Delineation of pulmonary alveolar macrophage subpopulations by flow cytometry in normal subjects and in patients with lung cancer

The glycophoryn A (GPA) assay evaluates somaticin vivomutations. It is considered a cumulative biodosimeter for genotoxic exposures and is under evaluation in cancer risk assessment.GPA, a polymorphic membrane protein of the erythrocytes, determines the MN blood groups. The N0 and NN variant frequencies (VF) may be detected in MN subjects (about 50% of the population) by flow cytometry using two differently labelled antibodies.We explored if GPA N0 and NN VF might be relevant to the assessment of individual lung cancer risk and susceptibility, in a small population with a high prevalence of heavy tobacco smokers: 8 lung cancer patients and 16 subjects with non‐malignant lung diseases associated with increased risk of lung cancer.There was a wide interindividual variability and complete overlap between non‐neoplastic and neoplastic patients. A significant positive correlation was seen with smoking duration in N0 VF (p=0.04, age‐adjusted). Current smokers (n=12) displayed higher N0 values than never (n=1) or ex‐smokers (n=11), 36.3±18.2 and 21.0±13.2, respectively (p< 0:01). No association was shown with occupational exposure.The present exploratory study suggests that assessment of individual lung cancer risk and susceptibility by the GPA assay does not seem to be feasible. The assay appears to provide a biomarker of longterm exposure to tobacco smoke.

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Rsf-1 Influences the Sensitivity of Non-Small Cell Lung Cancer to Paclitaxel by Regulating NF-κB Pathway and Its Downstream Proteins

Cellular Physiology and Biochemistry ◽

10.1159/000486116 ◽

2017 ◽

Vol 44 (6) ◽

pp. 2322-2336 ◽

Cited By ~ 18

Author(s):

Xitao Chen ◽

Xiaodi Sun ◽

Jingqian Guan ◽

Junda Gai ◽

Jilin Xing ◽

...

Keyword(s):

Lung Cancer ◽

Flow Cytometry ◽

Cell Proliferation ◽

Small Cell Lung Cancer ◽

Cell Apoptosis ◽

Small Cell ◽

Small Cell Lung ◽

Protein Levels ◽

H460 Cell ◽

Xenograft Mice

Background/Aims: The therapeutic efficacy of paclitaxel is hampered by chemotherapeutic resistance in non-small cell lung cancer (NSCLC). Rsf-1 enhanced paclitaxel resistance via nuclear factor-κB (NF-κB) in ovarian cancer cells and nasopharyngeal carcinoma. This study assessed the function of Rsf-1 in the modulation of the sensitivity of NSCLC to paclitaxel via the NF-κB pathway. Methods: The mRNA and protein levels of the related genes were quantified by RT-PCR and Western blotting. Rsf-1 silencing was achieved with CRISPR/Cas9 gene editing. Cell cycle, migration and proliferation were tested with flow cytometry, transwell test and CCK8 test. Cell apoptosis was analyzed with flow cytometry and quantification of C-capase3. The parameters of the tumors were measured in H460 cell xenograft mice. Results: Rsf-1 was highly expressed in H460 and H1299 cells. Rsf-1 knockout caused cell arrest at the G1 phase, increased cell apoptosis, and decreased migration and cell proliferation. Rsf-1 knockout increased the inhibition of cell proliferation, the reduction in cell migration and the augment in cell apoptosis in paclitaxel treated H460 and H1299 cells. Rsf-1 knockout further enhanced the paclitaxel-mediated decrease in the volume and weight of the tumors in H460 cell xenograft mice. Helenalin and Rsf-1 knockout decreased the protein levels of p-P65, BcL2, CFLAR, and XIAP; hSNF2H knockout decreased the protein level of NF-κB p-P65 without altering Rsf-1 and p65 protein levels, while Rsf-1 and hSNF2H double knockout decreased the level of NF-κB p-P65, in H1299 and H460 cells. Conclusion: These results demonstrate that Rsf-1 influences the sensitivity of NSCLC to paclitaxel via regulation of the NF-κB pathway and its downstream genes.

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Abstract 1671: Evaluation of total PD-1 expression using multi-color flow cytometry in metastatic non-small Cell lung cancer patients treated with multi-neoantigen vector (ADXS-503) alone and in combination of pembrolizumab to assess T-cell & T-cell memory subsets

10.1158/1538-7445.am2021-1671 ◽

2021 ◽

Author(s):

Venkat Mohanram ◽

Natalya Belkina ◽

Angelina R. Bisconte ◽

Jonathan W. Goldman ◽

Gregory J. Gerstner ◽

...

Keyword(s):

Lung Cancer ◽

Flow Cytometry ◽

T Cell ◽

Small Cell Lung Cancer ◽

Cancer Patients ◽

Small Cell ◽

T Cell Memory ◽

Small Cell Lung ◽

Color Flow ◽

Lung Cancer Patients

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Use of Allophycocyanin Allows Quantitative Description by Flow Cytometry of Alveolar Macrophage Surface Antigens Present in Low Numbers of Cells

American Review of Respiratory Disease ◽

10.1164/ajrccm/138.5.1124 ◽

1988 ◽

Vol 138 (5) ◽

pp. 1124-1128 ◽

Cited By ~ 13

Author(s):

Henry J. Fuchs ◽

Joan McDowell ◽

Judd E. Shellito

Keyword(s):

Flow Cytometry ◽

Alveolar Macrophage ◽

Quantitative Description ◽

Surface Antigens

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The Synergistic Anti-tumor Effect of Iodine-125 Low-dose-rate Brachytherapy and Anti-PD-1 Therapy on Lung Cancer in Mice

10.21203/rs.3.rs-102600/v1 ◽

2020 ◽

Author(s):

Yiyi Cao ◽

Wenbo Li ◽

Jia Li ◽

Yu Weng ◽

Chang Chen ◽

...

Keyword(s):

Lung Cancer ◽

Flow Cytometry ◽

Low Dose ◽

Immune Microenvironment ◽

Secondary Tumor ◽

Low Dose Rate ◽

Tumor Tissues ◽

Tumor Immune Microenvironment ◽

Iodine 125 ◽

Low Dose Rate Brachytherapy

Abstract Background: Radiotherapy (RT) when combined with anti-PD-1 therapy has a significant effect, but RT fractionation and dose impact the effects of this combined therapy. Iodine-125 particle implantation (125I RPI) is a hyperfractionated low-dose-rate brachytherapy. Its impact on the tumor immune microenvironment and the efficacy of 125I RPI combined with anti-PD-1 therapy are unknown. In this study, we evaluated the effectiveness of 125I RPI combined with anti-PD-1 therapy and their impact on tumor immunity. Methods: A Lewis lung cancer (LLC) mouse model was established and radioactive iodine-125 particles were implanted into the tumors. Tumor tissues were obtained six and 12 days after particle implantation, and the expression of PD-1/PD-L1 was detected by flow cytometry. On day 0, LLC cells were injected subcutaneously into the right hindlimb (primary tumor) and left flank (secondary tumor) of mice. On day 10, the mice were randomly divided into PBS, α-PD-1, 125I RPI, and α-PD-1+125I RPI groups. On day 22, tumor tissues were extracted from the mice. The proportion of immune cell subsets in the tumor immune microenvironment was detected by flow cytometry, and the primary and secondary tumor volumes were monitored. Results: After 125I RPI, the expression of PD-L1 on tumor cells was upregulated (P < 0.0001), and the proportion of CD8+ PD-1+ T cells also increased (P = 0.0001). 125I RPI combined with anti-PD-1 therapy synergistically inhibited primary (P = 0.0139) and secondary (P = 0.0494) tumor growth. The flow cytometry results showed that the combination therapy could increase the proportion of CD8+ T cells (P = 0.0055) and decrease the proportion of T regulatory cells (P < 0.0227) in the tumor microenvironment. Survival analysis shows that the sequence of 125I RPI and α-PD-1 affects efficacy, and early initiation of 125I RPI is more beneficial. Conclusion: 125I RPI combined with anti-PD-1 therapy can significantly inhibit tumor growth and activate anti-tumor immunity, which present a promising approach for the treatment of cancer.

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