Prodrug-Based Versatile Nanomedicine for Enhancing Cancer Immunotherapy by Increasing Immunogenic Cell Death.

Cholangiocarcinoma (CCA) is an aggressive epithelial malignancy of the bile ducts. Recent reports have suggested a rise in the incidence of CCA globally. Current treatments for CCA have limited effectiveness, with only 20-30% response rates in patients with advanced cancer. Thus, there is a growing need to investigate new therapeutic targets for CCA. The tumor microenvironment (TME) is a potential target for novel therapies. The TME plays an important role in progression and metastasis of cancer through facilitating immune escape. Recent studies have demonstrated potential roles for heparan sulfate proteoglycans (HSPGs) in tumorigenesis. Sulfatase 2 (SULF2) is a heparan sulfate editing sulfatase that removes the 6-O sulfate moiety within the heparan sulfate (HS) chains and alters the affinity of HS chains for growth factors and cytokines and their receptors, thus modulating HSPG function in cell signaling pathways. HSPG specific sulfatases have been recently identified as a potential therapeutic target in CCA. We aimed to assess the effects of anti-Sulfatase-2 antibody on immune cells in the tumor microenvironment of syngeneic CCA xenografts in C57BL6 mice. Mice bearing syngeneic CCA xenografts were treated with an anti-sulf2 antibody, and the tumors compared to untreated control xenografts. After treatment, the tumors were harvested and analyzed by Cytometry by time of flight (Cytof) to identify changes in the types and numbers of immune cells in the tumor xenografts. Treatment with anti-sulf2 antibody resulted in a decrease in immunosuppressive cells such as tumor associated macrophages (TAMs) and myeloid derived suppressor cells (MDSC) compared to the control. There was also an increase in anti-tumor cells such as T-cells and NK cells, suggesting that suppressing sulf-2 in the microenvironment can shift the TME toward an anti-tumorigenic rather than a pro-tumorigenic environment. In summary, our work demonstrates the potential to target Sulfatase 2 as a therapeutic mechanism for immunomodulation of the tumor microenvironment. Future experiments to determine the relationship between specific clinical phenotypes and the number, activities and types of immune cells seen in the TME, and experiments in other hepatobiliary cancers models, such as hepatocellular carcinoma and gallbladder cancer, will be conducted. Citation Format: Tayla R. Brooks, Nellie Campbell, Melanie Keim, Lewis R. Roberts. Modulating the immune response in cholangiocarcinoma: Targeting sulfatase-2 for therapeutic intervention [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6525.

In this work, we propose an adaptive robust control for a second order nonlinear model of the interaction between cancer and immune cells of the body to control the growth of cancer and maintain the number of immune cells in an appropriate level. Up to now, most of the control approaches are based on minimizing the drug dosage based on an optimal control structure. However, in many cases, measuring the exact quantity of the model parameters is not possible. This is due to limitation in measuring devices, variational and undetermined characteristics of micro-environmental factors and the variable nature of parameters during the growth and treatment phases of cancer. It is of great importance to present a control strategy that can deal with these variables and unknown factors in a nonlinear model. Adaptive control is a suitable choice to achieve this goal. We assume linear uncertainties for the model parameters and employ a sliding term for updating the estimated parameters and the control signals. Moreover, due to difficulties in measuring the number of immune cells in biological experiments, an estimation technique is applied to infer this value. The convergence of the estimated number of immune cells to the actual value is shown. The stability and convergence of the number of cancer and immune cells to the specified target values are also proved using a time-varying Lyapunov function. Finally, we have shown the performance of the proposed control strategy in the context of various computational results.

Introduction: Over the past decade, classifications using immune cell infiltration have been applied to many types of tumors; however, mesotheliomas have been less frequently evaluated. Methods: In this study, 60 well-characterized pleural mesotheliomas (PMs) were evaluated immunohistochemically for the characteristics of immune cells within tumor microenvironment (TME) using 10 immunohistochemical markers: CD3, CD4, CD8, CD56, CD68, CD163, FOXP3, CD27, PD-1, and TIM-3. For further characterization of PMs, hierarchical clustering analyses using these 10 markers were performed. Results: Among the immune cell markers, CD3 (p < 0.0001), CD4 (p = 0.0016), CD8 (p = 0.00094), CD163+ (p = 0.042), and FOXP3+ (p = 0.025) were significantly associated with an unfavorable clinical outcome. Immune checkpoint receptor expressions on tumor-infiltrating lymphocytes such as PD-1 (p = 0.050), CD27 (p = 0.014), and TIM-3 (p = 0.0098) were also associated with unfavorable survival. Hierarchical clustering analyses identified three groups showing specific characteristics and significant associations with patient survival (p = 0.016): the highest number of immune cells (ICHigh); the lowest number of immune cells, especially CD8+ and CD163+ cells (ICLow); and intermediate number of immune cells (ICInt). ICHigh tumors showed significantly higher expression of PD-L1 (p = 0.00038). Cox proportional hazard model identified ICHigh [hazard ratio (HR) = 2.90] and ICInt (HR = 2.97) as potential risk factors compared with ICLow. Tumor CD47 (HR = 2.36), tumor CD70 (HR = 3.04), and tumor PD-L1 (HR = 3.21) expressions were also identified as potential risk factors for PM patients. Conclusion: Our findings indicate immune checkpoint and/or immune cell-targeting therapies against CD70-CD27 and/or CD47-SIRPA axes may be applied for PM patients in combination with PD-L1-PD-1 targeting therapies in accordance with their tumor immune microenvironment characteristics.

The application of tumor immunotherapy to glioblastoma (GBM) is limited by an unprecedented degree of immune suppression due to factors that include high numbers of immune suppressive myeloid cells, the blood brain barrier, and T cell sequestration to the bone marrow. We previously identified an increase in immune suppressive myeloid-derived suppressor cells (MDSCs) in GBM patients, which correlated with poor prognosis and was dependent on macrophage migration inhibitory factor (MIF). Here we examine the MIF signaling axis in detail in murine MDSC models, GBM-educated MDSCs and human GBM. We found that the monocytic subset of MDSCs (M-MDSCs) expressed high levels of the MIF cognate receptor CD74 and was localized in the tumor microenvironment. In contrast, granulocytic MDSCs (G-MDSCs) expressed high levels of the MIF non-cognate receptor CXCR2 and showed minimal accumulation in the tumor microenvironment. Furthermore, targeting M-MDSCs with Ibudilast, a brain penetrant MIF-CD74 interaction inhibitor, reduced MDSC function and enhanced CD8 T cell activity in the tumor microenvironment. These findings demonstrate the MDSC subsets differentially express MIF receptors and may be leveraged for specific MDSC targeting.

Combinatorial immunotherapy using anti-programmed cell death 1 (PD-1) monoclonal antibody (mAb) is being developed to overcome the limited efficacy of monotherapy with anti-PD-1 mAb for patients with advanced gastric cancer (GC). Anti-PD-1 mAb exhibits clinical efficacy by enhancing the function of cytotoxic T lymphocyte (CTL) through the inhibition of the PD-1 pathway; however, there are various immunosuppressive mechanisms that inhibit CTL function, as well as the PD-1 pathway in the tumor microenvironment (TME). Immune suppressive cells and expression of the inhibitory immune checkpoint molecules are included as main inhibitory mechanisms against CTL in the TME. On the other hand, increasing the number of CTLs enhances the efficacy of anti-PD-1 mAb, and immunogenic tumor cell death (ICD) is crucial to induce CTL through the activation of the cancer immunity cycle. In the present review, we discuss the therapeutic potential of developing combinatorial immunotherapy focusing on the inhibitory immune checkpoint molecules and immune suppressive cells in the TME, as well as on the ICD induced by radiotherapy for patients with advanced GC.

[Background]Neutrophils are known as immune cells involved in biological defense against foreign substances. On the other hand, in the tumor microenvironment, it is said that neutrophils affect on tumor progression as a tumor-associated neutrophil (TAN). Although, TAN and other immune cells in the progression of HCC is unclear. [Method]One hundred twenty eight patients who underwent curative hepatectomy for HCC in our institution were enrolled. Immunohistochemistry was performed with CD66b and CD8 antibody to evaluate TAN and CD8+ T lymphocytes, respectively. Expression high or low was determined by cut-off value of each number of immune cells. We counted the number of each immune cells in both tumor (intra-tumor) and the tumor marginal area (extra-tumor). The relationship between the infiltration of each immune cell and clinicopathological factors was examined. [Result]Comparing the number of immune cells of intra-tumor and extra-tumor, there were significantly more cells in extra-tumor area (p&amp;lt;0.001). There was no difference in between background clinicopathological factors and the number of TAN or CD8+ T lymphocytes in either intra-tumor or extra-tumor areas. Overall survival was significantly better in the group with high CD8+ T lymphocytes in extra-tumor area compared to the low group (p = 0.0491), intriguingly though CD8+ T lymphocytes in intra-tumor area was not associated with prognosis. In the contrast, overall survival was significantly poor in the group with high TAN in extra-tumor area compared to the low group (p = 0.0296), though TAN in intra-tumor area was not associated with prognosis. [Conclusion]Infiltration of TAN and CD8+ T lymphocytes not in intra-tumor but in the extra-tumor area may be involved in the progression of HCC. Citation Format: Toshihiko Yusa, Hirohisa Okabe, Yo-ichi Yamashita, Takanobu Yamao, Naoki Umezaki, Tatsunori Miyata, Shigeki Nakagawa, Hiromitsu Hayashi, Katsunori Imai, Akira Chikamoto, Takatoshi Ishiko, Hideo Baba. The role of tumor-associated neutrophils (TAN) and CD8 positive T lymphocyte in the progression of hepatocellular carcinoma (HCC) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1084.

Epithelial cell layer thickness and immune cell populations in the normal human vagina at different stages of the menstrual cycle

Purpose/Objectives: Her2-neu positive breast cancers have a good overall prognosis with the advent of Her2-directed therapies such as trastuzumab. However, despite the increased efficacy with Her-2-directed therapies 20-30% of patients still have local and/or distant failure despite being Her-2 amplified on pathology. The etiology of this local failure remains unknown. Recent evidence has suggested that there may be immune factors that contribute to the progression of breast cancer and the response to therapy so we undertook a study to examine the relationship between immune-based markers and traditional pathologic and clinical markers of outcome. Materials/Methods: Paraffin-embedded sections were generation and clinical records were reviewed for 88 patients, age ≥ 18 years, with pathologically-proven Her2-neu+ breast cancer who were treated at a single institution from 01/2001 to 12/2013. Single-color immunohistochemical staining was performed for CK5/6, CK14 and EGFR and scored by a breast pathologist. Adjacent sections were also then stained for CD45, CD4, CD8 and CD68 using a multi-color immunohistochemical approach. Slides were scanned using the Vectra Automated Quantitative Pathology Imaging System and analyzed using an in-house algorithm to quantitate the number of immune cells within the tumor, tumor margin and within 2 mm outside the tumor. We then compared the level of CK5/6, CK14, EGFR with the number of immune cells. The number of different immune cells were also analyzed with respect to other clinical parameters including age, tumor size, nodal status, hormone receptor status, time to progression, progression-free survival and overall survival. Results: At a median follow-up of 66.5 months, 20 (22.7%) patients had progressed. We found that the number of CD45+ leukocytes at the margin correlates with the expression of CK5/6 (p = 0.015) which predicted for local failure. Further, we found that the ratio of CD8 to CD4 cells within the tumor and margin highly correlates with the expression of the hormone receptors (p = 0.01). Conclusions: Our preliminary results suggest that immune markers may be important predictors of a basal-like phenotype as defined by CK5/6 expression in Her2+ breast cancers which itself correlated with significantly higher local failure. Further higher CD8 to CD4 ratios were highly correlated with hormone receptor expression, particularly PR expression suggesting that in the Her2+ population the more favorable prognosis for the "triple-positive" subtype of Her2+ cancers may be in part due to a more favorable immune microenvironment. Citation Format: Shiao SL, Gertych A, Ma Z, Zhang X, Burnison CM, Mirhadi AJ, Giuliano A, Knudsen BS, Chung A. Quantitative analysis of T cell and macrophage immune markers in Her2-positive breast cancer. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P3-07-37.

&lt;div&gt;Abstract&lt;p&gt;High levels of IL1β can result in chronic inflammation, which in turn can promote tumor growth and metastasis. Inhibition of IL1β could therefore be a promising therapeutic option in the treatment of cancer. Here, the effects of IL1β blockade induced by the mAbs canakinumab and gevokizumab were evaluated alone or in combination with docetaxel, anti–programmed cell death protein 1 (anti–PD-1), anti-VEGFα, and anti-TGFβ treatment in syngeneic and humanized mouse models of cancers of different origin. Canakinumab and gevokizumab did not show notable efficacy as single-agent therapies; however, IL1β blockade enhanced the effectiveness of docetaxel and anti–PD-1. Accompanying these effects, blockade of IL1β alone or in combination induced significant remodeling of the tumor microenvironment (TME), with decreased numbers of immune suppressive cells and increased tumor infiltration by dendritic cells (DC) and effector T cells. Further investigation revealed that cancer-associated fibroblasts (CAF) were the cell type most affected by treatment with canakinumab or gevokizumab in terms of change in gene expression. IL1β inhibition drove phenotypic changes in CAF populations, particularly those with the ability to influence immune cell recruitment. These results suggest that the observed remodeling of the TME following IL1β blockade may stem from changes in CAF populations. Overall, the results presented here support the potential use of IL1β inhibition in cancer treatment. Further exploration in ongoing clinical studies will help identify the best combination partners for different cancer types, cancer stages, and lines of treatment.&lt;/p&gt;&lt;/div&gt;

&lt;div&gt;Abstract&lt;p&gt;High levels of IL1β can result in chronic inflammation, which in turn can promote tumor growth and metastasis. Inhibition of IL1β could therefore be a promising therapeutic option in the treatment of cancer. Here, the effects of IL1β blockade induced by the mAbs canakinumab and gevokizumab were evaluated alone or in combination with docetaxel, anti–programmed cell death protein 1 (anti–PD-1), anti-VEGFα, and anti-TGFβ treatment in syngeneic and humanized mouse models of cancers of different origin. Canakinumab and gevokizumab did not show notable efficacy as single-agent therapies; however, IL1β blockade enhanced the effectiveness of docetaxel and anti–PD-1. Accompanying these effects, blockade of IL1β alone or in combination induced significant remodeling of the tumor microenvironment (TME), with decreased numbers of immune suppressive cells and increased tumor infiltration by dendritic cells (DC) and effector T cells. Further investigation revealed that cancer-associated fibroblasts (CAF) were the cell type most affected by treatment with canakinumab or gevokizumab in terms of change in gene expression. IL1β inhibition drove phenotypic changes in CAF populations, particularly those with the ability to influence immune cell recruitment. These results suggest that the observed remodeling of the TME following IL1β blockade may stem from changes in CAF populations. Overall, the results presented here support the potential use of IL1β inhibition in cancer treatment. Further exploration in ongoing clinical studies will help identify the best combination partners for different cancer types, cancer stages, and lines of treatment.&lt;/p&gt;&lt;/div&gt;

Simple SummaryImmunotherapy may be an attractive treatment option to increase survival, and to reduce treatment-related side effects, for children with acute myeloid leukemia (AML). While immunotherapies have shown successes in many cancer types, the development and subsequent clinical implementation have proven difficult in pediatric AML. To expedite the development of immunotherapy, it will be crucial to understand which pediatric AML patients are likely to respond to immunotherapies. Emerging research in solid malignancies has shown that the number and phenotype of immune cells in the tumor microenvironment is predictive of response to several types of immunotherapies. Such a predictive model may also be applicable for AML and, thus, knowledge on the immune cells infiltrating the bone marrow environment is needed. Here, we discuss the current state of knowledge on these infiltrating immune cells in pediatric AML, as well as ongoing immunotherapy trials, and provide suggestions concerning the way forward.Immunotherapeutic agents may be an attractive option to further improve outcomes and to reduce treatment-related toxicity for pediatric AML. While improvements in outcome have been observed with immunotherapy in many cancer types, immunotherapy development and implementation into patient care for both adult and pediatric AML has been hampered by an incomplete understanding of the bone marrow environment and a paucity of tumor-specific antigens. Since only a minority of patients respond in most immunotherapy trials across different cancer types, it will be crucial to understand which children with AML are likely to respond to or may benefit from immunotherapies. Immune cell profiling efforts hold promise to answer this question, as illustrated by the development of predictive scores in solid cancers. Such information on the number and phenotype of immune cells during current treatment regimens will be pivotal to generate hypotheses on how and when to intervene with immunotherapy in pediatric AML. In this review, we discuss the current understanding of the number and phenotype of immune cells in the bone marrow in pediatric AML, ongoing immunotherapy trials and how comprehensive immune profiling efforts may pave the way for successful clinical trials (and, ultimately, implementation into patient care).

Atherosclerosis is a chronic inflammatory disease of the arterial wall characterized by lipid deposition, plaque formation, and immune cell infiltration. Innate and adaptive immune cells infiltrate the artery during development of the disease. Moreover, advanced disease leads to formation of artery tertiary lymphoid organs in the adventitia (Grabner et al., 2009; Hu et al., 2015). Various and diverse types of immune cells have been identified in the aorta adventitia vs atherosclerotic plaques (Elewa et al., 2016; Galkina et al., 2006; Lotzer et al., 2010; Mohanta et al., 2016; Mohanta et al., 2014; Moos et al., 2005; Srikakulapu et al., 2016; Zhao et al., 2004). There are conflicting reports on the number and subtypes of immune cells in the aorta depending on the age of the animals, the protocol that is used to obtain single cell suspensions, and the dietary conditions of the mice (Campbell et al., 2012; Clement et al., 2015; Galkina et al., 2006; Kyaw et al., 2012). The number of immune cells in the aorta differs as much as tenfold using different protocols (Butcher et al., 2012; Galkina et al., 2006; Gjurich et al., 2015; Grabner et al., 2009; Hu et al., 2015). These discrepant results call for a protocol that robustly documents bona fide aorta cells rather than those in the surrounding tissues or blood. Critical methodological hurdles include the removal of adjacent adipose tissue and small paraaortic lymph nodes lining the entire aortic tree that are not visible by the naked eye. A dissection microscope is therefore recommended. Moreover protocols of aorta preparations should ascertain that lymphocyte aggregates referred to as fat associated lymphoid clusters (FALCs) (Benezech et al., 2015; Elewa et al., 2015) that are often present at the border between the adipose tissue and the adventitia are removed before enzyme digestion. We propose - besides other approaches (Hu et al., 2015; Mohanta et al., 2014) - a combination of immunohistochemical staining and fluorescence activated cell sorter (FACS) analyses from single cell suspensions to quantify the cells of interest. This protocol describes isolation of single cells from mouse aorta for FACS and other analysis.

AIM: This study investigated the immune cells within in the tumor stroma of primary and metastatic gastrointestinal stromal tumors (GISTs) by immunohistochemistry. GISTs appear usually as histological homogeneous tumors with a dense cell structure. However, GISTs have been shown to have a tumor stroma, in which different immune cells of the innate and acquired immune system are embedded. The frequency and function of these cells in GISTs remain widely unknown. In this work the various immune cells are described morphologically and their frequency in primary GISTs as well as peritoneal and liver metastases is being compared. Furthermore, the criteria proliferation activity, morphology, tumor size, organ localization and malignancy are examined and compared with the number of immune cells in the primary and metastatic GISTs. In a small number of metastases real time RT-PCR was performed to gain an overview of the functionality of immune cells using RT-PCR.MATERIAL & METHODS: A total of 188 untreated primary GISTs and 52 untreated GIST metastases were immunohistochemically processed to determine the percentage of tissue-associated fibrohistiocytic cells (Kim-1P) and cells of macrophage lineage (CD68). In addition, the number of several lymphocytes (T-Lymphocytes: CD3, B-Lymphocytes: CD20, NK-cells: CD56) was studied in these samples. For this purpose punches of the paraffin blocks of resected specimen or biopsies were lined up site by site in 22 tissue microarryas (TMAs) using a manual tissue arrayer. The photographs of 3-6 punches have been analysed computer-assisted with subsequent statistical evaluation of the results. This analysis resulted in very precise numbers of the percentage of immune cells in the respective tissue preparations. Furthermore, reverse transcription and real-time RT-PCR were used to detect the expression of the proinflammatory cytokines interleukin 1β, interleukin 6 and tumor necrosis factor α in a small sample size of snap-frozen tissue samples of metastases.RESULTS: GISTs have a tumor stroma, in which many immune cells are embedded. In the primary GISTs Kim-1P+ cells (28,8% ±7,1) dominated, but also lymphocytic cells (CD3+(2,2% ±1,8), CD20+(0,6% ±0,7), CD56+(1,1% ±0,9)) and macrophages (CD68+(3,6% ±2,1)) were present. Interestingly, the number of immune cells differ in the metastases of GISTs. On the one hand metastases show more lymphocytic cells than the primary GISTs (CD3+: 7,3% ±2,3 (p<0,01); CD20+: 1,8% ±0,3 (p<0,05)), on the other hand, the peritoneal metastases differ from the liver metastases. Peritoneal metastases have a significant higher number of Kim-1P+ cells (31,8% ±7,5) than liver metastases (18,2% ±3,8; p<0,01), while the latter in turn contain more CD3+ T-lymphocytes (11,7% ±1,8) than the peritoneal metastases (4,4% ±2,6; p<0,01). At the same time, the two sites differ in proliferation activity. Liver metastases have a lower proliferation activity (12,9% ±8,2) than peritoneal metastases of GISTs (18,3% ±7,3; p<0,05). In the RT-PCR of fresh-frozen tissue from metastases of a small cohort, one patient with a high percentage of Kim 1P+ cells (49,5% ±17) did not only show a comparatively high expression of Il-6 (CT value 26,8) and Il-1β (CT value 26,1) but also the lowest expression of TNF (CT 34,1) as well as a rapid clinical progress.CONCLUSION: The different numbers of the various immune cells in primary GISTs compared with metastatic GISTs as well as within the two metastatic sites, suggest a location specific microenvironment, which may play a part in the tumor growth of primary and secondary GISTs. Further studies will be needed to understand the type of communication between the immune cells of the GIST stroma and the tumor itself. In the future, the individuality of this tumor will come to the fore, elucidating that tumors with a different composition of their stromal cells, for example immune cells, might also need a different and individual treatment. Thus, not only the description of the immune cells is necessary, but also the understanding of GISTs as a family of tumors, which should not be seen as uniform, homogeneous tumors.

BackgroundProgranulin (PGRN), as a multifunctional growth factor, is overexpressed in multiple tumors, but the role of PGRN on tumor immunity is still unclear. Here, we studied the effect of PGRN on breast cancer tumor immunity and its possible molecular mechanism.MethodsThe changes of macrophage phenotypes after PGRN treatment were detected by western blot, quantitative polymerase chain reaction (PCR) and flow cytometry. Western blot was used to study the signal molecular mechanism of PGRN regulating this process. The number and localization of immune cells in Wild-type (WT) and PGRN−/− breast cancer tissues were analyzed by immunohistochemical staining and immunofluorescence techniques. The activation and proliferation of CD8+ T cells were measured by flow cytometry.ResultsAfter being treated with PGRN, the expressions of M2 markers and programmed death ligand 1 (PD-L1) on macrophages increased significantly. Signal transducer and activator of transcription 3 (STAT3) signaling pathway inhibitor Stattic significantly inhibited the expression of PD-L1 and M2 related markers induced by PGRN. In WT group, CD8 were co-localized with macrophages and PD-L1, but not tumor cells. The number of immune cells in PGRN−/− breast cancer tissue increased, and their infiltration into tumor parenchyma was also enhanced. Moreover, in the co-culture system, WT peritoneal macrophages not only reduced the ratio of activated CD8+ T cells but also reduced the proportion of proliferating CD8+ T cells. The addition of programmed death receptor 1 (PD-1) and PD-L1 neutralizing antibodies effectively reversed this effect and restored the immune function of CD8+ T cells.ConclusionThese results demonstrate that PGRN promotes M2 polarization and PD-L1 expression by activating the STAT3 signaling pathway. Furthermore, through PD-1/PD-L1 interaction, PGRN can promote the breast tumor immune escape. Our research may provide new ideas and targets for clinical breast cancer immunotherapy.

BackgroundTumor microenvironment is composed of tumor cells, fibroblasts, endothelial cells, and infiltrating immune cells. Tumor-associated immune cells may inhibit or promote tumor growth and progression. This study was conducted to determine whether the number and microlocalization of macrophages, mature dendritic cells and cytotoxic T cells in non-small cell lung cancer are associated with patient's survival time.MethodsNinety-nine patients with non-small cell lung cancer (NSCLC) were included in this retrospective study. Paraffin-embedded NSCLC specimens and their clinicopathological data including up to 8-year follow-up information were used. Immunohistochemical staining for CD68 (marker for macrophages), CD83 (marker for mature dendritic cells), and CD8 (marker for cytotoxic T cells) was performed and evaluated in a blinded fashion. The numbers of immune cells in tumor islets and stroma, tumor islets, or tumor stroma were counted under a microscope. Correlation of the cell numbers and patient's survival time was analyzed using the Statistical Package for the Social Sciences (version 13.0).ResultsThe numbers of macrophages, mature dendritic cells and cytotoxic T cells were significantly more in the tumor stroma than in the tumor islets. The number of macrophages in the tumor islets was positively associated with patient's survival time, whereas the number of macrophages in the tumor stroma was negatively associated with patient's survival time in both univariate and multivariate analyses. The number of mature dendritic cells in the tumor islets and stroma, tumor islets only, or tumor stroma only was positively associated with patient's survival time in a univariate analysis but not in a multivariate analysis. The number of cytotoxic T cells in the tumor islets and stroma was positively associated with patient's survival time in a univariate analysis but not in a multivariate analysis. The number of cytotoxic T cells in the tumor islets only or stroma only was not associated with patient's survival time.ConclusionsThe number of macrophages in the tumor islets or stroma is an independent predictor of survival time in NSCLC patients. Counting macrophages in the tumor islets or stroma is more useful in predicting patient's survival time than counting mature dendritic cells or cytotoxic T cells.