Carboxymethylpachymaran enhances immunologic function of dendritic cells cultured in two kinds of hepatoma carcinoma cell line’s supernatant via nuclear factor κB/Rel pathway
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)...
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| Veröffentlicht in: | Chinese journal of integrative medicine 2012-03, Vol.18 (3), p.203-208 |
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| container_title | Chinese journal of integrative medicine |
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| creator | Chen, Zhuo Yu, Bin Wu, Xian-lin Dai, Cong-qi Qian, Guo-qiang Yu, Jian-zhong He, Hai-bin Wang, Zhi-xin Hou, Jun Chen, Xiao-yin |
| description | 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 |
| doi_str_mv | 10.1007/s11655-011-0943-4 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_963828248</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>963828248</sourcerecordid><originalsourceid>FETCH-LOGICAL-c343t-cebdfe09a67860db24fdf3a73ef56a034ff15a35697c52b595a21f135ab485953</originalsourceid><addsrcrecordid>eNp9kTtuFTEYhUcIREJgATTIHdUQP-dRwhUvKRISgtr6x2NnHDz2xY-E6dgGy6BlESyCleDLDZRUPrbPOfr1f03zmOBnBOP-PBHSCdFiQlo8ctbyO80pGUfWYo7p3aq7nlZNxEnzIKUrjEXfYXG_OaGUd93IxWnzfQdxCl-2Vedlc3tQy7ZCBI-0X8ArnZBd1-KDC5dWIVO8yjZ4FAyatZ-jzfVVaecSUsXlEvWMrEf5JqBP1s_pYFz0HnJYASmIyvo_qiaQs17_-votoVT2OnrI4DO6toB8UU5DRAZUDhH9_PHi_L12qLYsN7A9bO4ZcEk_uj3Pmo-vXn7YvWkv3r1-u3t-0SrGWW6Vnmaj8QhdP3R4nig3s2HQM21EB5hxY4gAJrqxV4JOYhRAiSFMwMSHemNnzdNj7z6Gz0WnLFebDoOD16EkOXZsoAPlQ3WSo1PFkFLURu6jrVvcJMHyAEoeQckKSh5ASV4zT27by7Tq-V_iL5lqoEdDql_-Ukd5FUrdkkv_af0NuISlFw</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>963828248</pqid></control><display><type>article</type><title>Carboxymethylpachymaran enhances immunologic function of dendritic cells cultured in two kinds of hepatoma carcinoma cell line’s supernatant via nuclear factor κB/Rel pathway</title><source>MEDLINE</source><source>SpringerNature Journals</source><source>Alma/SFX Local Collection</source><creator>Chen, Zhuo ; Yu, Bin ; Wu, Xian-lin ; Dai, Cong-qi ; Qian, Guo-qiang ; Yu, Jian-zhong ; He, Hai-bin ; Wang, Zhi-xin ; Hou, Jun ; Chen, Xiao-yin</creator><creatorcontrib>Chen, Zhuo ; Yu, Bin ; Wu, Xian-lin ; Dai, Cong-qi ; Qian, Guo-qiang ; Yu, Jian-zhong ; He, Hai-bin ; Wang, Zhi-xin ; Hou, Jun ; Chen, Xiao-yin</creatorcontrib><description>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.</description><identifier>ISSN: 1672-0415</identifier><identifier>EISSN: 1993-0402</identifier><identifier>DOI: 10.1007/s11655-011-0943-4</identifier><identifier>PMID: 22466945</identifier><language>eng</language><publisher>Heidelberg: Chinese Association of Traditional and Western Medicine</publisher><subject>Carcinoma, Hepatocellular - pathology ; Carcinoma, Hepatocellular - ultrastructure ; Cell Line, Tumor ; Cell Shape ; Dendritic Cells - drug effects ; Dendritic Cells - immunology ; Glucans - pharmacology ; Humans ; Immunophenotyping ; Interferon-gamma - metabolism ; Interleukin-12 - metabolism ; Liver Neoplasms - pathology ; Liver Neoplasms - ultrastructure ; Lymphocyte Culture Test, Mixed ; Medicine ; Medicine & Public Health ; Original Article ; Signal Transduction - drug effects ; Subcellular Fractions - drug effects ; Transcription Factor RelA - metabolism</subject><ispartof>Chinese journal of integrative medicine, 2012-03, Vol.18 (3), p.203-208</ispartof><rights>Chinese Association of the Integration of Traditional and Western Medicine and Springer-Verlag Berlin Heidelberg 2011</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c343t-cebdfe09a67860db24fdf3a73ef56a034ff15a35697c52b595a21f135ab485953</citedby><cites>FETCH-LOGICAL-c343t-cebdfe09a67860db24fdf3a73ef56a034ff15a35697c52b595a21f135ab485953</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s11655-011-0943-4$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s11655-011-0943-4$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27924,27925,41488,42557,51319</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/22466945$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Chen, Zhuo</creatorcontrib><creatorcontrib>Yu, Bin</creatorcontrib><creatorcontrib>Wu, Xian-lin</creatorcontrib><creatorcontrib>Dai, Cong-qi</creatorcontrib><creatorcontrib>Qian, Guo-qiang</creatorcontrib><creatorcontrib>Yu, Jian-zhong</creatorcontrib><creatorcontrib>He, Hai-bin</creatorcontrib><creatorcontrib>Wang, Zhi-xin</creatorcontrib><creatorcontrib>Hou, Jun</creatorcontrib><creatorcontrib>Chen, Xiao-yin</creatorcontrib><title>Carboxymethylpachymaran enhances immunologic function of dendritic cells cultured in two kinds of hepatoma carcinoma cell line’s supernatant via nuclear factor κB/Rel pathway</title><title>Chinese journal of integrative medicine</title><addtitle>Chin. J. Integr. Med</addtitle><addtitle>Chin J Integr Med</addtitle><description>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.</description><subject>Carcinoma, Hepatocellular - pathology</subject><subject>Carcinoma, Hepatocellular - ultrastructure</subject><subject>Cell Line, Tumor</subject><subject>Cell Shape</subject><subject>Dendritic Cells - drug effects</subject><subject>Dendritic Cells - immunology</subject><subject>Glucans - pharmacology</subject><subject>Humans</subject><subject>Immunophenotyping</subject><subject>Interferon-gamma - metabolism</subject><subject>Interleukin-12 - metabolism</subject><subject>Liver Neoplasms - pathology</subject><subject>Liver Neoplasms - ultrastructure</subject><subject>Lymphocyte Culture Test, Mixed</subject><subject>Medicine</subject><subject>Medicine & Public Health</subject><subject>Original Article</subject><subject>Signal Transduction - drug effects</subject><subject>Subcellular Fractions - drug effects</subject><subject>Transcription Factor RelA - metabolism</subject><issn>1672-0415</issn><issn>1993-0402</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kTtuFTEYhUcIREJgATTIHdUQP-dRwhUvKRISgtr6x2NnHDz2xY-E6dgGy6BlESyCleDLDZRUPrbPOfr1f03zmOBnBOP-PBHSCdFiQlo8ctbyO80pGUfWYo7p3aq7nlZNxEnzIKUrjEXfYXG_OaGUd93IxWnzfQdxCl-2Vedlc3tQy7ZCBI-0X8ArnZBd1-KDC5dWIVO8yjZ4FAyatZ-jzfVVaecSUsXlEvWMrEf5JqBP1s_pYFz0HnJYASmIyvo_qiaQs17_-votoVT2OnrI4DO6toB8UU5DRAZUDhH9_PHi_L12qLYsN7A9bO4ZcEk_uj3Pmo-vXn7YvWkv3r1-u3t-0SrGWW6Vnmaj8QhdP3R4nig3s2HQM21EB5hxY4gAJrqxV4JOYhRAiSFMwMSHemNnzdNj7z6Gz0WnLFebDoOD16EkOXZsoAPlQ3WSo1PFkFLURu6jrVvcJMHyAEoeQckKSh5ASV4zT27by7Tq-V_iL5lqoEdDql_-Ukd5FUrdkkv_af0NuISlFw</recordid><startdate>20120301</startdate><enddate>20120301</enddate><creator>Chen, Zhuo</creator><creator>Yu, Bin</creator><creator>Wu, Xian-lin</creator><creator>Dai, Cong-qi</creator><creator>Qian, Guo-qiang</creator><creator>Yu, Jian-zhong</creator><creator>He, Hai-bin</creator><creator>Wang, Zhi-xin</creator><creator>Hou, Jun</creator><creator>Chen, Xiao-yin</creator><general>Chinese Association of Traditional and Western Medicine</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>20120301</creationdate><title>Carboxymethylpachymaran enhances immunologic function of dendritic cells cultured in two kinds of hepatoma carcinoma cell line’s supernatant via nuclear factor κB/Rel pathway</title><author>Chen, Zhuo ; Yu, Bin ; Wu, Xian-lin ; Dai, Cong-qi ; Qian, Guo-qiang ; Yu, Jian-zhong ; He, Hai-bin ; Wang, Zhi-xin ; Hou, Jun ; Chen, Xiao-yin</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c343t-cebdfe09a67860db24fdf3a73ef56a034ff15a35697c52b595a21f135ab485953</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>Carcinoma, Hepatocellular - pathology</topic><topic>Carcinoma, Hepatocellular - ultrastructure</topic><topic>Cell Line, Tumor</topic><topic>Cell Shape</topic><topic>Dendritic Cells - drug effects</topic><topic>Dendritic Cells - immunology</topic><topic>Glucans - pharmacology</topic><topic>Humans</topic><topic>Immunophenotyping</topic><topic>Interferon-gamma - metabolism</topic><topic>Interleukin-12 - metabolism</topic><topic>Liver Neoplasms - pathology</topic><topic>Liver Neoplasms - ultrastructure</topic><topic>Lymphocyte Culture Test, Mixed</topic><topic>Medicine</topic><topic>Medicine & Public Health</topic><topic>Original Article</topic><topic>Signal Transduction - drug effects</topic><topic>Subcellular Fractions - drug effects</topic><topic>Transcription Factor RelA - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Chen, Zhuo</creatorcontrib><creatorcontrib>Yu, Bin</creatorcontrib><creatorcontrib>Wu, Xian-lin</creatorcontrib><creatorcontrib>Dai, Cong-qi</creatorcontrib><creatorcontrib>Qian, Guo-qiang</creatorcontrib><creatorcontrib>Yu, Jian-zhong</creatorcontrib><creatorcontrib>He, Hai-bin</creatorcontrib><creatorcontrib>Wang, Zhi-xin</creatorcontrib><creatorcontrib>Hou, Jun</creatorcontrib><creatorcontrib>Chen, Xiao-yin</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Chinese journal of integrative medicine</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Chen, Zhuo</au><au>Yu, Bin</au><au>Wu, Xian-lin</au><au>Dai, Cong-qi</au><au>Qian, Guo-qiang</au><au>Yu, Jian-zhong</au><au>He, Hai-bin</au><au>Wang, Zhi-xin</au><au>Hou, Jun</au><au>Chen, Xiao-yin</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Carboxymethylpachymaran enhances immunologic function of dendritic cells cultured in two kinds of hepatoma carcinoma cell line’s supernatant via nuclear factor κB/Rel pathway</atitle><jtitle>Chinese journal of integrative medicine</jtitle><stitle>Chin. J. Integr. Med</stitle><addtitle>Chin J Integr Med</addtitle><date>2012-03-01</date><risdate>2012</risdate><volume>18</volume><issue>3</issue><spage>203</spage><epage>208</epage><pages>203-208</pages><issn>1672-0415</issn><eissn>1993-0402</eissn><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.</abstract><cop>Heidelberg</cop><pub>Chinese Association of Traditional and Western Medicine</pub><pmid>22466945</pmid><doi>10.1007/s11655-011-0943-4</doi><tpages>6</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1672-0415 |
| ispartof | Chinese journal of integrative medicine, 2012-03, Vol.18 (3), p.203-208 |
| issn | 1672-0415 1993-0402 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_963828248 |
| source | MEDLINE; SpringerNature Journals; Alma/SFX Local Collection |
| subjects | Carcinoma, Hepatocellular - pathology Carcinoma, Hepatocellular - ultrastructure Cell Line, Tumor Cell Shape Dendritic Cells - drug effects Dendritic Cells - immunology Glucans - pharmacology Humans Immunophenotyping Interferon-gamma - metabolism Interleukin-12 - metabolism Liver Neoplasms - pathology Liver Neoplasms - ultrastructure Lymphocyte Culture Test, Mixed Medicine Medicine & Public Health Original Article Signal Transduction - drug effects Subcellular Fractions - drug effects Transcription Factor RelA - metabolism |
| title | 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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