Limits...
Both tumour cells and infiltrating T-cells in equine sarcoids express FOXP3 associated with an immune-supressed cytokine microenvironment.

Wilson AD, Hicks C - Vet. Res. (2016)

Bottom Line: Transcription of IL17, which has been shown to have a regulatory function in human papillomavirus-associated tumours, was also elevated in equine sarcoids compared to spleen.In contrast, the levels of mRNA transcripts for effector T cell cytokines IL2, IL4 and interferon-gamma (IFNγ) were not elevated in sarcoids compared to healthy skin or spleen.Similarly neither interferon-alpha (IFNα), interferon-beta (IFNβ) nor IL12 family members were elevated in sarcoids compared to normal skin.

View Article: PubMed Central - PubMed

Affiliation: School of Veterinary Sciences, University of Bristol, Langford, Bristol, BS40 5DU, UK. doug.wilson@bris.ac.uk.

ABSTRACT
Bovine papillomavirus (BPV) infections of equine species have a central role in the aetiology of equine sarcoids; a common benign skin tumour of horses, zebras and donkeys. Within the lesions, all of the early papillomavirus genes are expressed and promote the excessive replication of fibroblasts which characterise these tumours. Equine sarcoids differ from BPV induced fibro-papillomas of cattle (the natural host of BPV), in that they do not produce high amounts of virus particles, do not usually regress spontaneously and do not sero-convert to BPV; features which suggest that affected horses lack an effective anti-viral immune response to BPV. Equine sarcoids contain large numbers of CD4+ CD8+ dual positive T-cells which uniformly express FOXP3, the key transcription factor of regulatory T-cells, and FOXP3 is also expressed within the BPV infected fibroblasts. Compared to healthy skin, sarcoids showed increased mRNA transcription for FOXP3 and the regulatory cytokine TGFβ. Transcription of IL17, which has been shown to have a regulatory function in human papillomavirus-associated tumours, was also elevated in equine sarcoids compared to spleen. In contrast, the levels of mRNA transcripts for effector T cell cytokines IL2, IL4 and interferon-gamma (IFNγ) were not elevated in sarcoids compared to healthy skin or spleen. Similarly neither interferon-alpha (IFNα), interferon-beta (IFNβ) nor IL12 family members were elevated in sarcoids compared to normal skin. We suggest that the regulatory cytokine micro-environment within sarcoids enables the persistence of the lesions by preventing an effective anti-viral immune response.

No MeSH data available.


Related in: MedlinePlus

CD4 and CD8 expression on sarcoid T-cells. Cryostat sections from a lymphocyte rich area of sarcoid tissue dual stained with different combination of rabbit anti-CD3 and monoclonal mouse anti-equine CD4 or equine CD8α. A–C Frozen tissue sections dual labelled with (A) rabbit anti-CD3 (texas red), (B) mouse anti-CD4 (FITC) (C) combined image with DAPI nuclear counterstain, most cells express both CD4 and CD3 although some CD4 only cells are present (green arrow). D–F Frozen tissue sections dual stained with (D) rabbit anti-CD3 (texas red), (E) mouse anti-CD8 (FITC), (F) combined image with DAPI counterstain most cells express both CD3 and CD8 although some CD3 only cells are present (red arrow). G–I Frozen tissue sections dual labelled with (G) monoclonal anti-CD8 TRITC and (H) anti-CD4 FITC (F) combined image with DAPI nuclear counterstain, the majority of cells are CD4 CD8 dual positive with a few CD8 and CD4 single positive cells (red and green arrows respectively).
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4862206&req=5

Fig2: CD4 and CD8 expression on sarcoid T-cells. Cryostat sections from a lymphocyte rich area of sarcoid tissue dual stained with different combination of rabbit anti-CD3 and monoclonal mouse anti-equine CD4 or equine CD8α. A–C Frozen tissue sections dual labelled with (A) rabbit anti-CD3 (texas red), (B) mouse anti-CD4 (FITC) (C) combined image with DAPI nuclear counterstain, most cells express both CD4 and CD3 although some CD4 only cells are present (green arrow). D–F Frozen tissue sections dual stained with (D) rabbit anti-CD3 (texas red), (E) mouse anti-CD8 (FITC), (F) combined image with DAPI counterstain most cells express both CD3 and CD8 although some CD3 only cells are present (red arrow). G–I Frozen tissue sections dual labelled with (G) monoclonal anti-CD8 TRITC and (H) anti-CD4 FITC (F) combined image with DAPI nuclear counterstain, the majority of cells are CD4 CD8 dual positive with a few CD8 and CD4 single positive cells (red and green arrows respectively).

Mentions: To further characterise the phenotype of the tumour infiltrating T-cells, dual staining of sarcoid sections was carried out using rabbit anti-CD3 and donkey anti-rabbit texas red conjugate (Figure 2A) in combination with mouse monoclonal anti-equine CD4 and goat anti-mouse FITC conjugate (Figure 2B). Comparison of the combined images shows that the majority of cells express both CD-antigens. However, there was considerable variation in the intensity of staining, the combined image (Figure 2C) showed a small number of CD3hi CD4hi cells which appear yellow or orange, with the remaining cells appearing green indicating that they express relatively little CD3 in comparison to CD4 although this too appears to be low in intensity on most cells. Occasional CD4+ only cells were also present (green arrows Figures 2A and C).Figure 2


Both tumour cells and infiltrating T-cells in equine sarcoids express FOXP3 associated with an immune-supressed cytokine microenvironment.

Wilson AD, Hicks C - Vet. Res. (2016)

CD4 and CD8 expression on sarcoid T-cells. Cryostat sections from a lymphocyte rich area of sarcoid tissue dual stained with different combination of rabbit anti-CD3 and monoclonal mouse anti-equine CD4 or equine CD8α. A–C Frozen tissue sections dual labelled with (A) rabbit anti-CD3 (texas red), (B) mouse anti-CD4 (FITC) (C) combined image with DAPI nuclear counterstain, most cells express both CD4 and CD3 although some CD4 only cells are present (green arrow). D–F Frozen tissue sections dual stained with (D) rabbit anti-CD3 (texas red), (E) mouse anti-CD8 (FITC), (F) combined image with DAPI counterstain most cells express both CD3 and CD8 although some CD3 only cells are present (red arrow). G–I Frozen tissue sections dual labelled with (G) monoclonal anti-CD8 TRITC and (H) anti-CD4 FITC (F) combined image with DAPI nuclear counterstain, the majority of cells are CD4 CD8 dual positive with a few CD8 and CD4 single positive cells (red and green arrows respectively).
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4862206&req=5

Fig2: CD4 and CD8 expression on sarcoid T-cells. Cryostat sections from a lymphocyte rich area of sarcoid tissue dual stained with different combination of rabbit anti-CD3 and monoclonal mouse anti-equine CD4 or equine CD8α. A–C Frozen tissue sections dual labelled with (A) rabbit anti-CD3 (texas red), (B) mouse anti-CD4 (FITC) (C) combined image with DAPI nuclear counterstain, most cells express both CD4 and CD3 although some CD4 only cells are present (green arrow). D–F Frozen tissue sections dual stained with (D) rabbit anti-CD3 (texas red), (E) mouse anti-CD8 (FITC), (F) combined image with DAPI counterstain most cells express both CD3 and CD8 although some CD3 only cells are present (red arrow). G–I Frozen tissue sections dual labelled with (G) monoclonal anti-CD8 TRITC and (H) anti-CD4 FITC (F) combined image with DAPI nuclear counterstain, the majority of cells are CD4 CD8 dual positive with a few CD8 and CD4 single positive cells (red and green arrows respectively).
Mentions: To further characterise the phenotype of the tumour infiltrating T-cells, dual staining of sarcoid sections was carried out using rabbit anti-CD3 and donkey anti-rabbit texas red conjugate (Figure 2A) in combination with mouse monoclonal anti-equine CD4 and goat anti-mouse FITC conjugate (Figure 2B). Comparison of the combined images shows that the majority of cells express both CD-antigens. However, there was considerable variation in the intensity of staining, the combined image (Figure 2C) showed a small number of CD3hi CD4hi cells which appear yellow or orange, with the remaining cells appearing green indicating that they express relatively little CD3 in comparison to CD4 although this too appears to be low in intensity on most cells. Occasional CD4+ only cells were also present (green arrows Figures 2A and C).Figure 2

Bottom Line: Transcription of IL17, which has been shown to have a regulatory function in human papillomavirus-associated tumours, was also elevated in equine sarcoids compared to spleen.In contrast, the levels of mRNA transcripts for effector T cell cytokines IL2, IL4 and interferon-gamma (IFNγ) were not elevated in sarcoids compared to healthy skin or spleen.Similarly neither interferon-alpha (IFNα), interferon-beta (IFNβ) nor IL12 family members were elevated in sarcoids compared to normal skin.

View Article: PubMed Central - PubMed

Affiliation: School of Veterinary Sciences, University of Bristol, Langford, Bristol, BS40 5DU, UK. doug.wilson@bris.ac.uk.

ABSTRACT
Bovine papillomavirus (BPV) infections of equine species have a central role in the aetiology of equine sarcoids; a common benign skin tumour of horses, zebras and donkeys. Within the lesions, all of the early papillomavirus genes are expressed and promote the excessive replication of fibroblasts which characterise these tumours. Equine sarcoids differ from BPV induced fibro-papillomas of cattle (the natural host of BPV), in that they do not produce high amounts of virus particles, do not usually regress spontaneously and do not sero-convert to BPV; features which suggest that affected horses lack an effective anti-viral immune response to BPV. Equine sarcoids contain large numbers of CD4+ CD8+ dual positive T-cells which uniformly express FOXP3, the key transcription factor of regulatory T-cells, and FOXP3 is also expressed within the BPV infected fibroblasts. Compared to healthy skin, sarcoids showed increased mRNA transcription for FOXP3 and the regulatory cytokine TGFβ. Transcription of IL17, which has been shown to have a regulatory function in human papillomavirus-associated tumours, was also elevated in equine sarcoids compared to spleen. In contrast, the levels of mRNA transcripts for effector T cell cytokines IL2, IL4 and interferon-gamma (IFNγ) were not elevated in sarcoids compared to healthy skin or spleen. Similarly neither interferon-alpha (IFNα), interferon-beta (IFNβ) nor IL12 family members were elevated in sarcoids compared to normal skin. We suggest that the regulatory cytokine micro-environment within sarcoids enables the persistence of the lesions by preventing an effective anti-viral immune response.

No MeSH data available.


Related in: MedlinePlus