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The Cellular Localization of Human Cytomegalovirus Glycoprotein Expression Greatly Influences the Frequency and Functional Phenotype of Specific CD4+ T Cell Responses.

Pachnio A, Zuo J, Ryan GB, Begum J, Moss PA - J. Immunol. (2015)

Bottom Line: In this study, we examine and contrast the magnitude and phenotype of the T cell immune response against gB, gH, and gL within healthy donors. gB-specific CD4(+) T cells were found in 95% of donors, and 29 epitopes were defined with gB-specific response sizes ranging from 0.02 to 2.88% of the CD4(+) T cell pool.Glycoproteins were effectively presented following delivery to APCs but only gB-derived epitopes were presented following endogenous synthesis. gB expression was observed exclusively within vesicular structures colocalizing with HLA-DM whereas gH was distributed evenly throughout the cytoplasm.These results reveal that gB is a uniquely immunogenic CMV glycoprotein and this is likely to reflect its unique pattern of endogenous Ag presentation.

View Article: PubMed Central - PubMed

Affiliation: School of Cancer Sciences, College of Medicine and Dentistry, University of Birmingham, Birmingham B15 2TT, United Kingdom; and a.pachnio@bham.ac.uk p.moss@bham.ac.uk.

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Glycoprotein-specific T cells vary in their functional capacities. Functional characterization of glycoprotein-specific T cells was carried out following peptide stimulation of PBMCs using multicolor flow cytometric analysis, and a Boolean gating strategy was applied. Bar charts represent individual data for responses against each of the three glycoproteins studied. Shown are box-and-whisker plots representing median values with interqartile range and minimum and maximum values. Pie charts summarize the data for gB and gH and represent the proportion of responding cells with cytotoxic potential and/or single, double, or triple cytokine production capacity.
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fig03: Glycoprotein-specific T cells vary in their functional capacities. Functional characterization of glycoprotein-specific T cells was carried out following peptide stimulation of PBMCs using multicolor flow cytometric analysis, and a Boolean gating strategy was applied. Bar charts represent individual data for responses against each of the three glycoproteins studied. Shown are box-and-whisker plots representing median values with interqartile range and minimum and maximum values. Pie charts summarize the data for gB and gH and represent the proportion of responding cells with cytotoxic potential and/or single, double, or triple cytokine production capacity.

Mentions: gB-specific CD4+ T cells displayed a strongly cytotoxic phenotype (87% GzmB+) following peptide stimulation with very high levels of TNF-α and IFN-γ production (Fig. 3). Only 26% of responding cells expressed IL-2, although 16% exhibited a complete polyfunctional profile by expression of all four markers. In contrast, gH-specific T cells exhibited a markedly reduced cytotoxic profile, with GzmB expression observed in only 26% of cells, although it was noteworthy that 59% of gH-specific T cells retain the capacity to produce IL-2. T cell responses against the two gL-derived epitopes were comparable to those seen for gH-specific T cells (Fig. 3, bar charts).


The Cellular Localization of Human Cytomegalovirus Glycoprotein Expression Greatly Influences the Frequency and Functional Phenotype of Specific CD4+ T Cell Responses.

Pachnio A, Zuo J, Ryan GB, Begum J, Moss PA - J. Immunol. (2015)

Glycoprotein-specific T cells vary in their functional capacities. Functional characterization of glycoprotein-specific T cells was carried out following peptide stimulation of PBMCs using multicolor flow cytometric analysis, and a Boolean gating strategy was applied. Bar charts represent individual data for responses against each of the three glycoproteins studied. Shown are box-and-whisker plots representing median values with interqartile range and minimum and maximum values. Pie charts summarize the data for gB and gH and represent the proportion of responding cells with cytotoxic potential and/or single, double, or triple cytokine production capacity.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4592104&req=5

fig03: Glycoprotein-specific T cells vary in their functional capacities. Functional characterization of glycoprotein-specific T cells was carried out following peptide stimulation of PBMCs using multicolor flow cytometric analysis, and a Boolean gating strategy was applied. Bar charts represent individual data for responses against each of the three glycoproteins studied. Shown are box-and-whisker plots representing median values with interqartile range and minimum and maximum values. Pie charts summarize the data for gB and gH and represent the proportion of responding cells with cytotoxic potential and/or single, double, or triple cytokine production capacity.
Mentions: gB-specific CD4+ T cells displayed a strongly cytotoxic phenotype (87% GzmB+) following peptide stimulation with very high levels of TNF-α and IFN-γ production (Fig. 3). Only 26% of responding cells expressed IL-2, although 16% exhibited a complete polyfunctional profile by expression of all four markers. In contrast, gH-specific T cells exhibited a markedly reduced cytotoxic profile, with GzmB expression observed in only 26% of cells, although it was noteworthy that 59% of gH-specific T cells retain the capacity to produce IL-2. T cell responses against the two gL-derived epitopes were comparable to those seen for gH-specific T cells (Fig. 3, bar charts).

Bottom Line: In this study, we examine and contrast the magnitude and phenotype of the T cell immune response against gB, gH, and gL within healthy donors. gB-specific CD4(+) T cells were found in 95% of donors, and 29 epitopes were defined with gB-specific response sizes ranging from 0.02 to 2.88% of the CD4(+) T cell pool.Glycoproteins were effectively presented following delivery to APCs but only gB-derived epitopes were presented following endogenous synthesis. gB expression was observed exclusively within vesicular structures colocalizing with HLA-DM whereas gH was distributed evenly throughout the cytoplasm.These results reveal that gB is a uniquely immunogenic CMV glycoprotein and this is likely to reflect its unique pattern of endogenous Ag presentation.

View Article: PubMed Central - PubMed

Affiliation: School of Cancer Sciences, College of Medicine and Dentistry, University of Birmingham, Birmingham B15 2TT, United Kingdom; and a.pachnio@bham.ac.uk p.moss@bham.ac.uk.

Show MeSH
Related in: MedlinePlus