The construction of in vitro nasal cavity-mimic M-cell model, design of M cell-targeting nanoparticles and evaluation of mucosal vaccination by nasal administration

Abstract M cell targeting is one of the critical issues to develop efficient mucosal vaccine design. In this study, peptide ligands with high affinity to porcine TLR2, which is highly expressed in M cells and play an important role in mucosal immune responses in pigs, were identified through the cell-based phage display technique combined with high-throughput sequencing. A random phage-peptide library was applied to the porcine TLR2 overexpressing cell line and total 85, 557 unique peptide sequences were identified from approximately 9.0 × 107 reads after three rounds of both subtractive and non-subtractive biopanning via high-throughput sequencing. Among the unique sequences, three candidate peptide sequences, NAGHLSQ, VPSKPGL, and RANLDGQ, were selected based on their abundance in the third round of biopanning. Consequently, NAGHLSQ showed the highest affinity exclusively to porcine TLR2 compared with other candidates and its binding mechanism was inferred to be directly associated with ligand binding site of the TLR2 through the in vitro competitive analysis. The peptide identified in this research could be used in development of effective porcine mucosal vaccine as an M cell targeting moiety to enhance the transport of antigens into the Peyer's patch via oral route.

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Salmonella translocates across an in vitro M cell model independently of SPI-1 and SPI-2

Microbiology ◽

10.1099/mic.0.2008/021162-0 ◽

2008 ◽

Vol 154 (12) ◽

pp. 3887-3894 ◽

Cited By ~ 40

Author(s):

Isabel Martinez-Argudo ◽

Mark A. Jepson

Keyword(s):

Cell Model ◽

M Cell

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Transcytosis of Murine-Adapted Bovine Spongiform Encephalopathy Agents in an In Vitro Bovine M Cell Model

Journal of Virology ◽

10.1128/jvi.00969-10 ◽

2010 ◽

Vol 84 (23) ◽

pp. 12285-12291 ◽

Cited By ~ 20

Author(s):

Kohtaro Miyazawa ◽

Takashi Kanaya ◽

Ikuro Takakura ◽

Sachi Tanaka ◽

Tetsuya Hondo ◽

...

Keyword(s):

Bovine Spongiform Encephalopathy ◽

Cell Model ◽

Transmissible Spongiform Encephalopathies ◽

Epithelial Cell Line ◽

Spongiform Encephalopathy ◽

M Cells ◽

Intestinal Epithelial ◽

M Cell ◽

Spongiform Encephalopathies

ABSTRACT Transmissible spongiform encephalopathies (TSE), including bovine spongiform encephalopathy (BSE), are fatal neurodegenerative disorders in humans and animals. BSE appears to have spread to cattle through the consumption of feed contaminated with BSE/scrapie agents. In the case of an oral infection, the agents have to cross the gut-epithelial barrier. We recently established a bovine intestinal epithelial cell line (BIE cells) that can differentiate into the M cell type in vitro after lymphocytic stimulation (K. Miyazawa, T. Hondo, T. Kanaya, S. Tanaka, I. Takakura, W. Itani, M. T. Rose, H. Kitazawa, T. Yamaguchi, and H. Aso, Histochem. Cell Biol. 133:125-134, 2010). In this study, we evaluated the role of M cells in the intestinal invasion of the murine-adapted BSE (mBSE) agent using our in vitro bovine intestinal epithelial model. We demonstrate here that M cell-differentiated BIE cells are able to transport the mBSE agent without inactivation at least 30-fold more efficiently than undifferentiated BIE cells in our in vitro model. As M cells in the follicle-associated epithelium are known to have a high ability to transport a variety of macromolecules, viruses, and bacteria from gut lumen to mucosal immune cells, our results indicate the possibility that bovine M cells are able to deliver agents of TSE, not just the mBSE agent.

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Direct Delivery of ANA-TA9, a Peptide Capable of Aβ Hydrolysis, to the Brain by Intranasal Administration

Pharmaceutics ◽

10.3390/pharmaceutics13101673 ◽

2021 ◽

Vol 13 (10) ◽

pp. 1673

Author(s):

Yusuke Hatakawa ◽

Akiko Tanaka ◽

Tomoyuki Furubayashi ◽

Rina Nakamura ◽

Motomi Konishi ◽

...

Keyword(s):

Nasal Cavity ◽

Animal Studies ◽

Intraperitoneal Administration ◽

Systemic Circulation ◽

Brain Parenchyma ◽

Nasal Administration ◽

Nasal Application ◽

Direct Delivery ◽

The Brain

We have recently reported Catalytides (Catalytic peptides) JAL-TA9 (YKGSGFRMI) and ANA-TA9 (SKGQAYRMI), which are the first Catalytides found to cleave Aβ42. Although the Catalytides must be delivered to the brain parenchyma to treat Alzheimer’s disease, the blood–brain barrier (BBB) limits their entry into the brain from the systemic circulation. To avoid the BBB, the direct route from the nasal cavity to the brain was used in this study. The animal studies using rats and mice clarified that the plasma clearance of ANA-TA9 was more rapid than in vitro degradation in the plasma, whole blood, and the cerebrospinal fluid (CSF). The brain concentrations of ANA-TA9 were higher after nasal administration than those after intraperitoneal administration, despite a much lower plasma concentration after nasal administration, suggesting the direct delivery of ANA-TA9 to the brain from the nasal cavity. Similar findings were observed for its transport to CSF after nasal and intravenous administration. The concentration of ANA-TA9 in the olfactory bulb reached the peak at 5 min, whereas those in the frontal and occipital brains was 30 min, suggesting the sequential backward translocation of ANA-TA9 in the brain. In conclusion, ANA-TA9 was efficiently delivered to the brain by nasal application, as compared to other routes.

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