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Characterisation and comparison of adipose tissue macrophages from human subcutaneous, visceral and perivascular adipose tissue.

Kralova Lesna I, Kralova A, Cejkova S, Fronek J, Petras M, Sekerkova A, Thieme F, Janousek L, Poledne R - J Transl Med (2016)

Bottom Line: Subcutaneous, visceral and perivascular adipose tissues were obtained from 52 living kidney donors during live donor nephrectomy.This difference was substantially more pronounced in the postmenopausal women subgroup, consequentlly, the total difference was driven by this subgroup.The visceral and perivascular adipose tissues had substantially higher pro-inflammatory characteristics than the subcutaneous tissue.

View Article: PubMed Central - PubMed

Affiliation: Laboratory for Atherosclerosis Research, Centre for Experimental Medicine, Institute for Clinical and Experimental Medicine, Prague, Czech Republic. ivka@ikem.cz.

ABSTRACT

Background and aims: Macrophages play important roles in adipose tissue inflammation and its consequences. Unfortunately, a detailed description of the macrophage phenotypes in different human adipose tissues is not available.

Subjects and methods: Subcutaneous, visceral and perivascular adipose tissues were obtained from 52 living kidney donors during live donor nephrectomy. Stromal vascular fractions were isolated, and the macrophage phenotypes were analyzed by flow cytometry using surface markers (CD14, CD16, CD36, and CD163).

Results: In addition to CD16 positivity, pro-inflammatory macrophages also display high scavenger receptor CD36 expression. The great majority of CD16 negative macrophages express the anti-inflammatory CD163 marker. The presence of pro-inflammatory macrophages was almost twice as high in visceral (p < 0.0001) and perivascular (p < 0.0001) adipose tissues than in subcutaneous tissue. This difference was substantially more pronounced in the postmenopausal women subgroup, consequentlly, the total difference was driven by this subgroup.

Conclusion: We obtained detailed information about M1 and M2 macrophage phenotypes in human adipose tissue. The visceral and perivascular adipose tissues had substantially higher pro-inflammatory characteristics than the subcutaneous tissue. The higher proportion of pro-inflammatory macrophages in the visceral adipose tissue of postmenopausal women might be related to an increased cardiovascular risk.

No MeSH data available.


Example of the SVF flow cytometric analysis. a CD16 positive monocytes were first identified and delineated in the blood sample (left, CD16positive macrophages in the upper part). The setting was fixed and subsequently used for the SVF analysis (b). Total macrophages in the SVF were identified by positivity for CD14 (c). Based on the CD16 marker, two subpopulations were distinguished (b, CD16-positive macrophages in the upper part). d The CD16+ subpopulation was divided according to CD36 marker markers (d, left) and then the CD163 presence was determined within both the CD36 positive subpopulations [i.e., CD16+ CD36high (d, middle) and CD16+ CD36low (d, right)]. Similarly, the CD16 negative subpopulation was divided according to the CD36 marker (e, left) and then the CD163 presence was determined in both the CD16− CD36low (e, middle) and CD16− CD36− subpopulations (e, right)
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Fig1: Example of the SVF flow cytometric analysis. a CD16 positive monocytes were first identified and delineated in the blood sample (left, CD16positive macrophages in the upper part). The setting was fixed and subsequently used for the SVF analysis (b). Total macrophages in the SVF were identified by positivity for CD14 (c). Based on the CD16 marker, two subpopulations were distinguished (b, CD16-positive macrophages in the upper part). d The CD16+ subpopulation was divided according to CD36 marker markers (d, left) and then the CD163 presence was determined within both the CD36 positive subpopulations [i.e., CD16+ CD36high (d, middle) and CD16+ CD36low (d, right)]. Similarly, the CD16 negative subpopulation was divided according to the CD36 marker (e, left) and then the CD163 presence was determined in both the CD16− CD36low (e, middle) and CD16− CD36− subpopulations (e, right)

Mentions: Due to difficulties in delineating CD16 positive cells in the SVF, the CD16 positive monocytes were first identified and delineated in a blood sample where the CD16 positive subpopulation was clearly visible. The setting was fixed and subsequently used for the SVF analysis. The gating strategy used to identify the SVF macrophage subpopulations is shown in Fig. 1.Fig. 1


Characterisation and comparison of adipose tissue macrophages from human subcutaneous, visceral and perivascular adipose tissue.

Kralova Lesna I, Kralova A, Cejkova S, Fronek J, Petras M, Sekerkova A, Thieme F, Janousek L, Poledne R - J Transl Med (2016)

Example of the SVF flow cytometric analysis. a CD16 positive monocytes were first identified and delineated in the blood sample (left, CD16positive macrophages in the upper part). The setting was fixed and subsequently used for the SVF analysis (b). Total macrophages in the SVF were identified by positivity for CD14 (c). Based on the CD16 marker, two subpopulations were distinguished (b, CD16-positive macrophages in the upper part). d The CD16+ subpopulation was divided according to CD36 marker markers (d, left) and then the CD163 presence was determined within both the CD36 positive subpopulations [i.e., CD16+ CD36high (d, middle) and CD16+ CD36low (d, right)]. Similarly, the CD16 negative subpopulation was divided according to the CD36 marker (e, left) and then the CD163 presence was determined in both the CD16− CD36low (e, middle) and CD16− CD36− subpopulations (e, right)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4940901&req=5

Fig1: Example of the SVF flow cytometric analysis. a CD16 positive monocytes were first identified and delineated in the blood sample (left, CD16positive macrophages in the upper part). The setting was fixed and subsequently used for the SVF analysis (b). Total macrophages in the SVF were identified by positivity for CD14 (c). Based on the CD16 marker, two subpopulations were distinguished (b, CD16-positive macrophages in the upper part). d The CD16+ subpopulation was divided according to CD36 marker markers (d, left) and then the CD163 presence was determined within both the CD36 positive subpopulations [i.e., CD16+ CD36high (d, middle) and CD16+ CD36low (d, right)]. Similarly, the CD16 negative subpopulation was divided according to the CD36 marker (e, left) and then the CD163 presence was determined in both the CD16− CD36low (e, middle) and CD16− CD36− subpopulations (e, right)
Mentions: Due to difficulties in delineating CD16 positive cells in the SVF, the CD16 positive monocytes were first identified and delineated in a blood sample where the CD16 positive subpopulation was clearly visible. The setting was fixed and subsequently used for the SVF analysis. The gating strategy used to identify the SVF macrophage subpopulations is shown in Fig. 1.Fig. 1

Bottom Line: Subcutaneous, visceral and perivascular adipose tissues were obtained from 52 living kidney donors during live donor nephrectomy.This difference was substantially more pronounced in the postmenopausal women subgroup, consequentlly, the total difference was driven by this subgroup.The visceral and perivascular adipose tissues had substantially higher pro-inflammatory characteristics than the subcutaneous tissue.

View Article: PubMed Central - PubMed

Affiliation: Laboratory for Atherosclerosis Research, Centre for Experimental Medicine, Institute for Clinical and Experimental Medicine, Prague, Czech Republic. ivka@ikem.cz.

ABSTRACT

Background and aims: Macrophages play important roles in adipose tissue inflammation and its consequences. Unfortunately, a detailed description of the macrophage phenotypes in different human adipose tissues is not available.

Subjects and methods: Subcutaneous, visceral and perivascular adipose tissues were obtained from 52 living kidney donors during live donor nephrectomy. Stromal vascular fractions were isolated, and the macrophage phenotypes were analyzed by flow cytometry using surface markers (CD14, CD16, CD36, and CD163).

Results: In addition to CD16 positivity, pro-inflammatory macrophages also display high scavenger receptor CD36 expression. The great majority of CD16 negative macrophages express the anti-inflammatory CD163 marker. The presence of pro-inflammatory macrophages was almost twice as high in visceral (p < 0.0001) and perivascular (p < 0.0001) adipose tissues than in subcutaneous tissue. This difference was substantially more pronounced in the postmenopausal women subgroup, consequentlly, the total difference was driven by this subgroup.

Conclusion: We obtained detailed information about M1 and M2 macrophage phenotypes in human adipose tissue. The visceral and perivascular adipose tissues had substantially higher pro-inflammatory characteristics than the subcutaneous tissue. The higher proportion of pro-inflammatory macrophages in the visceral adipose tissue of postmenopausal women might be related to an increased cardiovascular risk.

No MeSH data available.