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Nanoscale Relationship Between CD4 and CD25 of T Cells Visualized with NSOM/QD-Based Dual-Color Imaging System.

Fan J, Lu X, Liu S, Zhong L - Nanoscale Res Lett (2015)

Bottom Line: This result presented that the formation of these CD25 nanodomains on the surface of CD4(+)CD25(high) T cells were not associated with the response of T cell receptor (TCR)/CD3-dependent signal transduction.Collectively, above these results exhibited that TCR/CD3-based microdomains were indeed required for TCR/CD3-mediated T cells activation and enhanced the immune activity of CD4(+)CD25(low) T cells or CD8(+)CD25(+) T cells.In particular, it was found that the formation of CD25 nanodomains and their segregation from TCR/CD3 microdomains were the intrinsic capability of CD4(+)CD25(high) T cells, suggesting this specific imaging feature of CD25 should be greatly associated with the regulatory activity of CD4(+)CD25(high) T cells.

View Article: PubMed Central - PubMed

Affiliation: Guangdong Provincial Laboratory of Nanophotonic Functional Materials and Devices, South China Normal University, Guangzhou, 510006, Guangdong, China.

ABSTRACT
In this study, by using of near-field scanning optical microscopy (NSOM)/immune-labeling quantum dot (QD)-based dual-color imaging system, we achieved the direct visualization of nanoscale profiles for distribution and organization of CD4 and CD25 molecules in T cells. A novel and interesting finding was that though CD25 clustering as nanodomains were observed on the surface of CD4(+)CD25(high) regulatory T cells, these CD25 nanodomains were not co-localized with CD4 nanodomains. This result presented that the formation of these CD25 nanodomains on the surface of CD4(+)CD25(high) T cells were not associated with the response of T cell receptor (TCR)/CD3-dependent signal transduction. In contrast, on the surface of CD4(+)CD25(low) T cells, CD25 molecules distributed randomly without forming nanodomains while CD4 clustering as nanodomains can be observed; on the surface of CD8(+)CD25(+) T cells, CD25 clustering as nanodomains and co-localization with CD8 nanodomains were observed. Collectively, above these results exhibited that TCR/CD3-based microdomains were indeed required for TCR/CD3-mediated T cells activation and enhanced the immune activity of CD4(+)CD25(low) T cells or CD8(+)CD25(+) T cells. In particular, it was found that the formation of CD25 nanodomains and their segregation from TCR/CD3 microdomains were the intrinsic capability of CD4(+)CD25(high) T cells, suggesting this specific imaging feature of CD25 should be greatly associated with the regulatory activity of CD4(+)CD25(high) T cells. Importantly, this novel NSOM/QD-based dual-color imaging system will provide a useful tool for the research of distribution-function relationship of cell-surface molecules.

No MeSH data available.


Related in: MedlinePlus

Simultaneous nanoscale dual-color imaging of CD4 and CD25 on the surface of CD8+CD25+ T cells though using NSOM/QD system. a T cell topography. b Fluorescence image of CD8 labeled with QD-655 (red). c Fluorescence image of CD25 labeled with QD-605 (blue). d Merge of CD8 and CD25 two color fluorescence images. e Merge of cell topography and two color fluorescence images. f–h Zoom images of the areas as indicated by the squares on (b–d), respectively. i The percentage numbers of CD8 or CD25 molecules arrayed to form nanodomains. j Molecule density of CD8 or CD25 nanodomains. Data were expressed as mean ± SEM in (i, j), *P < 0.02, **P < 0.01 (i) and *P < 0.02, **P < 0.005 (j) compared with control
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Fig5: Simultaneous nanoscale dual-color imaging of CD4 and CD25 on the surface of CD8+CD25+ T cells though using NSOM/QD system. a T cell topography. b Fluorescence image of CD8 labeled with QD-655 (red). c Fluorescence image of CD25 labeled with QD-605 (blue). d Merge of CD8 and CD25 two color fluorescence images. e Merge of cell topography and two color fluorescence images. f–h Zoom images of the areas as indicated by the squares on (b–d), respectively. i The percentage numbers of CD8 or CD25 molecules arrayed to form nanodomains. j Molecule density of CD8 or CD25 nanodomains. Data were expressed as mean ± SEM in (i, j), *P < 0.02, **P < 0.01 (i) and *P < 0.02, **P < 0.005 (j) compared with control

Mentions: Interestingly, after anti-CD3/anti-CD28 Abs co-stimulation of 48 h, both CD8 clustering and CD25 clustering as nanodomains of 200–300 nm can be observed on the surface of CD8+CD25+ T cells through using NSOM-QD-based dual-color system (Fig. 5a–c), moreover, all of CD25 nanodomains were co-localized with CD8 nanodomains. The statistical analysis showed that though the average molecule density of CD25 in nanodomains (434 ± 29/μm2) was nearly the same with CD8 in nanodomains (495 ± 41/μm2) (Fig. 5j), but the percentage of CD25 molecules that arrayed to form nanodomains (57.2 ± 4.1 %) was about two times bigger than that of CD8 molecules (27.6 ± 2.7 %) (Fig. 4i). Accordingly, as shown in Fig. 3a, b, the concentration of IL-2 production of CD8+CD25+ T cells (3579.2 ± 423.5 ng/L) was nearly 75 % higher than that of CD4+CD25high T cells (2054.8 ± 264.7 ng/L) while the concentration of IL-10 production of CD8+CD25+ T cells (228.6 ± 27.3 ng/L) was about 75 % lower than that of CD4+CD25high T cells (918.4 ± 86.7 ng/L). These results possibly exhibited that the formation of CD25 nanodomains and their co-localization with CD8 nanodomains were associated with TCR/CD3-mediated signaling, and this nanoscale imaging feature would play an important role to enhance the immune activity CD8+CD25+ T cells.Fig. 5


Nanoscale Relationship Between CD4 and CD25 of T Cells Visualized with NSOM/QD-Based Dual-Color Imaging System.

Fan J, Lu X, Liu S, Zhong L - Nanoscale Res Lett (2015)

Simultaneous nanoscale dual-color imaging of CD4 and CD25 on the surface of CD8+CD25+ T cells though using NSOM/QD system. a T cell topography. b Fluorescence image of CD8 labeled with QD-655 (red). c Fluorescence image of CD25 labeled with QD-605 (blue). d Merge of CD8 and CD25 two color fluorescence images. e Merge of cell topography and two color fluorescence images. f–h Zoom images of the areas as indicated by the squares on (b–d), respectively. i The percentage numbers of CD8 or CD25 molecules arrayed to form nanodomains. j Molecule density of CD8 or CD25 nanodomains. Data were expressed as mean ± SEM in (i, j), *P < 0.02, **P < 0.01 (i) and *P < 0.02, **P < 0.005 (j) compared with control
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Fig5: Simultaneous nanoscale dual-color imaging of CD4 and CD25 on the surface of CD8+CD25+ T cells though using NSOM/QD system. a T cell topography. b Fluorescence image of CD8 labeled with QD-655 (red). c Fluorescence image of CD25 labeled with QD-605 (blue). d Merge of CD8 and CD25 two color fluorescence images. e Merge of cell topography and two color fluorescence images. f–h Zoom images of the areas as indicated by the squares on (b–d), respectively. i The percentage numbers of CD8 or CD25 molecules arrayed to form nanodomains. j Molecule density of CD8 or CD25 nanodomains. Data were expressed as mean ± SEM in (i, j), *P < 0.02, **P < 0.01 (i) and *P < 0.02, **P < 0.005 (j) compared with control
Mentions: Interestingly, after anti-CD3/anti-CD28 Abs co-stimulation of 48 h, both CD8 clustering and CD25 clustering as nanodomains of 200–300 nm can be observed on the surface of CD8+CD25+ T cells through using NSOM-QD-based dual-color system (Fig. 5a–c), moreover, all of CD25 nanodomains were co-localized with CD8 nanodomains. The statistical analysis showed that though the average molecule density of CD25 in nanodomains (434 ± 29/μm2) was nearly the same with CD8 in nanodomains (495 ± 41/μm2) (Fig. 5j), but the percentage of CD25 molecules that arrayed to form nanodomains (57.2 ± 4.1 %) was about two times bigger than that of CD8 molecules (27.6 ± 2.7 %) (Fig. 4i). Accordingly, as shown in Fig. 3a, b, the concentration of IL-2 production of CD8+CD25+ T cells (3579.2 ± 423.5 ng/L) was nearly 75 % higher than that of CD4+CD25high T cells (2054.8 ± 264.7 ng/L) while the concentration of IL-10 production of CD8+CD25+ T cells (228.6 ± 27.3 ng/L) was about 75 % lower than that of CD4+CD25high T cells (918.4 ± 86.7 ng/L). These results possibly exhibited that the formation of CD25 nanodomains and their co-localization with CD8 nanodomains were associated with TCR/CD3-mediated signaling, and this nanoscale imaging feature would play an important role to enhance the immune activity CD8+CD25+ T cells.Fig. 5

Bottom Line: This result presented that the formation of these CD25 nanodomains on the surface of CD4(+)CD25(high) T cells were not associated with the response of T cell receptor (TCR)/CD3-dependent signal transduction.Collectively, above these results exhibited that TCR/CD3-based microdomains were indeed required for TCR/CD3-mediated T cells activation and enhanced the immune activity of CD4(+)CD25(low) T cells or CD8(+)CD25(+) T cells.In particular, it was found that the formation of CD25 nanodomains and their segregation from TCR/CD3 microdomains were the intrinsic capability of CD4(+)CD25(high) T cells, suggesting this specific imaging feature of CD25 should be greatly associated with the regulatory activity of CD4(+)CD25(high) T cells.

View Article: PubMed Central - PubMed

Affiliation: Guangdong Provincial Laboratory of Nanophotonic Functional Materials and Devices, South China Normal University, Guangzhou, 510006, Guangdong, China.

ABSTRACT
In this study, by using of near-field scanning optical microscopy (NSOM)/immune-labeling quantum dot (QD)-based dual-color imaging system, we achieved the direct visualization of nanoscale profiles for distribution and organization of CD4 and CD25 molecules in T cells. A novel and interesting finding was that though CD25 clustering as nanodomains were observed on the surface of CD4(+)CD25(high) regulatory T cells, these CD25 nanodomains were not co-localized with CD4 nanodomains. This result presented that the formation of these CD25 nanodomains on the surface of CD4(+)CD25(high) T cells were not associated with the response of T cell receptor (TCR)/CD3-dependent signal transduction. In contrast, on the surface of CD4(+)CD25(low) T cells, CD25 molecules distributed randomly without forming nanodomains while CD4 clustering as nanodomains can be observed; on the surface of CD8(+)CD25(+) T cells, CD25 clustering as nanodomains and co-localization with CD8 nanodomains were observed. Collectively, above these results exhibited that TCR/CD3-based microdomains were indeed required for TCR/CD3-mediated T cells activation and enhanced the immune activity of CD4(+)CD25(low) T cells or CD8(+)CD25(+) T cells. In particular, it was found that the formation of CD25 nanodomains and their segregation from TCR/CD3 microdomains were the intrinsic capability of CD4(+)CD25(high) T cells, suggesting this specific imaging feature of CD25 should be greatly associated with the regulatory activity of CD4(+)CD25(high) T cells. Importantly, this novel NSOM/QD-based dual-color imaging system will provide a useful tool for the research of distribution-function relationship of cell-surface molecules.

No MeSH data available.


Related in: MedlinePlus