Limits...
Urothelial antigen-specific CD4+ T cells function as direct effector cells and induce bladder autoimmune inflammation independent of CD8+ T cells.

Liu W, Chen X, Evanoff DP, Luo Y - Mucosal Immunol (2011)

Bottom Line: Adoptive transfer of naive OVA-specific CD4(+) T cells led to exogenous T-cell proliferation, activation, and bladder infiltration but no inflammatory induction.In contrast, adoptive transfer of preactivated OVA-specific CD4(+) T cells induced bladder inflammation.These results indicate that urothelial Ag-specific CD4(+) T cells can function as direct effector cells to induce bladder autoimmune inflammation independent of CD8(+) T cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, University of Iowa, Roy J. and Lucille A. Carver College of Medicine, Iowa City, Iowa, USA.

ABSTRACT
The role of CD4(+) T cells in bladder autoimmune inflammation has not been identified because of the lack of a proper animal model. We investigated CD4(+) T-cell responses to bladder urothelial ovalbumin (OVA), a model self-antigen (Ag), in transgenic URO-OVA mice. The expression of bladder urothelial OVA rendered mice unresponsive to OVA and resulted in quick clearance of Ag-specific CD4(+) T cells. Adoptive transfer of naive OVA-specific CD4(+) T cells led to exogenous T-cell proliferation, activation, and bladder infiltration but no inflammatory induction. In contrast, adoptive transfer of preactivated OVA-specific CD4(+) T cells induced bladder inflammation. Studies further demonstrated that CD4(+) T cells induced bladder inflammation in URO-OVA mice depleted of CD8(+) T cells or deficient in the recombinase activating gene-1 (Rag-1(-/-)). These results indicate that urothelial Ag-specific CD4(+) T cells can function as direct effector cells to induce bladder autoimmune inflammation independent of CD8(+) T cells.

Show MeSH

Related in: MedlinePlus

OVA-specific CD4+ T cells infiltrate into the bladder urothelium of URO-OVA/Rag-1−/− mice. The bladders of URO-OVA/Rag-1−/− mice were collected 10 days after adoptive transfer of pre-activated OT-II CD4+ T cells (1 × 107 cells/mouse). Bladder sections were prepared and analyzed by IHC using control IgG2a (a) or anti-mouse I-Ab Ab (b-d). Brown staining indicates the urothelial cell expression of I-Ab. Lymphocytes in the urothelium and submucosa are indicated by arrows.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3118865&req=5

Figure 6: OVA-specific CD4+ T cells infiltrate into the bladder urothelium of URO-OVA/Rag-1−/− mice. The bladders of URO-OVA/Rag-1−/− mice were collected 10 days after adoptive transfer of pre-activated OT-II CD4+ T cells (1 × 107 cells/mouse). Bladder sections were prepared and analyzed by IHC using control IgG2a (a) or anti-mouse I-Ab Ab (b-d). Brown staining indicates the urothelial cell expression of I-Ab. Lymphocytes in the urothelium and submucosa are indicated by arrows.

Mentions: To verify the effector role of OVA-specific CD4+ T cells, we generated URO-OVA/Rag-1−/− mice, a crossed line of URO-OVA mice and Rag-1−/− mice, that retained the bladder urothelial OVA expression but lacked endogenous T cells. Both URO-OVA (Thy1.2) and URO-OVA/Rag-1−/− mice (Thy1.2) were transferred i.v. with pre-activated OT-II CD4+ T cells (Thy1.1) for cystitis induction at day 0 and sacrificed for analysis at day 10. Despite the lack of endogenous T cells, URO-OVA/Rag-1−/− mice developed histological bladder inflammation with severer cellular infiltration (Figure 5a, right panel) than URO-OVA mice (Figure 5a, middle panel). Flow cytometric analysis of bladder infiltrating T cells showed the presence of both CD4+ (Figure 5b) and Thy1.1+ T cells (Figure 5c) in the inflamed bladders. However, in the bladders of URO-OVA/Rag-1−/− mice, the number of CD4+ T cells matched the number of Thy1.1+ T cells (48,220 cells vs. 48,104 cells), suggesting that they were all transferred exogenous OT-II CD4+Thy1.1+ T cells. In contrast, the bladders of URO-OVA mice contained more CD4+ T cells than Thy1.1+ cells (21,236 cells vs. 16,012 cells), suggesting that endogenous CD4+ T cells were also recruited during bladder inflammatory responses. Consistent with histological H&E staining, flow cytometry showed 2 fold higher CD4+ T cells (48,220 cells vs. 21,236 cells) and 3 fold higher Thy1.1+ cells (48,104 cells vs. 16,012 cells) in the bladders of URO-OVA/Rag-1−/− mice than in the bladders of URO-OVA mice. In addition, similar to the inflamed bladders of URO-OVA mice, the inflamed bladders of URO-OVA/Rag-1−/− mice expressed increased mRNAs of T cell effector molecules IFN-γ, perforin and FasL (Figure 5d). Immunohistochemistry (IHC) analysis further revealed the expression of OVA and I-Ab on the bladder urothelium of URO-OVA/Rag-1−/− mice that received pre-activated OT-II CD4+ T cells (Figure 6; OVA not shown). Numerous T cells were found in the urothelial layer other than submucosal regions, suggesting that OVA-specific CD4+ T cells interacted with I-Ab/OVA323-339 expressing urothelial cells. These observations supported that urothelial Ag-specific CD4+ T cells could function as direct effector cells and induce bladder autoimmune inflammation without CD8+ T cells.


Urothelial antigen-specific CD4+ T cells function as direct effector cells and induce bladder autoimmune inflammation independent of CD8+ T cells.

Liu W, Chen X, Evanoff DP, Luo Y - Mucosal Immunol (2011)

OVA-specific CD4+ T cells infiltrate into the bladder urothelium of URO-OVA/Rag-1−/− mice. The bladders of URO-OVA/Rag-1−/− mice were collected 10 days after adoptive transfer of pre-activated OT-II CD4+ T cells (1 × 107 cells/mouse). Bladder sections were prepared and analyzed by IHC using control IgG2a (a) or anti-mouse I-Ab Ab (b-d). Brown staining indicates the urothelial cell expression of I-Ab. Lymphocytes in the urothelium and submucosa are indicated by arrows.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3118865&req=5

Figure 6: OVA-specific CD4+ T cells infiltrate into the bladder urothelium of URO-OVA/Rag-1−/− mice. The bladders of URO-OVA/Rag-1−/− mice were collected 10 days after adoptive transfer of pre-activated OT-II CD4+ T cells (1 × 107 cells/mouse). Bladder sections were prepared and analyzed by IHC using control IgG2a (a) or anti-mouse I-Ab Ab (b-d). Brown staining indicates the urothelial cell expression of I-Ab. Lymphocytes in the urothelium and submucosa are indicated by arrows.
Mentions: To verify the effector role of OVA-specific CD4+ T cells, we generated URO-OVA/Rag-1−/− mice, a crossed line of URO-OVA mice and Rag-1−/− mice, that retained the bladder urothelial OVA expression but lacked endogenous T cells. Both URO-OVA (Thy1.2) and URO-OVA/Rag-1−/− mice (Thy1.2) were transferred i.v. with pre-activated OT-II CD4+ T cells (Thy1.1) for cystitis induction at day 0 and sacrificed for analysis at day 10. Despite the lack of endogenous T cells, URO-OVA/Rag-1−/− mice developed histological bladder inflammation with severer cellular infiltration (Figure 5a, right panel) than URO-OVA mice (Figure 5a, middle panel). Flow cytometric analysis of bladder infiltrating T cells showed the presence of both CD4+ (Figure 5b) and Thy1.1+ T cells (Figure 5c) in the inflamed bladders. However, in the bladders of URO-OVA/Rag-1−/− mice, the number of CD4+ T cells matched the number of Thy1.1+ T cells (48,220 cells vs. 48,104 cells), suggesting that they were all transferred exogenous OT-II CD4+Thy1.1+ T cells. In contrast, the bladders of URO-OVA mice contained more CD4+ T cells than Thy1.1+ cells (21,236 cells vs. 16,012 cells), suggesting that endogenous CD4+ T cells were also recruited during bladder inflammatory responses. Consistent with histological H&E staining, flow cytometry showed 2 fold higher CD4+ T cells (48,220 cells vs. 21,236 cells) and 3 fold higher Thy1.1+ cells (48,104 cells vs. 16,012 cells) in the bladders of URO-OVA/Rag-1−/− mice than in the bladders of URO-OVA mice. In addition, similar to the inflamed bladders of URO-OVA mice, the inflamed bladders of URO-OVA/Rag-1−/− mice expressed increased mRNAs of T cell effector molecules IFN-γ, perforin and FasL (Figure 5d). Immunohistochemistry (IHC) analysis further revealed the expression of OVA and I-Ab on the bladder urothelium of URO-OVA/Rag-1−/− mice that received pre-activated OT-II CD4+ T cells (Figure 6; OVA not shown). Numerous T cells were found in the urothelial layer other than submucosal regions, suggesting that OVA-specific CD4+ T cells interacted with I-Ab/OVA323-339 expressing urothelial cells. These observations supported that urothelial Ag-specific CD4+ T cells could function as direct effector cells and induce bladder autoimmune inflammation without CD8+ T cells.

Bottom Line: Adoptive transfer of naive OVA-specific CD4(+) T cells led to exogenous T-cell proliferation, activation, and bladder infiltration but no inflammatory induction.In contrast, adoptive transfer of preactivated OVA-specific CD4(+) T cells induced bladder inflammation.These results indicate that urothelial Ag-specific CD4(+) T cells can function as direct effector cells to induce bladder autoimmune inflammation independent of CD8(+) T cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, University of Iowa, Roy J. and Lucille A. Carver College of Medicine, Iowa City, Iowa, USA.

ABSTRACT
The role of CD4(+) T cells in bladder autoimmune inflammation has not been identified because of the lack of a proper animal model. We investigated CD4(+) T-cell responses to bladder urothelial ovalbumin (OVA), a model self-antigen (Ag), in transgenic URO-OVA mice. The expression of bladder urothelial OVA rendered mice unresponsive to OVA and resulted in quick clearance of Ag-specific CD4(+) T cells. Adoptive transfer of naive OVA-specific CD4(+) T cells led to exogenous T-cell proliferation, activation, and bladder infiltration but no inflammatory induction. In contrast, adoptive transfer of preactivated OVA-specific CD4(+) T cells induced bladder inflammation. Studies further demonstrated that CD4(+) T cells induced bladder inflammation in URO-OVA mice depleted of CD8(+) T cells or deficient in the recombinase activating gene-1 (Rag-1(-/-)). These results indicate that urothelial Ag-specific CD4(+) T cells can function as direct effector cells to induce bladder autoimmune inflammation independent of CD8(+) T cells.

Show MeSH
Related in: MedlinePlus