Pal Sukumar, Tifrea Delia F, Follmann Frank, Andersen Peter, de la Maza Luis M
Department of Pathology and Laboratory Medicine, Medical Sciences I, Room D440, University of California, Irvine, Irvine, CA 92697-4800, USA.
Department of Infectious Disease Immunology, Adjuvant Research, Staten Serum Institute, Copenhagen, Denmark.
Vaccine. 2017 Mar 23;35(13):1705-1711. doi: 10.1016/j.vaccine.2017.02.020. Epub 2017 Feb 24.
Two cationic liposomal adjuvants CAF01 and CAF09 were formulated with the native or the recombinant Chlamydia muridarum major outer membrane protein (nMOMP and rMOMP). BALB/c mice were immunized with the four vaccine formulations using the subcutaneous followed by the intranasal (i.n.) routes. As positive controls mice were inoculated i.n. with live C. muridarum and negative controls received i.n. minimal essential medium (MEM). Four weeks after the last immunization mice were challenged i.n. with 10 inclusion forming units (IFU) of C. muridarum. Following the challenge the mice were weighed daily. At 10days post-challenge the mice were euthanized, their lungs weighed and the number of C. muridarum IFU determined. Serum collected the day before the challenge showed that all four groups of mice immunized with CAF01, or CAF09 and MOMP had significant C. muridarum-specific antibody titers. As determined by a T-cell lymphoproliferative assay, these four groups of mice also mounted robust cell mediated immune responses with high production of IFN-γ and IL17 and low levels of IL-4. Following the challenge the four groups of mice lost significantly less body weight than the MEM-immunized group. Lungs of mice vaccinated with CAF01, or CAF09, and nMOMP were significantly lighter than those from mice immunized using rMOMP. The number of IFU recovered from the lungs of mice vaccinated with CAF01, or CAF09, and nMOMP was similar to the number of IFU recovered from mice immunized with live EB. Mice that received rMOMP had significantly higher numbers of IFU than other groups. In conclusion, CAF01 and CAF09 elicited very robust protective humoral and cellular immune responses and were equally effective at adjuntavizing the C. muridarum MOMP. Mice vaccinated with nMOMP were significantly better protected than those immunized with rMOMP, indicative of the importance of the structural conformation of this antigen in protection.
两种阳离子脂质体佐剂CAF01和CAF09分别与天然或重组的鼠衣原体主要外膜蛋白（nMOMP和rMOMP）配制而成。采用皮下接种随后鼻内（i.n.）接种的方式，用这四种疫苗制剂对BALB/c小鼠进行免疫。作为阳性对照，小鼠经鼻内接种活的鼠衣原体，阴性对照经鼻内接种最低限度基本培养基（MEM）。最后一次免疫四周后，小鼠经鼻内接种10个鼠衣原体包涵体形成单位（IFU）。攻毒后，每天对小鼠称重。攻毒后10天，对小鼠实施安乐死，称取其肺脏重量并测定鼠衣原体IFU的数量。攻毒前一天采集的血清显示，用CAF01、CAF09和MOMP免疫的所有四组小鼠均具有显著的鼠衣原体特异性抗体滴度。通过T细胞淋巴细胞增殖试验测定，这四组小鼠还产生了强烈的细胞介导免疫反应，IFN-γ和IL17产生量高，IL-4水平低。攻毒后，这四组小鼠体重减轻明显少于接种MEM的组。接种CAF01、CAF09和nMOMP的小鼠的肺脏明显轻于接种rMOMP的小鼠。从接种CAF01、CAF09和nMOMP的小鼠肺脏中回收的IFU数量与接种活EB的小鼠中回收的IFU数量相似。接受rMOMP的小鼠的IFU数量明显高于其他组。总之，CAF01和CAF09引发了非常强烈的保护性体液免疫和细胞免疫反应，在辅助鼠衣原体MOMP方面同样有效。接种nMOMP的小鼠比接种rMOMP的小鼠受到的保护明显更好，这表明该抗原的结构构象在保护中的重要性。
