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Differential expression of CC chemokines (CCLs) and receptors (CCRs) by human T lymphocytes in response to different Aggregatibacter actinomycetemcomitans serotypes.

Alvarez C, Benítez A, Rojas L, Pujol M, Carvajal P, Díaz-Zúñiga J, Vernal R - J Appl Oral Sci (2015)

Bottom Line: In Aggregatibacter actinomycetemcomitans, different serotypes have been described based on LPS antigenicity.The expression levels for the chemokines CCL1, CCL2, CCL3, CCL5, CCL11, CCL17, CCL20, CCL21, CCL25, and CCL28, as well as the chemokine receptors CCR1, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, and CCR10 were quantified by qPCR.Similarly, the expression levels for the transcription factors T-bet, GATA-3, RORC2, and Foxp3 were quantified and correlated with the CCL and CCR expression levels.Results Higher expression levels of CCL2, CCL3, CCL5, CCL20, CCL21, CCL28, CCR1, CCR2, CCR5, CCR6, CCR7, and CCR9 were detected in T lymphocytes stimulated with the serotype b of A. actinomycetemcomitans compared with the other serotypes.

View Article: PubMed Central - PubMed

Affiliation: Laboratorio de Biología Periodontal, Facultad de Odontología, Universidad de Chile, Santiago, Chile.

ABSTRACT
In Aggregatibacter actinomycetemcomitans, different serotypes have been described based on LPS antigenicity. Recently, our research group has reported a differential immunogenicity when T lymphocytes were stimulated with these different serotypes. In particular, it was demonstrated that the serotype b of A. actinomycetemcomitans has a stronger capacity to trigger Th1- and Th17-type cytokine production.Objective This study aimed to quantify the expression of different CC chemokines (CCLs) and receptors (CCRs) in T lymphocytes stimulated with the differentA. actinomycetemcomitans serotypes. In addition, the expression of the transcription factors T-bet, GATA-3, RORC2, and Foxp3, master-switch genes implied in the Th1, Th2, Th17, and T-regulatory differentiation, respectively, was analysed in order to determine T-cell phenotype-specific patterns of CCL and CCR expression upon A. actinomycetemcomitans stimulation.Material and Methods Human naïve CD4+ T lymphocytes were obtained from healthy subjects and stimulated with autologous dendritic cells primed with the differentA. actinomycetemcomitans serotypes. The expression levels for the chemokines CCL1, CCL2, CCL3, CCL5, CCL11, CCL17, CCL20, CCL21, CCL25, and CCL28, as well as the chemokine receptors CCR1, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, and CCR10 were quantified by qPCR. Similarly, the expression levels for the transcription factors T-bet, GATA-3, RORC2, and Foxp3 were quantified and correlated with the CCL and CCR expression levels.Results Higher expression levels of CCL2, CCL3, CCL5, CCL20, CCL21, CCL28, CCR1, CCR2, CCR5, CCR6, CCR7, and CCR9 were detected in T lymphocytes stimulated with the serotype b of A. actinomycetemcomitans compared with the other serotypes. In addition, these higher expression levels of CCLs and CCRs positively correlated with the increased levels of T-bet and RORC2 when T lymphocytes were stimulated with the serotype b.Conclusion A T-lymphocyte response biased towards a Th1- and Th17-pattern of CCL and CCR expression was detected under stimulation with the serotype b ofA. actinomycetemcomitans.

No MeSH data available.


CC chemokines (CCL) expression by A. actinomycetemcomitans-induced T lymphocytes. CCL mRNA expression in T lymphocytes activated by dendritic cells primed at a MOI=102 with the A. actinomycetemcomitans strains ATCC® 43717™ (serotype a), ATCC® 43718™ (serotype b), and ATCC® 43719™ (serotype c). For relative expression, the CCL mRNA expression in T lymphocytes exposed to non-induced dendritic cells was considered as 1, as a reference for fold-change in expression (n.i). Data are represented as fold-change for 8 independent experiments. Each experiment was performed in duplicate. Comparisons were done between the different A. actinomycetemcomitans serotypes (*p<0.05)
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f03: CC chemokines (CCL) expression by A. actinomycetemcomitans-induced T lymphocytes. CCL mRNA expression in T lymphocytes activated by dendritic cells primed at a MOI=102 with the A. actinomycetemcomitans strains ATCC® 43717™ (serotype a), ATCC® 43718™ (serotype b), and ATCC® 43719™ (serotype c). For relative expression, the CCL mRNA expression in T lymphocytes exposed to non-induced dendritic cells was considered as 1, as a reference for fold-change in expression (n.i). Data are represented as fold-change for 8 independent experiments. Each experiment was performed in duplicate. Comparisons were done between the different A. actinomycetemcomitans serotypes (*p<0.05)

Mentions: The mRNA expression for the analyzed chemokines was determined by qPCR and represented as fold-change for each condition (Figure 3). When the strain ATCC® 43718™ belonging to the serotype b of A. actinomycetemcomitans was used for T-lymphocyte activation, higher expression levels of CCL2 (p=0.025 and p=0.024), CCL3 (p=0.003 and p=0.005), CCL5 (p=0.004 and p=0.013), CCL20 (p=0.05 and p=0.02), CCL21 (p=0.001 and p=0.001), and CCL28 (p=0.004 and p=0.008) were detected, when compared with the strains ATCC® 43717™ and ATCC® 43719™ belonging to the serotypes a or c, respectively. Conversely, when the serotype a of A. actinomycetemcomitans was used for T-lymphocyte activation, higher expression levels of CCL1 (p=0.008 and p=0.009) and CCL17 (p>0.05 and p>0.05) were detected, when compared with the serotypes b or c, respectively. CCL11 and CCL25 were not over-expressed in any experimental condition.


Differential expression of CC chemokines (CCLs) and receptors (CCRs) by human T lymphocytes in response to different Aggregatibacter actinomycetemcomitans serotypes.

Alvarez C, Benítez A, Rojas L, Pujol M, Carvajal P, Díaz-Zúñiga J, Vernal R - J Appl Oral Sci (2015)

CC chemokines (CCL) expression by A. actinomycetemcomitans-induced T lymphocytes. CCL mRNA expression in T lymphocytes activated by dendritic cells primed at a MOI=102 with the A. actinomycetemcomitans strains ATCC® 43717™ (serotype a), ATCC® 43718™ (serotype b), and ATCC® 43719™ (serotype c). For relative expression, the CCL mRNA expression in T lymphocytes exposed to non-induced dendritic cells was considered as 1, as a reference for fold-change in expression (n.i). Data are represented as fold-change for 8 independent experiments. Each experiment was performed in duplicate. Comparisons were done between the different A. actinomycetemcomitans serotypes (*p<0.05)
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4621946&req=5

f03: CC chemokines (CCL) expression by A. actinomycetemcomitans-induced T lymphocytes. CCL mRNA expression in T lymphocytes activated by dendritic cells primed at a MOI=102 with the A. actinomycetemcomitans strains ATCC® 43717™ (serotype a), ATCC® 43718™ (serotype b), and ATCC® 43719™ (serotype c). For relative expression, the CCL mRNA expression in T lymphocytes exposed to non-induced dendritic cells was considered as 1, as a reference for fold-change in expression (n.i). Data are represented as fold-change for 8 independent experiments. Each experiment was performed in duplicate. Comparisons were done between the different A. actinomycetemcomitans serotypes (*p<0.05)
Mentions: The mRNA expression for the analyzed chemokines was determined by qPCR and represented as fold-change for each condition (Figure 3). When the strain ATCC® 43718™ belonging to the serotype b of A. actinomycetemcomitans was used for T-lymphocyte activation, higher expression levels of CCL2 (p=0.025 and p=0.024), CCL3 (p=0.003 and p=0.005), CCL5 (p=0.004 and p=0.013), CCL20 (p=0.05 and p=0.02), CCL21 (p=0.001 and p=0.001), and CCL28 (p=0.004 and p=0.008) were detected, when compared with the strains ATCC® 43717™ and ATCC® 43719™ belonging to the serotypes a or c, respectively. Conversely, when the serotype a of A. actinomycetemcomitans was used for T-lymphocyte activation, higher expression levels of CCL1 (p=0.008 and p=0.009) and CCL17 (p>0.05 and p>0.05) were detected, when compared with the serotypes b or c, respectively. CCL11 and CCL25 were not over-expressed in any experimental condition.

Bottom Line: In Aggregatibacter actinomycetemcomitans, different serotypes have been described based on LPS antigenicity.The expression levels for the chemokines CCL1, CCL2, CCL3, CCL5, CCL11, CCL17, CCL20, CCL21, CCL25, and CCL28, as well as the chemokine receptors CCR1, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, and CCR10 were quantified by qPCR.Similarly, the expression levels for the transcription factors T-bet, GATA-3, RORC2, and Foxp3 were quantified and correlated with the CCL and CCR expression levels.Results Higher expression levels of CCL2, CCL3, CCL5, CCL20, CCL21, CCL28, CCR1, CCR2, CCR5, CCR6, CCR7, and CCR9 were detected in T lymphocytes stimulated with the serotype b of A. actinomycetemcomitans compared with the other serotypes.

View Article: PubMed Central - PubMed

Affiliation: Laboratorio de Biología Periodontal, Facultad de Odontología, Universidad de Chile, Santiago, Chile.

ABSTRACT
In Aggregatibacter actinomycetemcomitans, different serotypes have been described based on LPS antigenicity. Recently, our research group has reported a differential immunogenicity when T lymphocytes were stimulated with these different serotypes. In particular, it was demonstrated that the serotype b of A. actinomycetemcomitans has a stronger capacity to trigger Th1- and Th17-type cytokine production.Objective This study aimed to quantify the expression of different CC chemokines (CCLs) and receptors (CCRs) in T lymphocytes stimulated with the differentA. actinomycetemcomitans serotypes. In addition, the expression of the transcription factors T-bet, GATA-3, RORC2, and Foxp3, master-switch genes implied in the Th1, Th2, Th17, and T-regulatory differentiation, respectively, was analysed in order to determine T-cell phenotype-specific patterns of CCL and CCR expression upon A. actinomycetemcomitans stimulation.Material and Methods Human naïve CD4+ T lymphocytes were obtained from healthy subjects and stimulated with autologous dendritic cells primed with the differentA. actinomycetemcomitans serotypes. The expression levels for the chemokines CCL1, CCL2, CCL3, CCL5, CCL11, CCL17, CCL20, CCL21, CCL25, and CCL28, as well as the chemokine receptors CCR1, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, and CCR10 were quantified by qPCR. Similarly, the expression levels for the transcription factors T-bet, GATA-3, RORC2, and Foxp3 were quantified and correlated with the CCL and CCR expression levels.Results Higher expression levels of CCL2, CCL3, CCL5, CCL20, CCL21, CCL28, CCR1, CCR2, CCR5, CCR6, CCR7, and CCR9 were detected in T lymphocytes stimulated with the serotype b of A. actinomycetemcomitans compared with the other serotypes. In addition, these higher expression levels of CCLs and CCRs positively correlated with the increased levels of T-bet and RORC2 when T lymphocytes were stimulated with the serotype b.Conclusion A T-lymphocyte response biased towards a Th1- and Th17-pattern of CCL and CCR expression was detected under stimulation with the serotype b ofA. actinomycetemcomitans.

No MeSH data available.