Movatterモバイル変換

[0]ホーム
URL:

Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
Thehttps:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

NIH NLM Logo
Log inShow account info
Access keysNCBI HomepageMyNCBI HomepageMain ContentMain Navigation
pubmed logo
Advanced Clipboard
User Guide

Full text links

Public Library of Science full text link Public Library of Science Free PMC article
Full text links

Actions

Share

doi: 10.1371/journal.pone.0047155. Epub 2012 Oct 16.

High-throughput 13-parameter immunophenotyping identifies shifts in the circulating T-cell compartment following reperfusion in patients with acute myocardial infarction

Affiliations

High-throughput 13-parameter immunophenotyping identifies shifts in the circulating T-cell compartment following reperfusion in patients with acute myocardial infarction

Jedrzej Hoffmann et al. PLoS One.2012.

Abstract

Rationale: With the advent of primary PCI (PPCI), reperfusion is achieved in almost all patients presenting with acute myocardial infarction. However, despite multiple trials, reperfusion injury has not been successfully dealt with so far. In mouse models, CD4(+) T lymphocytes (T cells) have been shown to be crucial instigators of reperfusion injury.

Objective: Our goal was to investigate the role of CD4(+) T cells during myocardial reperfusion following PPCI by developing a protocol for high-throughput multiplexed flow cytometric analysis and multivariate flow clustering.

Methods and results: 13-parameter immunophenotyping and hierarchical cluster analysis (HCA) identified a unique CD4(+)CD57(+) T-cell population in PPCI patients that reflected acute proliferation in the CD4(+) T-cell compartment. CD4(+)CCR7(+) T cells were specifically depleted from peripheral blood during the first 30 min of myocardial reperfusion after PPCI, suggesting a potential role for the chemokine receptor CCR7 in T-cell redistribution to either peripheral tissues or migration to the infarcted heart during ischemia/reperfusion following PPCI.

Conclusions: High-throughput polychromatic flow cytometry and HCA are capable of objective, time and cost efficient assessment of the individual T-cell immune profile in different stages of coronary heart disease and have broad applications in clinical trials.

PubMed Disclaimer

Conflict of interest statement

Competing Interests:The authors have declared that no competing interests exist.

Figures

Figure 1
Figure 1. Peripheral leukocytosis after primary percutaneus coronary intervention (PPCI).
Absolute number of total leukocytes, neutrophil granulocytes and monocytes (A), as well as lymphocytes and the main T-cell subsets (B) in peripheral blood. Concentration of leukocyte populations are displayed for patients 24 h following PPCI in STEMI (n = 31), 3 months post PPCI (n = 24), and in age-matched healthy Controls (n = 18). 1-way ANOVA with Tukeys' post-hoc test was performed. *p<0.05, ***p<0.001, ns not significant.
Figure 2
Figure 2. Basic gating strategy for CD4+ T cells.
Viable cells, as distiguished on the basis of Aqua dye staining were gated and CD4+ T cells were selected based on their light scatter properties and CD3 and CD4 co-expression. CD4+ T-cell subpopulations were separated into main subsets based on the CD45RA/CCR7 expression properties, as previously described by Sallusto et al.: NAIVE (CD45RA+CCR7+), CENTRAL MEMORY (CD45RA−CCR7+), EFFECTOR MEMORY (CD45RA−CCR7−) and CD45RA reexpressing „revertant” memory CD4+ T cells (T effector memory CD45RA+ cells, TEMRA). Further sequential gating analysis were perfomed based on the coexpression of co-stimulatory markers CD27/CD28 as well as proliferation/senescence markers CD57, KLRG1, and PD1.
Figure 3
Figure 3. Distribution of main CD4 T-cell subsets.
Frequencies of CD4+ naive (CD45RA+CCR7+), central memory (CD45RA−CCR7+), effector memory (CD45RA−CCR7−) and terminally differentiated effector memory TEMRA cells (CD45RA+CCR7−) are shown. Groups are identical to Figure 1. 1-way ANOVA with Tukeys' post-hoc test was performed. *p<0.05, ***p<0.001, ns not significant.
Figure 4
Figure 4. Hierarchical clustering analysis of 13-parameter flow cytometry data.
Dendrogram with heatmap-HCA of 104 events (gated on viable CD3+ T cells) acquired from PBMCs of healthy control and STEMI patient (24 h after PPCI). Heatmap shows relative levels of all 13 parameters (columns) in all 104 events (rows) in color coding (blue, low expression; red, high expression). Dendrogram shows the hierarchy of T cells based on their similarity in all parameters measured. Colored branches of the dendrogram are selected clusters (A). The T-cell subsets from healthy controls (as identified by HCA) are displayed on conventional scatter dot plots (B). HCA revealed that the proliferation marker CD57 is strongly expressed on CD4+ CM T cells in STEMI patients (zoom on cluster of interest in C).
Figure 5
Figure 5. Increased expression of CD57 in central memory cells indicates acute proliferation in the CD4 T-cell compartment.
Conventional gating analysis confirmed increased expression of proliferation associated markers CD57 on CM CD4+ T cells of STEMI pts (MFI – mean fluorescence intensity). This was reflected by significantly increased frequencies of specific KLRG1−CD57+ CD4 CM+ T cells (A–C). The validity of CD57, PD-1 and KLRG1 as proliferation-associated markers was determined in healthy controls according to Sallusto's model of T-cell differentiation (D). Mean telomere length (mTL) in CD4+ T cells is reduced in patients with STEMI when compared to age-matched controls (E). *p<0.05, **p<0.01, ***p<0.001, ns not significant.
Figure 6
Figure 6. CD4+CCR7+ T cells are specifically depleted from peripheral blood during myocardial reperfusion.
Time course pilot study showed no significant changes in numbers of monocytes (A) and granulocytes (B) following PPCI. In contrast, T cells decreased by 35% during the first 30 min following reperfusion showing significant relative increase within next 24 hours (C). The absolute cell count analysis revealed a significant decrease in CD4+ T cells during the first 60 min following PPCI (D). This was mainly due to the specific depletion of circulating CD4+CCR7+ cells within the first 30 min following reperfusion (E). *p<0.05, **p<0.01, ns not significant.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Wu KC, Zerhouni EA, Judd RM, Lugo-Olivieri CH, Barouch LA, et al. (1998) Prognostic significance of microvascular obstruction by magnetic resonance imaging in patients with acute myocardial infarction. Circulation 97: 765–772. - PubMed
    1. van 't Hof AW, Liem A, Suryapranata H, Hoorntje JC, de Boer MJ, et al. (1998) Angiographic assessment of myocardial reperfusion in patients treated with primary angioplasty for acute myocardial infarction: myocardial blush grade. Zwolle Myocardial Infarction Study Group. Circulation 97: 2302–2306. - PubMed
    1. Claeys MJ, Bosmans J, Veenstra L, Jorens P, De R, et al. (1999) Determinants and prognostic implications of persistent ST-segment elevation after primary angioplasty for acute myocardial infarction: importance of microvascular reperfusion injury on clinical outcome. Circulation 99: 1972–1977. - PubMed
    1. Angeja BG, Gunda M, Murphy SA, Sobel BE, Rundle AC, et al. (2002) TIMI myocardial perfusion grade and ST segment resolution: association with infarct size as assessed by single photon emission computed tomography imaging. Circulation 105: 282–285. - PubMed
    1. Yokota N, Burne-Taney M, Racusen L, Rabb H (2003) Contrasting roles for STAT4 and STAT6 signal transduction pathways in murine renal ischemia-reperfusion injury. Am J Physiol Renal Physiol 285: F319–25. - PubMed

Publication types

MeSH terms

Substances

Related information

Grants and funding

LinkOut - more resources

Full text links
Public Library of Science full text link Public Library of Science Free PMC article
Cite
Send To

NCBI Literature Resources

MeSHPMCBookshelfDisclaimer

The PubMed wordmark and PubMed logo are registered trademarks of the U.S. Department of Health and Human Services (HHS). Unauthorized use of these marks is strictly prohibited.

[8]ページ先頭
©2009-2025 Movatter.jp