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Induced bronchus-associated lymphoid tissue serves as a general priming site for T cells and is maintained by dendritic cells.

Halle S, Dujardin HC, Bakocevic N, Fleige H, Danzer H, Willenzon S, Suezer Y, Hämmerling G, Garbi N, Sutter G, Worbs T, Förster R - J. Exp. Med. (2009)

Bottom Line: After intratracheal application, in vitro-differentiated, antigen-loaded DCs rapidly migrate into preformed BALT and efficiently activate antigen-specific T cells, as revealed by two-photon microscopy.Furthermore, the lung-specific depletion of DCs in mice that express the diphtheria toxin receptor under the control of the CD11c promoter interferes with BALT maintenance.Collectively, these data identify BALT as tertiary lymphoid structures supporting the efficient priming of T cell responses directed against unrelated airborne antigens while crucially requiring DCs for its sustained presence.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Immunology, Hannover Medical School, 30625 Hannover, Germany.

ABSTRACT
Mucosal vaccination via the respiratory tract can elicit protective immunity in animal infection models, but the underlying mechanisms are still poorly understood. We show that a single intranasal application of the replication-deficient modified vaccinia virus Ankara, which is widely used as a recombinant vaccination vector, results in prominent induction of bronchus-associated lymphoid tissue (BALT). Although initial peribronchiolar infiltrations, characterized by the presence of dendritic cells (DCs) and few lymphocytes, can be found 4 d after virus application, organized lymphoid structures with segregated B and T cell zones are first observed at day 8. After intratracheal application, in vitro-differentiated, antigen-loaded DCs rapidly migrate into preformed BALT and efficiently activate antigen-specific T cells, as revealed by two-photon microscopy. Furthermore, the lung-specific depletion of DCs in mice that express the diphtheria toxin receptor under the control of the CD11c promoter interferes with BALT maintenance. Collectively, these data identify BALT as tertiary lymphoid structures supporting the efficient priming of T cell responses directed against unrelated airborne antigens while crucially requiring DCs for its sustained presence.

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i.n. vaccination with recombinant MVA induces a distinct population of antigen-specific cytotoxic T cells in the lung. (A) Mice i.n. received 107 IU MVA-Cy5, and 3 d later the phenotype of Cy5+ cells in the draining LNs was determined by flow cytometry. (B) Expression of CD86 on CD11c+MHCII+ brLN DCs before and 2 d after i.n. instillation of MVA (shaded area, isotype control). (C–G) 1 d after the i.v. transfer of 200 CD45.1+ CD8+ OT-I T cells into CD45.2+ recipients, mice were i.n. or i.m. infected with 107 IU MVA-OVA. At the indicated time points (days after infection) the absolute number of OT-I cells was determined in the brLNs (C), BAL (D), lung (E), and spleen (F). (G) The percentage of OT-I cells expressing CD69 in the lung and spleen. (H) Expression of CD3 and CD69 in BALT at 12 d after infection. Bar, 50 µm. Data in A and B are representative of four mice analyzed in two independent experiments. Data shown in C–G are pooled from two independent experiments with two to three mice per time point (means + SD). *, P < 0.05; ***, P < 0.001.
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fig3: i.n. vaccination with recombinant MVA induces a distinct population of antigen-specific cytotoxic T cells in the lung. (A) Mice i.n. received 107 IU MVA-Cy5, and 3 d later the phenotype of Cy5+ cells in the draining LNs was determined by flow cytometry. (B) Expression of CD86 on CD11c+MHCII+ brLN DCs before and 2 d after i.n. instillation of MVA (shaded area, isotype control). (C–G) 1 d after the i.v. transfer of 200 CD45.1+ CD8+ OT-I T cells into CD45.2+ recipients, mice were i.n. or i.m. infected with 107 IU MVA-OVA. At the indicated time points (days after infection) the absolute number of OT-I cells was determined in the brLNs (C), BAL (D), lung (E), and spleen (F). (G) The percentage of OT-I cells expressing CD69 in the lung and spleen. (H) Expression of CD3 and CD69 in BALT at 12 d after infection. Bar, 50 µm. Data in A and B are representative of four mice analyzed in two independent experiments. Data shown in C–G are pooled from two independent experiments with two to three mice per time point (means + SD). *, P < 0.05; ***, P < 0.001.

Mentions: After i.n. instillation of MVA-Cy5, we could readily detect Cy5+ cells in the lung-draining bronchial LNs (brLNs). These cells were primarily DCs expressing high levels of CD11c and MHCII (Fig. 3 A). The majority of these DCs were found to express either CD103 or CD11b (Fig. 3 A). These observations, together with the finding that brLN DCs of i.n. MVA-infected mice express higher levels of CD86 than those of noninfected mice (Fig. 3 B), indicate that the i.n. vaccination with MVA can elicit a productive immune response in the lung as well as in the lung-draining brLNs. We therefore hypothesized that the quality of an adaptive immune response directed against the model antigen OVA in the lung might differ between mucosal (i.n.) and systemic (i.m.) application of MVA encoding chicken OVA (MVA-OVA). To closely mirror the physiological situation of a small starting population of naive T cells being specific for a given antigen, we adoptively transferred by i.v. injection 200 OT-I T cells (CD45.1+) that recognize the SIINFEKL peptide derived from OVA in an MHC class I–restricted manner into CD45.2 recipients 1 d before infection with MVA-OVA via the i.n. or i.m. route.


Induced bronchus-associated lymphoid tissue serves as a general priming site for T cells and is maintained by dendritic cells.

Halle S, Dujardin HC, Bakocevic N, Fleige H, Danzer H, Willenzon S, Suezer Y, Hämmerling G, Garbi N, Sutter G, Worbs T, Förster R - J. Exp. Med. (2009)

i.n. vaccination with recombinant MVA induces a distinct population of antigen-specific cytotoxic T cells in the lung. (A) Mice i.n. received 107 IU MVA-Cy5, and 3 d later the phenotype of Cy5+ cells in the draining LNs was determined by flow cytometry. (B) Expression of CD86 on CD11c+MHCII+ brLN DCs before and 2 d after i.n. instillation of MVA (shaded area, isotype control). (C–G) 1 d after the i.v. transfer of 200 CD45.1+ CD8+ OT-I T cells into CD45.2+ recipients, mice were i.n. or i.m. infected with 107 IU MVA-OVA. At the indicated time points (days after infection) the absolute number of OT-I cells was determined in the brLNs (C), BAL (D), lung (E), and spleen (F). (G) The percentage of OT-I cells expressing CD69 in the lung and spleen. (H) Expression of CD3 and CD69 in BALT at 12 d after infection. Bar, 50 µm. Data in A and B are representative of four mice analyzed in two independent experiments. Data shown in C–G are pooled from two independent experiments with two to three mice per time point (means + SD). *, P < 0.05; ***, P < 0.001.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC2806625&req=5

fig3: i.n. vaccination with recombinant MVA induces a distinct population of antigen-specific cytotoxic T cells in the lung. (A) Mice i.n. received 107 IU MVA-Cy5, and 3 d later the phenotype of Cy5+ cells in the draining LNs was determined by flow cytometry. (B) Expression of CD86 on CD11c+MHCII+ brLN DCs before and 2 d after i.n. instillation of MVA (shaded area, isotype control). (C–G) 1 d after the i.v. transfer of 200 CD45.1+ CD8+ OT-I T cells into CD45.2+ recipients, mice were i.n. or i.m. infected with 107 IU MVA-OVA. At the indicated time points (days after infection) the absolute number of OT-I cells was determined in the brLNs (C), BAL (D), lung (E), and spleen (F). (G) The percentage of OT-I cells expressing CD69 in the lung and spleen. (H) Expression of CD3 and CD69 in BALT at 12 d after infection. Bar, 50 µm. Data in A and B are representative of four mice analyzed in two independent experiments. Data shown in C–G are pooled from two independent experiments with two to three mice per time point (means + SD). *, P < 0.05; ***, P < 0.001.
Mentions: After i.n. instillation of MVA-Cy5, we could readily detect Cy5+ cells in the lung-draining bronchial LNs (brLNs). These cells were primarily DCs expressing high levels of CD11c and MHCII (Fig. 3 A). The majority of these DCs were found to express either CD103 or CD11b (Fig. 3 A). These observations, together with the finding that brLN DCs of i.n. MVA-infected mice express higher levels of CD86 than those of noninfected mice (Fig. 3 B), indicate that the i.n. vaccination with MVA can elicit a productive immune response in the lung as well as in the lung-draining brLNs. We therefore hypothesized that the quality of an adaptive immune response directed against the model antigen OVA in the lung might differ between mucosal (i.n.) and systemic (i.m.) application of MVA encoding chicken OVA (MVA-OVA). To closely mirror the physiological situation of a small starting population of naive T cells being specific for a given antigen, we adoptively transferred by i.v. injection 200 OT-I T cells (CD45.1+) that recognize the SIINFEKL peptide derived from OVA in an MHC class I–restricted manner into CD45.2 recipients 1 d before infection with MVA-OVA via the i.n. or i.m. route.

Bottom Line: After intratracheal application, in vitro-differentiated, antigen-loaded DCs rapidly migrate into preformed BALT and efficiently activate antigen-specific T cells, as revealed by two-photon microscopy.Furthermore, the lung-specific depletion of DCs in mice that express the diphtheria toxin receptor under the control of the CD11c promoter interferes with BALT maintenance.Collectively, these data identify BALT as tertiary lymphoid structures supporting the efficient priming of T cell responses directed against unrelated airborne antigens while crucially requiring DCs for its sustained presence.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Immunology, Hannover Medical School, 30625 Hannover, Germany.

ABSTRACT
Mucosal vaccination via the respiratory tract can elicit protective immunity in animal infection models, but the underlying mechanisms are still poorly understood. We show that a single intranasal application of the replication-deficient modified vaccinia virus Ankara, which is widely used as a recombinant vaccination vector, results in prominent induction of bronchus-associated lymphoid tissue (BALT). Although initial peribronchiolar infiltrations, characterized by the presence of dendritic cells (DCs) and few lymphocytes, can be found 4 d after virus application, organized lymphoid structures with segregated B and T cell zones are first observed at day 8. After intratracheal application, in vitro-differentiated, antigen-loaded DCs rapidly migrate into preformed BALT and efficiently activate antigen-specific T cells, as revealed by two-photon microscopy. Furthermore, the lung-specific depletion of DCs in mice that express the diphtheria toxin receptor under the control of the CD11c promoter interferes with BALT maintenance. Collectively, these data identify BALT as tertiary lymphoid structures supporting the efficient priming of T cell responses directed against unrelated airborne antigens while crucially requiring DCs for its sustained presence.

Show MeSH
Related in: MedlinePlus