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Intranasal peptide-induced tolerance and linked suppression: consequences of complement deficiency.

Fossati-Jimack L, Ling GS, Baudino L, Szajna M, Manivannan K, Zhao JC, Midgley R, Chai JG, Simpson E, Botto M, Scott D - Immunology (2015)

Bottom Line: The mechanisms whereby complement can mediate these diverse regulatory effects are poorly understood.This was related to the inability of C3-deficient DC to up-regulate the arginine-consuming enzyme, inducible nitric oxide synthase (Nos-2), in the presence of antigen-specific Treg cells and peptide, leading to reduced Treg cell generation.Our findings demonstrate that the classical pathway and C3 play a critical role in the peptide-mediated induction of tolerance to HY by modulating DC function.

View Article: PubMed Central - PubMed

Affiliation: Centre for Complement and Inflammation Research, Imperial College London, London, UK.

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Related in: MedlinePlus

C3 deficiency does not alter the distribution of the peptide. B6 wild-type (WT; n = 3) and B6.C3–/– (n = 3) female mice were given the HY-AbDby peptide twice (100 μg at time 0 and 6 hr later) intranasally, control mice received PBS. Twenty-four hours after the first peptide dose, splenic dendritic cells (DC) were isolated and cultured with Marilyn T cells (25 000 T cells/well) at the ratios indicated. Proliferation was assessed by pulsing with [3H]thymidine for the final 18 hr of the 72 hr incubation. Data are shown as mean ± SEM, Student's t-test. (DC : T ratio 2 : 1 P = 0·9704, 1 : 1 P = 0·5030).
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fig02: C3 deficiency does not alter the distribution of the peptide. B6 wild-type (WT; n = 3) and B6.C3–/– (n = 3) female mice were given the HY-AbDby peptide twice (100 μg at time 0 and 6 hr later) intranasally, control mice received PBS. Twenty-four hours after the first peptide dose, splenic dendritic cells (DC) were isolated and cultured with Marilyn T cells (25 000 T cells/well) at the ratios indicated. Proliferation was assessed by pulsing with [3H]thymidine for the final 18 hr of the 72 hr incubation. Data are shown as mean ± SEM, Student's t-test. (DC : T ratio 2 : 1 P = 0·9704, 1 : 1 P = 0·5030).

Mentions: The initial phase in this model of tolerance induction is the presentation of the peptide by antigen-presenting cells, most likely DC. Twenty-four hours after the i.n. administration the peptide is found in CD11c-positive DC in spleen, and in peripheral and cervical draining lymph nodes.24 To determine whether complement affected the peptide dissemination and/or presentation, B6 WT and B6.C3–/– female mice were given i.n. HY-AbDby peptide. Twenty-four hours later splenic DC were isolated and used to stimulate HY-AbDby-specific CD4+ T cells from TCR transgenic Marilyn mice (Marilyn T cells). The purity, percentage of CD8+/CD8− DC and expression levels of CD40, CD86 and H2-Ab were similar (data not shown). Regardless of their complement status splenic, DC from the peptide-treated mice were equally able to stimulate the proliferation of the Marilyn T cells (Fig.2); hence complement does not influence the distribution or initial presentation of the peptide.


Intranasal peptide-induced tolerance and linked suppression: consequences of complement deficiency.

Fossati-Jimack L, Ling GS, Baudino L, Szajna M, Manivannan K, Zhao JC, Midgley R, Chai JG, Simpson E, Botto M, Scott D - Immunology (2015)

C3 deficiency does not alter the distribution of the peptide. B6 wild-type (WT; n = 3) and B6.C3–/– (n = 3) female mice were given the HY-AbDby peptide twice (100 μg at time 0 and 6 hr later) intranasally, control mice received PBS. Twenty-four hours after the first peptide dose, splenic dendritic cells (DC) were isolated and cultured with Marilyn T cells (25 000 T cells/well) at the ratios indicated. Proliferation was assessed by pulsing with [3H]thymidine for the final 18 hr of the 72 hr incubation. Data are shown as mean ± SEM, Student's t-test. (DC : T ratio 2 : 1 P = 0·9704, 1 : 1 P = 0·5030).
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4264918&req=5

fig02: C3 deficiency does not alter the distribution of the peptide. B6 wild-type (WT; n = 3) and B6.C3–/– (n = 3) female mice were given the HY-AbDby peptide twice (100 μg at time 0 and 6 hr later) intranasally, control mice received PBS. Twenty-four hours after the first peptide dose, splenic dendritic cells (DC) were isolated and cultured with Marilyn T cells (25 000 T cells/well) at the ratios indicated. Proliferation was assessed by pulsing with [3H]thymidine for the final 18 hr of the 72 hr incubation. Data are shown as mean ± SEM, Student's t-test. (DC : T ratio 2 : 1 P = 0·9704, 1 : 1 P = 0·5030).
Mentions: The initial phase in this model of tolerance induction is the presentation of the peptide by antigen-presenting cells, most likely DC. Twenty-four hours after the i.n. administration the peptide is found in CD11c-positive DC in spleen, and in peripheral and cervical draining lymph nodes.24 To determine whether complement affected the peptide dissemination and/or presentation, B6 WT and B6.C3–/– female mice were given i.n. HY-AbDby peptide. Twenty-four hours later splenic DC were isolated and used to stimulate HY-AbDby-specific CD4+ T cells from TCR transgenic Marilyn mice (Marilyn T cells). The purity, percentage of CD8+/CD8− DC and expression levels of CD40, CD86 and H2-Ab were similar (data not shown). Regardless of their complement status splenic, DC from the peptide-treated mice were equally able to stimulate the proliferation of the Marilyn T cells (Fig.2); hence complement does not influence the distribution or initial presentation of the peptide.

Bottom Line: The mechanisms whereby complement can mediate these diverse regulatory effects are poorly understood.This was related to the inability of C3-deficient DC to up-regulate the arginine-consuming enzyme, inducible nitric oxide synthase (Nos-2), in the presence of antigen-specific Treg cells and peptide, leading to reduced Treg cell generation.Our findings demonstrate that the classical pathway and C3 play a critical role in the peptide-mediated induction of tolerance to HY by modulating DC function.

View Article: PubMed Central - PubMed

Affiliation: Centre for Complement and Inflammation Research, Imperial College London, London, UK.

Show MeSH
Related in: MedlinePlus