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Influenza virus-induced dendritic cell maturation is associated with the induction of strong T cell immunity to a coadministered, normally nonimmunogenic protein.

Brimnes MK, Bonifaz L, Steinman RM, Moran TM - J. Exp. Med. (2003)

Bottom Line: In its absence, OVA failed to induce B and T cell responses and even tolerized, but with influenza, OVA-specific antibodies and CD8+ cytolytic T lymphocytes developed.The relatively slow (2-3 d) kinetics of maturation correlated closely to the time at which OVA inhalation elicited specific antibodies.Therefore respiratory infection can induce DC maturation and simultaneously B and T cell immunity to an innocuous antigen inhaled concurrently.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Mount Sinai School of Medicine, New York, 10029 NY, USA.

ABSTRACT
We evaluated the proposal that during microbial infection, dendritic cells (DCs) undergo maturation and present a mixture of peptides derived from the microbe as well as harmless environmental antigens. Mice were exposed to an aerosol of endotoxin free ovalbumin (OVA) in the absence or presence of influenza virus. In its absence, OVA failed to induce B and T cell responses and even tolerized, but with influenza, OVA-specific antibodies and CD8+ cytolytic T lymphocytes developed. With or without infection, OVA was presented selectively in the draining mediastinal lymph nodes, as assessed by the comparable proliferation of infused, CD8+ and CD4+, TCR transgenic T cells. In the absence of influenza, these OVA-specific T cells produced little IL-2, IL-4, IL-10, and IFN-gamma, but with infection, both CD4+ and CD8+ T cells made high levels of IL-2 and IFN-gamma. The OVA plus influenza-treated mice also showed accelerated recovery to a challenge with recombinant vaccinia OVA virus. CD11c+ DCs from the mediastinal lymph nodes of infected mice selectively stimulated both OVA- and influenza-specific T cells and underwent maturation, with higher levels of MHC class II, CD80, and CD86 molecules. The relatively slow (2-3 d) kinetics of maturation correlated closely to the time at which OVA inhalation elicited specific antibodies. Therefore respiratory infection can induce DC maturation and simultaneously B and T cell immunity to an innocuous antigen inhaled concurrently.

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Generation of antibody and CTL responses in naive animals following coadministration of OVA and influenza virus. (A and B) B6 mice were infected with aerosolized X-31 on day 1 followed by egg OVA (A) or LPS OVA (B) on days 2, 3, and 4. 14 d after infection, serum levels of OVA-specific antibodies were determined by ELISA (mean ± SD from four individual mice; representative of two or more experiments. (C) B6 mice were infected with aerosolized X-31 on day 1 and given LPS OVA or egg OVA on days 2, 3, and 4. 8 d after infection the mediastinal lymph nodes were harvested and the lymph node cells were restimulated with SIINFEKL in vitro for 5 d. Data are means ± SD from groups of four mice and representative of three experiments. (D) B6 mice were given egg OVA on days –5, −3, and –1. On day 1, the mice were infected with X-31 and on days 2, 3, and 4 they were administered LPS OVA (egg OVA, then influenza + LPS OVA). A positive control group which was infected with X-31 on day 1 and administered LPS OVA on day 2, 3, and 4 was also included (influenza + LPS OVA). Data are means ± SD from groups of four mice and representative of two experiments.
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fig1: Generation of antibody and CTL responses in naive animals following coadministration of OVA and influenza virus. (A and B) B6 mice were infected with aerosolized X-31 on day 1 followed by egg OVA (A) or LPS OVA (B) on days 2, 3, and 4. 14 d after infection, serum levels of OVA-specific antibodies were determined by ELISA (mean ± SD from four individual mice; representative of two or more experiments. (C) B6 mice were infected with aerosolized X-31 on day 1 and given LPS OVA or egg OVA on days 2, 3, and 4. 8 d after infection the mediastinal lymph nodes were harvested and the lymph node cells were restimulated with SIINFEKL in vitro for 5 d. Data are means ± SD from groups of four mice and representative of three experiments. (D) B6 mice were given egg OVA on days –5, −3, and –1. On day 1, the mice were infected with X-31 and on days 2, 3, and 4 they were administered LPS OVA (egg OVA, then influenza + LPS OVA). A positive control group which was infected with X-31 on day 1 and administered LPS OVA on day 2, 3, and 4 was also included (influenza + LPS OVA). Data are means ± SD from groups of four mice and representative of two experiments.

Mentions: To identify an abundant source of an endotoxin free and harmless protein antigen, we used egg white as a source of OVA. We compared the endotoxin levels in egg white OVA and commercially available OVA using the Limulus Amebocyte Lysate assay. The egg white OVA was endotoxin free (below detection level <0.1 EU/ml) at a concentration as high as 10 mg/ml OVA. In contrast, the commercial source of OVA was contaminated with endotoxin, as 100 μg/ml OVA contained 2 EU/ml. When administered to B6 (Fig. 1) or BALB/C (unpublished data) mice via the airway, the commercial “LPS OVA” induced anti-OVA antibody responses primarily of the IgG1 and IgG2b isotypes, whereas the LPS-free “egg OVA” did not induce antibodies (Fig. 1, A and B). To assess the response to the two different forms of OVA at the T cell level, we measured the generation of CD8+ CTL, but neither preparation of OVA was immunogenic (Fig. 1 C). We therefore prioritized the use of egg OVA as a harmless protein that is by itself nonimmunogenic via the airway.


Influenza virus-induced dendritic cell maturation is associated with the induction of strong T cell immunity to a coadministered, normally nonimmunogenic protein.

Brimnes MK, Bonifaz L, Steinman RM, Moran TM - J. Exp. Med. (2003)

Generation of antibody and CTL responses in naive animals following coadministration of OVA and influenza virus. (A and B) B6 mice were infected with aerosolized X-31 on day 1 followed by egg OVA (A) or LPS OVA (B) on days 2, 3, and 4. 14 d after infection, serum levels of OVA-specific antibodies were determined by ELISA (mean ± SD from four individual mice; representative of two or more experiments. (C) B6 mice were infected with aerosolized X-31 on day 1 and given LPS OVA or egg OVA on days 2, 3, and 4. 8 d after infection the mediastinal lymph nodes were harvested and the lymph node cells were restimulated with SIINFEKL in vitro for 5 d. Data are means ± SD from groups of four mice and representative of three experiments. (D) B6 mice were given egg OVA on days –5, −3, and –1. On day 1, the mice were infected with X-31 and on days 2, 3, and 4 they were administered LPS OVA (egg OVA, then influenza + LPS OVA). A positive control group which was infected with X-31 on day 1 and administered LPS OVA on day 2, 3, and 4 was also included (influenza + LPS OVA). Data are means ± SD from groups of four mice and representative of two experiments.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2196079&req=5

fig1: Generation of antibody and CTL responses in naive animals following coadministration of OVA and influenza virus. (A and B) B6 mice were infected with aerosolized X-31 on day 1 followed by egg OVA (A) or LPS OVA (B) on days 2, 3, and 4. 14 d after infection, serum levels of OVA-specific antibodies were determined by ELISA (mean ± SD from four individual mice; representative of two or more experiments. (C) B6 mice were infected with aerosolized X-31 on day 1 and given LPS OVA or egg OVA on days 2, 3, and 4. 8 d after infection the mediastinal lymph nodes were harvested and the lymph node cells were restimulated with SIINFEKL in vitro for 5 d. Data are means ± SD from groups of four mice and representative of three experiments. (D) B6 mice were given egg OVA on days –5, −3, and –1. On day 1, the mice were infected with X-31 and on days 2, 3, and 4 they were administered LPS OVA (egg OVA, then influenza + LPS OVA). A positive control group which was infected with X-31 on day 1 and administered LPS OVA on day 2, 3, and 4 was also included (influenza + LPS OVA). Data are means ± SD from groups of four mice and representative of two experiments.
Mentions: To identify an abundant source of an endotoxin free and harmless protein antigen, we used egg white as a source of OVA. We compared the endotoxin levels in egg white OVA and commercially available OVA using the Limulus Amebocyte Lysate assay. The egg white OVA was endotoxin free (below detection level <0.1 EU/ml) at a concentration as high as 10 mg/ml OVA. In contrast, the commercial source of OVA was contaminated with endotoxin, as 100 μg/ml OVA contained 2 EU/ml. When administered to B6 (Fig. 1) or BALB/C (unpublished data) mice via the airway, the commercial “LPS OVA” induced anti-OVA antibody responses primarily of the IgG1 and IgG2b isotypes, whereas the LPS-free “egg OVA” did not induce antibodies (Fig. 1, A and B). To assess the response to the two different forms of OVA at the T cell level, we measured the generation of CD8+ CTL, but neither preparation of OVA was immunogenic (Fig. 1 C). We therefore prioritized the use of egg OVA as a harmless protein that is by itself nonimmunogenic via the airway.

Bottom Line: In its absence, OVA failed to induce B and T cell responses and even tolerized, but with influenza, OVA-specific antibodies and CD8+ cytolytic T lymphocytes developed.The relatively slow (2-3 d) kinetics of maturation correlated closely to the time at which OVA inhalation elicited specific antibodies.Therefore respiratory infection can induce DC maturation and simultaneously B and T cell immunity to an innocuous antigen inhaled concurrently.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Mount Sinai School of Medicine, New York, 10029 NY, USA.

ABSTRACT
We evaluated the proposal that during microbial infection, dendritic cells (DCs) undergo maturation and present a mixture of peptides derived from the microbe as well as harmless environmental antigens. Mice were exposed to an aerosol of endotoxin free ovalbumin (OVA) in the absence or presence of influenza virus. In its absence, OVA failed to induce B and T cell responses and even tolerized, but with influenza, OVA-specific antibodies and CD8+ cytolytic T lymphocytes developed. With or without infection, OVA was presented selectively in the draining mediastinal lymph nodes, as assessed by the comparable proliferation of infused, CD8+ and CD4+, TCR transgenic T cells. In the absence of influenza, these OVA-specific T cells produced little IL-2, IL-4, IL-10, and IFN-gamma, but with infection, both CD4+ and CD8+ T cells made high levels of IL-2 and IFN-gamma. The OVA plus influenza-treated mice also showed accelerated recovery to a challenge with recombinant vaccinia OVA virus. CD11c+ DCs from the mediastinal lymph nodes of infected mice selectively stimulated both OVA- and influenza-specific T cells and underwent maturation, with higher levels of MHC class II, CD80, and CD86 molecules. The relatively slow (2-3 d) kinetics of maturation correlated closely to the time at which OVA inhalation elicited specific antibodies. Therefore respiratory infection can induce DC maturation and simultaneously B and T cell immunity to an innocuous antigen inhaled concurrently.

Show MeSH
Related in: MedlinePlus