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Phenotypic and functional characterization of the major lymphocyte populations in the fruit-eating bat Pteropus alecto

View Article: PubMed Central - PubMed

ABSTRACT

The unique ability of bats to act as reservoir for viruses that are highly pathogenic to humans suggests unique properties and functional characteristics of their immune system. However, the lack of bat specific reagents, in particular antibodies, has limited our knowledge of bat’s immunity. Using cross-reactive antibodies, we report the phenotypic and functional characterization of T cell subsets, B and NK cells in the fruit-eating bat Pteropus alecto. Our findings indicate the predominance of CD8+ T cells in the spleen from wild-caught bats that may reflect either the presence of viruses in this organ or predominance of this cell subset at steady state. Instead majority of T cells in circulation, lymph nodes and bone marrow (BM) were CD4+ subsets. Interestingly, 40% of spleen T cells expressed constitutively IL-17, IL-22 and TGF-β mRNA, which may indicate a strong bias towards the Th17 and regulatory T cell subsets. Furthermore, the unexpected high number of T cells in bats BM could suggest an important role in T cell development. Finally, mitogenic stimulation induced proliferation and production of effector molecules by bats immune cells. This work contributes to a better understanding of bat’s immunity, opening up new perspectives of therapeutic interventions for humans.

No MeSH data available.


Production of cytokines by B and NK cells upon mitogenic stimulation.Bat splenocytes or PBMCs were stimulated for 4 h with PDBu/ionomycin or media in the presence of brefeldin A and monensin. Cells were gated on CD3− MHCII+ (B cells) (a,b) or CD3− Tbet+ Eomes+ (NK cells). Production of intracellular TNF (a,c), IL-10 (b), IFN-γ (d) was done at the protein level, and production of granzyme B and perforin (e) was performed at the mRNA level by Flow-FISH. Dot plots from one bat are shown and are representative of the results obtained with 2–3 different bats.
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f6: Production of cytokines by B and NK cells upon mitogenic stimulation.Bat splenocytes or PBMCs were stimulated for 4 h with PDBu/ionomycin or media in the presence of brefeldin A and monensin. Cells were gated on CD3− MHCII+ (B cells) (a,b) or CD3− Tbet+ Eomes+ (NK cells). Production of intracellular TNF (a,c), IL-10 (b), IFN-γ (d) was done at the protein level, and production of granzyme B and perforin (e) was performed at the mRNA level by Flow-FISH. Dot plots from one bat are shown and are representative of the results obtained with 2–3 different bats.

Mentions: Both B cells and NK cells responded to PDBu/Iono stimulation (Fig. 6). Of the B cells, 20% and 8.8% produced TNF and IL-10, respectively (Fig. 6a,b). Of the NK cells, 66.7% and 33.9% produced TNF and IFN-γ, respectively (Fig. 6c,d). Furthermore, 46.5% of NK cells were also found to produce perforin and 24.7% produced both perforin and granzyme B (Fig. 6e).


Phenotypic and functional characterization of the major lymphocyte populations in the fruit-eating bat Pteropus alecto
Production of cytokines by B and NK cells upon mitogenic stimulation.Bat splenocytes or PBMCs were stimulated for 4 h with PDBu/ionomycin or media in the presence of brefeldin A and monensin. Cells were gated on CD3− MHCII+ (B cells) (a,b) or CD3− Tbet+ Eomes+ (NK cells). Production of intracellular TNF (a,c), IL-10 (b), IFN-γ (d) was done at the protein level, and production of granzyme B and perforin (e) was performed at the mRNA level by Flow-FISH. Dot plots from one bat are shown and are representative of the results obtained with 2–3 different bats.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5121612&req=5

f6: Production of cytokines by B and NK cells upon mitogenic stimulation.Bat splenocytes or PBMCs were stimulated for 4 h with PDBu/ionomycin or media in the presence of brefeldin A and monensin. Cells were gated on CD3− MHCII+ (B cells) (a,b) or CD3− Tbet+ Eomes+ (NK cells). Production of intracellular TNF (a,c), IL-10 (b), IFN-γ (d) was done at the protein level, and production of granzyme B and perforin (e) was performed at the mRNA level by Flow-FISH. Dot plots from one bat are shown and are representative of the results obtained with 2–3 different bats.
Mentions: Both B cells and NK cells responded to PDBu/Iono stimulation (Fig. 6). Of the B cells, 20% and 8.8% produced TNF and IL-10, respectively (Fig. 6a,b). Of the NK cells, 66.7% and 33.9% produced TNF and IFN-γ, respectively (Fig. 6c,d). Furthermore, 46.5% of NK cells were also found to produce perforin and 24.7% produced both perforin and granzyme B (Fig. 6e).

View Article: PubMed Central - PubMed

ABSTRACT

The unique ability of bats to act as reservoir for viruses that are highly pathogenic to humans suggests unique properties and functional characteristics of their immune system. However, the lack of bat specific reagents, in particular antibodies, has limited our knowledge of bat’s immunity. Using cross-reactive antibodies, we report the phenotypic and functional characterization of T cell subsets, B and NK cells in the fruit-eating bat Pteropus alecto. Our findings indicate the predominance of CD8+ T cells in the spleen from wild-caught bats that may reflect either the presence of viruses in this organ or predominance of this cell subset at steady state. Instead majority of T cells in circulation, lymph nodes and bone marrow (BM) were CD4+ subsets. Interestingly, 40% of spleen T cells expressed constitutively IL-17, IL-22 and TGF-β mRNA, which may indicate a strong bias towards the Th17 and regulatory T cell subsets. Furthermore, the unexpected high number of T cells in bats BM could suggest an important role in T cell development. Finally, mitogenic stimulation induced proliferation and production of effector molecules by bats immune cells. This work contributes to a better understanding of bat’s immunity, opening up new perspectives of therapeutic interventions for humans.

No MeSH data available.