Limits...
Goblet cells deliver luminal antigen to CD103+ dendritic cells in the small intestine.

McDole JR, Wheeler LW, McDonald KG, Wang B, Konjufca V, Knoop KA, Newberry RD, Miller MJ - Nature (2012)

Bottom Line: The intestinal immune system is exposed to a mixture of foreign antigens from diet, commensal flora and potential pathogens.The lamina propria (LP) underlies the expansive single-cell absorptive villous epithelium and contains a large population of DCs (CD11c(+) CD11b(+) MHCII(+) cells) comprised of two predominant subsets: CD103(+) CX(3)CR1(-) DCs, which promote IgA production, imprint gut homing on lymphocytes and induce the development of regulatory T cells, and CD103(-) CX(3)CR1(+) DCs (with features of macrophages), which promote tumour necrosis factor-α (TNF-α) production, colitis, and the development of T(H)17 T cells.Using a minimally disruptive in vivo imaging approach we show that in the steady state, small intestine goblet cells (GCs) function as passages delivering low molecular weight soluble antigens from the intestinal lumen to underlying CD103(+) LP-DCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St Louis, Missouri 63110, USA.

Show MeSH

Related in: MedlinePlus

GCs are associated with the trans-epithelial passage of luminal material(a) PAS staining of GCs and (b) dextran columns visualized by 2P microscopy displayed similar morphology and (c) dimensions. (d) Dextran columns were often associated with a nucleus (white arrow). Dextran columns co-localized with GC markers (e) cytokeratin 18 (cyt18, white) and (f) MUC2. (g) Cytokeratin 18 positive cells (white arrows) did not co-localize with the M-cell marker GP2 (yellow arrows). Dextran columns were present in (h) healthy human small intestine and (i) stained positive for cytokeratin 18. Scale bars = 30μm (a, b, and h), 20 μm (d, e, g, and i), and 10μm (f). Error bars = SD.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3313460&req=5

Figure 2: GCs are associated with the trans-epithelial passage of luminal material(a) PAS staining of GCs and (b) dextran columns visualized by 2P microscopy displayed similar morphology and (c) dimensions. (d) Dextran columns were often associated with a nucleus (white arrow). Dextran columns co-localized with GC markers (e) cytokeratin 18 (cyt18, white) and (f) MUC2. (g) Cytokeratin 18 positive cells (white arrows) did not co-localize with the M-cell marker GP2 (yellow arrows). Dextran columns were present in (h) healthy human small intestine and (i) stained positive for cytokeratin 18. Scale bars = 30μm (a, b, and h), 20 μm (d, e, g, and i), and 10μm (f). Error bars = SD.

Mentions: Periodic acid-Schiff (PAS) staining of mucin in sections of small intestine (Fig. 2a) produced a goblet cell (GC) staining pattern similar in frequency, distribution, and dimensions to the dextran columns identified by 2P microscopy (Fig. 2b and c). Furthermore, in contrast to the acellular and impermeable discontinuities seen in the small intestine epithelium11, dextran columns were associated with a nucleus (Fig. 2d; Supplementary Fig. 2a; Supplementary Movie 3). To determine if the dextran filled cells were in fact GCs, sections of intestine from mice given lysine-fixable dextran were stained with antibodies to mucin 2 (MUC2) and cytokeratin 18, which are both highly expressed by GCs12. Dextran columns , showed near prefect co-localization with MUC2+ and cytokeratin 18+ epithelial cells displaying GC morphology (Fig. 2e, f). Therefore, we term this phenomenon “goblet cell-associated antigen passages” (GAPs). To address the possibility that GAPs are apoptotic GCs, we co-stained for various markers of apoptosis including cleaved cytokeratin 18, cleaved caspase 3, and TUNEL (Supplementary Fig 3 a–i). In all cases, we found no association between apoptotic GCs and GAPs. Moreover, GAPs are distinct from villous M-cells, since they did not co-localize with the M-cell marker glycoprotein 2 (GP2) (Fig. 2g)13. The frequency and distribution of GAPs assessed by 2P microscopy was similar in all strains of specific-pathogen-free (SPF) mice examined (supplementary Fig. 2 b–d), with a non-significant trend more GAPs detected in the terminal ileum (supplementary Fig. 2h). GAPS were also evident in human jejunum resection specimens (Fig. 2h, i), suggesting that GAPs are a general phenomenon of the healthy small intestine. We examined the frequency of GAPs in C3H/HejBir IL-10−/− mice14, which develop spontaneous intestinal inflammation with GC loss, and in germ-free (GF) mice that lack normal gut flora. The number of GAPs and GCs correlated strongly; GAPs and GCs were significantly more numerous in GF mice (supplementary Fig. 2 e and g) and significantly fewer in IL-10−/− mice (supplementary Fig. 2 f and g).


Goblet cells deliver luminal antigen to CD103+ dendritic cells in the small intestine.

McDole JR, Wheeler LW, McDonald KG, Wang B, Konjufca V, Knoop KA, Newberry RD, Miller MJ - Nature (2012)

GCs are associated with the trans-epithelial passage of luminal material(a) PAS staining of GCs and (b) dextran columns visualized by 2P microscopy displayed similar morphology and (c) dimensions. (d) Dextran columns were often associated with a nucleus (white arrow). Dextran columns co-localized with GC markers (e) cytokeratin 18 (cyt18, white) and (f) MUC2. (g) Cytokeratin 18 positive cells (white arrows) did not co-localize with the M-cell marker GP2 (yellow arrows). Dextran columns were present in (h) healthy human small intestine and (i) stained positive for cytokeratin 18. Scale bars = 30μm (a, b, and h), 20 μm (d, e, g, and i), and 10μm (f). Error bars = SD.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3313460&req=5

Figure 2: GCs are associated with the trans-epithelial passage of luminal material(a) PAS staining of GCs and (b) dextran columns visualized by 2P microscopy displayed similar morphology and (c) dimensions. (d) Dextran columns were often associated with a nucleus (white arrow). Dextran columns co-localized with GC markers (e) cytokeratin 18 (cyt18, white) and (f) MUC2. (g) Cytokeratin 18 positive cells (white arrows) did not co-localize with the M-cell marker GP2 (yellow arrows). Dextran columns were present in (h) healthy human small intestine and (i) stained positive for cytokeratin 18. Scale bars = 30μm (a, b, and h), 20 μm (d, e, g, and i), and 10μm (f). Error bars = SD.
Mentions: Periodic acid-Schiff (PAS) staining of mucin in sections of small intestine (Fig. 2a) produced a goblet cell (GC) staining pattern similar in frequency, distribution, and dimensions to the dextran columns identified by 2P microscopy (Fig. 2b and c). Furthermore, in contrast to the acellular and impermeable discontinuities seen in the small intestine epithelium11, dextran columns were associated with a nucleus (Fig. 2d; Supplementary Fig. 2a; Supplementary Movie 3). To determine if the dextran filled cells were in fact GCs, sections of intestine from mice given lysine-fixable dextran were stained with antibodies to mucin 2 (MUC2) and cytokeratin 18, which are both highly expressed by GCs12. Dextran columns , showed near prefect co-localization with MUC2+ and cytokeratin 18+ epithelial cells displaying GC morphology (Fig. 2e, f). Therefore, we term this phenomenon “goblet cell-associated antigen passages” (GAPs). To address the possibility that GAPs are apoptotic GCs, we co-stained for various markers of apoptosis including cleaved cytokeratin 18, cleaved caspase 3, and TUNEL (Supplementary Fig 3 a–i). In all cases, we found no association between apoptotic GCs and GAPs. Moreover, GAPs are distinct from villous M-cells, since they did not co-localize with the M-cell marker glycoprotein 2 (GP2) (Fig. 2g)13. The frequency and distribution of GAPs assessed by 2P microscopy was similar in all strains of specific-pathogen-free (SPF) mice examined (supplementary Fig. 2 b–d), with a non-significant trend more GAPs detected in the terminal ileum (supplementary Fig. 2h). GAPS were also evident in human jejunum resection specimens (Fig. 2h, i), suggesting that GAPs are a general phenomenon of the healthy small intestine. We examined the frequency of GAPs in C3H/HejBir IL-10−/− mice14, which develop spontaneous intestinal inflammation with GC loss, and in germ-free (GF) mice that lack normal gut flora. The number of GAPs and GCs correlated strongly; GAPs and GCs were significantly more numerous in GF mice (supplementary Fig. 2 e and g) and significantly fewer in IL-10−/− mice (supplementary Fig. 2 f and g).

Bottom Line: The intestinal immune system is exposed to a mixture of foreign antigens from diet, commensal flora and potential pathogens.The lamina propria (LP) underlies the expansive single-cell absorptive villous epithelium and contains a large population of DCs (CD11c(+) CD11b(+) MHCII(+) cells) comprised of two predominant subsets: CD103(+) CX(3)CR1(-) DCs, which promote IgA production, imprint gut homing on lymphocytes and induce the development of regulatory T cells, and CD103(-) CX(3)CR1(+) DCs (with features of macrophages), which promote tumour necrosis factor-α (TNF-α) production, colitis, and the development of T(H)17 T cells.Using a minimally disruptive in vivo imaging approach we show that in the steady state, small intestine goblet cells (GCs) function as passages delivering low molecular weight soluble antigens from the intestinal lumen to underlying CD103(+) LP-DCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St Louis, Missouri 63110, USA.

Show MeSH
Related in: MedlinePlus