scholarly journals Fluorescent Multiplex Immunohistochemistry Coupled With Other State-Of-The-Art Techniques to Systematically Characterize the Tumor Immune Microenvironment

2021 ◽  
Vol 8 ◽  
Author(s):  
Anaïs Boisson ◽  
Grégory Noël ◽  
Manuel Saiselet ◽  
Joël Rodrigues-Vitória ◽  
Noémie Thomas ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Immune Microenvironment ◽  
Cancer Treatments ◽  
Luminal B ◽  
Her2 Breast Cancer ◽  
Technological Advances ◽  
Tumor Immune Microenvironment ◽  
Tertiary Lymphoid Structures ◽  
Treatment Responses ◽  
Lymphoid Structures

Our expanding knowledge of the interactions between tumor cells and their microenvironment has helped to revolutionize cancer treatments, including the more recent development of immunotherapies. Immune cells are an important component of the tumor microenvironment that influence progression and treatment responses, particularly to the new immunotherapies. Technological advances that help to decipher the complexity and diversity of the tumor immune microenvironment (TIME) are increasingly used in translational research and biomarker studies. Current techniques that facilitate TIME evaluation include flow cytometry, multiplex bead-based immunoassays, chromogenic immunohistochemistry (IHC), fluorescent multiplex IHC, immunofluorescence, and spatial transcriptomics. This article offers an overview of our representative data, discusses the application of each approach to studies of the TIME, including their advantages and challenges, and reviews the potential clinical applications. Flow cytometry and chromogenic and fluorescent multiplex IHC were used to immune profile a HER2+ breast cancer, illustrating some points. Spatial transcriptomic analysis of a luminal B breast tumor demonstrated that important additional insight can be gained from this new technique. Finally, the development of a multiplex panel to identify proliferating B cells, Tfh, and Tfr cells on the same tissue section demonstrates their co-localization in tertiary lymphoid structures.

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

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.

scholarly journals Developing mammary terminal duct lobular units have a dynamic mucosal and stromal immune microenvironment

2020 ◽  
Author(s):  
Dorottya Nagy ◽  
Clare M. C. Gillis ◽  
Katie Davies ◽  
Abigail L. Fowden ◽  
Paul Rees ◽  
...  
Keyword(s):  
Mammary Gland ◽  
Leading Edge ◽  
Mammary Epithelial ◽  
Breast Development ◽  
Growth Phases ◽  
Immune Microenvironment ◽  
Ki67 Expression ◽  
Tertiary Lymphoid Structures ◽  
Summary Statement ◽  
Lymphoid Structures

AbstractThe human breast and ovine mammary gland undergo a striking degree of postnatal development, leading to formation of terminal duct lobular units (TDLUs). In this study we interrogated aspects of sheep TDLU growth to increase understanding of ovine mammogenesis and as a model for the study of breast development. Mammary epithelial proliferation is significantly higher in lambs less than two months old than in peri-pubertal animals. Ki67 expression is polarized to the leading edge of the developing TDLUs. Intraepithelial ductal macrophages exhibit striking periodicity and significantly increased density in lambs approaching puberty. Stromal macrophages are more abundant centrally than peripherally. The developing ovine mammary gland is infiltrated by intraepithelial and stromal T lymphocytes that are significantly more numerous in older lambs. In the stroma, hotspots of Ki67 expression colocalize with large aggregates of lymphocytes and macrophages. Multifocally these aggregates exhibit distinct organization consistent with tertiary lymphoid structures. The lamb mammary gland thus exhibits a dynamic mucosal and stromal immune microenvironment and, as such, constitutes a valuable model system that provides new insights into postnatal breast development.Summary statementDevelopment of terminal duct lobular units in the sheep mammary gland involves distinct growth phases and macrophage and lymphocyte fluxes. Tertiary lymphoid structures are present subjacent to the mucosal epithelium.

scholarly journals Prognostic Significance of Gene Signature of Tertiary Lymphoid Structures in Patients With Lung Adenocarcinoma

2021 ◽  
Vol 11 ◽  
Author(s):  
Hong Feng ◽  
Fujun Yang ◽  
Lihong Qiao ◽  
Kai Zhou ◽  
Junfei Wang ◽  
...  
Keyword(s):  
Overall Survival ◽  
Lung Adenocarcinoma ◽  
Immune Cell ◽  
Prognostic Significance ◽  
Gene Signature ◽  
Driver Gene ◽  
Immune Microenvironment ◽  
High Group ◽  
Tertiary Lymphoid Structures ◽  
Lymphoid Structures

BackgroundLung adenocarcinoma (LUAD) is a highly mortal cancer. Tertiary lymphoid structures (TLS) are ectopic lymphoid organs with similar morphological and molecular characters to secondary lymphoid organ. The aim of this study is to investigate the prognostic effect of a gene signature associated with TLSs, including B-cell-specific genes.MethodsClinical data of 515 LUAD patients in the TGCA cohort were used to examine the relationship of TLS signature with immune microenvironment, tumor mutational burden (TMB), and driver gene mutations. Patients were divided into the TLS signature high group and TLS signature low group, and comparative analysis of survival and its influencing factors between the two groups was performed. The resulting data were then validated in the GSE37745 cohort.ResultsTLS signature high group had significantly better overall survival (OS) and progression-free interval (PFI) as well as significantly higher infiltration of immune cell subsets, cancer immune cycle (CIC) signature except for immunogram score2 (IGS2), and expression of major checkpoint genes than the TLS signature low group. Notably, while TLS signature was not markedly associated with TMB and mutation frequencies of driver genes, there were significant differences in overall survival of patients with given mutation status of EGFR, KRAS, BRAF and TP53 genes between the TLS signature high and low groups.ConclusionThis study provided evidence that LUAD patients with high TLS signature had a favorable immune microenvironment and better prognosis, suggesting that TLS signature is an independent positive prognostic factor for LUAD patients.

scholarly journals Developing ovine mammary terminal duct lobular units have a dynamic mucosal and stromal immune microenvironment

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Dorottya Nagy ◽  
Clare M. C. Gillis ◽  
Katie Davies ◽  
Abigail L. Fowden ◽  
Paul Rees ◽  
...  
Keyword(s):  
Mammary Gland ◽  
Immune Cell ◽  
Leading Edge ◽  
Breast Development ◽  
Cell Aggregates ◽  
Human Breast ◽  
Immune Microenvironment ◽  
Ki67 Expression ◽  
Tertiary Lymphoid Structures ◽  
Lymphoid Structures

AbstractThe human breast and ovine mammary gland undergo striking levels of postnatal development, leading to formation of terminal duct lobular units (TDLUs). Here we interrogate aspects of sheep TDLU growth as a model of breast development and to increase understanding of ovine mammogenesis. The distributions of epithelial nuclear Ki67 positivity differ significantly between younger and older lambs. Ki67 expression is polarised to the leading edge of the developing TDLUs. Intraepithelial ductal macrophages exhibit periodicity and considerably increased density in lambs approaching puberty. Stromal macrophages are more abundant centrally than peripherally. Intraepithelial T lymphocytes are more numerous in older lambs. Stromal hotspots of Ki67 expression colocalize with immune cell aggregates that exhibit distinct organisation consistent with tertiary lymphoid structures. The lamb mammary gland thus exhibits a dynamic mucosal and stromal immune microenvironment and constitutes a valuable model system that provides new insights into postnatal breast development.

scholarly journals The synergistic anti-tumor effect of iodine-125 low-dose-rate brachytherapy and anti-PD-1 therapy on lung cancer in mice

2021 ◽  
Author(s):  
Yiyi Cao ◽  
Wenbo Li ◽  
Jia Li ◽  
Yu Weng ◽  
Chang Chen ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Low Dose ◽  
Immune Microenvironment ◽  
Secondary Tumor ◽  
Low Dose Rate ◽  
Tumor Tissues ◽  
Tumor Immune Microenvironment ◽  
Iodine 125 ◽  
The Right ◽  
Low Dose Rate Brachytherapy

Abstract Background: Radiotherapy (RT) when combined with anti-PD-1 therapy can have a significant anti-tumor 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 in the right hindlimb 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. LLC cells were injected subcutaneously into the right hindlimb (primary tumor) on day 0 and into the left flank (secondary tumor) on day 3 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, primary 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 presents a promising approach for the treatment of cancer.

scholarly journals The Human Microbiomes in Pancreatic Cancer: Towards Evidence-Based Manipulation Strategies?

2021 ◽  
Vol 22 (18) ◽  
pp. 9914
Author(s):  
Giovanni Brandi ◽  
Silvia Turroni ◽  
Florencia McAllister ◽  
Giorgio Frega
Keyword(s):  
Pancreatic Cancer ◽  
Therapeutic Approaches ◽  
Immune Microenvironment ◽  
Essential Components ◽  
Tumor Immune Microenvironment ◽  
Current Therapies ◽  
Treatment Responses ◽  
The One ◽  
Resistance To Chemotherapy

Recent pieces of evidence have emerged on the relevance of microorganisms in modulating responses to anticancer treatments and reshaping the tumor-immune microenvironment. On the one hand, many studies have addressed the role of the gut microbiota, providing interesting correlative findings with respect to etiopathogenesis and treatment responses. On the other hand, intra-tumoral bacteria are being recognized as intrinsic and essential components of the cancer microenvironment, able to promote a plethora of tumor-related aspects from cancer growth to resistance to chemotherapy. These elements will be probably more and more valuable in the coming years in early diagnosis and risk stratification. Furthermore, microbial-targeted intervention strategies may be used as adjuvants to current therapies to improve therapeutic responses and overall survival. This review focuses on new insights and therapeutic approaches that are dawning against pancreatic cancer: a neoplasm that arises in a central metabolic “hub” interfaced between the gut and the host.

scholarly journals Tumor Immune Microenvironment Characteristics and Their Prognostic Value in Non-Small-Cell Lung Cancer

2021 ◽  
Vol 11 ◽  
Author(s):  
Dan Su ◽  
Gao Wu ◽  
Ran Xiong ◽  
Xiangxiang Sun ◽  
Meiqing Xu ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Flow Cytometry ◽  
T Cells ◽  
Immune Cells ◽  
Small Cell ◽  
Normal Lung ◽  
Small Cell Lung ◽  
Immune Microenvironment ◽  
Tumor Immune Microenvironment ◽  
Nsclc Patients

IntroductionCancer progression is determined not only by the malignant behavior of tumors but also by the immune microenvironment. The tumor immune microenvironment also plays a pivotal role in determining the clinical response of non-small-cell lung cancer (NSCLC) to immunotherapies. To understand the possible mechanisms and explore new targets in lung cancer immunotherapy, we characterized the immune profiles in NSCLC patients.MethodsSeventy-one NSCLC patients who underwent radical resection were selected. The immune cell composition in paired tumor and adjacent normal lung tissues was tested by flow cytometry. The associations of tumor immune microenvironment characteristics with clinicopathological factors and overall survival were analyzed. Kaplan–Meier curves and Cox proportional hazards models were used to determine differences in survival.ResultsCompared with adjacent normal lung tissues, an increased proportion of CD45+ hematopoietic-derived cells, CD4+ T cell subtypes, Tregs and B cells was observed in tumor samples with a reduced frequency of myeloid cell populations. There was no significant increase in total CD8+ T cells, but both PD1+ and CD38+ CD8+ T cells were significantly enriched in tumor samples and statistically significantly associated with tumor size. In addition, positive CD38 expression was highly correlated with PD1 positivity. A high proportion of CD8+ T cells and a low percentage of PD1+ CD8+ T cells were statistically significantly associated with better survival in stage II and III patients, whereas a low frequency of CD38+ CD8+ T cells was statistically significantly associated with better survival in all patients and identified as an independent prognostic factor (p=0.049).ConclusionWe profiled the immune cells in the tumor tissues of NSCLC patients using flow cytometry. The results revealed significant enrichment of infiltrating immune cells. A strong correlation was identified between CD38 and PD-1 expression on CD8+ T cells in tumors. CD8+ T cells and their subtypes play a critical role in the prediction of prognosis.

NIR-active Nanoterminator with Mature Dendritic Cell Acitivities for Immuno-Priming Mild Photothermal Cancer Therapy

2020 ◽  
Author(s):  
zhihong sun ◽  
Guanjun Deng ◽  
Xinghua Peng ◽  
Xiuli Xu ◽  
Lanlan Liu ◽  
...  
Keyword(s):  
Dendritic Cell ◽  
Photothermal Therapy ◽  
Whole Body ◽  
Mature Dendritic Cell ◽  
Immune Microenvironment ◽  
Highly Efficient ◽  
Tumor Immune Microenvironment ◽  
Shock Proteins ◽  
Mild Temperature ◽  
Broad Interest

Recently, photothermal-immuno synergistic therapy under mild temperature (~ 45 °C) has got broad interest in cancer treatment. Inhibition the intratumorally HSPs production is the key to accomplish highly efficient and mild photothermal therapy. In this work, we developed biomimetic nanoterminators with mature DCs functions by coating the mature dendritic cell membrane on photothermal nanoagents. As-prepared nanoterminators could automatically locate on T cell in the complex tumor-immune microenvironment and promote the T cells proliferation, activation and cytokine secretion, which could not only inhibit the expression of heat shock proteins to cooperate on highly efficient mild photothermal therapy (~42°C), but also promote tumor apoptosis during the treatment. More importantly, this nanoterminator could serve as vaccine to trigger anti-tumor immune response of the whole body, which would be promising to long-life tumor inhibition and termination.

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