Generation of Large Numbers of Antigen-Expressing Human Dendritic Cells Using CD14-ML Technology.
We previously reported a method to expand human monocytes through lentivirus-mediated introduction of cMYC and BMI1, and we named the monocyte-derived proliferating cells, CD14-ML. CD14-ML differentiated into functional DC (CD14-ML-DC) upon addition of IL-4, resulting in the generation of a large nu...
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2016-01-01
|
| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4822879?pdf=render |
| _version_ | 1818418526895996928 |
|---|---|
| author | Yuya Imamura Miwa Haruta Yusuke Tomita Keiko Matsumura Tokunori Ikeda Akira Yuno Masatoshi Hirayama Hideki Nakayama Hiroshi Mizuta Yasuharu Nishimura Satoru Senju |
| author_facet | Yuya Imamura Miwa Haruta Yusuke Tomita Keiko Matsumura Tokunori Ikeda Akira Yuno Masatoshi Hirayama Hideki Nakayama Hiroshi Mizuta Yasuharu Nishimura Satoru Senju |
| author_sort | Yuya Imamura |
| collection | DOAJ |
| description | We previously reported a method to expand human monocytes through lentivirus-mediated introduction of cMYC and BMI1, and we named the monocyte-derived proliferating cells, CD14-ML. CD14-ML differentiated into functional DC (CD14-ML-DC) upon addition of IL-4, resulting in the generation of a large number of DC. One drawback of this method was the extensive donor-dependent variation in proliferation efficiency. In the current study, we found that introduction of BCL2 or LYL1 along with cMYC and BMI1 was beneficial. Using the improved method, we obtained CD14-ML from all samples, regardless of whether the donors were healthy individuals or cancer patients. In vitro stimulation of peripheral blood T cells with CD14-ML-DC that were loaded with cancer antigen-derived peptides led to the establishment of CD4+ and CD8+ T cell lines that recognized the peptides. Since CD14-ML was propagated for more than 1 month, we could readily conduct genetic modification experiments. To generate CD14-ML-DC that expressed antigenic proteins, we introduced lentiviral antigen-expression vectors and subjected the cells to 2 weeks of culture for drug-selection and expansion. The resulting antigen-expressing CD14-ML-DC successfully induced CD8+ T cell lines that were reactive to CMVpp65 or MART1/MelanA, suggesting an application in vaccination therapy. Thus, this improved method enables the generation of a sufficient number of DC for vaccination therapy from a small amount of peripheral blood from cancer patients. Information on T cell epitopes is not necessary in vaccination with cancer antigen-expressing CD14-ML-DC; therefore, all patients, irrespective of HLA type, will benefit from anti-cancer therapy based on this technology. |
| first_indexed | 2024-12-14T12:24:05Z |
| format | Article |
| id | doaj.art-a32997cad22043bcb23f127d278245a2 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-14T12:24:05Z |
| publishDate | 2016-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-a32997cad22043bcb23f127d278245a22022-12-21T23:01:24ZengPublic Library of Science (PLoS)PLoS ONE1932-62032016-01-01114e015238410.1371/journal.pone.0152384Generation of Large Numbers of Antigen-Expressing Human Dendritic Cells Using CD14-ML Technology.Yuya ImamuraMiwa HarutaYusuke TomitaKeiko MatsumuraTokunori IkedaAkira YunoMasatoshi HirayamaHideki NakayamaHiroshi MizutaYasuharu NishimuraSatoru SenjuWe previously reported a method to expand human monocytes through lentivirus-mediated introduction of cMYC and BMI1, and we named the monocyte-derived proliferating cells, CD14-ML. CD14-ML differentiated into functional DC (CD14-ML-DC) upon addition of IL-4, resulting in the generation of a large number of DC. One drawback of this method was the extensive donor-dependent variation in proliferation efficiency. In the current study, we found that introduction of BCL2 or LYL1 along with cMYC and BMI1 was beneficial. Using the improved method, we obtained CD14-ML from all samples, regardless of whether the donors were healthy individuals or cancer patients. In vitro stimulation of peripheral blood T cells with CD14-ML-DC that were loaded with cancer antigen-derived peptides led to the establishment of CD4+ and CD8+ T cell lines that recognized the peptides. Since CD14-ML was propagated for more than 1 month, we could readily conduct genetic modification experiments. To generate CD14-ML-DC that expressed antigenic proteins, we introduced lentiviral antigen-expression vectors and subjected the cells to 2 weeks of culture for drug-selection and expansion. The resulting antigen-expressing CD14-ML-DC successfully induced CD8+ T cell lines that were reactive to CMVpp65 or MART1/MelanA, suggesting an application in vaccination therapy. Thus, this improved method enables the generation of a sufficient number of DC for vaccination therapy from a small amount of peripheral blood from cancer patients. Information on T cell epitopes is not necessary in vaccination with cancer antigen-expressing CD14-ML-DC; therefore, all patients, irrespective of HLA type, will benefit from anti-cancer therapy based on this technology.http://europepmc.org/articles/PMC4822879?pdf=render |
| spellingShingle | Yuya Imamura Miwa Haruta Yusuke Tomita Keiko Matsumura Tokunori Ikeda Akira Yuno Masatoshi Hirayama Hideki Nakayama Hiroshi Mizuta Yasuharu Nishimura Satoru Senju Generation of Large Numbers of Antigen-Expressing Human Dendritic Cells Using CD14-ML Technology. PLoS ONE |
| title | Generation of Large Numbers of Antigen-Expressing Human Dendritic Cells Using CD14-ML Technology. |
| title_full | Generation of Large Numbers of Antigen-Expressing Human Dendritic Cells Using CD14-ML Technology. |
| title_fullStr | Generation of Large Numbers of Antigen-Expressing Human Dendritic Cells Using CD14-ML Technology. |
| title_full_unstemmed | Generation of Large Numbers of Antigen-Expressing Human Dendritic Cells Using CD14-ML Technology. |
| title_short | Generation of Large Numbers of Antigen-Expressing Human Dendritic Cells Using CD14-ML Technology. |
| title_sort | generation of large numbers of antigen expressing human dendritic cells using cd14 ml technology |
| url | http://europepmc.org/articles/PMC4822879?pdf=render |
| work_keys_str_mv | AT yuyaimamura generationoflargenumbersofantigenexpressinghumandendriticcellsusingcd14mltechnology AT miwaharuta generationoflargenumbersofantigenexpressinghumandendriticcellsusingcd14mltechnology AT yusuketomita generationoflargenumbersofantigenexpressinghumandendriticcellsusingcd14mltechnology AT keikomatsumura generationoflargenumbersofantigenexpressinghumandendriticcellsusingcd14mltechnology AT tokunoriikeda generationoflargenumbersofantigenexpressinghumandendriticcellsusingcd14mltechnology AT akirayuno generationoflargenumbersofantigenexpressinghumandendriticcellsusingcd14mltechnology AT masatoshihirayama generationoflargenumbersofantigenexpressinghumandendriticcellsusingcd14mltechnology AT hidekinakayama generationoflargenumbersofantigenexpressinghumandendriticcellsusingcd14mltechnology AT hiroshimizuta generationoflargenumbersofantigenexpressinghumandendriticcellsusingcd14mltechnology AT yasuharunishimura generationoflargenumbersofantigenexpressinghumandendriticcellsusingcd14mltechnology AT satorusenju generationoflargenumbersofantigenexpressinghumandendriticcellsusingcd14mltechnology |