Immunomodulatory effects of β-defensin 2 on macrophages induced immuno-upregulation and their antitumor function in breast cancer
Abstract Background Macrophages are mononuclear CD34+ antigen-presenting cells of defense mechanism and play dual roles in tumor burden. The immunomodulatory and their antitumor function of β-defensin 2 is still unclear, despite the accumulating evidence of the response in infection. So, the aim of...
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| Format: | Article |
| Language: | English |
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BMC
2022-11-01
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| Series: | BMC Immunology |
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| Online Access: | https://doi.org/10.1186/s12865-022-00527-y |
| _version_ | 1798044405920169984 |
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| author | Sonam Agarwal Anita Chauhan Khushwant Singh Kunal Kumar Rupinder Kaur Marilyn Masih Pramod Kumar Gautam |
| author_facet | Sonam Agarwal Anita Chauhan Khushwant Singh Kunal Kumar Rupinder Kaur Marilyn Masih Pramod Kumar Gautam |
| author_sort | Sonam Agarwal |
| collection | DOAJ |
| description | Abstract Background Macrophages are mononuclear CD34+ antigen-presenting cells of defense mechanism and play dual roles in tumor burden. The immunomodulatory and their antitumor function of β-defensin 2 is still unclear, despite the accumulating evidence of the response in infection. So, the aim of present study is to elucidate the role of β-defensin 2 on the level of ROS, cytokines, chemokine expression in macrophages and antitumor function in breast cancer. Method Swiss albino mice were used to harvest PEC macrophages and C127i breast cancer cells line for tumor model was used in this study. Macrophages were harvested and characterized by flow-cytometry using F4/80 and CD11c antibodies. MTT was performed to estimate cytotoxicity and dose optimization of β-defensin 2. Oxidative stress was analyzed by H2O2 and NO estimation followed by iNOS quantified by q-PCR. Cytokines and chemokines estimation was done using q-PCR. Co-culture experiment was performed to study anti-tumor function using PI for cell cycle, Annexin –V and CFSE analysis for cell proliferation. Results PEC harvested macrophages were characterized by flow-cytometry using F4/80 and CD11c antibodies with the purity of 8% pure population of macrophages. It was found that 99% of cells viable at the maximum dose of 100 ng/ml of β-defensin 2 in MTT. Levels of NO and H2O2 were found to be decreased in β-defensin 2 as compared to control. Expression of cytokines of IFN-γ, IL-1α, TNF-α, TGF-βwas found to be increased while IL-3 was decreased in β-defensin 2 group as compared to control. Levels of chemokines CXCL-1, CXCL-5 and CCL5 increased in treated macrophages while CCL24 and CXCL-15 expression decreased. Adhesion receptor (CD32) and fusion receptor (CD204) were decreased in the β-defensin 2 group as compared to control. Anti-tumor experiment was performed using co-culture experiment apoptosis (Annexin-V) was induced, cell cycle arrest in phage and cell proliferation of C127i cells was decreased. Conclusion This is the first report of β-defensin 2 modulates macrophage immunomodulatory and their antitumor function in breast cancer. β-defensin 2 as a new therapeutic target for immunotherapy as an adjuvant in vaccines. |
| first_indexed | 2024-04-11T23:03:16Z |
| format | Article |
| id | doaj.art-ec199212603642ba98d477de1eab15bb |
| institution | Directory Open Access Journal |
| issn | 1471-2172 |
| language | English |
| last_indexed | 2024-04-11T23:03:16Z |
| publishDate | 2022-11-01 |
| publisher | BMC |
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| series | BMC Immunology |
| spelling | doaj.art-ec199212603642ba98d477de1eab15bb2022-12-22T03:58:04ZengBMCBMC Immunology1471-21722022-11-0123111210.1186/s12865-022-00527-yImmunomodulatory effects of β-defensin 2 on macrophages induced immuno-upregulation and their antitumor function in breast cancerSonam Agarwal0Anita Chauhan1Khushwant Singh2Kunal Kumar3Rupinder Kaur4Marilyn Masih5Pramod Kumar Gautam6Department of Biochemistry, All India Institute of Medical SciencesDepartment of Biochemistry, All India Institute of Medical SciencesDepartment of Biochemistry, All India Institute of Medical SciencesDepartment of Biochemistry, All India Institute of Medical SciencesDepartment of Biochemistry, All India Institute of Medical SciencesDepartment of Biochemistry, All India Institute of Medical SciencesDepartment of Biochemistry, All India Institute of Medical SciencesAbstract Background Macrophages are mononuclear CD34+ antigen-presenting cells of defense mechanism and play dual roles in tumor burden. The immunomodulatory and their antitumor function of β-defensin 2 is still unclear, despite the accumulating evidence of the response in infection. So, the aim of present study is to elucidate the role of β-defensin 2 on the level of ROS, cytokines, chemokine expression in macrophages and antitumor function in breast cancer. Method Swiss albino mice were used to harvest PEC macrophages and C127i breast cancer cells line for tumor model was used in this study. Macrophages were harvested and characterized by flow-cytometry using F4/80 and CD11c antibodies. MTT was performed to estimate cytotoxicity and dose optimization of β-defensin 2. Oxidative stress was analyzed by H2O2 and NO estimation followed by iNOS quantified by q-PCR. Cytokines and chemokines estimation was done using q-PCR. Co-culture experiment was performed to study anti-tumor function using PI for cell cycle, Annexin –V and CFSE analysis for cell proliferation. Results PEC harvested macrophages were characterized by flow-cytometry using F4/80 and CD11c antibodies with the purity of 8% pure population of macrophages. It was found that 99% of cells viable at the maximum dose of 100 ng/ml of β-defensin 2 in MTT. Levels of NO and H2O2 were found to be decreased in β-defensin 2 as compared to control. Expression of cytokines of IFN-γ, IL-1α, TNF-α, TGF-βwas found to be increased while IL-3 was decreased in β-defensin 2 group as compared to control. Levels of chemokines CXCL-1, CXCL-5 and CCL5 increased in treated macrophages while CCL24 and CXCL-15 expression decreased. Adhesion receptor (CD32) and fusion receptor (CD204) were decreased in the β-defensin 2 group as compared to control. Anti-tumor experiment was performed using co-culture experiment apoptosis (Annexin-V) was induced, cell cycle arrest in phage and cell proliferation of C127i cells was decreased. Conclusion This is the first report of β-defensin 2 modulates macrophage immunomodulatory and their antitumor function in breast cancer. β-defensin 2 as a new therapeutic target for immunotherapy as an adjuvant in vaccines.https://doi.org/10.1186/s12865-022-00527-yMacrophagesβ-defensin 2Cytokine profilingAnti-tumor functionOxidative stress |
| spellingShingle | Sonam Agarwal Anita Chauhan Khushwant Singh Kunal Kumar Rupinder Kaur Marilyn Masih Pramod Kumar Gautam Immunomodulatory effects of β-defensin 2 on macrophages induced immuno-upregulation and their antitumor function in breast cancer BMC Immunology Macrophages β-defensin 2 Cytokine profiling Anti-tumor function Oxidative stress |
| title | Immunomodulatory effects of β-defensin 2 on macrophages induced immuno-upregulation and their antitumor function in breast cancer |
| title_full | Immunomodulatory effects of β-defensin 2 on macrophages induced immuno-upregulation and their antitumor function in breast cancer |
| title_fullStr | Immunomodulatory effects of β-defensin 2 on macrophages induced immuno-upregulation and their antitumor function in breast cancer |
| title_full_unstemmed | Immunomodulatory effects of β-defensin 2 on macrophages induced immuno-upregulation and their antitumor function in breast cancer |
| title_short | Immunomodulatory effects of β-defensin 2 on macrophages induced immuno-upregulation and their antitumor function in breast cancer |
| title_sort | immunomodulatory effects of β defensin 2 on macrophages induced immuno upregulation and their antitumor function in breast cancer |
| topic | Macrophages β-defensin 2 Cytokine profiling Anti-tumor function Oxidative stress |
| url | https://doi.org/10.1186/s12865-022-00527-y |
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