Lussow A R, Barrios C, van Embden J, Van der Zee R, Verdini A S, Pessi A, Louis J A, Lambert P H, Del Giudice G
World Health Organization-Immunology Research and Training Center, Department of Pathology, University of Geneva, Switzerland.
Eur J Immunol. 1991 Oct;21(10):2297-302. doi: 10.1002/eji.1830211002.
We have previously shown that the priming of mice with live Mycobacterium tuberculosis var. bovis (Bacillus Calmette-Guérin, BCG) and immunization with the repetitive malaria synthetic peptide (NANP)40 conjugated to purified protein derivative (PPD), led to the induction of high and long-lasting titers of anti-peptide IgG antibodies, overcoming the requirement of adjuvants and the genetic restriction of the antibody response to the peptide (Lussow et al., Proc. Natl. Acad. Sci. USA 1990. 87:2960). This initial work led us to the following observations. BCG had to be live for priming to lead to the induction of anti-peptide antibodies. Surprisingly, priming with other living microorganisms which chronically infect the macrophage (e.g. Salmonella typhimurium and Leishmania major) also induced anti-peptide antibodies in mice immunized with PPD-(NANP)40 conjugate. It was, thus, hypothesized that molecules expressed during active infection and also known to be highly conserved between species, namely the heat-shock proteins (hsp), could mediate the T cell sensitization required for the production of anti-peptide antibodies. In fact, when the PPD protion of the conjugate was replaced by a highly purified recombinant protein corresponding to the 65-kDa (GroEL-type) hsp of M. bovis, this resulted in the production of anti-(NANP) IgG antibodies in BCG-primed mice, irrespective of the major histocompatibility complex-controlled responsiveness to the (NANP) sequence itself. Further, similar induction of anti-peptide antibody response was also obtained with a recombinant 70-kDa (DnaK-type) hsp of M. tuberculosis, but not with a small molecular mass (18 kDa) of M. leprae. Finally, an adjuvant-free carrier effect for anti-peptide IgG antibody production in BCG-primed mice, was also exerted by the GroEL hsp of Escherichia coli. This finding that hsp can act as carrier molecules without requiring conventional adjuvants is of potential importance in the development of vaccine strategies.
我们之前已经表明，用活的牛分枝杆菌（卡介苗，BCG）对小鼠进行致敏，并使用与纯化蛋白衍生物（PPD）偶联的重复疟疾合成肽（NANP）40进行免疫，可诱导产生高滴度且持久的抗肽IgG抗体，克服了对佐剂的需求以及抗体对该肽反应的遗传限制（Lussow等人，《美国国家科学院院刊》1990年。87:2960）。这项初步工作使我们得出以下观察结果。BCG必须是活的才能致敏以诱导抗肽抗体的产生。令人惊讶的是，用其他长期感染巨噬细胞的活微生物（如鼠伤寒沙门氏菌和硕大利什曼原虫）进行致敏，也能在接种PPD-(NANP)40偶联物的小鼠中诱导产生抗肽抗体。因此，有人推测在活跃感染期间表达且已知在物种间高度保守的分子，即热休克蛋白（hsp），可能介导产生抗肽抗体所需的T细胞致敏。事实上，当偶联物的PPD部分被对应于牛分枝杆菌65 kDa（GroEL型）hsp的高度纯化重组蛋白取代时，这导致在BCG致敏的小鼠中产生抗-(NANP) IgG抗体，而与主要组织相容性复合体对（NANP）序列本身的反应性控制无关。此外，用结核分枝杆菌的重组70 kDa（DnaK型）hsp也获得了类似的抗肽抗体反应诱导，但用麻风分枝杆菌的小分子量（18 kDa）hsp则未获得。最后，大肠杆菌的GroEL hsp对BCG致敏小鼠中抗肽IgG抗体的产生也发挥了无佐剂的载体效应。热休克蛋白无需传统佐剂即可作为载体分子这一发现，在疫苗策略的开发中具有潜在重要性。
