Nag Dhrubajyoti, Sinha Ritam, Mitra Soma, Barman Soumik, Takeda Yoshifumi, Shinoda Sumio, Chakrabarti M K, Koley Hemanta
Division of Bacteriology, National Institute of Cholera and Enteric Diseases, Kolkata, India.
Collaborative Research Centre of Okayama University for Infectious Diseases, National Institute of Cholera and Enteric Diseases, Kolkata, India.
Immunobiology. 2015 Nov;220(11):1275-83. doi: 10.1016/j.imbio.2015.07.002. Epub 2015 Jul 17.
Recently we have shown the homologous protective efficacy of heat killed multi-serotype Shigella (HKMS) immunogens in a guinea pig colitis model. In our present study, we have advanced our research by immunizing rabbits with a reduced number of oral doses and evaluating the host's adaptive immune responses. The duration of immunogenicity and subsequently protective efficacy was determined against wild type heterologous Shigella strains in a rabbit luminal model. After three successive oral immunizations with HKMS immunogens, serum and lymphocyte supernatant antibody titer against the heterologous shigellae were reciprocally increased and remained at an elevated level up to 180 days. Serogroup and serotype specific O-antigen of lipopolysaccharide and immunogenic proteins of heterologous challenge strains were detected by immunoblot assay. Up-regulation of IL-12p35, IFN-γ and IL-10 mRNA expression was detected in immunized rabbit peripheral blood mononuclear cells (PBMC) after stimulation with HKMS in vitro. HKMS-specific plasma cell response was confirmed by production of a relatively higher level of HKMS-specific IgG in immunized PBMC supernatant compared to control group. Furthermore, the immunized groups of rabbits exhibited complete protection against wild type heterologous shigellae challenge. Thus HKMS immunogens induced humoral and Th1-mediated adaptive immunity and provided complete protection in a rabbit model. These immunogens could be a broad spectrum non-living vaccine candidate for human use in the near future.
最近,我们已在豚鼠结肠炎模型中证实了热灭活多血清型志贺氏菌(HKMS)免疫原具有同源保护效力。在本研究中,我们通过减少口服剂量免疫兔子并评估宿主的适应性免疫反应,推进了我们的研究。在兔肠腔模型中,确定了针对野生型异源志贺氏菌菌株的免疫原性持续时间以及随后的保护效力。用HKMS免疫原连续进行三次口服免疫后,针对异源志贺氏菌的血清和淋巴细胞上清液抗体滴度相互升高,并在长达180天的时间内保持在较高水平。通过免疫印迹分析检测了异源攻击菌株脂多糖的血清群和血清型特异性O抗原以及免疫原性蛋白。在体外经HKMS刺激后,在免疫兔子外周血单核细胞(PBMC)中检测到IL-12p35、IFN-γ和IL-10 mRNA表达上调。与对照组相比,免疫PBMC上清液中产生相对较高水平的HKMS特异性IgG,证实了HKMS特异性浆细胞反应。此外,免疫兔子组对野生型异源志贺氏菌攻击表现出完全保护作用。因此,HKMS免疫原诱导了体液免疫和Th1介导的适应性免疫,并在兔模型中提供了完全保护。这些免疫原可能在不久的将来成为一种可供人类使用的广谱非活性疫苗候选物。