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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.

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URO-OVA mice show no responsiveness to OVA stimulation. Both URO-OVA mice and B6 mice were immunized by s.c. injection of 100 μg of OVA323-339 peptide emulsified with CFA at day 0 and sacrificed for analysis at day 14. Splenocytes were prepared and cultured in the absence or presence of PMA plus ionomycin (PI; 100 ng/ml and 1,500 ng/ml, respectively), a control ras peptide (CP; 10 μg/ml), or OVA323-339 peptide (10 μg/ml) for 3 days. The production of IFN-γ in culture supernatants was then analyzed by ELISA and presented as mean ± s.d. from triplicate determinations. *p<0.01 by Student t-Test.
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Figure 1: URO-OVA mice show no responsiveness to OVA stimulation. Both URO-OVA mice and B6 mice were immunized by s.c. injection of 100 μg of OVA323-339 peptide emulsified with CFA at day 0 and sacrificed for analysis at day 14. Splenocytes were prepared and cultured in the absence or presence of PMA plus ionomycin (PI; 100 ng/ml and 1,500 ng/ml, respectively), a control ras peptide (CP; 10 μg/ml), or OVA323-339 peptide (10 μg/ml) for 3 days. The production of IFN-γ in culture supernatants was then analyzed by ELISA and presented as mean ± s.d. from triplicate determinations. *p<0.01 by Student t-Test.

Mentions: To determine the impact of the expression of bladder urothelial OVA on host immune responses to OVA Ag, we immunized URO-OVA mice with OVA323-339 peptide emulsified with complete Freund’s adjuvant (CFA). Sex- and age-matched C57BL/6 (B6) mice were immunized as a control. After 14 days splenocytes were prepared, restimulated with OVA323-339 peptide in vitro, and analyzed for IFN-γ production by enzyme-linked immunosorbent assay (ELISA) (Figure 1). Compared to B6 splenocytes, splenocytes from URO-OVA mice produced no IFN-γ in response to OVA323-339 peptide restimulation, although both splenocytes produced a similar level of IFN-γ in response to phorbol myristate acetate (PMA)-ionomycin, two known positive T cell stimulators. We further challenged both OVA-immunized URO-OVA and B6 mice intravesically with OVA323-339 peptide at day 14 and found that only B6 mice but not URO-OVA mice developed histological bladder inflammation (data not shown). These observations suggested that constitutive expression of bladder urothelial OVA rendered mice unresponsive to OVA Ag.


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)

URO-OVA mice show no responsiveness to OVA stimulation. Both URO-OVA mice and B6 mice were immunized by s.c. injection of 100 μg of OVA323-339 peptide emulsified with CFA at day 0 and sacrificed for analysis at day 14. Splenocytes were prepared and cultured in the absence or presence of PMA plus ionomycin (PI; 100 ng/ml and 1,500 ng/ml, respectively), a control ras peptide (CP; 10 μg/ml), or OVA323-339 peptide (10 μg/ml) for 3 days. The production of IFN-γ in culture supernatants was then analyzed by ELISA and presented as mean ± s.d. from triplicate determinations. *p<0.01 by Student t-Test.
© Copyright Policy
Related In: Results  -  Collection

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Figure 1: URO-OVA mice show no responsiveness to OVA stimulation. Both URO-OVA mice and B6 mice were immunized by s.c. injection of 100 μg of OVA323-339 peptide emulsified with CFA at day 0 and sacrificed for analysis at day 14. Splenocytes were prepared and cultured in the absence or presence of PMA plus ionomycin (PI; 100 ng/ml and 1,500 ng/ml, respectively), a control ras peptide (CP; 10 μg/ml), or OVA323-339 peptide (10 μg/ml) for 3 days. The production of IFN-γ in culture supernatants was then analyzed by ELISA and presented as mean ± s.d. from triplicate determinations. *p<0.01 by Student t-Test.
Mentions: To determine the impact of the expression of bladder urothelial OVA on host immune responses to OVA Ag, we immunized URO-OVA mice with OVA323-339 peptide emulsified with complete Freund’s adjuvant (CFA). Sex- and age-matched C57BL/6 (B6) mice were immunized as a control. After 14 days splenocytes were prepared, restimulated with OVA323-339 peptide in vitro, and analyzed for IFN-γ production by enzyme-linked immunosorbent assay (ELISA) (Figure 1). Compared to B6 splenocytes, splenocytes from URO-OVA mice produced no IFN-γ in response to OVA323-339 peptide restimulation, although both splenocytes produced a similar level of IFN-γ in response to phorbol myristate acetate (PMA)-ionomycin, two known positive T cell stimulators. We further challenged both OVA-immunized URO-OVA and B6 mice intravesically with OVA323-339 peptide at day 14 and found that only B6 mice but not URO-OVA mice developed histological bladder inflammation (data not shown). These observations suggested that constitutive expression of bladder urothelial OVA rendered mice unresponsive to OVA Ag.

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