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[Blocking p38MAPK pathway inhibits the proliferation of OT-II cells mediated by splenic dendritic cells]
Ling Han1, Yuechen Luo2, Wantong Wu2
1Department of Biotechnology, Jinan University, Guangzhou 510632, China.
Objective:
To investigate the effect of blocking p38 mitogen-activated protein kinase (p38MAPK) pathway on the proliferation of OT-II cells mediated by splenic dendritic cells (DCs).
Methods:
Splenic DCs of C57BL/6 mice were purified with anti-CD11c immunomagnetic beads, and OT-II cells were isolated from the splenic of CD4(+)-ovalbumin transgenic mice (OT-IItransgenic mice) by mouse CD4 T cell isolation kits. After being pretreated with SB203580, an inhibitor of p38MAPK, DCs were stimulated with lipopolysaccharides (LPS). Then the expression levels of co-stimulatory molecules (CD80, CD86) and MHCII in DCs, and antigen-presenting ability of DCs treated with the 52-68 fragment of the E alpha-chain of I-E class II molecules (Eα52-68 peptide) were detected by flow cytometry. The protein levels of tumor necrosis factor α (TNF-α), interleukin 1α (IL-1α), interleukin 6 (IL-6) and transforming growth factor β (TGF-β) in the culture supernatants were measured by ELISA. The proliferation of OT-II cells which were co-cultured with OVA323-339-treated DCs was analyzed by flow cytometry.
Results:
The purity of both DCs and OT-II cells reached over 90% after isolation. SB203580 downregulated the expressions of CD80, CD86 and MHC II, and suppressed the antigen-presenting ability of DCs. The expressions of TNF-α, IL-1α and IL-6 were downregulated, while the expression of TGF-β was raised. Finally, SB203580 inhibited DCs-mediated proliferation of OT-II cells.
Conclusion:
Blocking p38MAPK pathway with SB203580 could inhibit DCs-mediated proliferation of OT-II cells, which might be involved in modulating the expressions of CD80, CD86 and MHC II, the antigen-presenting ability, as well as the expressions of pro-inflammatory and anti-inflammatory cytokines.
Insights
Blocking the p38 mitogen-activated protein kinase (p38MAPK) pathway with SB203580 inhibits dendritic cell-mediated OT-II cell proliferation. This effect involves altered co-stimulatory molecule expression and cytokine profiles, impacting T cell activation.
Area of Science:
- Immunology
- Cell Biology
- Molecular Biology
Background:
- Dendritic cells (DCs) are crucial for T cell activation.
- The p38 mitogen-activated protein kinase (p38MAPK) pathway plays a role in immune cell function.
- Understanding DC-T cell interactions is vital for immune response modulation.
Purpose of the Study:
- To investigate the impact of inhibiting the p38MAPK pathway on dendritic cell (DC)-mediated OT-II cell proliferation.
- To determine how p38MAPK inhibition affects DC co-stimulatory molecule expression and cytokine production.
- To elucidate the role of p38MAPK in regulating T cell activation by DCs.
Main Methods:
- Purification of splenic DCs and OT-II T cells from C57BL/6 and OT-II transgenic mice, respectively.
- Treatment of DCs with SB203580 (p38MAPK inhibitor) and lipopolysaccharides (LPS).
- Flow cytometry analysis of DC co-stimulatory molecules (CD80, CD86), MHC II, and OT-II cell proliferation; ELISA for cytokine levels (TNF-α, IL-1α, IL-6, TGF-β).
Main Results:
- SB203580 treatment downregulated CD80, CD86, and MHC II expression on DCs.
- Inhibition of p38MAPK suppressed DC antigen-presenting capacity and pro-inflammatory cytokine (TNF-α, IL-1α, IL-6) production.
- TGF-β expression was increased, and OT-II cell proliferation was significantly inhibited.
Conclusions:
- Blocking the p38MAPK pathway with SB203580 effectively inhibits DC-mediated OT-II cell proliferation.
- The mechanism involves modulation of DC co-stimulatory molecules, antigen presentation, and cytokine balance.
- This finding suggests p38MAPK as a potential target for modulating immune responses involving T cell activation.
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