Institute for Pharmacy and Foods, University of Havana, Cuba.
BMC Immunol. 2013;14 Suppl 1(Suppl 1):S12. doi: 10.1186/1471-2172-14-S1-S12. Epub 2013 Feb 25.
Proteoliposomes purified from the Outer Membrane of Neisseria meningitidis B, have been successfully used as core for adjuvants and vaccine formulations. We have tried to increase their structural definition and to conserve their efficacy and stability avoiding the addition of the aluminum hydroxide to the final formulation. Liposomal particle systems were prepared from components of defined molecular structure, such as a Neisseria meningitidis B protein complex, extracted and purified without forming vesicle structures. Liposomes were prepared from a mixture of dioleoyl phosphatidyl serine and cholesterol, using the classical dehydration-rehydration method. Transmission Electron Microscopy (TEM) was used to characterize the liposomes. BALB/c mice were used for animal testing procedures. Analysis of specific IgG response, serum bactericidal activity as well as DTH reaction was carried out. Isolation and purification of mRNA and real-time PCR, was performed to determine the dominating Th lymphokine pattern. The new antimeningococcal formulation without aluminum hydroxide prepared with components of defined molecular structure assembled itself into Neoproteoliposomes (NPL) ranging from 50 to 70 nm in diameter. The extraction and purification of selected membrane proteins to provide the antigen for this new formulation (PD-Tp), as well as the NPL-formulation favors a Th1 response pattern, suggested by the higher percentages of DTH, increased expression of proinflamatory lymphokine mRNAs when administered by intramuscular and intranasal routes. It stimulates a systemic bactericidal antibody response against Neisseria meningitidis B and immunologic memory similar to the Cuban VA-MENGOC-BC vaccine, even at lower dosages and is less reactogenic at the injection site in comparison with the formulation with aluminum hydroxide. This new adjuvant formulation could be applicable to the development of new and improved vaccines against meningococcal disease, and eventually as modulators of the immune response against other diseases.
从脑膜炎奈瑟菌 B 的外膜中纯化的蛋白脂囊泡已成功用作佐剂和疫苗制剂的核心。我们试图增加它们的结构清晰度,并在避免向最终制剂中添加氢氧化铝的情况下保持其功效和稳定性。脂质体颗粒系统由具有明确分子结构的成分组成,例如从脑膜炎奈瑟菌 B 中提取和纯化的蛋白质复合物,而不形成囊泡结构。使用经典的脱水-再水合方法从二油酰基磷脂酰丝氨酸和胆固醇的混合物中制备脂质体。使用透射电子显微镜(TEM)对脂质体进行了表征。使用 BALB/c 小鼠进行了动物测试程序。进行了特异性 IgG 反应、血清杀菌活性以及 DTH 反应的分析。分离和纯化 mRNA 以及实时 PCR,以确定主导 Th 淋巴因子模式。使用具有明确分子结构的组件制备的不含氢氧化铝的新型抗脑膜炎球菌制剂,自行组装成直径为 50 至 70nm 的新型 Neoproteoliposomes(NPL)。为这种新制剂(PD-Tp)提供抗原的选定膜蛋白的提取和纯化,以及 NPL 制剂有利于 Th1 反应模式,这表明 DTH 的百分比更高,炎症淋巴因子 mRNAs 的表达增加,这是通过肌内和鼻内途径给药时。它刺激针对脑膜炎奈瑟菌 B 的全身性杀菌抗体反应和免疫记忆,类似于古巴 VA-MENGOC-BC 疫苗,即使在较低剂量下,与含有氢氧化铝的制剂相比,在注射部位的反应性也较低。这种新的佐剂制剂可适用于开发针对脑膜炎球菌病的新型和改进疫苗,最终可作为针对其他疾病的免疫反应调节剂。