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Carboxymethylpachymaran enhances immunologic function of dendritic cells cultured in two kinds of hepatoma carcinoma cell line’s supernatant via nuclear factor κB/Rel pathway

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Abstract

Objective

To study the immunologic function of dendritic cells (DCs) cultured in two kinds of hepatoma cell line’s supernatant and the enhancing effects of carboxymethylpachymaran (CMP) on DCs.

Methods

DCs were harvested after stimulation by granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-4 from umbilical cord blood using density-gradient centrifugation method. Cultured supernatant of two hepatoma cell lines (HepG2 and HepG2.2.15) were collected for condition medium (CM) according to a volume ratio of supernatant to incomplete RPMI-1640 medium, which was 3:1. CMP was dissolved in incomplete RPMI-1640 medium. Experimental groups were divided according to the culture medium, either CM or with CMP in it. DCs subsets CD83, CD86, CD1a, and d-related human leukocyte antigens (HLA-DR) were analyzed by flow cytometry. The proliferation ability of allogeneic T cells in mixed lymphocyte reaction (MLR) stimulated by DCs was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) analysis. IL-12p70, interferon-γ (IFN-γ), and nuclear factor κB (NF-κB) were detected by enzyme-linked immunosorbent assay analysis.

Results

The proliferation of lymphocytes and secreting level of IL-12 and expression of phenotype of DCs cultured in two kinds of CM were lower than those of normal group (P <0.01). Compared with the normal group, groups treated with CMP showed a higher expression level of DCs subsets, lymphocyte reproductive activity, as well as IL-12 and IFN-γ secretion levels. Groups treated with CMP also demonstrated higher levels of DCs phenotype expression and IL-12 and IFN-γ secretion in supernatant of MLR and higher lymphocyte reproductive activity compared with CM group (P <0.05). Compared with the normal group, the expression level of NF-κB in DCs nuclear was higher in CMP groups but lower in two CM groups (P <0.05). After CMP was added, the NF-κB expression levels of two CM groups were increased compared with levels before CMP was added (P <0.05). However, there was no significant difference between the two CM groups (P >0.05).

Conclusions

Two kinds of hepatoma cell line’s supernatant can inhibit the immunologic function of DCs. This suppressive effect may be related to the inhibition of NF-κB/Rel pathway. CMP may up-regulate the DCs function by activating the NF-κB/Rel pathway.

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Authors and Affiliations

  1. Medical College, Jinan University, Guangzhou, 510632, China

    Zhuo Chen  (陈 卓), Bin Yu  (于 斌), Xian-lin Wu  (吴先林), Cong-qi Dai  (戴聪奇) & Xiao-yin Chen  (陈孝银)

  2. Nursing Department, Huanghuai University, Zhumadian, Henan Province, 463000, China

    Guo-qiang Qian  (钱国强)

  3. Tongrentang Hospital, Beijing, 10051, China

    Jian-zhong Yu  (于建中)

  4. Heshan People Hospital, Guangzhou, 529700, China

    Hai-bin He  (何海滨)

  5. Dongpu Branch of the First Affiliated Hospital of Jinan University, Guangzhou, 511630, China

    Zhi-xin Wang  (王志新) & Jun Hou  (侯 军)

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  1. Zhuo Chen  (陈 卓)
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  2. Bin Yu  (于 斌)
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  5. Guo-qiang Qian  (钱国强)
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  6. Jian-zhong Yu  (于建中)
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  7. Hai-bin He  (何海滨)
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  8. Zhi-xin Wang  (王志新)
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  9. Jun Hou  (侯 军)
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  10. Xiao-yin Chen  (陈孝银)
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Corresponding author

Correspondence to Xiao-yin Chen  (陈孝银).

Additional information

Supported by the National Natural Science Foundation of China (No. 30973693) and the Research Fund for the Doctoral Program of Higher Education of China (No. 20104401110003)

These authors contributed equally to this work

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Chen, Z., Yu, B., Wu, Xl. et al. Carboxymethylpachymaran enhances immunologic function of dendritic cells cultured in two kinds of hepatoma carcinoma cell line’s supernatant via nuclear factor κB/Rel pathway. Chin. J. Integr. Med. 18, 203–208 (2012). https://doi.org/10.1007/s11655-011-0943-4

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  • DOI: https://doi.org/10.1007/s11655-011-0943-4

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