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壳聚糖盐酸盐稳定化乳液的制备及其免疫刺激作用

[Preparation of chitosan hydrochloride stabilized emulsion and its immunostimulatory effect].

作者信息

Wang Danyang, Wu Jie, Wang Ning

机构信息

School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China.

Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2023 Jan 25;39(1):262-274. doi: 10.13345/j.cjb.220276.

Abstract

In order to increase the ability of oil-emulsion adjuvant to stimulate cellular immunity, chitosan hydrochloride with positive charge was selected to stabilize oil-in-water emulsion (CHE). In this paper, model antigen ovalbumin was selected to prepare vaccines with emulsion adjuvant, commercial adjuvant or no adjuvant. The emulsion was characterized by measuring the particle size, electric potential and antigen adsorption rate. BALB/c mice were immunized by intramuscular injection. Serum antibody levels, the numbers of IL-4-secreting cells in splenocytes, cytotoxic T lymphocyte (CTL) response, and the expression of central memory T cells were measured to evaluate the immunostimulatory effect. The results showed that chitosan hydrochloride can effectively stabilize the emulsion. The emulsion size is about 600 nm, and the antigen adsorption rate is more than 90%. After immunization, CHE could increase serum antibodies levels and increase IL-4 secretion. Expression of CTL surface activation molecules was also increased to stimulate CTL response further and to increase the CD44CD62L in T cells proportion. CHE as adjuvant can stimulate humoral and cellular immunity more efficiently, and is expected to extend the duration of protection.

摘要

为了提高油乳佐剂刺激细胞免疫的能力,选择带正电荷的壳聚糖盐酸盐来稳定水包油乳液(CHE)。本文选用模型抗原卵清蛋白,分别用乳液佐剂、市售佐剂或无佐剂制备疫苗。通过测量粒径、电位和抗原吸附率对乳液进行表征。通过肌肉注射对BALB/c小鼠进行免疫。检测血清抗体水平、脾细胞中分泌IL-4的细胞数量、细胞毒性T淋巴细胞(CTL)反应以及中央记忆T细胞的表达,以评估免疫刺激效果。结果表明,壳聚糖盐酸盐能有效稳定乳液。乳液粒径约为600 nm,抗原吸附率超过90%。免疫后,CHE可提高血清抗体水平并增加IL-4分泌。CTL表面活化分子的表达也增加,进一步刺激CTL反应,并增加T细胞中CD44CD62L的比例。CHE作为佐剂能更有效地刺激体液免疫和细胞免疫,有望延长保护期。

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