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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

TMEV-activated NKDCs are poor APCs but efficient killers. FACS-purified NK cells, DCs, and NKDCs from TMEV-infected B6 mice were tested for APCs and killing function cocultured with target cells (T cells or YAC-1 cells). (A and B) Splenic DCs and NKDCs were FACS sorted from B6 mice 24 h post-TMEV infection and cocultured at a 1:1 ratio with CFSE-labeled pan-splenic T cells from VP425-38- or VP2121-130-primed mice ± 2 µM cognate peptide. Seventy-two hours later, CD3+ T cells were analyzed for proliferation via CFSE dilution (A) and activation marker expression via expression of CD44 and CD69 (B). (C) YAC-1 target cells were cocultured with FACS-purified NK cells, DCs, and NKDCs from TMEV-infected B6 mice at the indicated effector-to-target ratios for 4 h and assayed for LDH release as a marker of cytotoxicity. Data are representative of 3 independent experiments.
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fig4: TMEV-activated NKDCs are poor APCs but efficient killers. FACS-purified NK cells, DCs, and NKDCs from TMEV-infected B6 mice were tested for APCs and killing function cocultured with target cells (T cells or YAC-1 cells). (A and B) Splenic DCs and NKDCs were FACS sorted from B6 mice 24 h post-TMEV infection and cocultured at a 1:1 ratio with CFSE-labeled pan-splenic T cells from VP425-38- or VP2121-130-primed mice ± 2 µM cognate peptide. Seventy-two hours later, CD3+ T cells were analyzed for proliferation via CFSE dilution (A) and activation marker expression via expression of CD44 and CD69 (B). (C) YAC-1 target cells were cocultured with FACS-purified NK cells, DCs, and NKDCs from TMEV-infected B6 mice at the indicated effector-to-target ratios for 4 h and assayed for LDH release as a marker of cytotoxicity. Data are representative of 3 independent experiments.

Mentions: Based on both the accumulation of NKDCs in the CNS and their upregulation of classical APC molecules following TMEV infection, we hypothesized that NKDCs may functionally serve as APCs, playing a potential role in the initiation of the antiviral adaptive immune response. To assess their APC capacity in comparison to conventional DCs, fluorescence-activated cell sorter (FACS)-sorted NKDCs (CD11c+ NK1.1+) and DCs (CD11c+ NK1.1−) from the spleens of B6 animals 24 h post-TMEV infection were cocultured with carboxyfluorescein succinimidyl ester (CFSE)-labeled T cells isolated from naive B6 animals primed with either VP425-38 or VP2121-130, which are the dominant TMEV CD4+ and CD8+ T cell epitopes, respectively. While DCs were considerably more efficient at inducing proliferation of both CD4+ and CD8+ T cells, NKDCs were still however able to induce T cell proliferation, as assessed by CFSE dilution (Fig. 4A). In terms of activation marker (CD44 and CD69) expression on the CD4+ and CD8+ T cells (Fig. 4B), there was a significant background in the cultures lacking peptide as the cells came from primed mice and the DCs and NKDCs likely were presenting endogenous viral peptides as they were purified from TMEV-infected B6 mice. Upregulation of CD44 on CD4+ T cells cocultured with VP425-28 was observed with both NKDCs (60%) and DCs (64%) as stimulators. However, there was no CD44hi population in cultures with NKDCs. Interestingly, the percentages of CD69+ CD4+ T cells were greater after culture with NKDCS (62.5%) versus DCs (50.8%). In both cases, most of the CD69+ CD4+ T cells were also CD44+.


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

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

TMEV-activated NKDCs are poor APCs but efficient killers. FACS-purified NK cells, DCs, and NKDCs from TMEV-infected B6 mice were tested for APCs and killing function cocultured with target cells (T cells or YAC-1 cells). (A and B) Splenic DCs and NKDCs were FACS sorted from B6 mice 24 h post-TMEV infection and cocultured at a 1:1 ratio with CFSE-labeled pan-splenic T cells from VP425-38- or VP2121-130-primed mice ± 2 µM cognate peptide. Seventy-two hours later, CD3+ T cells were analyzed for proliferation via CFSE dilution (A) and activation marker expression via expression of CD44 and CD69 (B). (C) YAC-1 target cells were cocultured with FACS-purified NK cells, DCs, and NKDCs from TMEV-infected B6 mice at the indicated effector-to-target ratios for 4 h and assayed for LDH release as a marker of cytotoxicity. Data are representative of 3 independent experiments.
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Related In: Results  -  Collection

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fig4: TMEV-activated NKDCs are poor APCs but efficient killers. FACS-purified NK cells, DCs, and NKDCs from TMEV-infected B6 mice were tested for APCs and killing function cocultured with target cells (T cells or YAC-1 cells). (A and B) Splenic DCs and NKDCs were FACS sorted from B6 mice 24 h post-TMEV infection and cocultured at a 1:1 ratio with CFSE-labeled pan-splenic T cells from VP425-38- or VP2121-130-primed mice ± 2 µM cognate peptide. Seventy-two hours later, CD3+ T cells were analyzed for proliferation via CFSE dilution (A) and activation marker expression via expression of CD44 and CD69 (B). (C) YAC-1 target cells were cocultured with FACS-purified NK cells, DCs, and NKDCs from TMEV-infected B6 mice at the indicated effector-to-target ratios for 4 h and assayed for LDH release as a marker of cytotoxicity. Data are representative of 3 independent experiments.
Mentions: Based on both the accumulation of NKDCs in the CNS and their upregulation of classical APC molecules following TMEV infection, we hypothesized that NKDCs may functionally serve as APCs, playing a potential role in the initiation of the antiviral adaptive immune response. To assess their APC capacity in comparison to conventional DCs, fluorescence-activated cell sorter (FACS)-sorted NKDCs (CD11c+ NK1.1+) and DCs (CD11c+ NK1.1−) from the spleens of B6 animals 24 h post-TMEV infection were cocultured with carboxyfluorescein succinimidyl ester (CFSE)-labeled T cells isolated from naive B6 animals primed with either VP425-38 or VP2121-130, which are the dominant TMEV CD4+ and CD8+ T cell epitopes, respectively. While DCs were considerably more efficient at inducing proliferation of both CD4+ and CD8+ T cells, NKDCs were still however able to induce T cell proliferation, as assessed by CFSE dilution (Fig. 4A). In terms of activation marker (CD44 and CD69) expression on the CD4+ and CD8+ T cells (Fig. 4B), there was a significant background in the cultures lacking peptide as the cells came from primed mice and the DCs and NKDCs likely were presenting endogenous viral peptides as they were purified from TMEV-infected B6 mice. Upregulation of CD44 on CD4+ T cells cocultured with VP425-28 was observed with both NKDCs (60%) and DCs (64%) as stimulators. However, there was no CD44hi population in cultures with NKDCs. Interestingly, the percentages of CD69+ CD4+ T cells were greater after culture with NKDCS (62.5%) versus DCs (50.8%). In both cases, most of the CD69+ CD4+ T cells were also CD44+.

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