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Characterization of immune cells in psoriatic adipose tissue.

Rose S, Stansky E, Dagur PK, Samsel L, Weiner E, Jahanshad A, Doveikis J, Naik HB, Playford MP, McCoy JP, Mehta NN - J Transl Med (2014)

Bottom Line: Rarely have these populations been verified with confirmatory methodologies or functional studies.Further, both CD16+CD56(Lo) and CD16-CD56(Hi) NK cells were found to correlate inversely with body mass index.The relationship between the predominant CD16+CD56(Lo) NK cell population and body mass index persisted after adjusting for age, sex, diabetes, and tobacco use.

View Article: PubMed Central - PubMed

ABSTRACT

Background: Adipose tissue normally contains immune cells that regulate adipocyte function and contribute to metabolic disorders including obesity and diabetes mellitus. Psoriasis is associated with increased risk for metabolic disease, which may in part be due to adipose dysfunction, which has not been investigated in psoriasis. There is currently no standardized method for immunophenotyping human adipose tissue. In prior studies, characteristic phenotypic markers of immune cell populations identified in animal models or in other human tissues have been applied in a similar manner to human adipose tissue. Rarely have these populations been verified with confirmatory methodologies or functional studies. Thus, we performed a comprehensive phenotypic and functional analysis of immune cell populations in psoriatic adipose tissue.

Methods: Conventional and imaging flow cytometry were used to define immune cell populations in biopsy specimens of psoriatic adipose tissue (n = 30) including T cells, B cells, NK cells, NKT cells, neutrophils, and macrophages. Relationships between adipose immune cell types and body mass index were determined using Spearman regression analysis, and multivariate linear regression analysis was performed to adjust for cardiometabolic disease risk factors.

Results: These analyses revealed a wide range of cell surface receptors on adipose tissue macrophages, which may serve a dual purpose in immunity and metabolism. Further, both CD16+CD56(Lo) and CD16-CD56(Hi) NK cells were found to correlate inversely with body mass index. The relationship between the predominant CD16+CD56(Lo) NK cell population and body mass index persisted after adjusting for age, sex, diabetes, and tobacco use.

Conclusions: Together, these studies enhance our understanding of adipose immune cell phenotype and function, and demonstrate that examination of adipose tissue may provide greater insight into cardiometabolic pathophysiology in psoriasis.

No MeSH data available.


Related in: MedlinePlus

Flow cytometric characterization of B cells, T cells, NK cells, NKT cells, and neutrophils in a representative sample of psoriatic adipose tissue. After exclusion of debris, doublets, and non-viable cells, CD16-CD19+ B cells, CD3+CD56- T cells, CD3-CD16-CD56Hi NK cells, CD3+CD56+ NKT cells, and total CD16+ cells were identified. CD16+ cells were divided into neutrophils (CD16+SSCHi) and NK cells (CD16+CD56LoSSCLo) based on SSC properties. CD3+ T cells were further gated into naïve and effector and/or memory (eff/mem) αβ T cell subsets, CD4+CD8- T cells, CD4-CD8+ T cells, γδ T cells, and T regulatory (FoxP3+) cells. All cell populations are presented as percentages of viable cells except for granzyme B, IFN-γ, and TNF-α expressing cells, which are reported as percentages of the parent population. Positive gating for each fluorochrome parameter was established using FMO controls.
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Fig2: Flow cytometric characterization of B cells, T cells, NK cells, NKT cells, and neutrophils in a representative sample of psoriatic adipose tissue. After exclusion of debris, doublets, and non-viable cells, CD16-CD19+ B cells, CD3+CD56- T cells, CD3-CD16-CD56Hi NK cells, CD3+CD56+ NKT cells, and total CD16+ cells were identified. CD16+ cells were divided into neutrophils (CD16+SSCHi) and NK cells (CD16+CD56LoSSCLo) based on SSC properties. CD3+ T cells were further gated into naïve and effector and/or memory (eff/mem) αβ T cell subsets, CD4+CD8- T cells, CD4-CD8+ T cells, γδ T cells, and T regulatory (FoxP3+) cells. All cell populations are presented as percentages of viable cells except for granzyme B, IFN-γ, and TNF-α expressing cells, which are reported as percentages of the parent population. Positive gating for each fluorochrome parameter was established using FMO controls.

Mentions: Multi-parameter flow cytometry was applied to psoriatic adipose tissue, and gating strategies for cell identification (Figures 1, 2) and immune cell frequencies (Additional file 2: Table S1) are presented. Together, ATM (CD3-CD14+CD15-CD16-CD19-CD56-, Figure 1) were the most numerous immune cell type, comprising ~10% of viable cells. ATM made the prototypical cytokines IL-1β and IL-8 (Figure 1). Three populations of CD14+ ATM were identified based on HLADRII and CD206 expression (Figure 1). The HLADRII+CD206- subset was more abundant than the HLADRII-CD206- and HLADRII+CD206+ subsets. Frequencies of IL-1β and IL-8 expressing cells were not statistically different among the ATM subsets, except that the percentage of HLADRII+CD206-IL-8+ cells was significantly greater (p < 0.05) than HLADRII-CD206-IL-8+ cells (Additional file 3: Table S2). T cells (CD3+CD14-CD15-CD16-CD19-CD56-, Figure 2) collectively made up ~4.5% of viable cells, and were primarily αβ T cells with a predominance of CD4+ T cells. αβ T cells were largely either effector or memory cells (CD45RA-, Figure 2) and made the T cell cytokines IFN-γ and TNF-α (Figure 2). CD8+ T cells, FoxP3+ Tregs (predominately CD4+, Additional file 4: Figure S1), γδ T cells, and NKT cells were also identifiable (Figure 2). Confirmation of NKT cell (CD3+CD14-CD15-CD16-CD19-CD56+SSClo, Figure 2) phenotype was attempted using alpha-gal-cer loaded CD1d tetramers, but the scarcity of this population precluded definitive characterization. NK cells (CD3-CD14-CD15-CD16+/−CD19-CD56Hi/LoSSClo, Figures 1, 2) comprised ~1.5% of viable cells, were largely CD16+CD56Lo versus CD16-CD56Hi, and expressed the characteristic protease granzyme B (Figure 2). B cells (CD3-CD14-CD15-CD16-CD19+CD56-PD-1+, Figure 2, Additional file 5: Figure S2) made up <1% of viable cells. Neutrophils (CD3-CD14-CD15+CD16 + CD19-CD56-SSCHi, Figures 1, 2) were also infrequent and expressed the characteristic marker granzyme B (Figure 2).Figure 1


Characterization of immune cells in psoriatic adipose tissue.

Rose S, Stansky E, Dagur PK, Samsel L, Weiner E, Jahanshad A, Doveikis J, Naik HB, Playford MP, McCoy JP, Mehta NN - J Transl Med (2014)

Flow cytometric characterization of B cells, T cells, NK cells, NKT cells, and neutrophils in a representative sample of psoriatic adipose tissue. After exclusion of debris, doublets, and non-viable cells, CD16-CD19+ B cells, CD3+CD56- T cells, CD3-CD16-CD56Hi NK cells, CD3+CD56+ NKT cells, and total CD16+ cells were identified. CD16+ cells were divided into neutrophils (CD16+SSCHi) and NK cells (CD16+CD56LoSSCLo) based on SSC properties. CD3+ T cells were further gated into naïve and effector and/or memory (eff/mem) αβ T cell subsets, CD4+CD8- T cells, CD4-CD8+ T cells, γδ T cells, and T regulatory (FoxP3+) cells. All cell populations are presented as percentages of viable cells except for granzyme B, IFN-γ, and TNF-α expressing cells, which are reported as percentages of the parent population. Positive gating for each fluorochrome parameter was established using FMO controls.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4197293&req=5

Fig2: Flow cytometric characterization of B cells, T cells, NK cells, NKT cells, and neutrophils in a representative sample of psoriatic adipose tissue. After exclusion of debris, doublets, and non-viable cells, CD16-CD19+ B cells, CD3+CD56- T cells, CD3-CD16-CD56Hi NK cells, CD3+CD56+ NKT cells, and total CD16+ cells were identified. CD16+ cells were divided into neutrophils (CD16+SSCHi) and NK cells (CD16+CD56LoSSCLo) based on SSC properties. CD3+ T cells were further gated into naïve and effector and/or memory (eff/mem) αβ T cell subsets, CD4+CD8- T cells, CD4-CD8+ T cells, γδ T cells, and T regulatory (FoxP3+) cells. All cell populations are presented as percentages of viable cells except for granzyme B, IFN-γ, and TNF-α expressing cells, which are reported as percentages of the parent population. Positive gating for each fluorochrome parameter was established using FMO controls.
Mentions: Multi-parameter flow cytometry was applied to psoriatic adipose tissue, and gating strategies for cell identification (Figures 1, 2) and immune cell frequencies (Additional file 2: Table S1) are presented. Together, ATM (CD3-CD14+CD15-CD16-CD19-CD56-, Figure 1) were the most numerous immune cell type, comprising ~10% of viable cells. ATM made the prototypical cytokines IL-1β and IL-8 (Figure 1). Three populations of CD14+ ATM were identified based on HLADRII and CD206 expression (Figure 1). The HLADRII+CD206- subset was more abundant than the HLADRII-CD206- and HLADRII+CD206+ subsets. Frequencies of IL-1β and IL-8 expressing cells were not statistically different among the ATM subsets, except that the percentage of HLADRII+CD206-IL-8+ cells was significantly greater (p < 0.05) than HLADRII-CD206-IL-8+ cells (Additional file 3: Table S2). T cells (CD3+CD14-CD15-CD16-CD19-CD56-, Figure 2) collectively made up ~4.5% of viable cells, and were primarily αβ T cells with a predominance of CD4+ T cells. αβ T cells were largely either effector or memory cells (CD45RA-, Figure 2) and made the T cell cytokines IFN-γ and TNF-α (Figure 2). CD8+ T cells, FoxP3+ Tregs (predominately CD4+, Additional file 4: Figure S1), γδ T cells, and NKT cells were also identifiable (Figure 2). Confirmation of NKT cell (CD3+CD14-CD15-CD16-CD19-CD56+SSClo, Figure 2) phenotype was attempted using alpha-gal-cer loaded CD1d tetramers, but the scarcity of this population precluded definitive characterization. NK cells (CD3-CD14-CD15-CD16+/−CD19-CD56Hi/LoSSClo, Figures 1, 2) comprised ~1.5% of viable cells, were largely CD16+CD56Lo versus CD16-CD56Hi, and expressed the characteristic protease granzyme B (Figure 2). B cells (CD3-CD14-CD15-CD16-CD19+CD56-PD-1+, Figure 2, Additional file 5: Figure S2) made up <1% of viable cells. Neutrophils (CD3-CD14-CD15+CD16 + CD19-CD56-SSCHi, Figures 1, 2) were also infrequent and expressed the characteristic marker granzyme B (Figure 2).Figure 1

Bottom Line: Rarely have these populations been verified with confirmatory methodologies or functional studies.Further, both CD16+CD56(Lo) and CD16-CD56(Hi) NK cells were found to correlate inversely with body mass index.The relationship between the predominant CD16+CD56(Lo) NK cell population and body mass index persisted after adjusting for age, sex, diabetes, and tobacco use.

View Article: PubMed Central - PubMed

ABSTRACT

Background: Adipose tissue normally contains immune cells that regulate adipocyte function and contribute to metabolic disorders including obesity and diabetes mellitus. Psoriasis is associated with increased risk for metabolic disease, which may in part be due to adipose dysfunction, which has not been investigated in psoriasis. There is currently no standardized method for immunophenotyping human adipose tissue. In prior studies, characteristic phenotypic markers of immune cell populations identified in animal models or in other human tissues have been applied in a similar manner to human adipose tissue. Rarely have these populations been verified with confirmatory methodologies or functional studies. Thus, we performed a comprehensive phenotypic and functional analysis of immune cell populations in psoriatic adipose tissue.

Methods: Conventional and imaging flow cytometry were used to define immune cell populations in biopsy specimens of psoriatic adipose tissue (n = 30) including T cells, B cells, NK cells, NKT cells, neutrophils, and macrophages. Relationships between adipose immune cell types and body mass index were determined using Spearman regression analysis, and multivariate linear regression analysis was performed to adjust for cardiometabolic disease risk factors.

Results: These analyses revealed a wide range of cell surface receptors on adipose tissue macrophages, which may serve a dual purpose in immunity and metabolism. Further, both CD16+CD56(Lo) and CD16-CD56(Hi) NK cells were found to correlate inversely with body mass index. The relationship between the predominant CD16+CD56(Lo) NK cell population and body mass index persisted after adjusting for age, sex, diabetes, and tobacco use.

Conclusions: Together, these studies enhance our understanding of adipose immune cell phenotype and function, and demonstrate that examination of adipose tissue may provide greater insight into cardiometabolic pathophysiology in psoriasis.

No MeSH data available.


Related in: MedlinePlus