Effect of dendritic cells loaded with HBsAg on the differentiation of HBsAg-specific T helper 1 cells in patients with chronic hepatitis B

Qiong Wu, Xiao Ying Wang, Xiao Rong Zhou, Wei Tang, Li Su

Research output: Contribution to journalArticle

Abstract

AIM: To investigate the influence of dendritic cells (DCs) loaded with HBsAg from peripheral blood monocytes of patients with chronic hepatitis B (CHB) on the differentiation of autologous T helper 1 (Th1). METHODS: Mononuclear cells were isolated from CHB patients using the methods of density gradient centrifugation (Ficoll-Hypaque) and adherence. The adherent monocytes were incubated with recombinant human GM-CSF and interleukin-4 (IL-4) to induce DC generation. The phenotypes on DCs including CD80, CD86, CD40 and HLA-DR were determined by flow cytometry (FCM). DCs' ability to stimulate allogenic mixed lymphocyte reaction (AMLR) was detected by CCK-8 assay. The concentration of IL-12 in DC supernate was tested by enzyme linked immunosorbent assay (ELISA). CD4+ Th cell subpopulation in peripheral blood of CHB patients was isolated and purified by immunomagnetic beads. Subsequently, purified CD4 + Th cells were co-cultured with autologous mature DCs. The differentiation of Th1/Th2 was detected by FCM using intracellular staining of interferon-γ (IFN-γ) and IL-4. ELISA assay was also used to quantify the IFN-γ and IL-4 in co-culture supernate. RESULTS: In comparison with those in the control group, the expression levels of CD80, CD86, CD40 and HLA-DR and the concentration of IL-12 in DC supernate of the IFN-γ group, HBsAg group and HBsAg + IFN-γ group were higher, and the capability of DCs to stimulate allogenic lymphocytes to proliferate in the three groups was also stronger. Meanwhile, the percentages of Th1 cells in CD4+ T cells and the level of IFN-γ in co-culture supernate of the IFN-γ group, HBsAg group and HBsAg + IFN-γ group were higher (Th1 cells: 10.76% ± 3.98%, 11.43% ± 4.32%, 15.28% ± 4.73% vs 7.84% ± 3.10%, P < 0.01; IFN-γ: 578 ± 47 mg/L, 496 ± 92 mg/L, 784 ± 97 mg/L vs 342 ± 34 mg/L, P < 0.05). However, the percentages of Th2 cells in CD4+ T cells and the level of IL-4 in the three groups were lower (Th2 cells: 1.43% ± 0.96%, 1.68% ± 0.16%, 0.92% ± 0.21% vs 2.61% ±1.27%, P < 0.01; IL-4:187 ± 52 mg/L, 169 ± 38 mg/L, 89 ± 37 mg/L vs 226 ± 48 mg/L, P < 0.05). The most significant effect was observed in the HBsAg + IFN-γ group. CONCLUSION: DCs loaded with HBsAg can improve the insufficient differentiation of Th1 cells resulted from dysfunction of DCs in vivo in CHB patients.

Original languageEnglish (US)
Pages (from-to)1344-1349
Number of pages6
JournalWorld Chinese Journal of Digestology
Volume16
Issue number12
StatePublished - Apr 2008

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Th1 Cells
Chronic Hepatitis B
Hepatitis B Surface Antigens
Dendritic Cells
Interferons
Interleukin-4
Th2 Cells
HLA-DR Antigens
Interleukin-12
Coculture Techniques
Monocytes
Flow Cytometry
Enzyme-Linked Immunosorbent Assay
Diatrizoate
T-Lymphocytes
Sincalide
Ficoll
Mixed Lymphocyte Culture Test
Density Gradient Centrifugation
Granulocyte-Macrophage Colony-Stimulating Factor

Keywords

  • Chronic hepatitis B
  • Cytokine
  • Dendritic cell
  • Enzyme linked immunosorbent assay
  • Flow cytometry
  • T helper 1/T helper 2

ASJC Scopus subject areas

  • Gastroenterology

Cite this

Effect of dendritic cells loaded with HBsAg on the differentiation of HBsAg-specific T helper 1 cells in patients with chronic hepatitis B. / Wu, Qiong; Wang, Xiao Ying; Zhou, Xiao Rong; Tang, Wei; Su, Li.

In: World Chinese Journal of Digestology, Vol. 16, No. 12, 04.2008, p. 1344-1349.

Research output: Contribution to journalArticle

Wu, Qiong ; Wang, Xiao Ying ; Zhou, Xiao Rong ; Tang, Wei ; Su, Li. / Effect of dendritic cells loaded with HBsAg on the differentiation of HBsAg-specific T helper 1 cells in patients with chronic hepatitis B. In: World Chinese Journal of Digestology. 2008 ; Vol. 16, No. 12. pp. 1344-1349.
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abstract = "AIM: To investigate the influence of dendritic cells (DCs) loaded with HBsAg from peripheral blood monocytes of patients with chronic hepatitis B (CHB) on the differentiation of autologous T helper 1 (Th1). METHODS: Mononuclear cells were isolated from CHB patients using the methods of density gradient centrifugation (Ficoll-Hypaque) and adherence. The adherent monocytes were incubated with recombinant human GM-CSF and interleukin-4 (IL-4) to induce DC generation. The phenotypes on DCs including CD80, CD86, CD40 and HLA-DR were determined by flow cytometry (FCM). DCs' ability to stimulate allogenic mixed lymphocyte reaction (AMLR) was detected by CCK-8 assay. The concentration of IL-12 in DC supernate was tested by enzyme linked immunosorbent assay (ELISA). CD4+ Th cell subpopulation in peripheral blood of CHB patients was isolated and purified by immunomagnetic beads. Subsequently, purified CD4 + Th cells were co-cultured with autologous mature DCs. The differentiation of Th1/Th2 was detected by FCM using intracellular staining of interferon-γ (IFN-γ) and IL-4. ELISA assay was also used to quantify the IFN-γ and IL-4 in co-culture supernate. RESULTS: In comparison with those in the control group, the expression levels of CD80, CD86, CD40 and HLA-DR and the concentration of IL-12 in DC supernate of the IFN-γ group, HBsAg group and HBsAg + IFN-γ group were higher, and the capability of DCs to stimulate allogenic lymphocytes to proliferate in the three groups was also stronger. Meanwhile, the percentages of Th1 cells in CD4+ T cells and the level of IFN-γ in co-culture supernate of the IFN-γ group, HBsAg group and HBsAg + IFN-γ group were higher (Th1 cells: 10.76{\%} ± 3.98{\%}, 11.43{\%} ± 4.32{\%}, 15.28{\%} ± 4.73{\%} vs 7.84{\%} ± 3.10{\%}, P < 0.01; IFN-γ: 578 ± 47 mg/L, 496 ± 92 mg/L, 784 ± 97 mg/L vs 342 ± 34 mg/L, P < 0.05). However, the percentages of Th2 cells in CD4+ T cells and the level of IL-4 in the three groups were lower (Th2 cells: 1.43{\%} ± 0.96{\%}, 1.68{\%} ± 0.16{\%}, 0.92{\%} ± 0.21{\%} vs 2.61{\%} ±1.27{\%}, P < 0.01; IL-4:187 ± 52 mg/L, 169 ± 38 mg/L, 89 ± 37 mg/L vs 226 ± 48 mg/L, P < 0.05). The most significant effect was observed in the HBsAg + IFN-γ group. CONCLUSION: DCs loaded with HBsAg can improve the insufficient differentiation of Th1 cells resulted from dysfunction of DCs in vivo in CHB patients.",
keywords = "Chronic hepatitis B, Cytokine, Dendritic cell, Enzyme linked immunosorbent assay, Flow cytometry, T helper 1/T helper 2",
author = "Qiong Wu and Wang, {Xiao Ying} and Zhou, {Xiao Rong} and Wei Tang and Li Su",
year = "2008",
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language = "English (US)",
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pages = "1344--1349",
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TY - JOUR

T1 - Effect of dendritic cells loaded with HBsAg on the differentiation of HBsAg-specific T helper 1 cells in patients with chronic hepatitis B

AU - Wu, Qiong

AU - Wang, Xiao Ying

AU - Zhou, Xiao Rong

AU - Tang, Wei

AU - Su, Li

PY - 2008/4

Y1 - 2008/4

N2 - AIM: To investigate the influence of dendritic cells (DCs) loaded with HBsAg from peripheral blood monocytes of patients with chronic hepatitis B (CHB) on the differentiation of autologous T helper 1 (Th1). METHODS: Mononuclear cells were isolated from CHB patients using the methods of density gradient centrifugation (Ficoll-Hypaque) and adherence. The adherent monocytes were incubated with recombinant human GM-CSF and interleukin-4 (IL-4) to induce DC generation. The phenotypes on DCs including CD80, CD86, CD40 and HLA-DR were determined by flow cytometry (FCM). DCs' ability to stimulate allogenic mixed lymphocyte reaction (AMLR) was detected by CCK-8 assay. The concentration of IL-12 in DC supernate was tested by enzyme linked immunosorbent assay (ELISA). CD4+ Th cell subpopulation in peripheral blood of CHB patients was isolated and purified by immunomagnetic beads. Subsequently, purified CD4 + Th cells were co-cultured with autologous mature DCs. The differentiation of Th1/Th2 was detected by FCM using intracellular staining of interferon-γ (IFN-γ) and IL-4. ELISA assay was also used to quantify the IFN-γ and IL-4 in co-culture supernate. RESULTS: In comparison with those in the control group, the expression levels of CD80, CD86, CD40 and HLA-DR and the concentration of IL-12 in DC supernate of the IFN-γ group, HBsAg group and HBsAg + IFN-γ group were higher, and the capability of DCs to stimulate allogenic lymphocytes to proliferate in the three groups was also stronger. Meanwhile, the percentages of Th1 cells in CD4+ T cells and the level of IFN-γ in co-culture supernate of the IFN-γ group, HBsAg group and HBsAg + IFN-γ group were higher (Th1 cells: 10.76% ± 3.98%, 11.43% ± 4.32%, 15.28% ± 4.73% vs 7.84% ± 3.10%, P < 0.01; IFN-γ: 578 ± 47 mg/L, 496 ± 92 mg/L, 784 ± 97 mg/L vs 342 ± 34 mg/L, P < 0.05). However, the percentages of Th2 cells in CD4+ T cells and the level of IL-4 in the three groups were lower (Th2 cells: 1.43% ± 0.96%, 1.68% ± 0.16%, 0.92% ± 0.21% vs 2.61% ±1.27%, P < 0.01; IL-4:187 ± 52 mg/L, 169 ± 38 mg/L, 89 ± 37 mg/L vs 226 ± 48 mg/L, P < 0.05). The most significant effect was observed in the HBsAg + IFN-γ group. CONCLUSION: DCs loaded with HBsAg can improve the insufficient differentiation of Th1 cells resulted from dysfunction of DCs in vivo in CHB patients.

AB - AIM: To investigate the influence of dendritic cells (DCs) loaded with HBsAg from peripheral blood monocytes of patients with chronic hepatitis B (CHB) on the differentiation of autologous T helper 1 (Th1). METHODS: Mononuclear cells were isolated from CHB patients using the methods of density gradient centrifugation (Ficoll-Hypaque) and adherence. The adherent monocytes were incubated with recombinant human GM-CSF and interleukin-4 (IL-4) to induce DC generation. The phenotypes on DCs including CD80, CD86, CD40 and HLA-DR were determined by flow cytometry (FCM). DCs' ability to stimulate allogenic mixed lymphocyte reaction (AMLR) was detected by CCK-8 assay. The concentration of IL-12 in DC supernate was tested by enzyme linked immunosorbent assay (ELISA). CD4+ Th cell subpopulation in peripheral blood of CHB patients was isolated and purified by immunomagnetic beads. Subsequently, purified CD4 + Th cells were co-cultured with autologous mature DCs. The differentiation of Th1/Th2 was detected by FCM using intracellular staining of interferon-γ (IFN-γ) and IL-4. ELISA assay was also used to quantify the IFN-γ and IL-4 in co-culture supernate. RESULTS: In comparison with those in the control group, the expression levels of CD80, CD86, CD40 and HLA-DR and the concentration of IL-12 in DC supernate of the IFN-γ group, HBsAg group and HBsAg + IFN-γ group were higher, and the capability of DCs to stimulate allogenic lymphocytes to proliferate in the three groups was also stronger. Meanwhile, the percentages of Th1 cells in CD4+ T cells and the level of IFN-γ in co-culture supernate of the IFN-γ group, HBsAg group and HBsAg + IFN-γ group were higher (Th1 cells: 10.76% ± 3.98%, 11.43% ± 4.32%, 15.28% ± 4.73% vs 7.84% ± 3.10%, P < 0.01; IFN-γ: 578 ± 47 mg/L, 496 ± 92 mg/L, 784 ± 97 mg/L vs 342 ± 34 mg/L, P < 0.05). However, the percentages of Th2 cells in CD4+ T cells and the level of IL-4 in the three groups were lower (Th2 cells: 1.43% ± 0.96%, 1.68% ± 0.16%, 0.92% ± 0.21% vs 2.61% ±1.27%, P < 0.01; IL-4:187 ± 52 mg/L, 169 ± 38 mg/L, 89 ± 37 mg/L vs 226 ± 48 mg/L, P < 0.05). The most significant effect was observed in the HBsAg + IFN-γ group. CONCLUSION: DCs loaded with HBsAg can improve the insufficient differentiation of Th1 cells resulted from dysfunction of DCs in vivo in CHB patients.

KW - Chronic hepatitis B

KW - Cytokine

KW - Dendritic cell

KW - Enzyme linked immunosorbent assay

KW - Flow cytometry

KW - T helper 1/T helper 2

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JO - World Chinese Journal of Digestology

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