San Gil F, Turner B, Mullbacher A, Walker M J, Djordjevic S P, Eamens G J, Chin J C
NSW Agriculture, Elizabeth Macarthur Agricultural Institute, Camden, Australia.
Scand J Immunol. 1998 Mar;47(3):243-53. doi: 10.1046/j.1365-3083.1998.00304.x.
As it is not known what changes to leucocyte homeostasis are mandatory for effective adjuvant action, the biological relevance of systemic changes elicited by different vaccine formulations can only be interpreted in the context of the immunological outcomes. We used flow cytometry to quantify the changes in leucocyte subsets induced in mice intradermally immunized with SAMA4 (adjuvant group), outer membrane proteins (OMP) purified from Actinobacillus pleuropneumoniae (OMP antigen group), SAMA4 adjuvanted OMP (OMP vaccine group), or phosphate-buffered saline (PBS: control group). This approach allowed direct comparisons to be made between the effects of antigen, adjuvant or antigen-adjuvant complexes on immune effector cell populations. Antigens complexed with the liposome-iscom hybrid adjuvant, SAMA4, generated strong antibody responses and cytotoxic T-cell activity in animals immunized intradermally, reflecting remobilization and recruitment of specific cell populations. Splenomegaly, due to granulocytosis, monocytosis and megakaryocytosis, was most prominent in the OMP vaccine group. Histological examination of spleen sections confirmed that these changes were due primarily to splenic haematopoiesis. Circulating numbers of granulocytes and monocytes increased significantly (P < 0.05) in the blood of the OMP vaccine group, as did granulocyte numbers in the lungs (P < 0.05). No changes in T- and B-cell numbers were detected by flow cytometry in the spleens, lungs or blood over the 28-day period in any treatment group. Thymocyte numbers (predominantly CD4+CD8+ cells) in the OMP vaccine group fell by 95% within 3 days of immunization. Identical cellular responses were obtained when an innocuous antigen, ovalbumin, was complexed with SAMA4 instead of OMP, thus demonstrating that the adjuvant effects of SAMA4 were due to synergistic interaction between antigen and adjuvant and not due to the presence of toxic components. The association of strong adaptive immune responses with such complex changes in leucocyte homeostasis induced by complexing adjuvant and antigen suggested that the changes were important for effective vaccination and were not purely circumstantial.
由于尚不清楚白细胞稳态的哪些变化对于有效的佐剂作用是必不可少的，因此不同疫苗制剂引起的全身变化的生物学相关性只能在免疫结果的背景下进行解释。我们使用流式细胞术来量化在皮内接种SAMA4（佐剂组）、从胸膜肺炎放线杆菌纯化的外膜蛋白（OMP抗原组）、SAMA4佐剂化的OMP（OMP疫苗组）或磷酸盐缓冲盐水（PBS：对照组）的小鼠中诱导的白细胞亚群变化。这种方法允许直接比较抗原、佐剂或抗原-佐剂复合物对免疫效应细胞群体的影响。与脂质体-免疫刺激复合物杂交佐剂SAMA4复合的抗原，在皮内免疫的动物中产生了强烈的抗体反应和细胞毒性T细胞活性，反映了特定细胞群体的重新动员和募集。由于粒细胞增多、单核细胞增多和巨核细胞增多导致的脾肿大在OMP疫苗组中最为明显。脾脏切片的组织学检查证实这些变化主要是由于脾造血。OMP疫苗组血液中粒细胞和单核细胞的循环数量显著增加（P<0.05），肺部粒细胞数量也显著增加（P<0.05）。在任何治疗组的28天期间，通过流式细胞术未检测到脾脏、肺部或血液中T细胞和B细胞数量的变化。OMP疫苗组的胸腺细胞数量（主要是CD4+CD8+细胞）在免疫后3天内下降了95%。当无害抗原卵清蛋白与SAMA4而不是OMP复合时，获得了相同的细胞反应，因此证明SAMA4的佐剂作用是由于抗原和佐剂之间的协同相互作用，而不是由于有毒成分的存在。强大的适应性免疫反应与佐剂和抗原复合诱导的白细胞稳态的这种复杂变化之间的关联表明，这些变化对于有效的疫苗接种很重要，而不仅仅是偶然的。
