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Resident CD8(+) and migratory CD103(+) dendritic cells control CD8 T cell immunity during acute influenza infection.

Waithman J, Zanker D, Xiao K, Oveissi S, Wylie B, Ng R, Tögel L, Chen W - PLoS ONE (2013)

Bottom Line: This transcription factor is required for the development of lymph node resident CD8(+) and migratory CD103(+)CD11b(-) DCs and we found both of these subtypes could efficiently stimulate anti-IAV TCD8+.We postulate the differences reported can partially be explained by how DC are phenotyped, namely the use of MHC class II to segregate subtypes.Our results show that resident CD8(+) DC upregulate this marker during IAV infection and we advise against its use when isolating DC subtypes.

View Article: PubMed Central - PubMed

Affiliation: Telethon Institute for Child Health Research and Centre for Child Health Research, University of Western Australia, Perth, Western Australia, Australia. jwaithman@ichr.uwa.edu.au

ABSTRACT
The identification of the specific DC subsets providing a critical role in presenting influenza antigens to naïve T cell precursors remains contentious and under considerable debate. Here we show that CD8(+) T lymphocyte (TCD8+) responses are severely hampered in C57BL/6 mice deficient in the transcription factor Batf3 after intranasal challenge with influenza A virus (IAV). This transcription factor is required for the development of lymph node resident CD8(+) and migratory CD103(+)CD11b(-) DCs and we found both of these subtypes could efficiently stimulate anti-IAV TCD8+. Using a similar ex vivo approach, many publications on this subject matter excluded a role for resident, non-migratory CD8(+) DC. We postulate the differences reported can partially be explained by how DC are phenotyped, namely the use of MHC class II to segregate subtypes. Our results show that resident CD8(+) DC upregulate this marker during IAV infection and we advise against its use when isolating DC subtypes.

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PA224–33 TCD8+ can be generated in Batf3o/o mice.B6 or Batf3o/o mice were inoculated intraperitoneally with 2.5×106 LPS-treated, PA224–233 peptide-pulsed B6 bone-marrow-derived DC. 7 days later, the number of PA224–233 responding TCD8+ present in the spleen was determined by ICS. Average is taken from 6 mice per group over two independent experiments and the error shows the SEM.
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pone-0066136-g002: PA224–33 TCD8+ can be generated in Batf3o/o mice.B6 or Batf3o/o mice were inoculated intraperitoneally with 2.5×106 LPS-treated, PA224–233 peptide-pulsed B6 bone-marrow-derived DC. 7 days later, the number of PA224–233 responding TCD8+ present in the spleen was determined by ICS. Average is taken from 6 mice per group over two independent experiments and the error shows the SEM.

Mentions: TCD8+ responses were enumerated using intracellular cytokine staining (ICS) after being stimulated with 1 μM of the following antigenic peptides: IAV NP366–374 (ASNENMETM), PA224–233 (SSLENFRAYV), PB1F262–70 (LSLRNPIKV) and PB1703–711 (SSYRRPVGI) in the presence of brefeldin A (Sigma). Anti-CD8 and -IFNγ antibodies (BD) were used to identify positive cells in Fig 1–2. Flow cytometrically sorted DC subsets from influenza infected B6.Ly5.2 mice were co-cultured with a CD45.1+TCD8+ cell line specific to IAV NP366–374 for six hours in the presence of BFA. Anti-CD45.1 and -CD8 antibodies (BD) were used to identify the TCD8+ line and anti-IFNγ antibody (BD) was used to gauge responsiveness to stimulation.


Resident CD8(+) and migratory CD103(+) dendritic cells control CD8 T cell immunity during acute influenza infection.

Waithman J, Zanker D, Xiao K, Oveissi S, Wylie B, Ng R, Tögel L, Chen W - PLoS ONE (2013)

PA224–33 TCD8+ can be generated in Batf3o/o mice.B6 or Batf3o/o mice were inoculated intraperitoneally with 2.5×106 LPS-treated, PA224–233 peptide-pulsed B6 bone-marrow-derived DC. 7 days later, the number of PA224–233 responding TCD8+ present in the spleen was determined by ICS. Average is taken from 6 mice per group over two independent experiments and the error shows the SEM.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3672151&req=5

pone-0066136-g002: PA224–33 TCD8+ can be generated in Batf3o/o mice.B6 or Batf3o/o mice were inoculated intraperitoneally with 2.5×106 LPS-treated, PA224–233 peptide-pulsed B6 bone-marrow-derived DC. 7 days later, the number of PA224–233 responding TCD8+ present in the spleen was determined by ICS. Average is taken from 6 mice per group over two independent experiments and the error shows the SEM.
Mentions: TCD8+ responses were enumerated using intracellular cytokine staining (ICS) after being stimulated with 1 μM of the following antigenic peptides: IAV NP366–374 (ASNENMETM), PA224–233 (SSLENFRAYV), PB1F262–70 (LSLRNPIKV) and PB1703–711 (SSYRRPVGI) in the presence of brefeldin A (Sigma). Anti-CD8 and -IFNγ antibodies (BD) were used to identify positive cells in Fig 1–2. Flow cytometrically sorted DC subsets from influenza infected B6.Ly5.2 mice were co-cultured with a CD45.1+TCD8+ cell line specific to IAV NP366–374 for six hours in the presence of BFA. Anti-CD45.1 and -CD8 antibodies (BD) were used to identify the TCD8+ line and anti-IFNγ antibody (BD) was used to gauge responsiveness to stimulation.

Bottom Line: This transcription factor is required for the development of lymph node resident CD8(+) and migratory CD103(+)CD11b(-) DCs and we found both of these subtypes could efficiently stimulate anti-IAV TCD8+.We postulate the differences reported can partially be explained by how DC are phenotyped, namely the use of MHC class II to segregate subtypes.Our results show that resident CD8(+) DC upregulate this marker during IAV infection and we advise against its use when isolating DC subtypes.

View Article: PubMed Central - PubMed

Affiliation: Telethon Institute for Child Health Research and Centre for Child Health Research, University of Western Australia, Perth, Western Australia, Australia. jwaithman@ichr.uwa.edu.au

ABSTRACT
The identification of the specific DC subsets providing a critical role in presenting influenza antigens to naïve T cell precursors remains contentious and under considerable debate. Here we show that CD8(+) T lymphocyte (TCD8+) responses are severely hampered in C57BL/6 mice deficient in the transcription factor Batf3 after intranasal challenge with influenza A virus (IAV). This transcription factor is required for the development of lymph node resident CD8(+) and migratory CD103(+)CD11b(-) DCs and we found both of these subtypes could efficiently stimulate anti-IAV TCD8+. Using a similar ex vivo approach, many publications on this subject matter excluded a role for resident, non-migratory CD8(+) DC. We postulate the differences reported can partially be explained by how DC are phenotyped, namely the use of MHC class II to segregate subtypes. Our results show that resident CD8(+) DC upregulate this marker during IAV infection and we advise against its use when isolating DC subtypes.

Show MeSH
Related in: MedlinePlus