Fan Ai-Lin, Jian Wen, Shi Chang-Hong, Su Ming-Quan, Bai Yin-Lan, Ma Jing, Xu Zhi-Kai, Hao Xiao-Ke
Department of Clinical Laboratory, Fourth Military Medical University, Xi'an 710032, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2008 Jul;24(7):686-8.
To investigate the immunobiology of Rpf domain from Micrococcus luteus.
BALB/c mice were immunized with Rpf domain three times at 2-week interval. ELISA was used to detect the title of the anti-Rpf domain antibody titer in the immunized mice sera. The spleen lymphocytes of the immunized mice were separated and the stimulation index (SI) was measured by MTT colorimetry. The levels of secreted IFN-gamma, IL-10 and IL-12 upon specific antigen stimulation was detected by ELISA. The Rpf domain immunized BALB/c mice were intravenously infected with 10(5) CFU MTB H37Rv. The number of CFU in the spleens was determined four weeks after final injection.
The titer of the specific antibody in sera of the immunized BALB/c mice was 1:128 000. The SI of Rpf domain immunized group (2.10+/-0.12) was significantly higher than that of saline immunized group (0.90+/-0.21). The lever of IFN-gamma, IL-10 and IL-12 levels in culture supernatant of spleen lymphocytes from the fusion protein immunized mice was (1 126+/-36) ng/L, (368+/-13) ng/L and (289+/-14) ng/L, respectively, which was markedly higher than that of saline immunized group (P<0.01). Compared with normal saline immunized mice (6.64+/-0.13) four weeks after final injection, dramatic reduction in MTB replication was observed in the spleen (5.03+/-0.11) from the BALB/c mice immunized with fusion proteins.
Rpf domain can be used as a candidate for a new TB vaccine.
研究藤黄微球菌Rpf结构域的免疫生物学特性。
以2周的间隔对BALB/c小鼠进行3次Rpf结构域免疫。采用ELISA法检测免疫小鼠血清中抗Rpf结构域抗体的效价。分离免疫小鼠的脾淋巴细胞,用MTT比色法测定刺激指数(SI)。用ELISA法检测特异性抗原刺激后分泌的干扰素-γ、白细胞介素-10和白细胞介素-12水平。用10⁵CFU结核分枝杆菌H37Rv静脉感染经Rpf结构域免疫的BALB/c小鼠。末次注射4周后测定脾脏中的CFU数量。
免疫的BALB/c小鼠血清中特异性抗体效价为1:128 000。Rpf结构域免疫组的SI(2.10±0.12)显著高于生理盐水免疫组(0.90±0.21)。融合蛋白免疫小鼠脾淋巴细胞培养上清中干扰素-γ、白细胞介素-10和白细胞介素-12水平分别为(1 126±36)ng/L、(368±13)ng/L和(289±14)ng/L,明显高于生理盐水免疫组(P<0.01)。末次注射4周后,与生理盐水免疫小鼠(6.64±0.13)相比,用融合蛋白免疫的BALB/c小鼠脾脏中结核分枝杆菌的复制显著减少(5.03±0.11)。
Rpf结构域可作为新型结核疫苗的候选物。