摘要
目的 :筛选和鉴定结核分枝杆菌特异性和保护性抗原 ,研究结核杆菌的免疫反应特点 ,以探索结核病诊断和治疗的新途径。方法 :采用超声破碎和滤膜抽滤的方法分别得到菌体蛋白和滤液蛋白 ,通过Westernblot试验用结核杆菌的单克隆抗体及结核病人血清来检测蛋白样品 ,把发生阳性反应的蛋白在BECKMANLF32 0 0 多肽氨基酸序列测定仪上进行N末端序列分析 ,并用结核杆菌的单抗对自身抗原组蛋白进行了检测。结果 :结核杆菌的 31kD和 30kD蛋白与结核杆菌的单抗及病人血清反应均呈阳性 ,但与正常小鼠血清和健康人血清反应呈阴性。 31kD和 30kD蛋白的N末端序列分别为 :AlaGluValAspTrpLeuValPheAlaVal和PheSerArgProGlyLeuProValGluTyr。结核分枝杆菌的单抗与自身抗原组蛋白能发生免疫交叉反应。结论 :结核杆菌的 31kD和 30kD蛋白是免疫保护性抗原 ,对免疫交叉反应分子基础的进一步研究必将增加对结核免疫机理的了解。
Objective:To select and identify the specific and protective antigens of Mycobacterium tuberculosis for searching a new approach to diagnose and treat tuberculosis. Methods:Extract and culture filtrates of Mycobacterium tuberculosis were obtained by ultrasonic treatment and millipore membrane filtration, respectively. The protein samples were tested with monoclonal antibody (MAb) and patients sera. The proteins showing positive reaction were sequenced on Beckman /LF 3200/peptide amino acid sequencer. BCG was used as the contrast. Self-antigen histone was tested with MAb also. Results:Proteins of Mycobacterium tuberculosis with molecular weight of 31 kD and 30 kD showed positive results when reacted with anti-Mycobacterium tuberculosis MAb and sera of TB patients, but not with normal mouse serum and healthy human sera. N-terminal sequences of the 31 kD and 30 kD antigen are Ala Glu Val Asp Trp Leu Val Phe Ala Val and Phe Ser Arg Pro Gly Leu Pro Val Glu Tyr respectively. Immune cross-reaction of Mycobacterium tuberculosis MAb with self-antigen histone was demonstrated by Western blot.Conclusion:31 kD and 30 kD proteins of Mycobacterium tuberculosis are immune protective proteins. The discovery of immune cross-reaction induce a new idea about immunological mechanism of Mycobacterium tuberculosis. [
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2003年第5期309-312,共4页
Chinese Journal of Immunology
关键词
结核分枝杆菌
抗原分析
免疫交叉反应
研究
结核病
Mycobacterium tuberculosis
Antigen
Amino acid sequence
Monoclonal antibody(MAb)
