Three-Dimensional Reconstruction of Murine Peyer's Patches from Immunostained Cryosections

Ahlawat, Sarita; Jesus, Magdia De; Khare, Kedar; Cole, Richard W.; Mantis, Nicholas J.

doi:10.1017/s1431927613013640

Cited by 8 publications

(13 citation statements)

References 25 publications

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“…To test if there were changes in uptake of C. albicans by SED CD11c + DC stimulated with CT, we cogavaged C. albicans with CT and harvested PP after 4 h. The results revealed when compared to the control (Fig. 5A) that CT did in fact promote DC migration towards the FAE as previously reported [34,47]; however, CT did not affect sampling efficiency of C. albicans by SED DCs (Fig. 5B).…”

Section: Resultssupporting

confidence: 62%

“…We and others had previously shown that the potent non-inflammatory mucosal adjuvant CT stimulates migration of DCs into the FAE [34,47,48]. To test if there were changes in uptake of C. albicans by SED CD11c + DC stimulated with CT, we cogavaged C. albicans with CT and harvested PP after 4 h. The results revealed when compared to the control (Fig.…”

Section: Resultsmentioning

confidence: 93%

“…As needed, cholera toxin (10 g; CT) was coadministered with FITC-labeled C. albicans, C. tropicalis or GPs. PPs were collected 4 h later, then cryosectioned, immunostained and viewed by confocal laser scanning microscopy [34].…”

Section: Administration Of Lps and Ctmentioning

confidence: 99%

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Sampling of Candida albicans and Candida tropicalis by Langerin-positive dendritic cells in mouse Peyer’s patches

et al. 2015

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Section: Resultssupporting

confidence: 62%

Section: Resultsmentioning

confidence: 93%

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Sampling of Candida albicans and Candida tropicalis by Langerin-positive dendritic cells in mouse Peyer’s patches

et al. 2015

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“…As the inductive sites of intestinal immunity, PPs are the primary sites through which particulate antigens and microbes are sampled (11). The microarchitecture of PPs is quite unique, as they are interspersed along the antimesenteric side of the ileum and can be seen with the naked eye as aggregate lymphoid structures (11, 12). Each PP contains multiple follicles that are individually equipped with an epithelial barrier called the follicle-associated epithelium (FAE).…”

Section: Introductionmentioning

confidence: 99%

“…Below the FAE in the sub-epithelial dome (SED), there is an extensive network of at least five subsets of mononuclear phagocytes and a B-cell-rich germinal center (GC) that also contains follicular dendritic cells flanked by T-cell-rich interfollicular regions. In concert, these cells all play an important role in sampling and immune surveillance (11, 13). Because PPs can be a very delicate and technically difficult tissue to work with, many gut models have used cell lines such as the Caco-2 and FaDu to understand sampling and transcytosis (14, 15).…”

Section: Introductionmentioning

confidence: 99%

Setting off the Alarms:Candida albicansElicits Pro-Inflammatory Differential Gene Expression in Intestinal Peyer’s Patches

Singh

Song

et al. 2019

Preprint

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24Candida albicans has been associated with a number of human diseases that pertain to 25 the gastrointestinal (GI) tract. However, the details of how gut-associated lymphoid tissues 26 (GALT) such as Peyer's patches (PPs) in the small intestine play a role in immune surveillance 27 and microbial differentiation, and what mechanisms PP use to protect the mucosal barrier in 28 response to fungal organisms such as C. albicans, are still unclear. We particularly focus on PPs 29 as they are the immune sensors and inductive sites of the gut that influence inflammation and 30 tolerance. We have previously demonstrated that CD11c + phagocytes located in the sub-31 epithelial dome (SED) within PPs sample C. albicans. To gain insight on how specific cells 32within PPs sense and respond to the sampling of fungi, we gavaged mice with C. albicans strains 33 ATCC 18804 and SC5314 as well as Saccharomyces cerevisiae. We measured the differential 34 gene expression of sorted CD45 + B220 + B-cells, CD3 + T-cells, and CD11c + DCs within the first 35 24 hrs post-gavage using nanostring nCounter® technology. The results reveal that at 24 hrs, PP 36 phagocytes were the cell type that displayed differential gene expression. These phagocytes were 37 both able to sample C. albicans and able to discriminate between strains. In particular, strain 38 ATCC 18804 upregulated fungal specific pro-inflammatory genes in CD11c + phagocytes 39 pertaining to innate and adaptive immune responses. Interestingly, PP CD11c + phagocytes 40 differentially expressed genes in response to C. albicans that were important in the protection of 41 the mucosal barrier. These results highlight that the mucosal barrier not only responds to C. 42 albicans, but also aids in the protection of the host. 43 Importance 44The specific gene expression changes within PPs that send the warning signals when 45 encountering fungi, and how PPs can discriminate between innocuous S. cerevisiae or different 46 strains of C. albicans during early stages of sampling, have not been elucidated. Here we show 47 that within the first 24 hours of sampling, CD11c + phagocytes were not only important in 48 sampling, but they were the cell type that exhibited clear differential gene expression. These 49 differentially expressed genes play important dual roles in inflammation, chemotaxis, and fungal 50 specific recognition, as well as maintaining homeostasis and protection of the mucosal barrier. 51Using nanostring technology, we were also able to demonstrate that PPs can distinguish between 52 different strains of C. albicans and can "set off the alarms" when necessary. 53 54

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Candida albicans Elicits Pro-Inflammatory Differential Gene Expression in Intestinal Peyer’s Patches

et al. 2019

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Candida albicans has been associated with a number of human diseases that pertain to the gastrointestinal (GI) tract. However, the details of how gut-associated lymphoid tissues (GALT) such as Peyer's patches (PPs) in the small intestine play a role in immune surveillance and microbial differentiation, and what mechanisms PP use to protect the mucosal barrier in response to fungal organisms such as C. albicans, are still unclear. We particularly focus on PPs as they are the immune sensors and inductive sites of the gut that influence inflammation and tolerance. We have previously demonstrated that CD11c + phagocytes located in the subepithelial dome (SED) within PPs sample C. albicans. To gain insight on how specific cells within PPs sense and respond to the sampling of fungi, we gavaged mice with C. albicans strains ATCC 18804 and SC5314 as well as Saccharomyces cerevisiae. We measured the differential gene expression of sorted CD45 + B220 + B-cells, CD3 + T-cells, and CD11c + DCs within the first 24 hrs post-gavage using nanostring nCounter® technology. The results reveal that at 24 hrs, PP phagocytes were the cell type that displayed differential gene expression. These phagocytes were both able to sample C. albicans and able to discriminate between strains. In particular, strain ATCC 18804 upregulated fungal specific pro-inflammatory genes in CD11c + phagocytes pertaining to innate and adaptive immune responses. Interestingly, PP CD11c + phagocytes differentially expressed genes in response to C. albicans that were important in the protection of the mucosal barrier. These results highlight that the mucosal barrier not only responds to C. albicans, but also aids in the protection of the host. ImportanceThe specific gene expression changes within PPs that send the warning signals when encountering fungi, and how PPs can discriminate between innocuous S. cerevisiae or different

show abstract

Three-Dimensional Reconstruction of Murine Peyer's Patches from Immunostained Cryosections

Cited by 8 publications

References 25 publications

Sampling of Candida albicans and Candida tropicalis by Langerin-positive dendritic cells in mouse Peyer’s patches

Sampling of Candida albicans and Candida tropicalis by Langerin-positive dendritic cells in mouse Peyer’s patches

Setting off the Alarms:Candida albicansElicits Pro-Inflammatory Differential Gene Expression in Intestinal Peyer’s Patches

Candida albicans Elicits Pro-Inflammatory Differential Gene Expression in Intestinal Peyer’s Patches

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