000的区带显色较清晰；实验组和对照组中肠卵囊感染率分别为28.70%（62/216）和51.09%（47/92）（P＜0.05），中肠卵囊指数分别为14.14（1 541/109）和26.02（1 223/47），两者差异有统计学意义（P＜0.01）。 结论 斯氏按蚊Mr 38 000~50 000中肠蛋白有免疫活性；针对该中肠蛋白的抗体对约氏疟原虫卵囊的发育有明显的抑制作用。
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| 关 键 词： | 斯氏按蚊 中肠蛋白 抗体 卵囊 约氏疟原虫 抑制作用 |
| 文章编号： | 1000-7423（2006）-06-0441-04 |
| 收稿时间： | 2006-08-26 |
| 修稿时间： | 2006年8月26日 |
Effect of Anti-Midgut-Protein-Ingredient Antibodies of Anophelesstephensi on the Oocysts of Plasmodium yoelii |
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| WEI Qiu-fen,ZENG Ling-e,SUN Bao-qing,SHAO Chang-ling,WANG Feng-yun,ZHU Xin-ping.Effect of Anti-Midgut-Protein-Ingredient Antibodies of Anophelesstephensi on the Oocysts of Plasmodium yoelii[J].Chinese Journal of Parasitology and Parasitic Diseases,2006,24(6):441-444. |
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| Authors: | WEI Qiu-fen ZENG Ling-e SUN Bao-qing SHAO Chang-ling WANG Feng-yun ZHU Xin-ping |
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| Affiliation: | Yanjing School, Capital University of Medical Sciences, Beijing 101300, China. weiquif@tom.com |
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| Abstract: | OBJECTIVE: To observe the inhibitory effect of the antibodies against midgut-protein-ingredient of Anopheles stephensi on the oocysts of Plasmodium yoelii. METHODS: Female An. stephensi mosquitoes raised in laboratory were dissected and the midguts were collected. Eight BALB/c mice were immunized using midgut-protein (100 microg/mouse, 4 times with an interval of 7-10 day). Ten days after the last immunization, blood was taken from mice armpit artery and serum separated. The immune active antigen of the midgut protein was analyzed by Western blotting. Protein with Mr 38 000-50 000 was separated by sephadex filtering and used to immunize 12 BALB/c mice (100 microg/ mouse, 4 times with interval of 7-10 days). PBS control group was established. Seven days after the last immunization, serum antibody was detected by ELISA. When the antibody titer in immunized mice reached > or = 1:2 560, mice in both groups were infected by P. yoelii (about 2 x 10(7) plasmodium-infected RBC) by abdominal injection. The mosquitoes were fed on the infected mice when the number of female gametes was higher than 2 per 10 microscopical fields 3 days later. After 9 days, the mosquitoes were dissected and the amount of oocysts in midgut was counted. RESULTS: Eight protein bands were shown in midgut-protein of An. stephensi by Western blotting and the band of Mr 38 000-50 000-midgut-protein appeared clearer. The infection rate of oocysts in the experiment and control groups were 28.70% (62/216) and 51.09% (47/92) respectively (P<0.05), with an oocyst index of 14.14 (1 541/109) and 26.02 (1 223/47) respectively (P<0.01). CONCLUSIONS: The midgut protein of Anopheles stephensi with Mr 38 000-50 000 has immune activity, and the antibodies against this protein shows an inhibitory effect on the development of oocysts of Plasmodium yoelii. |
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| Keywords: | Anopheles stephensi Midgut protein Antibody Oocyst Plasmodium yoelii Inhibitory effect |
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