Mullerad Jacob, Michal Israel, Fishman Yolanta, Hovav Avi-Hai, Barletta Raúl G, Bercovier Herve
Department of Clinical Microbiology, Hadassah Medical School, The Hebrew University, Jerusalem, Israel.
Med Microbiol Immunol. 2002 Mar;190(4):179-87. doi: 10.1007/s00430-001-0104-z.
Mycobacterium paratuberculosis (MPT) is the etiological agent of paratuberculosis. The disease is prevalent throughout the world, and exacts a heavy financial toll. At present, the only means of controlling this disease are culling or vaccination. The existing vaccines are not very efficient and produce a long-lasting local reaction at the point of injection and induce anti-bodies/delayed-type hypersensitivity (DTH) reaction that cannot be differentiated from those of naturally infected animals. New potent acellular vaccines that allow discrimination between infected and vaccinated animals are necessary to improve the control of this disease. We have isolated, overexpressed and purified the 85B antigen of MPT, and characterized the immune response induced by this antigen in mice. Our results showed that the recombinant MPT 85B (rMPT 85B) antigen induced a high production of interferon (IFN)gamma, interleukin (IL)-6, IL-10 and nitric oxide (NO). Spleen cells from mice immunized with rMPT 85B in Ribi adjuvant produced a higher level of IL-10 and NO than spleen cells of mice immunized with rMPT 85B only. In contrast, the addition of Ribi to the immunization protocol resulted in a lower amount of IFNgamma released by spleen cells. The levels of spleen cells proliferation in mice vaccinated with the rMPT 85B protein alone or with rMPT 85B with Ribi adjuvant were, respectively, four times or five times greater than in the control mice. The Ribi adjuvant induced significantly higher anti-85B antibody production of all classes tested and increased the IgG1/IgG2a ratio. DTH responses in mice footpads were observed only in mice immunized with rMPT 85B emulsified in Ribi. rMPT 85B induced both a Th1 and Th2 type of immune response with the later slightly more pronounced when the vaccination protocol comprised Ribi as an adjuvant. The rMPT 85B antigen elicited a strong immune response and can be considered as a potential candidate for a future acellular vaccine.
副结核分枝杆菌（MPT）是副结核病的病原体。该病在全球普遍存在，造成了沉重的经济损失。目前，控制这种疾病的唯一方法是扑杀或接种疫苗。现有的疫苗效率不高，会在注射部位产生持久的局部反应，并诱导产生抗体/迟发型超敏反应（DTH），而这些反应无法与自然感染动物的反应区分开来。为了更好地控制这种疾病，需要新型高效的无细胞疫苗，以便区分感染动物和接种疫苗的动物。我们已经分离、过表达并纯化了MPT的85B抗原，并对该抗原在小鼠体内诱导的免疫反应进行了表征。我们的结果表明，重组MPT 85B（rMPT 85B）抗原可诱导产生高水平的干扰素（IFN）γ、白细胞介素（IL）-6、IL-10和一氧化氮（NO）。用Ribi佐剂免疫rMPT 85B的小鼠脾脏细胞产生的IL-10和NO水平高于仅用rMPT 85B免疫的小鼠脾脏细胞。相反，在免疫方案中添加Ribi会导致脾脏细胞释放的IFNγ量减少。单独接种rMPT 85B蛋白或接种含Ribi佐剂的rMPT 85B的小鼠脾脏细胞增殖水平分别比对照小鼠高四倍或五倍。Ribi佐剂诱导所有测试类别中抗85B抗体的产生显著增加，并提高了IgG1/IgG2a比值。仅在用Ribi乳化的rMPT 85B免疫的小鼠脚垫中观察到DTH反应。rMPT 85B诱导了Th1和Th2两种类型的免疫反应，当免疫方案包含Ribi作为佐剂时，后者反应稍明显。rMPT 85B抗原引发了强烈的免疫反应，可被视为未来无细胞疫苗的潜在候选物。
