Characterization of monoclonal antibodies to equine interleukin-10 and detection of T regulatory 1 cells in horses

Wagner, Bettina; Hillegas, Julia M.; Brinker, Danielle R.; Horohov, David W.; Antczak, Douglas F.

doi:10.1016/j.vetimm.2007.10.012

Cited by 53 publications

(27 citation statements)

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“…Monoclonal antibodies to equine IL-4 [42], equine IL-10 [43], and bovine IFN-γ (MorphoSys, AbD Serotec, Oxford, UK) were used for intracellular staining. The later antibody was previously found to cross-react with equine IFN-γ [32, 41].…”

Section: Methodsmentioning

confidence: 99%

“…During the past few years, reagents to detect the production of several equine cytokines became available including monoclonal antibodies (mAb) to equine IL-4 [42] and IL-10 [43]. In addition, various groups have previously shown that a mAb to bovine IFN-γ cross-reacts with equine IFN-γ [32, 41].…”

Section: Introductionmentioning

confidence: 99%

“…Although antibodies to CD3, CD4 and CD8 were developed and characterized by two workshops on equine leukocyte antigens [18, 21], reagents for T cell receptors, natural killer cells or CD25 to better identify Treg cells are still missing. Thus far, the only Treg cell subsets that were characterized in horses were (1) a population of CD4 + T cells that simultaneously produces IFN-γ and IL-10 [43], and (2) a population of CD4 + foxp3 expressing cells that was recently characterized in peripheral blood mononuclear cells (PBMC) and endometrial cup cells [8]. The CD4 + /IFN-γ + /IL-10 + Treg cell subset was named T R 1 cells according to a similar cell population identified in humans and mice [14].…”

Section: Introductionmentioning

confidence: 99%

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Interferon-gamma, interleukin-4 and interleukin-10 production by T helper cells reveals intact Th1 and regulatory T_R1 cell activation and a delay of the Th2 cell response in equine neonates and foals

2010

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Cytokines produced by T helper (Th) cells are important in orchestrating the immune response during health and disease. Recent reports indicated that cytokine mRNA expression in foals is often quantitatively lower than that of adult horses suggesting that foal T cells are not fully mature. Here, peripheral blood mononuclear cells from foals and adult horses were stimulated with phorbol 12-myristate 13-acetate and analyzed for intracellular interferon-gamma (IFN-γ), interleukin-4 (IL-4) and IL-10 production, representing the Th1, Th2 and regulatory TR1 cell phenotypes respectively, by flow cytometry. In agreement with previous reports, all three cytokines were quantitatively reduced in foals compared to adults. However, the balance between Th1 and Th2 cytokines (IFN-γ/IL-4 ratio) showed a clear Th1-biased response in foals by 6 and 12 weeks of life, while similar IFN-γ/IL-10 ratios were found in foals and adult horses. By day 5 after birth, intracellular IFN-γ production by foal CD4+ and CD8+ T cells resembled that in adults. Overall, IL-4 production was low in foals. IL-4+ cells peaked at day 5 of age when IL-4 was mainly produced by IgE+ cells. Relative percentages of IL-4+ Th2 cells were significantly lower in foals at all time points. The data suggested that equine neonates and young foals have an impaired Th2 response, that the immune response of foals is Th1 biased, that IFN-γ production by Th and cytotoxic T cells is qualitatively similar to adult horses, and regulatory IL-10 production by T cells is developmentally mature in foals during the first three months of life.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Interferon-gamma, interleukin-4 and interleukin-10 production by T helper cells reveals intact Th1 and regulatory T_R1 cell activation and a delay of the Th2 cell response in equine neonates and foals

2010

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“…22,23 Regulatory T cells may be broadly divided into naturally occurring subsets (nTreg), which develop in the thymus along a regulatory lineage (for example, the canonical CD4 + CD25 + Foxp3 + Treg cells); and induced, or 'adaptive', subsets, which arise in the periphery following activation of conventional T (Tcon) cells in a microenvironment rich in regulatory cytokines, or following the interaction of Tcon with nTreg cells in a process called infectious tolerance -for example, the Tr1, Th3 and 'induced Treg (iTreg)' cells. 24 44 However, relatively little is known about Treg cells in dogs, though indirect evidence for their existence has been available for several years. [45][46][47] We 48 and others [49][50][51][52][53][54] have used the anti-mouse/rat Foxp3 antibody clone FJK-16s to identify a population of canine CD4 + T cells that phenotypically resembles Treg cells, but direct evidence for regulatory activity has remained elusive.…”

Section: M M U N O L O G Y O R I G I N a L A R T I C L Ementioning

confidence: 99%

Phenotypic and functional characterization of a CD4+ CD25high FOXP3high regulatory T-cell population in the dog

et al. 2010

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Summary Relatively little is known about regulatory T (Treg) cells and their functional responses in dogs. We have used the cross‐reactive anti‐mouse/rat Foxp3 antibody clone FJK‐16s to identify a population of canine CD4+ FOXP3high T cells in both the peripheral blood (PB) and popliteal lymph node (LN). FOXP3+ cells in both PB and LN yielded positive staining with the newly developed anti‐murine/human Helios antibody clone 22F6, consistent with the notion that they were naturally occurring Treg cells. Stimulation of mononuclear cells of LN origin with concanavalin A (Con A) in vitro yielded increased proportions and median fluorescence intensity of FOXP3 expression by both CD4+ and CD8+ T cells. Removal of the Con A and continued culture disclosed a CD4+ FOXP3high population, distinct from the CD4+ FOXP3intermediate T cells; very few CD8+ FOXP3high T cells were observed, though CD8+ FOXP3intermediate cells were present in equal abundance to CD4+ FOXP3intermediate cells. The CD4+ FOXP3high T cells were thought to represent activated Treg cells, in contrast to the FOXP3intermediate cells, which were thought to be a more heterogeneous population comprising predominantly activated conventional T cells. Co‐staining with interferon‐γ (IFN‐γ) supported this notion, because the FOXP3high T cells were almost exclusively IFN‐γ−, whereas the FOXP3intermediate cells expressed a more heterogeneous IFN‐γ phenotype. Following activation of mononuclear cells with Con A and interleukin‐2, the 5% of CD4+ T cells showing the highest CD25 expression (CD4+ CD25high) were enriched in cells expressing FOXP3. These cells were anergic in vitro, in contrast to the 20% of CD4+ T cells with the lowest CD25 expression (CD4+ CD25−), which proliferated readily. The CD4+ CD25high FOXP3high T cells were able to suppress the proliferation of responder CD4+ T cells in vitro, in contrast to the CD4+ CD25− cells, which showed no regulatory properties.

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“…Following the generation of novel mAbs against equine IL-10, Wagner and colleagues defined a population of CD4 + IL-10 + IFN-γ + cells that were thought to represent Tr1 cells [372]. Further indirect evidence for the role of immunosuppressive cytokines was generated by cultures of peripheral blood mononuclear cells derived from Icelandic horses, which suggested that heavy helminth burdens imposed a suppressive effect on IL-4 production by inducing IL-10 and TGF-β synthesis [373].…”

Section: Regulatory T Cells In Other Domestic Animal Species -An Expamentioning

confidence: 99%

All creatures great and small: regulatory T cells in mice, humans, dogs and other domestic animal species

Garden

Pinheiro

Cunningham

2011

International Immunopharmacology

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Characterization of monoclonal antibodies to equine interleukin-10 and detection of T regulatory 1 cells in horses

Cited by 53 publications

References 19 publications

Interferon-gamma, interleukin-4 and interleukin-10 production by T helper cells reveals intact Th1 and regulatory T_R1 cell activation and a delay of the Th2 cell response in equine neonates and foals

Interferon-gamma, interleukin-4 and interleukin-10 production by T helper cells reveals intact Th1 and regulatory T_R1 cell activation and a delay of the Th2 cell response in equine neonates and foals

Phenotypic and functional characterization of a CD4+ CD25high FOXP3high regulatory T-cell population in the dog

All creatures great and small: regulatory T cells in mice, humans, dogs and other domestic animal species

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Characterization of monoclonal antibodies to equine interleukin-10 and detection of T regulatory 1 cells in horses

Cited by 53 publications

References 19 publications

Interferon-gamma, interleukin-4 and interleukin-10 production by T helper cells reveals intact Th1 and regulatory TR1 cell activation and a delay of the Th2 cell response in equine neonates and foals

Interferon-gamma, interleukin-4 and interleukin-10 production by T helper cells reveals intact Th1 and regulatory TR1 cell activation and a delay of the Th2 cell response in equine neonates and foals

Phenotypic and functional characterization of a CD4+ CD25high FOXP3high regulatory T-cell population in the dog

All creatures great and small: regulatory T cells in mice, humans, dogs and other domestic animal species

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Interferon-gamma, interleukin-4 and interleukin-10 production by T helper cells reveals intact Th1 and regulatory T_R1 cell activation and a delay of the Th2 cell response in equine neonates and foals

Interferon-gamma, interleukin-4 and interleukin-10 production by T helper cells reveals intact Th1 and regulatory T_R1 cell activation and a delay of the Th2 cell response in equine neonates and foals