In silico analysis and in planta production of recombinant ccl21/IL1[beta] protein and characterization of its in vitro anti-tumor and immunogenic activity
CCL21 has an essential role in anti-tumor immune activity. Epitopes of IL1[beta] have adjuvant activity without causing inflammatory responses. CCR7 and its ligands play a vital role in the immune balance; specifically, in transport of T lymphocytes and antigen-presenting cells such as dendritic cel...
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| Veröffentlicht in: | PloS one 2022-08, Vol.17 (8), p.e0261101 |
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| creator | Marashi, Hasan Beihaghi, Maria Chaboksavar, Masoud Khaksar, Samad Tehrani, Homan Abiri, Ardavan |
| description | CCL21 has an essential role in anti-tumor immune activity. Epitopes of IL1[beta] have adjuvant activity without causing inflammatory responses. CCR7 and its ligands play a vital role in the immune balance; specifically, in transport of T lymphocytes and antigen-presenting cells such as dendritic cells to the lymph nodes. This study aimed to produce epitopes of CCL21 and IL1[beta] as a recombinant protein and characterize its in vitro anti-tumor and immunogenic activity. A codon-optimized ccl21/IL1[beta] gene was designed and synthesized from human genes. Stability and binding affinity of CCL21/IL1[beta] protein and CCR7 receptor were examined through in silico analyses. The construct was introduced into N. tabacum to produce this recombinant protein and the structure and function of CCL21/IL1[beta] were examined. Purified protein from transgenic leaves generated a strong signal in SDS PAGE and western blotting assays. FTIR measurement and MALDI-TOF/TOF mass spectrography showed that ccl21/IL-1[beta] was correctly expressed in tobacco plants. Potential activity of purified CCL21/IL1[beta] in stimulating the proliferation and migration of MCF7 cancer cell line was investigated using the wound healing method. The results demonstrated a decrease in survival rate and metastasization of cancer cells in the presence of CCL21/IL1[beta], and IC.sub.50 of CCL21 on MCF7 cells was less than that of non-recombinant protein. Agarose assay on PBMCsCCR7+ showed that CCL21/IL1[beta] has biological activity and there is a distinguishable difference between chemokinetic (CCL21) and chemotactic (FBS) movements. Overall, the results suggest that CCL21/IL1[beta] could be considered an effective adjuvant in future in vivo and clinical tests. |
| doi_str_mv | 10.1371/journal.pone.0261101 |
| format | Article |
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Epitopes of IL1[beta] have adjuvant activity without causing inflammatory responses. CCR7 and its ligands play a vital role in the immune balance; specifically, in transport of T lymphocytes and antigen-presenting cells such as dendritic cells to the lymph nodes. This study aimed to produce epitopes of CCL21 and IL1[beta] as a recombinant protein and characterize its in vitro anti-tumor and immunogenic activity. A codon-optimized ccl21/IL1[beta] gene was designed and synthesized from human genes. Stability and binding affinity of CCL21/IL1[beta] protein and CCR7 receptor were examined through in silico analyses. The construct was introduced into N. tabacum to produce this recombinant protein and the structure and function of CCL21/IL1[beta] were examined. Purified protein from transgenic leaves generated a strong signal in SDS PAGE and western blotting assays. FTIR measurement and MALDI-TOF/TOF mass spectrography showed that ccl21/IL-1[beta] was correctly expressed in tobacco plants. Potential activity of purified CCL21/IL1[beta] in stimulating the proliferation and migration of MCF7 cancer cell line was investigated using the wound healing method. The results demonstrated a decrease in survival rate and metastasization of cancer cells in the presence of CCL21/IL1[beta], and IC.sub.50 of CCL21 on MCF7 cells was less than that of non-recombinant protein. Agarose assay on PBMCsCCR7+ showed that CCL21/IL1[beta] has biological activity and there is a distinguishable difference between chemokinetic (CCL21) and chemotactic (FBS) movements. Overall, the results suggest that CCL21/IL1[beta] could be considered an effective adjuvant in future in vivo and clinical tests.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0261101</identifier><language>eng</language><publisher>Public Library of Science</publisher><subject>Analysis ; Cytokines ; Dendritic cells ; Health aspects ; Immune response ; Recombinant proteins</subject><ispartof>PloS one, 2022-08, Vol.17 (8), p.e0261101</ispartof><rights>COPYRIGHT 2022 Public Library of Science</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,864,27924,27925</link.rule.ids></links><search><creatorcontrib>Marashi, Hasan</creatorcontrib><creatorcontrib>Beihaghi, Maria</creatorcontrib><creatorcontrib>Chaboksavar, Masoud</creatorcontrib><creatorcontrib>Khaksar, Samad</creatorcontrib><creatorcontrib>Tehrani, Homan</creatorcontrib><creatorcontrib>Abiri, Ardavan</creatorcontrib><title>In silico analysis and in planta production of recombinant ccl21/IL1[beta] protein and characterization of its in vitro anti-tumor and immunogenic activity</title><title>PloS one</title><description>CCL21 has an essential role in anti-tumor immune activity. Epitopes of IL1[beta] have adjuvant activity without causing inflammatory responses. CCR7 and its ligands play a vital role in the immune balance; specifically, in transport of T lymphocytes and antigen-presenting cells such as dendritic cells to the lymph nodes. This study aimed to produce epitopes of CCL21 and IL1[beta] as a recombinant protein and characterize its in vitro anti-tumor and immunogenic activity. A codon-optimized ccl21/IL1[beta] gene was designed and synthesized from human genes. Stability and binding affinity of CCL21/IL1[beta] protein and CCR7 receptor were examined through in silico analyses. The construct was introduced into N. tabacum to produce this recombinant protein and the structure and function of CCL21/IL1[beta] were examined. Purified protein from transgenic leaves generated a strong signal in SDS PAGE and western blotting assays. FTIR measurement and MALDI-TOF/TOF mass spectrography showed that ccl21/IL-1[beta] was correctly expressed in tobacco plants. Potential activity of purified CCL21/IL1[beta] in stimulating the proliferation and migration of MCF7 cancer cell line was investigated using the wound healing method. The results demonstrated a decrease in survival rate and metastasization of cancer cells in the presence of CCL21/IL1[beta], and IC.sub.50 of CCL21 on MCF7 cells was less than that of non-recombinant protein. Agarose assay on PBMCsCCR7+ showed that CCL21/IL1[beta] has biological activity and there is a distinguishable difference between chemokinetic (CCL21) and chemotactic (FBS) movements. Overall, the results suggest that CCL21/IL1[beta] could be considered an effective adjuvant in future in vivo and clinical tests.</description><subject>Analysis</subject><subject>Cytokines</subject><subject>Dendritic cells</subject><subject>Health aspects</subject><subject>Immune response</subject><subject>Recombinant proteins</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2022</creationdate><recordtype>article</recordtype><recordid>eNqNkF1LwzAUhosoOKf_wIuAIHjRLmnWdL0c4kdhMPDrRmQk6WmX0SajScX5V_yzpkxhAy8kFzmcPM97wgmCc4IjQlMyWpmu1byO1kZDhGNGCCYHwYBkNA5ZjOnhTn0cnFi7wjihE8YGwVeukVW1kgZxH7GxyvqiQEqjdc2142jdmqKTThmNTIlakKYRSvsnJGUdk1E-I68CHH_rSQde7H255C2XDlr1yX9d5Wyf-65c209zKnRdY9rtvKbptKlAK4m8pzy0OQ2OSl5bOPu5h8Hz7c3T9X04m9_l19NZWBHGkpBxIWnC0jHhAElKx5wWgpUsExPgWSbSMU6ySRpLKUosJMlSLLEkKYDIPJ_QYXCxza14DQulS-P83xtl5WKaEr8oSmhPRX9Q_hTQ-PVpKJXv7wlXe4JnHHy4infWLvLHh_-z85d99nKHXQKv3dKauuvXbHfBb0hpqAM</recordid><startdate>20220829</startdate><enddate>20220829</enddate><creator>Marashi, Hasan</creator><creator>Beihaghi, Maria</creator><creator>Chaboksavar, Masoud</creator><creator>Khaksar, Samad</creator><creator>Tehrani, Homan</creator><creator>Abiri, Ardavan</creator><general>Public Library of Science</general><scope>IOV</scope><scope>ISR</scope></search><sort><creationdate>20220829</creationdate><title>In silico analysis and in planta production of recombinant ccl21/IL1[beta] protein and characterization of its in vitro anti-tumor and immunogenic activity</title><author>Marashi, Hasan ; Beihaghi, Maria ; Chaboksavar, Masoud ; Khaksar, Samad ; Tehrani, Homan ; Abiri, Ardavan</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-g1665-6abc356741aee5734a3db6f69b8ea99b74059872ccbf0bc1970c0c17eeb9e5753</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2022</creationdate><topic>Analysis</topic><topic>Cytokines</topic><topic>Dendritic cells</topic><topic>Health aspects</topic><topic>Immune response</topic><topic>Recombinant proteins</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Marashi, Hasan</creatorcontrib><creatorcontrib>Beihaghi, Maria</creatorcontrib><creatorcontrib>Chaboksavar, Masoud</creatorcontrib><creatorcontrib>Khaksar, Samad</creatorcontrib><creatorcontrib>Tehrani, Homan</creatorcontrib><creatorcontrib>Abiri, Ardavan</creatorcontrib><collection>Gale In Context: Opposing Viewpoints</collection><collection>Gale In Context: Science</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Marashi, Hasan</au><au>Beihaghi, Maria</au><au>Chaboksavar, Masoud</au><au>Khaksar, Samad</au><au>Tehrani, Homan</au><au>Abiri, Ardavan</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>In silico analysis and in planta production of recombinant ccl21/IL1[beta] protein and characterization of its in vitro anti-tumor and immunogenic activity</atitle><jtitle>PloS one</jtitle><date>2022-08-29</date><risdate>2022</risdate><volume>17</volume><issue>8</issue><spage>e0261101</spage><pages>e0261101-</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>CCL21 has an essential role in anti-tumor immune activity. Epitopes of IL1[beta] have adjuvant activity without causing inflammatory responses. CCR7 and its ligands play a vital role in the immune balance; specifically, in transport of T lymphocytes and antigen-presenting cells such as dendritic cells to the lymph nodes. This study aimed to produce epitopes of CCL21 and IL1[beta] as a recombinant protein and characterize its in vitro anti-tumor and immunogenic activity. A codon-optimized ccl21/IL1[beta] gene was designed and synthesized from human genes. Stability and binding affinity of CCL21/IL1[beta] protein and CCR7 receptor were examined through in silico analyses. The construct was introduced into N. tabacum to produce this recombinant protein and the structure and function of CCL21/IL1[beta] were examined. Purified protein from transgenic leaves generated a strong signal in SDS PAGE and western blotting assays. FTIR measurement and MALDI-TOF/TOF mass spectrography showed that ccl21/IL-1[beta] was correctly expressed in tobacco plants. Potential activity of purified CCL21/IL1[beta] in stimulating the proliferation and migration of MCF7 cancer cell line was investigated using the wound healing method. The results demonstrated a decrease in survival rate and metastasization of cancer cells in the presence of CCL21/IL1[beta], and IC.sub.50 of CCL21 on MCF7 cells was less than that of non-recombinant protein. Agarose assay on PBMCsCCR7+ showed that CCL21/IL1[beta] has biological activity and there is a distinguishable difference between chemokinetic (CCL21) and chemotactic (FBS) movements. Overall, the results suggest that CCL21/IL1[beta] could be considered an effective adjuvant in future in vivo and clinical tests.</abstract><pub>Public Library of Science</pub><doi>10.1371/journal.pone.0261101</doi><tpages>e0261101</tpages></addata></record> |
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| source | DOAJ Directory of Open Access Journals; Public Library of Science (PLoS) Journals Open Access; EZB-FREE-00999 freely available EZB journals; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Analysis Cytokines Dendritic cells Health aspects Immune response Recombinant proteins |
| title | In silico analysis and in planta production of recombinant ccl21/IL1[beta] protein and characterization of its in vitro anti-tumor and immunogenic activity |
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