Real-Time Tracking of Ex Vivo-Expanded Natural Killer Cells Toward Human Triple-Negative Breast Cancers
IntroductionEx vivo-expanded natural killer (NK) cells are a potential candidate for cancer immunotherapy based on high cytotoxicity against malignant tumor cells. However, a limited understanding of the migration of activated NK cells toward solid tumors is a critical dilemma in the development of...
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Frontiers Media S.A.
2018-05-01
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| Series: | Frontiers in Immunology |
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| Online Access: | http://journal.frontiersin.org/article/10.3389/fimmu.2018.00825/full |
| _version_ | 1819195415243784192 |
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| author | Tung Nguyen Thanh Uong Tung Nguyen Thanh Uong Kyung-Hwa Lee Sung-Ja Ahn Kyung Won Kim Jung-Joon Min Hoon Hyun Mee Sun Yoon Mee Sun Yoon Mee Sun Yoon |
| author_facet | Tung Nguyen Thanh Uong Tung Nguyen Thanh Uong Kyung-Hwa Lee Sung-Ja Ahn Kyung Won Kim Jung-Joon Min Hoon Hyun Mee Sun Yoon Mee Sun Yoon Mee Sun Yoon |
| author_sort | Tung Nguyen Thanh Uong |
| collection | DOAJ |
| description | IntroductionEx vivo-expanded natural killer (NK) cells are a potential candidate for cancer immunotherapy based on high cytotoxicity against malignant tumor cells. However, a limited understanding of the migration of activated NK cells toward solid tumors is a critical dilemma in the development of effective and adoptive NK cell-based immunotherapy.MethodsEx vivo-expanded NK cells from healthy donors were stained with near-infrared fluorophores at different concentrations. NK cell proliferation and cytotoxicity were assessed using a WST-8 assay, while the expression levels of surface molecules were analyzed by flow cytometry. To investigate the biodistribution of NK cells in both normal and tumor-bearing NSG mice, NK cells labeled with ESNF13 were subjected to NIR fluorescence imaging using the Mini-FLARE imaging system. Finally, mice were sacrificed and histopathological tests were performed in resected organs.ResultsThe signal intensity of ESNF-stained NK cells was long-lasting at 72 h using concentrations as low as 0.04 µM. At a low dose range, ESNF13 did not affect NK cell purity, expression levels of surface receptors, or cytotoxic functions against MDA-MB-231 cancer cells. Ex vivo-expanded NK cells labeled with ESNF13 had a 4-h biodistribution in non-tumor-bearing NSG mice that mainly localized to the lungs immediately after injection and then fully migrated to the kidney after 4 h. In an MDA-MB-231 tumor-bearing NSG mice with extensive metastasis in both lungs, the fluorescence signal was dominant in both lungs and steady at 1, 2, and 4 h post-injection. In a early phase of tumor progression, administered NK cell migrated to the lungs and tumor sites within 30 min post-injection, the signal dominated the tumor site after 1 h, and remained steady at 4 h.ConclusionOptical imaging with NIR fluorophore ESNF13 is a highly sensitive, applicable, and inexpensive method for the real-time tracking of ex vivo-expanded NK cells both in vitro and in vivo. Administered NK cells had different patterns of NK cell distribution and accumulation to the tumor site according to tumor progression in triple-negative breast cancer xenograft models. |
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| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-12-23T02:12:24Z |
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| spelling | doaj.art-1e043e827e2b41d29361e9553f61c1f02022-12-21T18:03:46ZengFrontiers Media S.A.Frontiers in Immunology1664-32242018-05-01910.3389/fimmu.2018.00825349672Real-Time Tracking of Ex Vivo-Expanded Natural Killer Cells Toward Human Triple-Negative Breast CancersTung Nguyen Thanh Uong0Tung Nguyen Thanh Uong1Kyung-Hwa Lee2Sung-Ja Ahn3Kyung Won Kim4Jung-Joon Min5Hoon Hyun6Mee Sun Yoon7Mee Sun Yoon8Mee Sun Yoon9Department of Radiation Oncology, Chonnam National University Hwasun Hospital, Chonnam National University Medical School, Gwangju, South KoreaDepartment of Biomedical Science, Chonnam National University Graduate School, Gwangju, South KoreaDepartment of Pathology, Chonnam National University Hwasun Hospital, Chonnam National University Medical School, Gwangju, South KoreaDepartment of Radiation Oncology, Chonnam National University Hwasun Hospital, Chonnam National University Medical School, Gwangju, South KoreaDepartment of Biomedical Science, Chonnam National University Graduate School, Gwangju, South KoreaDepartment of Nuclear Medicine, Chonnam National University Hwasun Hospital, Hwasun, South KoreaDepartment of Biomedical Sciences, Chonnam National University Medical School, Gwangju, South KoreaDepartment of Radiation Oncology, Chonnam National University Hwasun Hospital, Chonnam National University Medical School, Gwangju, South KoreaDepartment of Biomedical Science, Chonnam National University Graduate School, Gwangju, South KoreaResearch Center for Cancer Immunotherapy, Chonnam National University Hwasun Hospital, Jeollanam-do, South KoreaIntroductionEx vivo-expanded natural killer (NK) cells are a potential candidate for cancer immunotherapy based on high cytotoxicity against malignant tumor cells. However, a limited understanding of the migration of activated NK cells toward solid tumors is a critical dilemma in the development of effective and adoptive NK cell-based immunotherapy.MethodsEx vivo-expanded NK cells from healthy donors were stained with near-infrared fluorophores at different concentrations. NK cell proliferation and cytotoxicity were assessed using a WST-8 assay, while the expression levels of surface molecules were analyzed by flow cytometry. To investigate the biodistribution of NK cells in both normal and tumor-bearing NSG mice, NK cells labeled with ESNF13 were subjected to NIR fluorescence imaging using the Mini-FLARE imaging system. Finally, mice were sacrificed and histopathological tests were performed in resected organs.ResultsThe signal intensity of ESNF-stained NK cells was long-lasting at 72 h using concentrations as low as 0.04 µM. At a low dose range, ESNF13 did not affect NK cell purity, expression levels of surface receptors, or cytotoxic functions against MDA-MB-231 cancer cells. Ex vivo-expanded NK cells labeled with ESNF13 had a 4-h biodistribution in non-tumor-bearing NSG mice that mainly localized to the lungs immediately after injection and then fully migrated to the kidney after 4 h. In an MDA-MB-231 tumor-bearing NSG mice with extensive metastasis in both lungs, the fluorescence signal was dominant in both lungs and steady at 1, 2, and 4 h post-injection. In a early phase of tumor progression, administered NK cell migrated to the lungs and tumor sites within 30 min post-injection, the signal dominated the tumor site after 1 h, and remained steady at 4 h.ConclusionOptical imaging with NIR fluorophore ESNF13 is a highly sensitive, applicable, and inexpensive method for the real-time tracking of ex vivo-expanded NK cells both in vitro and in vivo. Administered NK cells had different patterns of NK cell distribution and accumulation to the tumor site according to tumor progression in triple-negative breast cancer xenograft models.http://journal.frontiersin.org/article/10.3389/fimmu.2018.00825/fullnatural killer cellsESNF13near-infrared fluorophoresMDA-MB-231 tumor-bearing mouseoptical imagingin vivo tracking |
| spellingShingle | Tung Nguyen Thanh Uong Tung Nguyen Thanh Uong Kyung-Hwa Lee Sung-Ja Ahn Kyung Won Kim Jung-Joon Min Hoon Hyun Mee Sun Yoon Mee Sun Yoon Mee Sun Yoon Real-Time Tracking of Ex Vivo-Expanded Natural Killer Cells Toward Human Triple-Negative Breast Cancers Frontiers in Immunology natural killer cells ESNF13 near-infrared fluorophores MDA-MB-231 tumor-bearing mouse optical imaging in vivo tracking |
| title | Real-Time Tracking of Ex Vivo-Expanded Natural Killer Cells Toward Human Triple-Negative Breast Cancers |
| title_full | Real-Time Tracking of Ex Vivo-Expanded Natural Killer Cells Toward Human Triple-Negative Breast Cancers |
| title_fullStr | Real-Time Tracking of Ex Vivo-Expanded Natural Killer Cells Toward Human Triple-Negative Breast Cancers |
| title_full_unstemmed | Real-Time Tracking of Ex Vivo-Expanded Natural Killer Cells Toward Human Triple-Negative Breast Cancers |
| title_short | Real-Time Tracking of Ex Vivo-Expanded Natural Killer Cells Toward Human Triple-Negative Breast Cancers |
| title_sort | real time tracking of ex vivo expanded natural killer cells toward human triple negative breast cancers |
| topic | natural killer cells ESNF13 near-infrared fluorophores MDA-MB-231 tumor-bearing mouse optical imaging in vivo tracking |
| url | http://journal.frontiersin.org/article/10.3389/fimmu.2018.00825/full |
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