Coated probiotic-based drug carriers for oral delivery of tumor antigens

doi:10.12211/2096-8280.2022-010

Abstract

Abstract:

Cancer vaccines, as a form of cancer immunotherapy, trigger host anti-tumor immunity by delivering tumor antigens. Oral administration has the advantages of convenience, simplicity and high efficacy. However, oral tumor vaccines have been rarely reported and existing forms are solely suitable for gastrointestinal cancer. The situation is associated with lack of advanced oral delivery systems. In this study, yeast cell membrane-coated probiotics are developed as drug carriers for oral delivery of tumor antigens. Yeast cell membranes play a dual role in protecting probiotics and antigens and facilitating targeted delivery of tumor antigens to the gut lymphatic system, while probiotics serve as both immune adjuvant and antigen carrier. As a model antigen, ovalbumin (OVA) is loaded onto probiotic surface by complexing with polyethyleneimine and hyaluronic acid via electrostatic interaction. Subsequently, antigen-loaded probiotics are individually camouflaged with yeast membranes by physical extrusion through a porous membrane. As reflected by similar growth to uncoated probiotics, both the coating and preparation procedures have limited influences on bacterial viability. Experimental results show that, compared to unmodified bacteria, yeast cell membrane coated probiotics retain their ability to grow and divide after incubation in simulated gastrointestinal environments and exhibit an improved bioavailability following oral ingestion. The presence of embedded β-glucan on yeast membranes promotes the phagocytosis by microfold cells that locate in intestinal epithelium. After oral gavage, the coating facilitates the accumulation of wrapped probiotics in Peyer's patches and then in mesenteric lymph nodes. Furthermore, the maturation of dendritic cells and corresponding antigen presentation are enhanced by coated probiotics, which induce an OVA-specific immune response as manifested by an upgraded serum level of OVA-IgG in mice after oral administration. These results demonstrate that yeast cell membrane-coated probiotics show enhanced tolerance against insults in the gastrointestinal environment and improved delivery of antigens into the gut lymphatic system, hence activating anti-tumor immunity.

Key words: probiotics, cancer vaccines, oral vaccines, oral delivery system, nanobiotechnology

摘要：

肿瘤疫苗是一种通过递送肿瘤抗原进而激活自身特异性抗肿瘤免疫的肿瘤免疫疗法。口服具有方便、简单和高效的优势。然而现有的口服肿瘤疫苗有限，并且局限于胃肠道肿瘤的治疗。这主要与缺乏先进的口服递送载体有关。本研究设计了一种酵母细胞膜包覆的益生菌应用于口服递送系统。其中，酵母细胞膜发挥益生菌和抗原保护以及肠道淋巴系统靶向的作用；益生菌发挥搭载抗原和免疫佐剂的作用。结果显示，与裸菌相比，酵母细胞膜包覆能提高益生菌的生物利用度，增强其在派氏结的富集，促进树突状细胞的活化和抗原递呈以及小鼠血浆中OVA-IgG水平。实验证明该系统能够保护益生菌和抗原免受胃肠道环境刺激并进一步向肠道淋巴系统递送抗原，激活抗肿瘤免疫。

关键词: 益生菌, 肿瘤疫苗, 口服疫苗, 口服递送系统, 生物纳米技术

CLC Number:

R392

Sisi LIN, Chao PAN, Yifan ZHANG, Jinyao LIU. Coated probiotic-based drug carriers for oral delivery of tumor antigens[J]. Synthetic Biology Journal, 2022, 3(4): 810-820.

林思思, 潘超, 张一帆, 刘尽尧. 基于表面涂层益生菌的肿瘤抗原口服递送系统[J]. 合成生物学, 2022, 3(4): 810-820.

Figures/Tables 4

Fig. 1 Characterization of YM-EcN@OVA(a) Typical laser scanning confocal microscopy images of YM-EcN@OVA. Red, green and blue channels show EcN expressing mCherry, fluorescein isothiocyanate-OVA, and yeast cell membranes stained with calcofluor-white, respectively; Scale bar: 10 μm. (b) Representative transmission electron microscopy images of EcN, EcN@OVA, and YM-EcN@OVA, respectively. Scale bars: 500 nm and 200 nm for the first and second row of the figure, respectively; The outer edge and coating layers of bacteria are indicated by white dotted line and arrows, respectively. (c) Size of EcN, EcN@OVA, and YM-EcN@OVA, respectively; (d) Zeta potential of EcN, EcN@OVA, and YM-EcN@OVA, respectively; (e) Flow cytometric analysis of EcN@OVA. (f) Flow cytometric analysis of YM-EcN@OVA. Error bars represent standard deviation. **P<0.01

Fig. 2 Stability of YM-EcN@OVA in simulated gastrointestinal environments

Fig. 3 Quantification of accumulated EcN(PBS, EcN@OVA, and YM-EcN@OVA were injected into the intestinal lumen of mice, respectively. EcN used in the experiment was transfected with pBBR1MCS2-Tac-mCherry. At 1.5 h post injection, fluorescence signals of MLNs were captured by in vivo imaging system (IVIS). Homogenates of MLNs and PPs were separately prepared for bacterial culture. MLNs, mesenteric lymph nodes; PPs, Peyer's patches Error bars represent standard deviation. **P < 0.01)

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