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Phenotypic and functional separation of memory and effector human CD8+ T cells.

Hamann D, Baars PA, Rep MH, Hooibrink B, Kerkhof-Garde SR, Klein MR, van Lier RA - J. Exp. Med. (1997)

Bottom Line: These T cells do not secrete IL-2 or -4 but can produce IFN-gamma and TNF-alpha.Interestingly, CD8+CD45RA+CD27- cells parallel effector CTLs, as they abundantly express Fas-ligand mRNA, contain perforin and granzyme B, and have high cytolytic activity without in vitro prestimulation.Based on both phenotypic and functional properties, we conclude that memory- and effector-type T cells can be separated as distinct entities within the human CD8+ T cell subset.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Viro-Immunology, Central Laboratory of the Netherlands Red Cross Blood Transfusion Service, University of Amsterdam.

ABSTRACT
Human CD8+ memory- and effector-type T cells are poorly defined. We show here that, next to a naive compartment, two discrete primed subpopulations can be found within the circulating human CD8+ T cell subset. First, CD45RA-CD45R0(+) cells are reminiscent of memory-type T cells in that they express elevated levels of CD95 (Fas) and the integrin family members CD11a, CD18, CD29, CD49d, and CD49e, compared to naive CD8+ T cells, and are able to secrete not only interleukin (IL) 2 but also interferon gamma, tumor necrosis factor alpha, and IL-4. This subset does not exert cytolytic activity without prior in vitro stimulation but does contain virus-specific cytotoxic T lymphocyte (CTL) precursors. A second primed population is characterized by CD45RA expression with concomitant absence of expression of the costimulatory molecules CD27 and CD28. The CD8+CD45RA+CD27- population contains T cells expressing high levels of CD11a, CD11b, CD18, and CD49d, whereas CD62L (L-selectin) is not expressed. These T cells do not secrete IL-2 or -4 but can produce IFN-gamma and TNF-alpha. In accordance with this finding, cells contained within this subpopulation depend for proliferation on exogenous growth factors such as IL-2 and -15. Interestingly, CD8+CD45RA+CD27- cells parallel effector CTLs, as they abundantly express Fas-ligand mRNA, contain perforin and granzyme B, and have high cytolytic activity without in vitro prestimulation. Based on both phenotypic and functional properties, we conclude that memory- and effector-type T cells can be separated as distinct entities within the human CD8+ T cell subset.

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Expression of mediators of cytotoxicity in CD8+ T cell subsets. After surface staining of freshly isolated CD8+ cells with CD45RA  and CD27 mAbs, cells were fixated and permeabilized and intracellularly  present perforin and granzyme B were detected with specific mAbs. The  histograms show the staining for the intracellular proteins. Dotted lines  indicate the negative controls. The positive cell fraction is painted black.  The dot-plots show the distribution of positive cells (black) among the total cell population (gray). Note that the different dot-plot staining is due  to the use of either biotinylated CD27 mAb in combination with red 670  or FITC-labeled CD27 mAb.
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Figure 6: Expression of mediators of cytotoxicity in CD8+ T cell subsets. After surface staining of freshly isolated CD8+ cells with CD45RA and CD27 mAbs, cells were fixated and permeabilized and intracellularly present perforin and granzyme B were detected with specific mAbs. The histograms show the staining for the intracellular proteins. Dotted lines indicate the negative controls. The positive cell fraction is painted black. The dot-plots show the distribution of positive cells (black) among the total cell population (gray). Note that the different dot-plot staining is due to the use of either biotinylated CD27 mAb in combination with red 670 or FITC-labeled CD27 mAb.

Mentions: Fas–Fas ligand interaction is an important mechanism used by CTLs for cell-mediated cytotoxicity (39, 40). Next to this, CTL effectors can kill target cells by releasing a pore-forming protein (perforin) and serine proteases (granzymes), which are stored in intracellular granules, into the vicinity of target cell membranes (41, 42). We analyzed freshly isolated CD8+ cells for the presence of perforin and granzyme B. In accordance with the cytolytic capacities of the different subsets, nearly all cells of the CD45RA+CD27− population contained perforin and granzyme B. Unprimed CD45RA+CD27+ cells did not contain these enzymes, whereas low staining could be found within the CD45RA− population (Fig. 6 and Table 4). As already seen in the analysis of cytokine production, cells with dull CD27 expression showed a similar granzyme B and perforin expression pattern to CD27− cells.


Phenotypic and functional separation of memory and effector human CD8+ T cells.

Hamann D, Baars PA, Rep MH, Hooibrink B, Kerkhof-Garde SR, Klein MR, van Lier RA - J. Exp. Med. (1997)

Expression of mediators of cytotoxicity in CD8+ T cell subsets. After surface staining of freshly isolated CD8+ cells with CD45RA  and CD27 mAbs, cells were fixated and permeabilized and intracellularly  present perforin and granzyme B were detected with specific mAbs. The  histograms show the staining for the intracellular proteins. Dotted lines  indicate the negative controls. The positive cell fraction is painted black.  The dot-plots show the distribution of positive cells (black) among the total cell population (gray). Note that the different dot-plot staining is due  to the use of either biotinylated CD27 mAb in combination with red 670  or FITC-labeled CD27 mAb.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2199103&req=5

Figure 6: Expression of mediators of cytotoxicity in CD8+ T cell subsets. After surface staining of freshly isolated CD8+ cells with CD45RA and CD27 mAbs, cells were fixated and permeabilized and intracellularly present perforin and granzyme B were detected with specific mAbs. The histograms show the staining for the intracellular proteins. Dotted lines indicate the negative controls. The positive cell fraction is painted black. The dot-plots show the distribution of positive cells (black) among the total cell population (gray). Note that the different dot-plot staining is due to the use of either biotinylated CD27 mAb in combination with red 670 or FITC-labeled CD27 mAb.
Mentions: Fas–Fas ligand interaction is an important mechanism used by CTLs for cell-mediated cytotoxicity (39, 40). Next to this, CTL effectors can kill target cells by releasing a pore-forming protein (perforin) and serine proteases (granzymes), which are stored in intracellular granules, into the vicinity of target cell membranes (41, 42). We analyzed freshly isolated CD8+ cells for the presence of perforin and granzyme B. In accordance with the cytolytic capacities of the different subsets, nearly all cells of the CD45RA+CD27− population contained perforin and granzyme B. Unprimed CD45RA+CD27+ cells did not contain these enzymes, whereas low staining could be found within the CD45RA− population (Fig. 6 and Table 4). As already seen in the analysis of cytokine production, cells with dull CD27 expression showed a similar granzyme B and perforin expression pattern to CD27− cells.

Bottom Line: These T cells do not secrete IL-2 or -4 but can produce IFN-gamma and TNF-alpha.Interestingly, CD8+CD45RA+CD27- cells parallel effector CTLs, as they abundantly express Fas-ligand mRNA, contain perforin and granzyme B, and have high cytolytic activity without in vitro prestimulation.Based on both phenotypic and functional properties, we conclude that memory- and effector-type T cells can be separated as distinct entities within the human CD8+ T cell subset.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Viro-Immunology, Central Laboratory of the Netherlands Red Cross Blood Transfusion Service, University of Amsterdam.

ABSTRACT
Human CD8+ memory- and effector-type T cells are poorly defined. We show here that, next to a naive compartment, two discrete primed subpopulations can be found within the circulating human CD8+ T cell subset. First, CD45RA-CD45R0(+) cells are reminiscent of memory-type T cells in that they express elevated levels of CD95 (Fas) and the integrin family members CD11a, CD18, CD29, CD49d, and CD49e, compared to naive CD8+ T cells, and are able to secrete not only interleukin (IL) 2 but also interferon gamma, tumor necrosis factor alpha, and IL-4. This subset does not exert cytolytic activity without prior in vitro stimulation but does contain virus-specific cytotoxic T lymphocyte (CTL) precursors. A second primed population is characterized by CD45RA expression with concomitant absence of expression of the costimulatory molecules CD27 and CD28. The CD8+CD45RA+CD27- population contains T cells expressing high levels of CD11a, CD11b, CD18, and CD49d, whereas CD62L (L-selectin) is not expressed. These T cells do not secrete IL-2 or -4 but can produce IFN-gamma and TNF-alpha. In accordance with this finding, cells contained within this subpopulation depend for proliferation on exogenous growth factors such as IL-2 and -15. Interestingly, CD8+CD45RA+CD27- cells parallel effector CTLs, as they abundantly express Fas-ligand mRNA, contain perforin and granzyme B, and have high cytolytic activity without in vitro prestimulation. Based on both phenotypic and functional properties, we conclude that memory- and effector-type T cells can be separated as distinct entities within the human CD8+ T cell subset.

Show MeSH
Related in: MedlinePlus