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Deficient Natural Killer Dendritic Cell Responses Underlay the Induction of Theiler's Virus-Induced Autoimmunity.

Chastain EM, Getts DR, Miller SD - MBio (2015)

Bottom Line: Viral infection is an important cofactor, along with genetic susceptibility, in the initiation of a variety of organ-specific autoimmune diseases.Thus, in-depth understanding of how virus infections trigger autoimmunity may lead to novel ways to prevent or treat these diseases.Theiler's murine encephalitis virus-induced demyelinating disease (TMEV-IDD) serves as an important model for the human T cell-mediated autoimmune demyelinating disease multiple sclerosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology-Immunology and Interdepartmental Immunobiology Center, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USA.

No MeSH data available.


Related in: MedlinePlus

CNS TMEV infection induces proliferation and activation of splenic NKDCs. Wild-type B6 mice were i.c. infected with 5 × 106 PFU of TMEV or sham infected. Spleens were harvested during acute infection, and mononuclear cells were isolated and analyzed by flow cytometry. (A) Quantification of CD3− CD45+ CD11c+ NK1.1+ DX5+ spleen cells on days 1, 3, 5, and 7 p.i. in TMEV- or sham-infected B6 mice. (B) At 24 and 48 h p.i., mice were i.p. injected with 1 mg BrdU in 100 µl sterile PBS. At 96 h p.i., the percentage of BrdU-positive NKDCs from spleens of TMEV- or sham-infected mice was quantified. (C) Spleens were harvested from B6 mice at 24 and 72 h post-TMEV infection, and expression levels of I-Ab, CD40, CD86, and CD80 on CD3− CD45+ CD11c+ NK1.1+ DX5+ spleen cells were determined. (D) Quantification of I-Ab-, CD40-, CD86-, and CD80-positive CD3− CD45+ CD11c+ NK1.1+ DX5+ spleen cells on days 24 and 72 h p.i. in TMEV- or sham-infected B6 mice. For histogram overlays, the shaded portion represents the isotype control, and the black line represents antibody staining. Data are representative of 2 to 3 independent experiments with 5 mice per group. Error bars show standard deviations. **, P < 0.01; *, P < 0.05.
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fig2: CNS TMEV infection induces proliferation and activation of splenic NKDCs. Wild-type B6 mice were i.c. infected with 5 × 106 PFU of TMEV or sham infected. Spleens were harvested during acute infection, and mononuclear cells were isolated and analyzed by flow cytometry. (A) Quantification of CD3− CD45+ CD11c+ NK1.1+ DX5+ spleen cells on days 1, 3, 5, and 7 p.i. in TMEV- or sham-infected B6 mice. (B) At 24 and 48 h p.i., mice were i.p. injected with 1 mg BrdU in 100 µl sterile PBS. At 96 h p.i., the percentage of BrdU-positive NKDCs from spleens of TMEV- or sham-infected mice was quantified. (C) Spleens were harvested from B6 mice at 24 and 72 h post-TMEV infection, and expression levels of I-Ab, CD40, CD86, and CD80 on CD3− CD45+ CD11c+ NK1.1+ DX5+ spleen cells were determined. (D) Quantification of I-Ab-, CD40-, CD86-, and CD80-positive CD3− CD45+ CD11c+ NK1.1+ DX5+ spleen cells on days 24 and 72 h p.i. in TMEV- or sham-infected B6 mice. For histogram overlays, the shaded portion represents the isotype control, and the black line represents antibody staining. Data are representative of 2 to 3 independent experiments with 5 mice per group. Error bars show standard deviations. **, P < 0.01; *, P < 0.05.

Mentions: In the TMEV-IDD model, piercing of the blood-brain barrier (BBB) results from i.c. virus inoculation and results in peripheral viremia. We were thus interested in assessing how TMEV infection might influence peripherally located NKDCs. Activation was studied by examining proliferation of NKDCs as well as upregulation of key molecules associated with DC activation. Examination of NKDC kinetics in the spleen showed that the number and percentage of NKDCs in the spleen peaked at day 3 and returned to baseline levels at day 5 p.i. (Fig. 2A). Employing bromodeoxyuridine (BrdU) incorporation, we interestingly found that NKDCs, but not DCs or NK cells, underwent significant proliferation after virus infection (Fig. 2B; see Fig. S2 in the supplemental material), suggesting that the increase in the numbers of NKDCs in the spleen was due to active replication. Infection-induced NKDC activation was evidenced by upregulation of molecules associated with antiviral immunity, including MHC-II (I-Ab), CD40, CD80, and CD86. Interestingly, significant upregulation of these molecules was observed within 24 h p.i. compared to sham-infected animals (Fig. 2C and D). MHC class II expression was seen to kinetically increase on NKDCs in the CNS as well (not shown).


Deficient Natural Killer Dendritic Cell Responses Underlay the Induction of Theiler's Virus-Induced Autoimmunity.

Chastain EM, Getts DR, Miller SD - MBio (2015)

CNS TMEV infection induces proliferation and activation of splenic NKDCs. Wild-type B6 mice were i.c. infected with 5 × 106 PFU of TMEV or sham infected. Spleens were harvested during acute infection, and mononuclear cells were isolated and analyzed by flow cytometry. (A) Quantification of CD3− CD45+ CD11c+ NK1.1+ DX5+ spleen cells on days 1, 3, 5, and 7 p.i. in TMEV- or sham-infected B6 mice. (B) At 24 and 48 h p.i., mice were i.p. injected with 1 mg BrdU in 100 µl sterile PBS. At 96 h p.i., the percentage of BrdU-positive NKDCs from spleens of TMEV- or sham-infected mice was quantified. (C) Spleens were harvested from B6 mice at 24 and 72 h post-TMEV infection, and expression levels of I-Ab, CD40, CD86, and CD80 on CD3− CD45+ CD11c+ NK1.1+ DX5+ spleen cells were determined. (D) Quantification of I-Ab-, CD40-, CD86-, and CD80-positive CD3− CD45+ CD11c+ NK1.1+ DX5+ spleen cells on days 24 and 72 h p.i. in TMEV- or sham-infected B6 mice. For histogram overlays, the shaded portion represents the isotype control, and the black line represents antibody staining. Data are representative of 2 to 3 independent experiments with 5 mice per group. Error bars show standard deviations. **, P < 0.01; *, P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig2: CNS TMEV infection induces proliferation and activation of splenic NKDCs. Wild-type B6 mice were i.c. infected with 5 × 106 PFU of TMEV or sham infected. Spleens were harvested during acute infection, and mononuclear cells were isolated and analyzed by flow cytometry. (A) Quantification of CD3− CD45+ CD11c+ NK1.1+ DX5+ spleen cells on days 1, 3, 5, and 7 p.i. in TMEV- or sham-infected B6 mice. (B) At 24 and 48 h p.i., mice were i.p. injected with 1 mg BrdU in 100 µl sterile PBS. At 96 h p.i., the percentage of BrdU-positive NKDCs from spleens of TMEV- or sham-infected mice was quantified. (C) Spleens were harvested from B6 mice at 24 and 72 h post-TMEV infection, and expression levels of I-Ab, CD40, CD86, and CD80 on CD3− CD45+ CD11c+ NK1.1+ DX5+ spleen cells were determined. (D) Quantification of I-Ab-, CD40-, CD86-, and CD80-positive CD3− CD45+ CD11c+ NK1.1+ DX5+ spleen cells on days 24 and 72 h p.i. in TMEV- or sham-infected B6 mice. For histogram overlays, the shaded portion represents the isotype control, and the black line represents antibody staining. Data are representative of 2 to 3 independent experiments with 5 mice per group. Error bars show standard deviations. **, P < 0.01; *, P < 0.05.
Mentions: In the TMEV-IDD model, piercing of the blood-brain barrier (BBB) results from i.c. virus inoculation and results in peripheral viremia. We were thus interested in assessing how TMEV infection might influence peripherally located NKDCs. Activation was studied by examining proliferation of NKDCs as well as upregulation of key molecules associated with DC activation. Examination of NKDC kinetics in the spleen showed that the number and percentage of NKDCs in the spleen peaked at day 3 and returned to baseline levels at day 5 p.i. (Fig. 2A). Employing bromodeoxyuridine (BrdU) incorporation, we interestingly found that NKDCs, but not DCs or NK cells, underwent significant proliferation after virus infection (Fig. 2B; see Fig. S2 in the supplemental material), suggesting that the increase in the numbers of NKDCs in the spleen was due to active replication. Infection-induced NKDC activation was evidenced by upregulation of molecules associated with antiviral immunity, including MHC-II (I-Ab), CD40, CD80, and CD86. Interestingly, significant upregulation of these molecules was observed within 24 h p.i. compared to sham-infected animals (Fig. 2C and D). MHC class II expression was seen to kinetically increase on NKDCs in the CNS as well (not shown).

Bottom Line: Viral infection is an important cofactor, along with genetic susceptibility, in the initiation of a variety of organ-specific autoimmune diseases.Thus, in-depth understanding of how virus infections trigger autoimmunity may lead to novel ways to prevent or treat these diseases.Theiler's murine encephalitis virus-induced demyelinating disease (TMEV-IDD) serves as an important model for the human T cell-mediated autoimmune demyelinating disease multiple sclerosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology-Immunology and Interdepartmental Immunobiology Center, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USA.

No MeSH data available.


Related in: MedlinePlus