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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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Patterns of cytokine expression in the clusters associated with active andinactive periapical granulomas nature. Hierarchical analysis demonstrated thatthe inactivity pole was characterized by the highest OPG levels, sequentiallyfollowed in a downward way by FOXp3, IL-10, IL-9, IL-4 and IL-22. OPG and IL-22expression profiles were relatively stable within inactive clusters; FOXP3 andIL-10 levels prevail in the clusters located in the inactivity pole edge, whilea significant variation in expression of IL-4 and IL-9 was verified in specificclusters. On the other hand, the lesions activity pole was characterized by thehighest expression of TNF-α, downward followed by RANKL, IL-21, IL-17 andIFN-γ. High levels of TNF-α expression were a hallmark of all active clusters,and RANKL expression prevail in the clusters located in the activity pole edge;also, a fairly specificity in the IFN-γ, IL-17 and IL-21 expression peaks wasverified in definite clusters within active lesions. Interestingly, one clusterfrom inactive lesions subset, presented a relatively high expression of IL-17.The cytokines IL-23 and IL-33 were not significantly associated with lesions’status
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f03: Patterns of cytokine expression in the clusters associated with active andinactive periapical granulomas nature. Hierarchical analysis demonstrated thatthe inactivity pole was characterized by the highest OPG levels, sequentiallyfollowed in a downward way by FOXp3, IL-10, IL-9, IL-4 and IL-22. OPG and IL-22expression profiles were relatively stable within inactive clusters; FOXP3 andIL-10 levels prevail in the clusters located in the inactivity pole edge, whilea significant variation in expression of IL-4 and IL-9 was verified in specificclusters. On the other hand, the lesions activity pole was characterized by thehighest expression of TNF-α, downward followed by RANKL, IL-21, IL-17 andIFN-γ. High levels of TNF-α expression were a hallmark of all active clusters,and RANKL expression prevail in the clusters located in the activity pole edge;also, a fairly specificity in the IFN-γ, IL-17 and IL-21 expression peaks wasverified in definite clusters within active lesions. Interestingly, one clusterfrom inactive lesions subset, presented a relatively high expression of IL-17.The cytokines IL-23 and IL-33 were not significantly associated with lesions’status

Mentions: The subsequent hierarchical analysis (Table 3and Figure 3) ordered the samples regarding theactivity level, from inactivity to activity ends, and demonstrated that inactivitypole was characterized by the highest OPG levels, sequentially followed in a downwardway by FOXp3, IL-10, IL-9, IL-4 and IL-22. OPG and IL-22 expression profiles wererelatively stable within inactive clusters; FOXP3 and IL-10 levels prevail in theclusters located in the inactivity pole edge, while a significant variation in theexpression of IL-4 and IL-9 was verified in specific clusters (Table 3 and Figure 3). Onthe other hand, the lesion activity pole was characterized by the highest expressionof TNF-α, downward followed by RANKL, IL-21, IL-17 and IFN-γ (Table 3 and Figure 3). Highlevels of TNF-α expression were a hallmark of all active clusters, RANKL expressionprevails in the clusters located in the activity pole edge; also, a fairlyspecificity in the IFN-γ, IL-17 and IL-21 expression peaks was verified in definiteclusters within active lesions (Table 3 andFigure 3). Interestingly, one cluster frominactive lesions subset presented a relatively high expression of IL-17. In anintermediate level within inactivity and activity poles, the cytokines IL-23 andIL-33 were not significantly associated with lesions' status (Table 3 and Figure 3).


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)

Patterns of cytokine expression in the clusters associated with active andinactive periapical granulomas nature. Hierarchical analysis demonstrated thatthe inactivity pole was characterized by the highest OPG levels, sequentiallyfollowed in a downward way by FOXp3, IL-10, IL-9, IL-4 and IL-22. OPG and IL-22expression profiles were relatively stable within inactive clusters; FOXP3 andIL-10 levels prevail in the clusters located in the inactivity pole edge, whilea significant variation in expression of IL-4 and IL-9 was verified in specificclusters. On the other hand, the lesions activity pole was characterized by thehighest expression of TNF-α, downward followed by RANKL, IL-21, IL-17 andIFN-γ. High levels of TNF-α expression were a hallmark of all active clusters,and RANKL expression prevail in the clusters located in the activity pole edge;also, a fairly specificity in the IFN-γ, IL-17 and IL-21 expression peaks wasverified in definite clusters within active lesions. Interestingly, one clusterfrom inactive lesions subset, presented a relatively high expression of IL-17.The cytokines IL-23 and IL-33 were not significantly associated with lesions’status
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f03: Patterns of cytokine expression in the clusters associated with active andinactive periapical granulomas nature. Hierarchical analysis demonstrated thatthe inactivity pole was characterized by the highest OPG levels, sequentiallyfollowed in a downward way by FOXp3, IL-10, IL-9, IL-4 and IL-22. OPG and IL-22expression profiles were relatively stable within inactive clusters; FOXP3 andIL-10 levels prevail in the clusters located in the inactivity pole edge, whilea significant variation in expression of IL-4 and IL-9 was verified in specificclusters. On the other hand, the lesions activity pole was characterized by thehighest expression of TNF-α, downward followed by RANKL, IL-21, IL-17 andIFN-γ. High levels of TNF-α expression were a hallmark of all active clusters,and RANKL expression prevail in the clusters located in the activity pole edge;also, a fairly specificity in the IFN-γ, IL-17 and IL-21 expression peaks wasverified in definite clusters within active lesions. Interestingly, one clusterfrom inactive lesions subset, presented a relatively high expression of IL-17.The cytokines IL-23 and IL-33 were not significantly associated with lesions’status
Mentions: The subsequent hierarchical analysis (Table 3and Figure 3) ordered the samples regarding theactivity level, from inactivity to activity ends, and demonstrated that inactivitypole was characterized by the highest OPG levels, sequentially followed in a downwardway by FOXp3, IL-10, IL-9, IL-4 and IL-22. OPG and IL-22 expression profiles wererelatively stable within inactive clusters; FOXP3 and IL-10 levels prevail in theclusters located in the inactivity pole edge, while a significant variation in theexpression of IL-4 and IL-9 was verified in specific clusters (Table 3 and Figure 3). Onthe other hand, the lesion activity pole was characterized by the highest expressionof TNF-α, downward followed by RANKL, IL-21, IL-17 and IFN-γ (Table 3 and Figure 3). Highlevels of TNF-α expression were a hallmark of all active clusters, RANKL expressionprevails in the clusters located in the activity pole edge; also, a fairlyspecificity in the IFN-γ, IL-17 and IL-21 expression peaks was verified in definiteclusters within active lesions (Table 3 andFigure 3). Interestingly, one cluster frominactive lesions subset presented a relatively high expression of IL-17. In anintermediate level within inactivity and activity poles, the cytokines IL-23 andIL-33 were not significantly associated with lesions' status (Table 3 and Figure 3).

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