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Impaired Cellular Immunity in the Murine Neural Crest Conditional Deletion of Endothelin Receptor-B Model of Hirschsprung's Disease.

Gosain A, Barlow-Anacker AJ, Erickson CS, Pierre JF, Heneghan AF, Epstein ML, Kudsk KA - PLoS ONE (2015)

Bottom Line: We found that EdnrBNCC-/- display lymphopenia of their Peyer's Patches, the major inductive site of GI mucosal immunity.In the spleen, which is the primary source of IgA-producing Mature B-lymphocytes, EdnrBNCC-/- animals display decreased B-lymphocytes, but an increase in Mature B-lymphocytes.EdnrBNCC-/- spleens are also small and show altered architecture, with decreased red pulp and a paucity of B-lymphocytes in the germinal centers and marginal zone.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, University of Wisconsin-Madison School of Medicine and Public Health, Madison, Wisconsin, United States of America; Department of Neuroscience, University of Wisconsin-Madison School of Medicine and Public Health, Madison, Wisconsin, United States of America.

ABSTRACT
Hirschsprung's disease (HSCR) is characterized by aganglionosis from failure of neural crest cell (NCC) migration to the distal hindgut. Up to 40% of HSCR patients suffer Hirschsprung's-associated enterocolitis (HAEC), with an incidence that is unchanged from the pre-operative to the post-operative state. Recent reports indicate that signaling pathways involved in NCC migration may also be involved in the development of secondary lymphoid organs. We hypothesize that gastrointestinal (GI) mucosal immune defects occur in HSCR that may contribute to enterocolitis. EdnrB was deleted from the neural crest (EdnrBNCC-/-) resulting in mutants with defective NCC migration, distal colonic aganglionosis and the development of enterocolitis. The mucosal immune apparatus of these mice was interrogated at post-natal day (P) 21-24, prior to histological signs of enterocolitis. We found that EdnrBNCC-/- display lymphopenia of their Peyer's Patches, the major inductive site of GI mucosal immunity. EdnrBNCC-/- Peyer's Patches demonstrate decreased B-lymphocytes, specifically IgM+IgDhi (Mature) B-lymphocytes, which are normally activated and produce IgA following antigen presentation. EdnrBNCC-/- animals demonstrate decreased small intestinal secretory IgA, but unchanged nasal and bronchial airway secretory IgA, indicating a gut-specific defect in IgA production or secretion. In the spleen, which is the primary source of IgA-producing Mature B-lymphocytes, EdnrBNCC-/- animals display decreased B-lymphocytes, but an increase in Mature B-lymphocytes. EdnrBNCC-/- spleens are also small and show altered architecture, with decreased red pulp and a paucity of B-lymphocytes in the germinal centers and marginal zone. Taken together, these findings suggest impaired GI mucosal immunity in EdnrBNCC-/- animals, with the spleen as a potential site of the defect. These findings build upon the growing body of literature that suggests that intestinal defects in HSCR are not restricted to the aganglionic colon but extend proximally, even into the ganglionated small intestine and immune cells.

No MeSH data available.


Related in: MedlinePlus

The spleen of EdnrBNCC-/- animals is small in size and has abnormal architecture.(A) Gross photographs of representative spleens isolated from EdnrBNCC+/- and EdnrBNCC-/- animals demonstrating the size difference. (B) H&E staining of spleen cross-sections from EdnrBNCC+/- and EdnrBNCC-/- animals. EdnrBNCC-/- spleens demonstrate decreased red pulp (RP) and white pulp (WP). Scale bar = 600μm. (C) IHC of spleen cross-sections with B220 (green) and CD3 (blue). There are decreased B-cells in the germinal centers and a paucity of these cells within the marginal zone, located outside the ring of MOMA+ (red) staining. Scale bar = 200μm.
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pone.0128822.g005: The spleen of EdnrBNCC-/- animals is small in size and has abnormal architecture.(A) Gross photographs of representative spleens isolated from EdnrBNCC+/- and EdnrBNCC-/- animals demonstrating the size difference. (B) H&E staining of spleen cross-sections from EdnrBNCC+/- and EdnrBNCC-/- animals. EdnrBNCC-/- spleens demonstrate decreased red pulp (RP) and white pulp (WP). Scale bar = 600μm. (C) IHC of spleen cross-sections with B220 (green) and CD3 (blue). There are decreased B-cells in the germinal centers and a paucity of these cells within the marginal zone, located outside the ring of MOMA+ (red) staining. Scale bar = 200μm.

Mentions: In the mouse, mature B-lymphocytes in PP are primarily of splenic origin [45,46]. Initial observations by Cheng, et.al. indicated the presence of small spleens and splenic lymphopenia in the conventional EdnrB knockout model of HSCR [47], indicating that the immune system defects in this animal model may extend beyond the GI tract. We have extended their analysis, in our NCC-conditional deletion of EdnrB model, that the spleens in P21-24 EdnrBNCC-/- are smaller in size than those in the EdnrBNCC+/- (Fig 5). We found that the spleens of EdnrBNCC-/- weighed significantly less than those of the EdnrBNCC+/- as a proportion of the total body weight (EdnrBNCC+/- 0.67±0.05% vs. EdnrBNCC-/- 0.39±0.06%, p = 0.02, n = 10, Fig 6). We then examined splenic architecture and noted a decrease in the size of both the white and red pulp of the EdnrBNCC-/- spleen (Fig 5). We further examined architecture by immmunostaining (Fig 5) and observed a paucity of B-lymphocytes in the germinal centers of EdnrBNCC-/- spleens, and a marked decrease of B-lymphocytes within the marginal zones, suggesting potential defects in B-lymphocyte maturation [48].


Impaired Cellular Immunity in the Murine Neural Crest Conditional Deletion of Endothelin Receptor-B Model of Hirschsprung's Disease.

Gosain A, Barlow-Anacker AJ, Erickson CS, Pierre JF, Heneghan AF, Epstein ML, Kudsk KA - PLoS ONE (2015)

The spleen of EdnrBNCC-/- animals is small in size and has abnormal architecture.(A) Gross photographs of representative spleens isolated from EdnrBNCC+/- and EdnrBNCC-/- animals demonstrating the size difference. (B) H&E staining of spleen cross-sections from EdnrBNCC+/- and EdnrBNCC-/- animals. EdnrBNCC-/- spleens demonstrate decreased red pulp (RP) and white pulp (WP). Scale bar = 600μm. (C) IHC of spleen cross-sections with B220 (green) and CD3 (blue). There are decreased B-cells in the germinal centers and a paucity of these cells within the marginal zone, located outside the ring of MOMA+ (red) staining. Scale bar = 200μm.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4465674&req=5

pone.0128822.g005: The spleen of EdnrBNCC-/- animals is small in size and has abnormal architecture.(A) Gross photographs of representative spleens isolated from EdnrBNCC+/- and EdnrBNCC-/- animals demonstrating the size difference. (B) H&E staining of spleen cross-sections from EdnrBNCC+/- and EdnrBNCC-/- animals. EdnrBNCC-/- spleens demonstrate decreased red pulp (RP) and white pulp (WP). Scale bar = 600μm. (C) IHC of spleen cross-sections with B220 (green) and CD3 (blue). There are decreased B-cells in the germinal centers and a paucity of these cells within the marginal zone, located outside the ring of MOMA+ (red) staining. Scale bar = 200μm.
Mentions: In the mouse, mature B-lymphocytes in PP are primarily of splenic origin [45,46]. Initial observations by Cheng, et.al. indicated the presence of small spleens and splenic lymphopenia in the conventional EdnrB knockout model of HSCR [47], indicating that the immune system defects in this animal model may extend beyond the GI tract. We have extended their analysis, in our NCC-conditional deletion of EdnrB model, that the spleens in P21-24 EdnrBNCC-/- are smaller in size than those in the EdnrBNCC+/- (Fig 5). We found that the spleens of EdnrBNCC-/- weighed significantly less than those of the EdnrBNCC+/- as a proportion of the total body weight (EdnrBNCC+/- 0.67±0.05% vs. EdnrBNCC-/- 0.39±0.06%, p = 0.02, n = 10, Fig 6). We then examined splenic architecture and noted a decrease in the size of both the white and red pulp of the EdnrBNCC-/- spleen (Fig 5). We further examined architecture by immmunostaining (Fig 5) and observed a paucity of B-lymphocytes in the germinal centers of EdnrBNCC-/- spleens, and a marked decrease of B-lymphocytes within the marginal zones, suggesting potential defects in B-lymphocyte maturation [48].

Bottom Line: We found that EdnrBNCC-/- display lymphopenia of their Peyer's Patches, the major inductive site of GI mucosal immunity.In the spleen, which is the primary source of IgA-producing Mature B-lymphocytes, EdnrBNCC-/- animals display decreased B-lymphocytes, but an increase in Mature B-lymphocytes.EdnrBNCC-/- spleens are also small and show altered architecture, with decreased red pulp and a paucity of B-lymphocytes in the germinal centers and marginal zone.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, University of Wisconsin-Madison School of Medicine and Public Health, Madison, Wisconsin, United States of America; Department of Neuroscience, University of Wisconsin-Madison School of Medicine and Public Health, Madison, Wisconsin, United States of America.

ABSTRACT
Hirschsprung's disease (HSCR) is characterized by aganglionosis from failure of neural crest cell (NCC) migration to the distal hindgut. Up to 40% of HSCR patients suffer Hirschsprung's-associated enterocolitis (HAEC), with an incidence that is unchanged from the pre-operative to the post-operative state. Recent reports indicate that signaling pathways involved in NCC migration may also be involved in the development of secondary lymphoid organs. We hypothesize that gastrointestinal (GI) mucosal immune defects occur in HSCR that may contribute to enterocolitis. EdnrB was deleted from the neural crest (EdnrBNCC-/-) resulting in mutants with defective NCC migration, distal colonic aganglionosis and the development of enterocolitis. The mucosal immune apparatus of these mice was interrogated at post-natal day (P) 21-24, prior to histological signs of enterocolitis. We found that EdnrBNCC-/- display lymphopenia of their Peyer's Patches, the major inductive site of GI mucosal immunity. EdnrBNCC-/- Peyer's Patches demonstrate decreased B-lymphocytes, specifically IgM+IgDhi (Mature) B-lymphocytes, which are normally activated and produce IgA following antigen presentation. EdnrBNCC-/- animals demonstrate decreased small intestinal secretory IgA, but unchanged nasal and bronchial airway secretory IgA, indicating a gut-specific defect in IgA production or secretion. In the spleen, which is the primary source of IgA-producing Mature B-lymphocytes, EdnrBNCC-/- animals display decreased B-lymphocytes, but an increase in Mature B-lymphocytes. EdnrBNCC-/- spleens are also small and show altered architecture, with decreased red pulp and a paucity of B-lymphocytes in the germinal centers and marginal zone. Taken together, these findings suggest impaired GI mucosal immunity in EdnrBNCC-/- animals, with the spleen as a potential site of the defect. These findings build upon the growing body of literature that suggests that intestinal defects in HSCR are not restricted to the aganglionic colon but extend proximally, even into the ganglionated small intestine and immune cells.

No MeSH data available.


Related in: MedlinePlus