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Effects of histamine and its antagonists on murine T-cells and bone marrow-derived dendritic cells.

Hu X, Zafar MI, Gao F - Drug Des Devel Ther (2015)

Bottom Line: LPS increased IL-10 and decreased IL-12 levels.GM-CSF + IL-4-generated DCs had a stronger stimulatory effect on DO11.10 T-cell proliferation than GM-CSF-generated DCs.Inducible costimulator ligand expression was higher in GM-CSF + IL-4- than in GM-CSF-generated DC groups after 2 days of coculture, but decreased 4 days later.

View Article: PubMed Central - PubMed

Affiliation: Department of Paediatrics, Tongji Hospital, Huazhong University of Science and Technology, Wuhan, People's Republic of China.

ABSTRACT
We determined the effects of histamine and its antagonists on the surface marker expression of dendritic cells (DCs) and the influence of lipopolysaccharide (LPS), histamine, and histamine receptor antagonists on DCs and T-cells. The bone marrow was extracted from the femurs and tibiae of 6- to 8-week-old female Balb/c mice and cultured in medium containing penicillin, streptomycin, L-glutamine, fetal calf serum, or granulocyte macrophage colony-stimulating factor (GM-CSF) alone or with interleukin (IL)-4. The cells received three different doses of LPS and histamine, plus three different doses of descarboethoxyloratadine (DCL). We assayed the supernatant for various cytokines. The spleen cells of DO11.10 mice were examined by flow cytometry, which included labeling and sorting CD4+ T-cells, as well as coculture of DCs and T-cells with ovalbumin (OVA)323-339 peptide. Histamine or histamine plus DCL did not affect the expression of major histocompatibility complex class II, CD11c, CD11b, CD86, and CD80. However, GM-CSF increased the expression of all markers except CD80. Histamine increased interferon-γ production in GM-CSF + IL-4-cultured cells; it also enhanced IL-10 production, but suppressed IL-12 production in LPS-stimulated DCs with no DCL. Cimetidine inhibited IL-10 production and restored IL-12 secretion in LPS-treated DCs. LPS increased IL-10 and decreased IL-12 levels. GM-CSF + IL-4-generated DCs had a stronger stimulatory effect on DO11.10 T-cell proliferation than GM-CSF-generated DCs. Inducible costimulator ligand expression was higher in GM-CSF + IL-4- than in GM-CSF-generated DC groups after 2 days of coculture, but decreased 4 days later. IL-13 production was higher in bone marrow DCs generated with GM-CSF than in those generated with GM-CSF + IL-4. OVA-pulsed DCs and OVA-plus-DCL DCs showed increased IL-12 levels. OVA plus LPS increased both IL-10 and interferon-α. Although histamine or histamine receptor-1 antagonists did not influence DC LPS-driven maturation, they influenced cytokine production. LPS and GM-CSF influenced surface marker expression and cytokine production.

No MeSH data available.


Related in: MedlinePlus

Influence of DCL on the proliferation of DO11.10 T-cells through OVA-pulsed BMDCs.Notes: The BMDCs were generated by GM-CSF only or GM-CSF plus IL-4 and cocultivated with CFDA-labeled CD4+ DO11.10 cells for 2 or 4 days. The proliferation of DO11.10 T-cells was assayed by FACS staining.Abbreviations: BMDCs, bone marrow-derived dendritic cells; CFDA, carboxyfluocein diacetate; DCL, descarboethoxyloratadine; FACS, fluorescence-activated cell sorting; GM-CSF, granulocyte macrophage colony-stimulating factor; ICOS, inducible costimulator; IL, interleukin; OVA, ovalbumin.
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f11-dddt-9-4847: Influence of DCL on the proliferation of DO11.10 T-cells through OVA-pulsed BMDCs.Notes: The BMDCs were generated by GM-CSF only or GM-CSF plus IL-4 and cocultivated with CFDA-labeled CD4+ DO11.10 cells for 2 or 4 days. The proliferation of DO11.10 T-cells was assayed by FACS staining.Abbreviations: BMDCs, bone marrow-derived dendritic cells; CFDA, carboxyfluocein diacetate; DCL, descarboethoxyloratadine; FACS, fluorescence-activated cell sorting; GM-CSF, granulocyte macrophage colony-stimulating factor; ICOS, inducible costimulator; IL, interleukin; OVA, ovalbumin.

Mentions: To examine the effects of LPS, histamine, and histamine antagonists on the capability of DCs to induce proliferation of antigen-specific CD4+ T-cells, a T-cell proliferation assay was performed by using CD4+ DO11.10 T-cells as responder cells in the presence of the OVA323–339 peptide. FACS analysis showed that DCL had no effect on the proliferation of DO11.10 T-cells that had been cocultured with OVA-pulsed BMDCs for 2 days, but DCL stimulated their proliferation after coculture for 4 days (Figure 11). Moreover, GM-CSF + IL-4 caused DCs to exhibit a stronger stimulatory capacity in DO.11.10 T-cell proliferation than GM-CSF-generated DCs. Similarly, LPS, histamine, and cimetidine (Figure 12) all generated antigen-specific T-cell proliferation to an extent similar to DCL on day 4. The M1 fraction (cells that did not proliferate at the time point of measurement) was reduced after DCL stimulation compared with that in cells pulsed with OVA/LPS.


Effects of histamine and its antagonists on murine T-cells and bone marrow-derived dendritic cells.

Hu X, Zafar MI, Gao F - Drug Des Devel Ther (2015)

Influence of DCL on the proliferation of DO11.10 T-cells through OVA-pulsed BMDCs.Notes: The BMDCs were generated by GM-CSF only or GM-CSF plus IL-4 and cocultivated with CFDA-labeled CD4+ DO11.10 cells for 2 or 4 days. The proliferation of DO11.10 T-cells was assayed by FACS staining.Abbreviations: BMDCs, bone marrow-derived dendritic cells; CFDA, carboxyfluocein diacetate; DCL, descarboethoxyloratadine; FACS, fluorescence-activated cell sorting; GM-CSF, granulocyte macrophage colony-stimulating factor; ICOS, inducible costimulator; IL, interleukin; OVA, ovalbumin.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4552258&req=5

f11-dddt-9-4847: Influence of DCL on the proliferation of DO11.10 T-cells through OVA-pulsed BMDCs.Notes: The BMDCs were generated by GM-CSF only or GM-CSF plus IL-4 and cocultivated with CFDA-labeled CD4+ DO11.10 cells for 2 or 4 days. The proliferation of DO11.10 T-cells was assayed by FACS staining.Abbreviations: BMDCs, bone marrow-derived dendritic cells; CFDA, carboxyfluocein diacetate; DCL, descarboethoxyloratadine; FACS, fluorescence-activated cell sorting; GM-CSF, granulocyte macrophage colony-stimulating factor; ICOS, inducible costimulator; IL, interleukin; OVA, ovalbumin.
Mentions: To examine the effects of LPS, histamine, and histamine antagonists on the capability of DCs to induce proliferation of antigen-specific CD4+ T-cells, a T-cell proliferation assay was performed by using CD4+ DO11.10 T-cells as responder cells in the presence of the OVA323–339 peptide. FACS analysis showed that DCL had no effect on the proliferation of DO11.10 T-cells that had been cocultured with OVA-pulsed BMDCs for 2 days, but DCL stimulated their proliferation after coculture for 4 days (Figure 11). Moreover, GM-CSF + IL-4 caused DCs to exhibit a stronger stimulatory capacity in DO.11.10 T-cell proliferation than GM-CSF-generated DCs. Similarly, LPS, histamine, and cimetidine (Figure 12) all generated antigen-specific T-cell proliferation to an extent similar to DCL on day 4. The M1 fraction (cells that did not proliferate at the time point of measurement) was reduced after DCL stimulation compared with that in cells pulsed with OVA/LPS.

Bottom Line: LPS increased IL-10 and decreased IL-12 levels.GM-CSF + IL-4-generated DCs had a stronger stimulatory effect on DO11.10 T-cell proliferation than GM-CSF-generated DCs.Inducible costimulator ligand expression was higher in GM-CSF + IL-4- than in GM-CSF-generated DC groups after 2 days of coculture, but decreased 4 days later.

View Article: PubMed Central - PubMed

Affiliation: Department of Paediatrics, Tongji Hospital, Huazhong University of Science and Technology, Wuhan, People's Republic of China.

ABSTRACT
We determined the effects of histamine and its antagonists on the surface marker expression of dendritic cells (DCs) and the influence of lipopolysaccharide (LPS), histamine, and histamine receptor antagonists on DCs and T-cells. The bone marrow was extracted from the femurs and tibiae of 6- to 8-week-old female Balb/c mice and cultured in medium containing penicillin, streptomycin, L-glutamine, fetal calf serum, or granulocyte macrophage colony-stimulating factor (GM-CSF) alone or with interleukin (IL)-4. The cells received three different doses of LPS and histamine, plus three different doses of descarboethoxyloratadine (DCL). We assayed the supernatant for various cytokines. The spleen cells of DO11.10 mice were examined by flow cytometry, which included labeling and sorting CD4+ T-cells, as well as coculture of DCs and T-cells with ovalbumin (OVA)323-339 peptide. Histamine or histamine plus DCL did not affect the expression of major histocompatibility complex class II, CD11c, CD11b, CD86, and CD80. However, GM-CSF increased the expression of all markers except CD80. Histamine increased interferon-γ production in GM-CSF + IL-4-cultured cells; it also enhanced IL-10 production, but suppressed IL-12 production in LPS-stimulated DCs with no DCL. Cimetidine inhibited IL-10 production and restored IL-12 secretion in LPS-treated DCs. LPS increased IL-10 and decreased IL-12 levels. GM-CSF + IL-4-generated DCs had a stronger stimulatory effect on DO11.10 T-cell proliferation than GM-CSF-generated DCs. Inducible costimulator ligand expression was higher in GM-CSF + IL-4- than in GM-CSF-generated DC groups after 2 days of coculture, but decreased 4 days later. IL-13 production was higher in bone marrow DCs generated with GM-CSF than in those generated with GM-CSF + IL-4. OVA-pulsed DCs and OVA-plus-DCL DCs showed increased IL-12 levels. OVA plus LPS increased both IL-10 and interferon-α. Although histamine or histamine receptor-1 antagonists did not influence DC LPS-driven maturation, they influenced cytokine production. LPS and GM-CSF influenced surface marker expression and cytokine production.

No MeSH data available.


Related in: MedlinePlus