Establishment of shared antigen reactive cytotoxic T lymphocyte using co-stimulatory molecule introduced autologous cancer cells
Cytotoxic T lymphocytes (CTLs) play an essential role in immunological responses for tumor rejection. In the past decade, many tumor-associated antigens (TAAs) have been identified predominantly in melanomas. Several clinical trials based on such antigenic peptides with or without adjuvants brought...
Gespeichert in:
| Veröffentlicht in: | Experimental and molecular pathology 2010-02, Vol.88 (1), p.128-132 |
|---|---|
| Hauptverfasser: | , , , , , , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 132 |
|---|---|
| container_issue | 1 |
| container_start_page | 128 |
| container_title | Experimental and molecular pathology |
| container_volume | 88 |
| creator | Hirohashi, Yoshihiko Torigoe, Toshihiko Hirai, Itaru Tamura, Yasuaki Nakatsugawa, Munehide Inoue, Yuuji Kanaseki, Takayuki Kamiguchi, Kenjiro Ikeda, Hideyuki Sasaki, Aya Yamanaka, Noboru Sato, Noriyuki |
| description | Cytotoxic T lymphocytes (CTLs) play an essential role in immunological responses for tumor rejection. In the past decade, many tumor-associated antigens (TAAs) have been identified predominantly in melanomas. Several clinical trials based on such antigenic peptides with or without adjuvants brought about partially favorable results, suggesting that identification of more immunogenic TAAs is needed. We show here the successful establishment of human leukocyte antigen (HLA)-A24-restricted CTL (TcLHK2 line1) from a pleural effusion of lung cancer patient, using B7.1 (CD80) transduced autologous lung cancer cells as an antigen-presenting cell (APC). TcLHK2 line1 recognized autologous lung adenocarcinoma cell line LHK2 in an HLA-A24-restricted fashion. Moreover, this CTL line also recognized allogeneic HLA-A24-positive lung adenocarcinoma cell line, gastric carcinoma cell line and melanoma cell line. These data raise the possibility that co-stimulatory molecule B7.1 (CD80) plays important role to overcome the immunological tolerance. Furthermore, TcLHK2 line1 is a useful tool for the identification of widely expressed shared antigens restricted by HLA-A24. Further analysis of this CTL and autologous cancer cell line will bring about novel TAAs. |
| doi_str_mv | 10.1016/j.yexmp.2009.09.021 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_734266286</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0014480009001348</els_id><sourcerecordid>734266286</sourcerecordid><originalsourceid>FETCH-LOGICAL-c388t-eeed4fa6e25380c5d9846f111fad71c365d7535aa56dab961ff5cca84ff2e59e3</originalsourceid><addsrcrecordid>eNp9kE1rGzEQhkVpaNykv6BQdCk9rSNpV7L20EMJ6QcEeknOQpZGtox25UrakL31p0dbm_ZWGBgYnnl5eRB6T8maEipuDusZnofjmhHSr5dh9BVaUdKLhvQdf41WhNCu6SQhl-htzgdSQULZG3RJe0nlRogV-n2Xi94Gn_cDjAVHh_NeJ7BYj8XvYMQJtCn-CbCZSyzx2Rv8gMM8HPexXgBP2Y87bGKTix-moEtMMx5iADMFwH4sKdrJLIFTiSHu4pSx0aOBhA2EkK_RhdMhw7vzvkKPX-8ebr839z-__bj9ct-YVsrSAIDtnBbAeCuJ4baXnXCUUqfthppWcLvhLdeaC6u3vaDOcWO07JxjwHtor9CnU-4xxV8T5KIGn5cGeoTaSW3ajgnBpKhkeyJNijkncOqY_KDTrChRi3l1UH_Mq8W8WobR-vXhnD9tB7D_fs6qK_DxDOhsdHCpSvD5L8dYJzhnfeU-nzioNp48JJWNhyrM-gSmKBv9f4u8AJXgp4w</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>734266286</pqid></control><display><type>article</type><title>Establishment of shared antigen reactive cytotoxic T lymphocyte using co-stimulatory molecule introduced autologous cancer cells</title><source>MEDLINE</source><source>Access via ScienceDirect (Elsevier)</source><creator>Hirohashi, Yoshihiko ; Torigoe, Toshihiko ; Hirai, Itaru ; Tamura, Yasuaki ; Nakatsugawa, Munehide ; Inoue, Yuuji ; Kanaseki, Takayuki ; Kamiguchi, Kenjiro ; Ikeda, Hideyuki ; Sasaki, Aya ; Yamanaka, Noboru ; Sato, Noriyuki</creator><creatorcontrib>Hirohashi, Yoshihiko ; Torigoe, Toshihiko ; Hirai, Itaru ; Tamura, Yasuaki ; Nakatsugawa, Munehide ; Inoue, Yuuji ; Kanaseki, Takayuki ; Kamiguchi, Kenjiro ; Ikeda, Hideyuki ; Sasaki, Aya ; Yamanaka, Noboru ; Sato, Noriyuki</creatorcontrib><description>Cytotoxic T lymphocytes (CTLs) play an essential role in immunological responses for tumor rejection. In the past decade, many tumor-associated antigens (TAAs) have been identified predominantly in melanomas. Several clinical trials based on such antigenic peptides with or without adjuvants brought about partially favorable results, suggesting that identification of more immunogenic TAAs is needed. We show here the successful establishment of human leukocyte antigen (HLA)-A24-restricted CTL (TcLHK2 line1) from a pleural effusion of lung cancer patient, using B7.1 (CD80) transduced autologous lung cancer cells as an antigen-presenting cell (APC). TcLHK2 line1 recognized autologous lung adenocarcinoma cell line LHK2 in an HLA-A24-restricted fashion. Moreover, this CTL line also recognized allogeneic HLA-A24-positive lung adenocarcinoma cell line, gastric carcinoma cell line and melanoma cell line. These data raise the possibility that co-stimulatory molecule B7.1 (CD80) plays important role to overcome the immunological tolerance. Furthermore, TcLHK2 line1 is a useful tool for the identification of widely expressed shared antigens restricted by HLA-A24. Further analysis of this CTL and autologous cancer cell line will bring about novel TAAs.</description><identifier>ISSN: 0014-4800</identifier><identifier>EISSN: 1096-0945</identifier><identifier>DOI: 10.1016/j.yexmp.2009.09.021</identifier><identifier>PMID: 19818766</identifier><identifier>CODEN: EXMPA6</identifier><language>eng</language><publisher>Amsterdam: Elsevier Inc</publisher><subject>Adenocarcinoma ; Adenocarcinoma - immunology ; Adenocarcinoma - pathology ; Aged ; Antigen-Presenting Cells ; Antigens, Neoplasm - immunology ; B7-1 Antigen - immunology ; B7.1 (CD80) ; Biological and medical sciences ; Cell Line, Tumor ; CTL ; Cytotoxicity, Immunologic - immunology ; Epitopes - immunology ; HLA-A Antigens - immunology ; HLA-A24 ; HLA-A24 Antigen ; Humans ; Immune Tolerance ; Investigative techniques, diagnostic techniques (general aspects) ; Lung cancer ; Lung Neoplasms - immunology ; Lung Neoplasms - pathology ; Male ; Medical sciences ; Melanoma - immunology ; Melanoma - pathology ; Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques ; Pneumology ; Stomach Neoplasms - immunology ; Stomach Neoplasms - pathology ; T-Lymphocytes, Cytotoxic - cytology ; T-Lymphocytes, Cytotoxic - immunology ; Tolerance ; Tumors of the respiratory system and mediastinum</subject><ispartof>Experimental and molecular pathology, 2010-02, Vol.88 (1), p.128-132</ispartof><rights>2009 Elsevier Inc.</rights><rights>2015 INIST-CNRS</rights><rights>Copyright 2009 Elsevier Inc. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c388t-eeed4fa6e25380c5d9846f111fad71c365d7535aa56dab961ff5cca84ff2e59e3</citedby><cites>FETCH-LOGICAL-c388t-eeed4fa6e25380c5d9846f111fad71c365d7535aa56dab961ff5cca84ff2e59e3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.yexmp.2009.09.021$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=22465529$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/19818766$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Hirohashi, Yoshihiko</creatorcontrib><creatorcontrib>Torigoe, Toshihiko</creatorcontrib><creatorcontrib>Hirai, Itaru</creatorcontrib><creatorcontrib>Tamura, Yasuaki</creatorcontrib><creatorcontrib>Nakatsugawa, Munehide</creatorcontrib><creatorcontrib>Inoue, Yuuji</creatorcontrib><creatorcontrib>Kanaseki, Takayuki</creatorcontrib><creatorcontrib>Kamiguchi, Kenjiro</creatorcontrib><creatorcontrib>Ikeda, Hideyuki</creatorcontrib><creatorcontrib>Sasaki, Aya</creatorcontrib><creatorcontrib>Yamanaka, Noboru</creatorcontrib><creatorcontrib>Sato, Noriyuki</creatorcontrib><title>Establishment of shared antigen reactive cytotoxic T lymphocyte using co-stimulatory molecule introduced autologous cancer cells</title><title>Experimental and molecular pathology</title><addtitle>Exp Mol Pathol</addtitle><description>Cytotoxic T lymphocytes (CTLs) play an essential role in immunological responses for tumor rejection. In the past decade, many tumor-associated antigens (TAAs) have been identified predominantly in melanomas. Several clinical trials based on such antigenic peptides with or without adjuvants brought about partially favorable results, suggesting that identification of more immunogenic TAAs is needed. We show here the successful establishment of human leukocyte antigen (HLA)-A24-restricted CTL (TcLHK2 line1) from a pleural effusion of lung cancer patient, using B7.1 (CD80) transduced autologous lung cancer cells as an antigen-presenting cell (APC). TcLHK2 line1 recognized autologous lung adenocarcinoma cell line LHK2 in an HLA-A24-restricted fashion. Moreover, this CTL line also recognized allogeneic HLA-A24-positive lung adenocarcinoma cell line, gastric carcinoma cell line and melanoma cell line. These data raise the possibility that co-stimulatory molecule B7.1 (CD80) plays important role to overcome the immunological tolerance. Furthermore, TcLHK2 line1 is a useful tool for the identification of widely expressed shared antigens restricted by HLA-A24. Further analysis of this CTL and autologous cancer cell line will bring about novel TAAs.</description><subject>Adenocarcinoma</subject><subject>Adenocarcinoma - immunology</subject><subject>Adenocarcinoma - pathology</subject><subject>Aged</subject><subject>Antigen-Presenting Cells</subject><subject>Antigens, Neoplasm - immunology</subject><subject>B7-1 Antigen - immunology</subject><subject>B7.1 (CD80)</subject><subject>Biological and medical sciences</subject><subject>Cell Line, Tumor</subject><subject>CTL</subject><subject>Cytotoxicity, Immunologic - immunology</subject><subject>Epitopes - immunology</subject><subject>HLA-A Antigens - immunology</subject><subject>HLA-A24</subject><subject>HLA-A24 Antigen</subject><subject>Humans</subject><subject>Immune Tolerance</subject><subject>Investigative techniques, diagnostic techniques (general aspects)</subject><subject>Lung cancer</subject><subject>Lung Neoplasms - immunology</subject><subject>Lung Neoplasms - pathology</subject><subject>Male</subject><subject>Medical sciences</subject><subject>Melanoma - immunology</subject><subject>Melanoma - pathology</subject><subject>Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques</subject><subject>Pneumology</subject><subject>Stomach Neoplasms - immunology</subject><subject>Stomach Neoplasms - pathology</subject><subject>T-Lymphocytes, Cytotoxic - cytology</subject><subject>T-Lymphocytes, Cytotoxic - immunology</subject><subject>Tolerance</subject><subject>Tumors of the respiratory system and mediastinum</subject><issn>0014-4800</issn><issn>1096-0945</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kE1rGzEQhkVpaNykv6BQdCk9rSNpV7L20EMJ6QcEeknOQpZGtox25UrakL31p0dbm_ZWGBgYnnl5eRB6T8maEipuDusZnofjmhHSr5dh9BVaUdKLhvQdf41WhNCu6SQhl-htzgdSQULZG3RJe0nlRogV-n2Xi94Gn_cDjAVHh_NeJ7BYj8XvYMQJtCn-CbCZSyzx2Rv8gMM8HPexXgBP2Y87bGKTix-moEtMMx5iADMFwH4sKdrJLIFTiSHu4pSx0aOBhA2EkK_RhdMhw7vzvkKPX-8ebr839z-__bj9ct-YVsrSAIDtnBbAeCuJ4baXnXCUUqfthppWcLvhLdeaC6u3vaDOcWO07JxjwHtor9CnU-4xxV8T5KIGn5cGeoTaSW3ajgnBpKhkeyJNijkncOqY_KDTrChRi3l1UH_Mq8W8WobR-vXhnD9tB7D_fs6qK_DxDOhsdHCpSvD5L8dYJzhnfeU-nzioNp48JJWNhyrM-gSmKBv9f4u8AJXgp4w</recordid><startdate>20100201</startdate><enddate>20100201</enddate><creator>Hirohashi, Yoshihiko</creator><creator>Torigoe, Toshihiko</creator><creator>Hirai, Itaru</creator><creator>Tamura, Yasuaki</creator><creator>Nakatsugawa, Munehide</creator><creator>Inoue, Yuuji</creator><creator>Kanaseki, Takayuki</creator><creator>Kamiguchi, Kenjiro</creator><creator>Ikeda, Hideyuki</creator><creator>Sasaki, Aya</creator><creator>Yamanaka, Noboru</creator><creator>Sato, Noriyuki</creator><general>Elsevier Inc</general><general>Elsevier</general><scope>IQODW</scope><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>20100201</creationdate><title>Establishment of shared antigen reactive cytotoxic T lymphocyte using co-stimulatory molecule introduced autologous cancer cells</title><author>Hirohashi, Yoshihiko ; Torigoe, Toshihiko ; Hirai, Itaru ; Tamura, Yasuaki ; Nakatsugawa, Munehide ; Inoue, Yuuji ; Kanaseki, Takayuki ; Kamiguchi, Kenjiro ; Ikeda, Hideyuki ; Sasaki, Aya ; Yamanaka, Noboru ; Sato, Noriyuki</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c388t-eeed4fa6e25380c5d9846f111fad71c365d7535aa56dab961ff5cca84ff2e59e3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>Adenocarcinoma</topic><topic>Adenocarcinoma - immunology</topic><topic>Adenocarcinoma - pathology</topic><topic>Aged</topic><topic>Antigen-Presenting Cells</topic><topic>Antigens, Neoplasm - immunology</topic><topic>B7-1 Antigen - immunology</topic><topic>B7.1 (CD80)</topic><topic>Biological and medical sciences</topic><topic>Cell Line, Tumor</topic><topic>CTL</topic><topic>Cytotoxicity, Immunologic - immunology</topic><topic>Epitopes - immunology</topic><topic>HLA-A Antigens - immunology</topic><topic>HLA-A24</topic><topic>HLA-A24 Antigen</topic><topic>Humans</topic><topic>Immune Tolerance</topic><topic>Investigative techniques, diagnostic techniques (general aspects)</topic><topic>Lung cancer</topic><topic>Lung Neoplasms - immunology</topic><topic>Lung Neoplasms - pathology</topic><topic>Male</topic><topic>Medical sciences</topic><topic>Melanoma - immunology</topic><topic>Melanoma - pathology</topic><topic>Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques</topic><topic>Pneumology</topic><topic>Stomach Neoplasms - immunology</topic><topic>Stomach Neoplasms - pathology</topic><topic>T-Lymphocytes, Cytotoxic - cytology</topic><topic>T-Lymphocytes, Cytotoxic - immunology</topic><topic>Tolerance</topic><topic>Tumors of the respiratory system and mediastinum</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Hirohashi, Yoshihiko</creatorcontrib><creatorcontrib>Torigoe, Toshihiko</creatorcontrib><creatorcontrib>Hirai, Itaru</creatorcontrib><creatorcontrib>Tamura, Yasuaki</creatorcontrib><creatorcontrib>Nakatsugawa, Munehide</creatorcontrib><creatorcontrib>Inoue, Yuuji</creatorcontrib><creatorcontrib>Kanaseki, Takayuki</creatorcontrib><creatorcontrib>Kamiguchi, Kenjiro</creatorcontrib><creatorcontrib>Ikeda, Hideyuki</creatorcontrib><creatorcontrib>Sasaki, Aya</creatorcontrib><creatorcontrib>Yamanaka, Noboru</creatorcontrib><creatorcontrib>Sato, Noriyuki</creatorcontrib><collection>Pascal-Francis</collection><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>Experimental and molecular pathology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Hirohashi, Yoshihiko</au><au>Torigoe, Toshihiko</au><au>Hirai, Itaru</au><au>Tamura, Yasuaki</au><au>Nakatsugawa, Munehide</au><au>Inoue, Yuuji</au><au>Kanaseki, Takayuki</au><au>Kamiguchi, Kenjiro</au><au>Ikeda, Hideyuki</au><au>Sasaki, Aya</au><au>Yamanaka, Noboru</au><au>Sato, Noriyuki</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Establishment of shared antigen reactive cytotoxic T lymphocyte using co-stimulatory molecule introduced autologous cancer cells</atitle><jtitle>Experimental and molecular pathology</jtitle><addtitle>Exp Mol Pathol</addtitle><date>2010-02-01</date><risdate>2010</risdate><volume>88</volume><issue>1</issue><spage>128</spage><epage>132</epage><pages>128-132</pages><issn>0014-4800</issn><eissn>1096-0945</eissn><coden>EXMPA6</coden><abstract>Cytotoxic T lymphocytes (CTLs) play an essential role in immunological responses for tumor rejection. In the past decade, many tumor-associated antigens (TAAs) have been identified predominantly in melanomas. Several clinical trials based on such antigenic peptides with or without adjuvants brought about partially favorable results, suggesting that identification of more immunogenic TAAs is needed. We show here the successful establishment of human leukocyte antigen (HLA)-A24-restricted CTL (TcLHK2 line1) from a pleural effusion of lung cancer patient, using B7.1 (CD80) transduced autologous lung cancer cells as an antigen-presenting cell (APC). TcLHK2 line1 recognized autologous lung adenocarcinoma cell line LHK2 in an HLA-A24-restricted fashion. Moreover, this CTL line also recognized allogeneic HLA-A24-positive lung adenocarcinoma cell line, gastric carcinoma cell line and melanoma cell line. These data raise the possibility that co-stimulatory molecule B7.1 (CD80) plays important role to overcome the immunological tolerance. Furthermore, TcLHK2 line1 is a useful tool for the identification of widely expressed shared antigens restricted by HLA-A24. Further analysis of this CTL and autologous cancer cell line will bring about novel TAAs.</abstract><cop>Amsterdam</cop><pub>Elsevier Inc</pub><pmid>19818766</pmid><doi>10.1016/j.yexmp.2009.09.021</doi><tpages>5</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0014-4800 |
| ispartof | Experimental and molecular pathology, 2010-02, Vol.88 (1), p.128-132 |
| issn | 0014-4800 1096-0945 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_734266286 |
| source | MEDLINE; Access via ScienceDirect (Elsevier) |
| subjects | Adenocarcinoma Adenocarcinoma - immunology Adenocarcinoma - pathology Aged Antigen-Presenting Cells Antigens, Neoplasm - immunology B7-1 Antigen - immunology B7.1 (CD80) Biological and medical sciences Cell Line, Tumor CTL Cytotoxicity, Immunologic - immunology Epitopes - immunology HLA-A Antigens - immunology HLA-A24 HLA-A24 Antigen Humans Immune Tolerance Investigative techniques, diagnostic techniques (general aspects) Lung cancer Lung Neoplasms - immunology Lung Neoplasms - pathology Male Medical sciences Melanoma - immunology Melanoma - pathology Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques Pneumology Stomach Neoplasms - immunology Stomach Neoplasms - pathology T-Lymphocytes, Cytotoxic - cytology T-Lymphocytes, Cytotoxic - immunology Tolerance Tumors of the respiratory system and mediastinum |
| title | Establishment of shared antigen reactive cytotoxic T lymphocyte using co-stimulatory molecule introduced autologous cancer cells |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-23T01%3A56%3A47IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Establishment%20of%20shared%20antigen%20reactive%20cytotoxic%20T%20lymphocyte%20using%20co-stimulatory%20molecule%20introduced%20autologous%20cancer%20cells&rft.jtitle=Experimental%20and%20molecular%20pathology&rft.au=Hirohashi,%20Yoshihiko&rft.date=2010-02-01&rft.volume=88&rft.issue=1&rft.spage=128&rft.epage=132&rft.pages=128-132&rft.issn=0014-4800&rft.eissn=1096-0945&rft.coden=EXMPA6&rft_id=info:doi/10.1016/j.yexmp.2009.09.021&rft_dat=%3Cproquest_cross%3E734266286%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=734266286&rft_id=info:pmid/19818766&rft_els_id=S0014480009001348&rfr_iscdi=true |