Establishment of a 21-color Panel for the Detection of Immune Cell Subsets in Human Non-small Cell Lung Cancer Tumor Tissues with Flow Cytometry
Background and objective With the rise of multicolor flow cytometry, flow cytometry has become an important means to detect the immune microenvironment of lung cancer, but most of them are used to detect the proportion of cell subsets or the function of major cell subsets, and they cannot be detecte...
| Main Authors: | , |
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| Format: | Article |
| Language: | zho |
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Chinese Anti-Cancer Association; Chinese Antituberculosis Association
2024-01-01
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| Series: | Chinese Journal of Lung Cancer |
| Subjects: | |
| Online Access: | http://dx.doi.org/10.3779/j.issn.1009-3419.2024.102.02 |
| _version_ | 1797337354542776320 |
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| author | Tingting GUO Hongguan XIE |
| author_facet | Tingting GUO Hongguan XIE |
| author_sort | Tingting GUO |
| collection | DOAJ |
| description | Background and objective With the rise of multicolor flow cytometry, flow cytometry has become an important means to detect the immune microenvironment of lung cancer, but most of them are used to detect the proportion of cell subsets or the function of major cell subsets, and they cannot be detected at the same time. Therefore, a reliable 21-color flow cytometry protocol was established to detect the immune cell subsets in human non-small cell lung cancer (NSCLC) tumor tissues. Methods Cell membrane surface antibodies cluster of differentiation (CD)45, CD3, CD19, CD4, CD8, programmed cell death 1 (PD-1), CD39, CD103, CD25, CD127, chemokine receptor 8 (CCR8), CD56, CD11c, human leukocyte antigen (HLA)-DR, CD38, CD27, CD69, CD62L, CD45RA, CCR7 and nucleic acid dye L/D were used to develop the protocol. Firstly, antibody titration experiments, voltage optimization, subtraction of one color staining and single color staining experiments were carried out for each antibody, and after the experimental conditions and detection schemes were determined, the feasibility of the scheme was verified by using peripheral blood mononuclear cells (PBMCs) specimens of six healthy adult volunteers. Tumor tissue samples from 6 NSCLC patients were tested and analyzed. Results The established 21-color flow cytometry protocol was used to detect the tumor tissue samples of 6 NSCLC patients, and the proportion of each cell subset in lung cancer tissue, as well as the immunophenotype and differentiation of the main cell population, were analyzed. Conclusion The successfully established 21-color flow cytometry protocol is suitable for the detection of PBMCs and NSCLC tissue samples, which provides an effective new idea for monitoring the immune microenvironment status in lung cancer. |
| first_indexed | 2024-03-08T09:08:23Z |
| format | Article |
| id | doaj.art-5f1b515cfbfd4338bd9dab9609a3a782 |
| institution | Directory Open Access Journal |
| issn | 1009-3419 1999-6187 |
| language | zho |
| last_indexed | 2024-03-08T09:08:23Z |
| publishDate | 2024-01-01 |
| publisher | Chinese Anti-Cancer Association; Chinese Antituberculosis Association |
| record_format | Article |
| series | Chinese Journal of Lung Cancer |
| spelling | doaj.art-5f1b515cfbfd4338bd9dab9609a3a7822024-02-01T01:54:02ZzhoChinese Anti-Cancer Association; Chinese Antituberculosis AssociationChinese Journal of Lung Cancer1009-34191999-61872024-01-01271566410.3779/j.issn.1009-3419.2024.102.02Establishment of a 21-color Panel for the Detection of Immune Cell Subsets
in Human Non-small Cell Lung Cancer Tumor Tissues with Flow CytometryTingting GUO0Hongguan XIE1College of Ecology and Environment, Chengdu University of Technology, Chengdu 610059, ChinaCollege of Ecology and Environment, Chengdu University of Technology, Chengdu 610059, ChinaBackground and objective With the rise of multicolor flow cytometry, flow cytometry has become an important means to detect the immune microenvironment of lung cancer, but most of them are used to detect the proportion of cell subsets or the function of major cell subsets, and they cannot be detected at the same time. Therefore, a reliable 21-color flow cytometry protocol was established to detect the immune cell subsets in human non-small cell lung cancer (NSCLC) tumor tissues. Methods Cell membrane surface antibodies cluster of differentiation (CD)45, CD3, CD19, CD4, CD8, programmed cell death 1 (PD-1), CD39, CD103, CD25, CD127, chemokine receptor 8 (CCR8), CD56, CD11c, human leukocyte antigen (HLA)-DR, CD38, CD27, CD69, CD62L, CD45RA, CCR7 and nucleic acid dye L/D were used to develop the protocol. Firstly, antibody titration experiments, voltage optimization, subtraction of one color staining and single color staining experiments were carried out for each antibody, and after the experimental conditions and detection schemes were determined, the feasibility of the scheme was verified by using peripheral blood mononuclear cells (PBMCs) specimens of six healthy adult volunteers. Tumor tissue samples from 6 NSCLC patients were tested and analyzed. Results The established 21-color flow cytometry protocol was used to detect the tumor tissue samples of 6 NSCLC patients, and the proportion of each cell subset in lung cancer tissue, as well as the immunophenotype and differentiation of the main cell population, were analyzed. Conclusion The successfully established 21-color flow cytometry protocol is suitable for the detection of PBMCs and NSCLC tissue samples, which provides an effective new idea for monitoring the immune microenvironment status in lung cancer.http://dx.doi.org/10.3779/j.issn.1009-3419.2024.102.02lung neoplasmsperipheral blood mononuclear cellsimmunophenotypemulticolor flow cytometry |
| spellingShingle | Tingting GUO Hongguan XIE Establishment of a 21-color Panel for the Detection of Immune Cell Subsets in Human Non-small Cell Lung Cancer Tumor Tissues with Flow Cytometry Chinese Journal of Lung Cancer lung neoplasms peripheral blood mononuclear cells immunophenotype multicolor flow cytometry |
| title | Establishment of a 21-color Panel for the Detection of Immune Cell Subsets
in Human Non-small Cell Lung Cancer Tumor Tissues with Flow Cytometry |
| title_full | Establishment of a 21-color Panel for the Detection of Immune Cell Subsets
in Human Non-small Cell Lung Cancer Tumor Tissues with Flow Cytometry |
| title_fullStr | Establishment of a 21-color Panel for the Detection of Immune Cell Subsets
in Human Non-small Cell Lung Cancer Tumor Tissues with Flow Cytometry |
| title_full_unstemmed | Establishment of a 21-color Panel for the Detection of Immune Cell Subsets
in Human Non-small Cell Lung Cancer Tumor Tissues with Flow Cytometry |
| title_short | Establishment of a 21-color Panel for the Detection of Immune Cell Subsets
in Human Non-small Cell Lung Cancer Tumor Tissues with Flow Cytometry |
| title_sort | establishment of a 21 color panel for the detection of immune cell subsets
in human non small cell lung cancer tumor tissues with flow cytometry |
| topic | lung neoplasms peripheral blood mononuclear cells immunophenotype multicolor flow cytometry |
| url | http://dx.doi.org/10.3779/j.issn.1009-3419.2024.102.02 |
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