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Simultaneous analysis of T helper subsets (Th1, Th2, Th9, Th17, Th22, Tfh, Tr1 and Tregs) markers expression in periapical lesions reveals multiple cytokine clusters accountable for lesions activity and inactivity status.

Araujo-Pires AC, Francisconi CF, Biguetti CC, Cavalla F, Aranha AM, Letra A, Trombone AP, Faveri M, Silva RM, Garlet GP - J Appl Oral Sci (2014 Jul-Aug)

Bottom Line: Five clusters were identified in inactive lesion groups, being the variance in the expression levels of IL-17, IL-10, FOXp3, IFN-γ, IL-9, IL-33 and IL-4 statistically significant (KW p<0.05).There is a clear dichotomy in the profile of cytokine expression in inactive and active periapical lesions.While the widespread cytokine expression seems to be a feature of chronic lesions, hierarchical cluster analysis demonstrates the association of TNF-α, IL-21, IL-17 and IFN-γ with lesions activity, and the association of FOXP3, IL-10, IL-9, IL-4 and IL-22 with lesions inactivity.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Bauru School of Dentistry, University of São Paulo, Bauru, SP, Brazil.

ABSTRACT

Unlabelled: Previous studies demonstrate that the balance between pro- and anti-inflammatory mediators determines the stable or progressive nature of periapical granulomas by modulating the balance of the osteoclastogenic factor RANKL and its antagonist OPG. However, the cytokine networks operating in the development of periapical lesions are quite more complex than what the simple pro- versus anti-inflammatory mediators' paradigm suggests. Here we simultaneously investigated the patterns of Th1, Th2, Th9, Th17, Th22, Thf, Tr1 and Tregs cytokines/markers expression in human periapical granulomas.

Methods: The expression of TNF-α, IFN-γ, IL-17A, IL23, IL21, IL-33, IL-10, IL-4, IL-9, IL-22, FOXp3 markers (via RealTimePCR array) was accessed in active/progressive (N=40) versus inactive/stable (N=70) periapical granulomas (as determined by RANKL/OPG expression ratio), and also to compare these samples with a panel of control specimens (N=26). A cluster analysis of 13 cytokine levels was performed to examine possible clustering between the cytokines in a total of 110 granulomas.

Results: The expression of all target cytokines was higher in the granulomas than in control samples. TNF-α, IFN-γ, IL-17A and IL-21 mRNA levels were significantly higher in active granulomas, while in inactive lesions the expression levels of IL-4, IL-9, IL-10, IL-22 and FOXp3 were higher than in active granulomas. Five clusters were identified in inactive lesion groups, being the variance in the expression levels of IL-17, IL-10, FOXp3, IFN-γ, IL-9, IL-33 and IL-4 statistically significant (KW p<0.05). Three clusters were identified in active lesions, being the variance in the expression levels of IL-22, IL-10, IFN-γ, IL-17, IL-33, FOXp3, IL-21 and RANKL statistically significant (KW p<0.05).

Conclusion: There is a clear dichotomy in the profile of cytokine expression in inactive and active periapical lesions. While the widespread cytokine expression seems to be a feature of chronic lesions, hierarchical cluster analysis demonstrates the association of TNF-α, IL-21, IL-17 and IFN-γ with lesions activity, and the association of FOXP3, IL-10, IL-9, IL-4 and IL-22 with lesions inactivity.

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Expression of individual mRNAs, with normalization to housekeeping genes, inperiapical granulomas. Total RNA was extracted from periapical granulomas(experimental groups, N=110) and periodontal ligament (control group, N=26),and levels of RANKL and OPG mRNA were measured quantitatively by RealTimePCRusing TaqMan chemistry. Based in profile of RANKL/OPG expression29 the lesionswere then categorized into active (RANKL>OPG) or inactive (RANKL»OPG andRANKL<OPG). Different letters (a, b, c) represent statistically significantdifferences among the respective groups (P<0.05; One-way ANOVA, Bonferronipost-test); *P<0.05 (unpaired t-test) represent statistically significantdifferences between controls and periapical lesions (active + inactive)
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f01: Expression of individual mRNAs, with normalization to housekeeping genes, inperiapical granulomas. Total RNA was extracted from periapical granulomas(experimental groups, N=110) and periodontal ligament (control group, N=26),and levels of RANKL and OPG mRNA were measured quantitatively by RealTimePCRusing TaqMan chemistry. Based in profile of RANKL/OPG expression29 the lesionswere then categorized into active (RANKL>OPG) or inactive (RANKL»OPG andRANKL<OPG). Different letters (a, b, c) represent statistically significantdifferences among the respective groups (P<0.05; One-way ANOVA, Bonferronipost-test); *P<0.05 (unpaired t-test) represent statistically significantdifferences between controls and periapical lesions (active + inactive)

Mentions: The mRNA levels expression of all targets investigated was found to be higher intotal periapical granulomas when compared to controls (Figure 1). When lesions were compared based in the RANKL/OPG expressionpattern29, 40 samples werefound to be active (RANKL>OPG), while 70 presented an inactive lesion profile(RANKL≤OPG) (Figure 1). When active andinactive lesions were compared, TNF-α, IFN-γ, IL-17A and IL-21 mRNA levels weresignificantly higher in active granulomas (Figure1), while in inactive lesions the expression levels of IL-4, IL-9, IL-10,IL-22 and FOXp3 were higher than in active granulomas (Figure 1). The levels of IL-23 and IL-33 in active and inactive lesionswere similar from a statistical viewpoint (Figure1).


Simultaneous analysis of T helper subsets (Th1, Th2, Th9, Th17, Th22, Tfh, Tr1 and Tregs) markers expression in periapical lesions reveals multiple cytokine clusters accountable for lesions activity and inactivity status.

Araujo-Pires AC, Francisconi CF, Biguetti CC, Cavalla F, Aranha AM, Letra A, Trombone AP, Faveri M, Silva RM, Garlet GP - J Appl Oral Sci (2014 Jul-Aug)

Expression of individual mRNAs, with normalization to housekeeping genes, inperiapical granulomas. Total RNA was extracted from periapical granulomas(experimental groups, N=110) and periodontal ligament (control group, N=26),and levels of RANKL and OPG mRNA were measured quantitatively by RealTimePCRusing TaqMan chemistry. Based in profile of RANKL/OPG expression29 the lesionswere then categorized into active (RANKL>OPG) or inactive (RANKL»OPG andRANKL<OPG). Different letters (a, b, c) represent statistically significantdifferences among the respective groups (P<0.05; One-way ANOVA, Bonferronipost-test); *P<0.05 (unpaired t-test) represent statistically significantdifferences between controls and periapical lesions (active + inactive)
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4126831&req=5

f01: Expression of individual mRNAs, with normalization to housekeeping genes, inperiapical granulomas. Total RNA was extracted from periapical granulomas(experimental groups, N=110) and periodontal ligament (control group, N=26),and levels of RANKL and OPG mRNA were measured quantitatively by RealTimePCRusing TaqMan chemistry. Based in profile of RANKL/OPG expression29 the lesionswere then categorized into active (RANKL>OPG) or inactive (RANKL»OPG andRANKL<OPG). Different letters (a, b, c) represent statistically significantdifferences among the respective groups (P<0.05; One-way ANOVA, Bonferronipost-test); *P<0.05 (unpaired t-test) represent statistically significantdifferences between controls and periapical lesions (active + inactive)
Mentions: The mRNA levels expression of all targets investigated was found to be higher intotal periapical granulomas when compared to controls (Figure 1). When lesions were compared based in the RANKL/OPG expressionpattern29, 40 samples werefound to be active (RANKL>OPG), while 70 presented an inactive lesion profile(RANKL≤OPG) (Figure 1). When active andinactive lesions were compared, TNF-α, IFN-γ, IL-17A and IL-21 mRNA levels weresignificantly higher in active granulomas (Figure1), while in inactive lesions the expression levels of IL-4, IL-9, IL-10,IL-22 and FOXp3 were higher than in active granulomas (Figure 1). The levels of IL-23 and IL-33 in active and inactive lesionswere similar from a statistical viewpoint (Figure1).

Bottom Line: Five clusters were identified in inactive lesion groups, being the variance in the expression levels of IL-17, IL-10, FOXp3, IFN-γ, IL-9, IL-33 and IL-4 statistically significant (KW p<0.05).There is a clear dichotomy in the profile of cytokine expression in inactive and active periapical lesions.While the widespread cytokine expression seems to be a feature of chronic lesions, hierarchical cluster analysis demonstrates the association of TNF-α, IL-21, IL-17 and IFN-γ with lesions activity, and the association of FOXP3, IL-10, IL-9, IL-4 and IL-22 with lesions inactivity.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Bauru School of Dentistry, University of São Paulo, Bauru, SP, Brazil.

ABSTRACT

Unlabelled: Previous studies demonstrate that the balance between pro- and anti-inflammatory mediators determines the stable or progressive nature of periapical granulomas by modulating the balance of the osteoclastogenic factor RANKL and its antagonist OPG. However, the cytokine networks operating in the development of periapical lesions are quite more complex than what the simple pro- versus anti-inflammatory mediators' paradigm suggests. Here we simultaneously investigated the patterns of Th1, Th2, Th9, Th17, Th22, Thf, Tr1 and Tregs cytokines/markers expression in human periapical granulomas.

Methods: The expression of TNF-α, IFN-γ, IL-17A, IL23, IL21, IL-33, IL-10, IL-4, IL-9, IL-22, FOXp3 markers (via RealTimePCR array) was accessed in active/progressive (N=40) versus inactive/stable (N=70) periapical granulomas (as determined by RANKL/OPG expression ratio), and also to compare these samples with a panel of control specimens (N=26). A cluster analysis of 13 cytokine levels was performed to examine possible clustering between the cytokines in a total of 110 granulomas.

Results: The expression of all target cytokines was higher in the granulomas than in control samples. TNF-α, IFN-γ, IL-17A and IL-21 mRNA levels were significantly higher in active granulomas, while in inactive lesions the expression levels of IL-4, IL-9, IL-10, IL-22 and FOXp3 were higher than in active granulomas. Five clusters were identified in inactive lesion groups, being the variance in the expression levels of IL-17, IL-10, FOXp3, IFN-γ, IL-9, IL-33 and IL-4 statistically significant (KW p<0.05). Three clusters were identified in active lesions, being the variance in the expression levels of IL-22, IL-10, IFN-γ, IL-17, IL-33, FOXp3, IL-21 and RANKL statistically significant (KW p<0.05).

Conclusion: There is a clear dichotomy in the profile of cytokine expression in inactive and active periapical lesions. While the widespread cytokine expression seems to be a feature of chronic lesions, hierarchical cluster analysis demonstrates the association of TNF-α, IL-21, IL-17 and IFN-γ with lesions activity, and the association of FOXP3, IL-10, IL-9, IL-4 and IL-22 with lesions inactivity.

Show MeSH
Related in: MedlinePlus