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Mast cells and dendritic cells form synapses that facilitate antigen transfer for T cell activation.

Carroll-Portillo A, Cannon JL, te Riet J, Holmes A, Kawakami Y, Kawakami T, Cambi A, Lidke DS - J. Cell Biol. (2015)

Bottom Line: Direct cellular contact differentially regulates the secreted cytokine profile, indicating that MC modulation of DC populations is influenced by the nature of their interaction.Synapse formation requires integrin engagement and facilitates the transfer of internalized MC-specific antigen from MCs to DCs.The transferred material is ultimately processed and presented by DCs and can activate T cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathology, The University of New Mexico School of Medicine, Albuquerque, NM 87131.

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DC interactions with actMCs alter cytokine secretion. ELISA results for supernatants from BMMC-imDC experiments at 2 and/or 12 h comparing secretion of IL-1ra, MIP-1α, MCP-1, and TNF under different conditions were plotted. Results are representative of data from ≥3 independent experiments for each cytokine tested. Samples are as follows: DC, DCs alone; DC+DNP, DCs with DNP-BSA; MC, MC alone; actMC, MCs activated with DNP-BSA; DC+MC, co-culture of MC and DCs; DC+actMC, co-culture of DNP-BSA activated MCs and DCs; DC+MC_TW, DCs and MCs separated by a transwell; and DC+actMC_TW, activated MCs separated from DCs by a transwell. Bonferroni’s multiple comparison tests were performed to determine significance. P-value is relative to the number of asterisks (*, P < 0.01; ***, P < 0.001; ****, P < 0.0001). For IL-1ra, changes are compared with DC sample and for MIP-1α, and MCP-1 changes are compared with actMC sample. (bottom right) TNF secretion shows the expected increase with MC activation and is not altered by DC interactions. Error bars are SEM.
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fig2: DC interactions with actMCs alter cytokine secretion. ELISA results for supernatants from BMMC-imDC experiments at 2 and/or 12 h comparing secretion of IL-1ra, MIP-1α, MCP-1, and TNF under different conditions were plotted. Results are representative of data from ≥3 independent experiments for each cytokine tested. Samples are as follows: DC, DCs alone; DC+DNP, DCs with DNP-BSA; MC, MC alone; actMC, MCs activated with DNP-BSA; DC+MC, co-culture of MC and DCs; DC+actMC, co-culture of DNP-BSA activated MCs and DCs; DC+MC_TW, DCs and MCs separated by a transwell; and DC+actMC_TW, activated MCs separated from DCs by a transwell. Bonferroni’s multiple comparison tests were performed to determine significance. P-value is relative to the number of asterisks (*, P < 0.01; ***, P < 0.001; ****, P < 0.0001). For IL-1ra, changes are compared with DC sample and for MIP-1α, and MCP-1 changes are compared with actMC sample. (bottom right) TNF secretion shows the expected increase with MC activation and is not altered by DC interactions. Error bars are SEM.

Mentions: As immune modulators, both MCs and DCs are capable of secreting a wide variety of cytokines to stimulate or repress immune responses. To determine whether direct cell–cell interactions might influence cellular responses, cytokine secretion was measured using ELISA. MCs and imDCs were co-incubated to allow for either direct or indirect (with transwell separation) interactions. To activate MCs, FcεRI signaling was initiated by the addition of the multivalent antigen, DNP-BSA, which cross-links the DNP-specific IgE bound to the receptor. Supernatants from 2- and 12-h co-cultures were collected for ELISAs. Of the cytokines tested (IL1ra, MIP1α, MCP-1, IL-4, IL-6, IL-10, IL-12, TNF, and TIMP-1), IL-1ra, MIP1α, and MCP-1 showed consistent changes during the co-incubation (Fig. 2). We found several different modes by which direct MC–DC contact can modulate cytokine responses. IL-1ra secretion by DCs is significantly up-regulated only when DCs were allowed to be in direct contact with actMCs (Fig. 2, left, compare DC with DC+actMC). On the other hand, MCP-1 secreted by actMCs was down-regulated when MCs and DCs were not in contact, but direct MC–DC interaction prevented this down-regulation (Fig. 2, top right, compare DC+actMC with DC+actMC_TW). MIPα secretion by actMCs was down-regulated in the presence of DCs, but direct MC–DC interaction delayed this down-regulation (Fig. 2, center, compare 2 h and 12 h DC+actMC). We note that TNF consistently increased with activation of MCs as expected (Fig. 2, bottom, right graph). These data indicate that cytokine production by MCs and DCs is differentially regulated depending on whether direct cellular interaction occurs. We postulate that the formation of cell–cell contacts alters cell signaling, likely through activation of adhesion molecules, and leads to changes in the quality and the timing of the cytokine response.


Mast cells and dendritic cells form synapses that facilitate antigen transfer for T cell activation.

Carroll-Portillo A, Cannon JL, te Riet J, Holmes A, Kawakami Y, Kawakami T, Cambi A, Lidke DS - J. Cell Biol. (2015)

DC interactions with actMCs alter cytokine secretion. ELISA results for supernatants from BMMC-imDC experiments at 2 and/or 12 h comparing secretion of IL-1ra, MIP-1α, MCP-1, and TNF under different conditions were plotted. Results are representative of data from ≥3 independent experiments for each cytokine tested. Samples are as follows: DC, DCs alone; DC+DNP, DCs with DNP-BSA; MC, MC alone; actMC, MCs activated with DNP-BSA; DC+MC, co-culture of MC and DCs; DC+actMC, co-culture of DNP-BSA activated MCs and DCs; DC+MC_TW, DCs and MCs separated by a transwell; and DC+actMC_TW, activated MCs separated from DCs by a transwell. Bonferroni’s multiple comparison tests were performed to determine significance. P-value is relative to the number of asterisks (*, P < 0.01; ***, P < 0.001; ****, P < 0.0001). For IL-1ra, changes are compared with DC sample and for MIP-1α, and MCP-1 changes are compared with actMC sample. (bottom right) TNF secretion shows the expected increase with MC activation and is not altered by DC interactions. Error bars are SEM.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4555818&req=5

fig2: DC interactions with actMCs alter cytokine secretion. ELISA results for supernatants from BMMC-imDC experiments at 2 and/or 12 h comparing secretion of IL-1ra, MIP-1α, MCP-1, and TNF under different conditions were plotted. Results are representative of data from ≥3 independent experiments for each cytokine tested. Samples are as follows: DC, DCs alone; DC+DNP, DCs with DNP-BSA; MC, MC alone; actMC, MCs activated with DNP-BSA; DC+MC, co-culture of MC and DCs; DC+actMC, co-culture of DNP-BSA activated MCs and DCs; DC+MC_TW, DCs and MCs separated by a transwell; and DC+actMC_TW, activated MCs separated from DCs by a transwell. Bonferroni’s multiple comparison tests were performed to determine significance. P-value is relative to the number of asterisks (*, P < 0.01; ***, P < 0.001; ****, P < 0.0001). For IL-1ra, changes are compared with DC sample and for MIP-1α, and MCP-1 changes are compared with actMC sample. (bottom right) TNF secretion shows the expected increase with MC activation and is not altered by DC interactions. Error bars are SEM.
Mentions: As immune modulators, both MCs and DCs are capable of secreting a wide variety of cytokines to stimulate or repress immune responses. To determine whether direct cell–cell interactions might influence cellular responses, cytokine secretion was measured using ELISA. MCs and imDCs were co-incubated to allow for either direct or indirect (with transwell separation) interactions. To activate MCs, FcεRI signaling was initiated by the addition of the multivalent antigen, DNP-BSA, which cross-links the DNP-specific IgE bound to the receptor. Supernatants from 2- and 12-h co-cultures were collected for ELISAs. Of the cytokines tested (IL1ra, MIP1α, MCP-1, IL-4, IL-6, IL-10, IL-12, TNF, and TIMP-1), IL-1ra, MIP1α, and MCP-1 showed consistent changes during the co-incubation (Fig. 2). We found several different modes by which direct MC–DC contact can modulate cytokine responses. IL-1ra secretion by DCs is significantly up-regulated only when DCs were allowed to be in direct contact with actMCs (Fig. 2, left, compare DC with DC+actMC). On the other hand, MCP-1 secreted by actMCs was down-regulated when MCs and DCs were not in contact, but direct MC–DC interaction prevented this down-regulation (Fig. 2, top right, compare DC+actMC with DC+actMC_TW). MIPα secretion by actMCs was down-regulated in the presence of DCs, but direct MC–DC interaction delayed this down-regulation (Fig. 2, center, compare 2 h and 12 h DC+actMC). We note that TNF consistently increased with activation of MCs as expected (Fig. 2, bottom, right graph). These data indicate that cytokine production by MCs and DCs is differentially regulated depending on whether direct cellular interaction occurs. We postulate that the formation of cell–cell contacts alters cell signaling, likely through activation of adhesion molecules, and leads to changes in the quality and the timing of the cytokine response.

Bottom Line: Direct cellular contact differentially regulates the secreted cytokine profile, indicating that MC modulation of DC populations is influenced by the nature of their interaction.Synapse formation requires integrin engagement and facilitates the transfer of internalized MC-specific antigen from MCs to DCs.The transferred material is ultimately processed and presented by DCs and can activate T cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathology, The University of New Mexico School of Medicine, Albuquerque, NM 87131.

Show MeSH
Related in: MedlinePlus