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Mesenchymal stem cells promote the sustained expression of CD69 on activated T lymphocytes: roles of canonical and non-canonical NF-κB signalling.

Saldanha-Araujo F, Haddad R, Farias KC, Souza Ade P, Palma PV, Araujo AG, Orellana MD, Voltarelli JC, Covas DT, Zago MA, Panepucci RA - J. Cell. Mol. Med. (2012)

Bottom Line: The frequency of CD69(+) cells among lymphocytes cultured alone progressively decreased after activation.In contrast, the frequency of CD69(+) cells increased significantly following activation in lymphocytes co-cultured with MSCs.These results indicate that the canonical NF-κB pathway controls the early expression of CD69 after activation; however, in an immunoregulatory context, late and sustained CD69 expression is promoted by the non-canonical pathway and is inhibited by canonical NF-κB signalling.

View Article: PubMed Central - PubMed

Affiliation: National Institute of Science and Technology in Stem Cell and Cell Therapy, Center for Cell Therapy, Regional Blood Center and Faculty of Medicine, University of São Paulo (FMRP-USP), Ribeirão Preto, Brazil.

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Gene expression analysis of selected transcripts. PB CD3+ T cells from four individuals were activated with anti-CD2/CD3/CD28 beads and cultured alone (L) or in the presence of MSCs (L/M). On the fifth day following activation, CD4+ T cells were immunomagnetically purified and profiled by real-time PCR to evaluate gene expression in regulatory cells (A) and selected components of NF-κB signalling (B). The relative fold change values were calculated by the formula 2ΔΔCt using the median Ct value from the samples of lymphocytes cultured alone as a reference. Statistical significance was evaluated using a one-tailed paired t-test.
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fig04: Gene expression analysis of selected transcripts. PB CD3+ T cells from four individuals were activated with anti-CD2/CD3/CD28 beads and cultured alone (L) or in the presence of MSCs (L/M). On the fifth day following activation, CD4+ T cells were immunomagnetically purified and profiled by real-time PCR to evaluate gene expression in regulatory cells (A) and selected components of NF-κB signalling (B). The relative fold change values were calculated by the formula 2ΔΔCt using the median Ct value from the samples of lymphocytes cultured alone as a reference. Statistical significance was evaluated using a one-tailed paired t-test.

Mentions: There are two NF-κB signalling pathways that are associated with different roles in the immune system: the classic or canonical pathway (mediated mainly by heterodimers encoded by RELA and NFKB1) and the non-canonical or developmental pathway (mediated by heterodimers encoded by RELB and NFKB2); reviewed in [17, 54-57]. Real-time PCR revealed that transcript levels of NFKB2, RELB and RELA (but not NFKB1) were all significantly elevated in CD4+ lymphocytes that were co-cultured with MSCs over those in CD4+ lymphocytes cultured alone (Fig. 4).


Mesenchymal stem cells promote the sustained expression of CD69 on activated T lymphocytes: roles of canonical and non-canonical NF-κB signalling.

Saldanha-Araujo F, Haddad R, Farias KC, Souza Ade P, Palma PV, Araujo AG, Orellana MD, Voltarelli JC, Covas DT, Zago MA, Panepucci RA - J. Cell. Mol. Med. (2012)

Gene expression analysis of selected transcripts. PB CD3+ T cells from four individuals were activated with anti-CD2/CD3/CD28 beads and cultured alone (L) or in the presence of MSCs (L/M). On the fifth day following activation, CD4+ T cells were immunomagnetically purified and profiled by real-time PCR to evaluate gene expression in regulatory cells (A) and selected components of NF-κB signalling (B). The relative fold change values were calculated by the formula 2ΔΔCt using the median Ct value from the samples of lymphocytes cultured alone as a reference. Statistical significance was evaluated using a one-tailed paired t-test.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3823077&req=5

fig04: Gene expression analysis of selected transcripts. PB CD3+ T cells from four individuals were activated with anti-CD2/CD3/CD28 beads and cultured alone (L) or in the presence of MSCs (L/M). On the fifth day following activation, CD4+ T cells were immunomagnetically purified and profiled by real-time PCR to evaluate gene expression in regulatory cells (A) and selected components of NF-κB signalling (B). The relative fold change values were calculated by the formula 2ΔΔCt using the median Ct value from the samples of lymphocytes cultured alone as a reference. Statistical significance was evaluated using a one-tailed paired t-test.
Mentions: There are two NF-κB signalling pathways that are associated with different roles in the immune system: the classic or canonical pathway (mediated mainly by heterodimers encoded by RELA and NFKB1) and the non-canonical or developmental pathway (mediated by heterodimers encoded by RELB and NFKB2); reviewed in [17, 54-57]. Real-time PCR revealed that transcript levels of NFKB2, RELB and RELA (but not NFKB1) were all significantly elevated in CD4+ lymphocytes that were co-cultured with MSCs over those in CD4+ lymphocytes cultured alone (Fig. 4).

Bottom Line: The frequency of CD69(+) cells among lymphocytes cultured alone progressively decreased after activation.In contrast, the frequency of CD69(+) cells increased significantly following activation in lymphocytes co-cultured with MSCs.These results indicate that the canonical NF-κB pathway controls the early expression of CD69 after activation; however, in an immunoregulatory context, late and sustained CD69 expression is promoted by the non-canonical pathway and is inhibited by canonical NF-κB signalling.

View Article: PubMed Central - PubMed

Affiliation: National Institute of Science and Technology in Stem Cell and Cell Therapy, Center for Cell Therapy, Regional Blood Center and Faculty of Medicine, University of São Paulo (FMRP-USP), Ribeirão Preto, Brazil.

Show MeSH
Related in: MedlinePlus