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Characterization of CD8+ T cell differentiation following SIVΔnef vaccination by transcription factor expression profiling.

Billingsley JM, Rajakumar PA, Connole MA, Salisch NC, Adnan S, Kuzmichev YV, Hong HS, Reeves RK, Kang HJ, Li W, Li Q, Haase AT, Johnson RP - PLoS Pathog. (2015)

Bottom Line: The onset of protective immunity against pathogenic SIV challenge in SIVΔnef-vaccinated macaques is delayed for 15-20 weeks, a process that is related to qualitative changes in CD8+ T cell responses induced by SIVΔnef.Expression of transcription factors associated with effector differentiation, such as ID2 and RUNX3, were decreased over time, while expression of transcription factors associated with quiescence or memory differentiation, such as TCF7, BCOR and EOMES, increased.These data suggest transcription factor expression profiling is a novel method that can provide additional data complementary to the analysis of memory cell differentiation based on classical phenotypic markers.

View Article: PubMed Central - PubMed

Affiliation: Division of Immunology, New England Primate Research Center, Harvard Medical School, Southborough, Massachusetts, United States of America; Yerkes National Primate Research Center, Emory University, Atlanta, Georgia, United States of America.

ABSTRACT
The onset of protective immunity against pathogenic SIV challenge in SIVΔnef-vaccinated macaques is delayed for 15-20 weeks, a process that is related to qualitative changes in CD8+ T cell responses induced by SIVΔnef. As a novel approach to characterize cell differentiation following vaccination, we used multi-target qPCR to measure transcription factor expression in naïve and memory subsets of CD8++ T cells, and in SIV-specific CD8+ T cells obtained from SIVΔnef-vaccinated or wild type SIVmac239-infected macaques. Unsupervised clustering of expression profiles organized naïve and memory CD8+ T cells into groups concordant with cell surface phenotype. Transcription factor expression patterns in SIV-specific CD8+ T cells in SIVΔnef-vaccinated animals were distinct from those observed in purified CD8+ T cell subsets obtained from naïve animals, and were intermediate to expression profiles of purified central memory and effector memory T cells. Expression of transcription factors elicited by SIVΔnef vaccination also varied over time: cells obtained at later time points, temporally associated with greater protection, appeared more central-memory like than cells obtained at earlier time points, which appeared more effector memory-like. Expression of transcription factors associated with effector differentiation, such as ID2 and RUNX3, were decreased over time, while expression of transcription factors associated with quiescence or memory differentiation, such as TCF7, BCOR and EOMES, increased. CD8+ T cells specific for a more conserved epitope expressed higher levels of TBX21 and BATF, and appeared more effector-like than cells specific for an escaped epitope, consistent with continued activation by replicating vaccine virus. These data suggest transcription factor expression profiling is a novel method that can provide additional data complementary to the analysis of memory cell differentiation based on classical phenotypic markers. Additionally, these data support the hypothesis that ongoing stimulation by SIVΔnef promotes a distinct protective balance of CD8+ T cell differentiation and activation states.

No MeSH data available.


Related in: MedlinePlus

Segregation of sorted CD8+ T cell subsets by unsupervised hierarchical clustering.Heat map expression values were transcript mean centered and represent expression relative to endogenous controls. Hierarchical clustering was performed using Euclidean distance and complete linkage methods.
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ppat.1004740.g001: Segregation of sorted CD8+ T cell subsets by unsupervised hierarchical clustering.Heat map expression values were transcript mean centered and represent expression relative to endogenous controls. Hierarchical clustering was performed using Euclidean distance and complete linkage methods.

Mentions: To determine if transcription factor expression profiling can be used to identify distinct stages of CD8+ T cell differentiation, we initially sorted highly purified populations of naïve, central memory, transitional memory, and effector memory CD8+ T cells from five healthy unvaccinated uninfected rhesus macaques based on differential surface expression of CD28, CD95 and CCR7 (S1 Fig., Fig. 1). We then measured the expression of the transcription factors included in our panel by multi-target qPCR, and used agglomerative unsupervised hierarchical clustering of the expression data to organize the samples. The sample dendrogram (Fig. 1) demonstrates clear segregation of samples by cell differentiation stage, with the three memory CD8+ T cell subsets segregating from naïve cells.


Characterization of CD8+ T cell differentiation following SIVΔnef vaccination by transcription factor expression profiling.

Billingsley JM, Rajakumar PA, Connole MA, Salisch NC, Adnan S, Kuzmichev YV, Hong HS, Reeves RK, Kang HJ, Li W, Li Q, Haase AT, Johnson RP - PLoS Pathog. (2015)

Segregation of sorted CD8+ T cell subsets by unsupervised hierarchical clustering.Heat map expression values were transcript mean centered and represent expression relative to endogenous controls. Hierarchical clustering was performed using Euclidean distance and complete linkage methods.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4358830&req=5

ppat.1004740.g001: Segregation of sorted CD8+ T cell subsets by unsupervised hierarchical clustering.Heat map expression values were transcript mean centered and represent expression relative to endogenous controls. Hierarchical clustering was performed using Euclidean distance and complete linkage methods.
Mentions: To determine if transcription factor expression profiling can be used to identify distinct stages of CD8+ T cell differentiation, we initially sorted highly purified populations of naïve, central memory, transitional memory, and effector memory CD8+ T cells from five healthy unvaccinated uninfected rhesus macaques based on differential surface expression of CD28, CD95 and CCR7 (S1 Fig., Fig. 1). We then measured the expression of the transcription factors included in our panel by multi-target qPCR, and used agglomerative unsupervised hierarchical clustering of the expression data to organize the samples. The sample dendrogram (Fig. 1) demonstrates clear segregation of samples by cell differentiation stage, with the three memory CD8+ T cell subsets segregating from naïve cells.

Bottom Line: The onset of protective immunity against pathogenic SIV challenge in SIVΔnef-vaccinated macaques is delayed for 15-20 weeks, a process that is related to qualitative changes in CD8+ T cell responses induced by SIVΔnef.Expression of transcription factors associated with effector differentiation, such as ID2 and RUNX3, were decreased over time, while expression of transcription factors associated with quiescence or memory differentiation, such as TCF7, BCOR and EOMES, increased.These data suggest transcription factor expression profiling is a novel method that can provide additional data complementary to the analysis of memory cell differentiation based on classical phenotypic markers.

View Article: PubMed Central - PubMed

Affiliation: Division of Immunology, New England Primate Research Center, Harvard Medical School, Southborough, Massachusetts, United States of America; Yerkes National Primate Research Center, Emory University, Atlanta, Georgia, United States of America.

ABSTRACT
The onset of protective immunity against pathogenic SIV challenge in SIVΔnef-vaccinated macaques is delayed for 15-20 weeks, a process that is related to qualitative changes in CD8+ T cell responses induced by SIVΔnef. As a novel approach to characterize cell differentiation following vaccination, we used multi-target qPCR to measure transcription factor expression in naïve and memory subsets of CD8++ T cells, and in SIV-specific CD8+ T cells obtained from SIVΔnef-vaccinated or wild type SIVmac239-infected macaques. Unsupervised clustering of expression profiles organized naïve and memory CD8+ T cells into groups concordant with cell surface phenotype. Transcription factor expression patterns in SIV-specific CD8+ T cells in SIVΔnef-vaccinated animals were distinct from those observed in purified CD8+ T cell subsets obtained from naïve animals, and were intermediate to expression profiles of purified central memory and effector memory T cells. Expression of transcription factors elicited by SIVΔnef vaccination also varied over time: cells obtained at later time points, temporally associated with greater protection, appeared more central-memory like than cells obtained at earlier time points, which appeared more effector memory-like. Expression of transcription factors associated with effector differentiation, such as ID2 and RUNX3, were decreased over time, while expression of transcription factors associated with quiescence or memory differentiation, such as TCF7, BCOR and EOMES, increased. CD8+ T cells specific for a more conserved epitope expressed higher levels of TBX21 and BATF, and appeared more effector-like than cells specific for an escaped epitope, consistent with continued activation by replicating vaccine virus. These data suggest transcription factor expression profiling is a novel method that can provide additional data complementary to the analysis of memory cell differentiation based on classical phenotypic markers. Additionally, these data support the hypothesis that ongoing stimulation by SIVΔnef promotes a distinct protective balance of CD8+ T cell differentiation and activation states.

No MeSH data available.


Related in: MedlinePlus