Tumor-Associated Macrophages and Myeloid-Derived Suppressor Cells as Immunosuppressive Mechanism in Ovarian Cancer Patients: Progress and Challenges

Cancers are complex masses of malignant cells and nonmalignant cells that create the tumor microenvironment (TME). Non-transformed cells of the TME such as tumor-associated macrophages (TAMs) and myeloid-derived suppressor cells (MDSCs) have been observed in the TME of ovarian cancer (OC) patients. Although these subsets may contribute to each step of carcinogenesis and are commonly associated with poor prognosis, still little is known about creation of the protumor microenvironment in OC. In this review, we focused on the nature and prognostic significance of TAMs and MDSCs in OC patients. Moreover, we discuss the main problems and challenges that must be overcome by researchers and clinicians to enrich our knowledge about the immunosuppressive microenvironment of cancers.     

Minireview: Regulatory T Cells and Ovarian Cancer

ABSTRACT

In the last 15 years, it has become apparent that ovarian cancer is recognized by the immune system, taking into account that T cell infiltration can be associated with increased overall survival. Several studies indicate that a correct combination of cluster of differentiation 8 and cluster of differentiation 4 T cells is key to fight tumor progression and that the presence of regulatory T cells (Tregs) infiltrating ovarian solid tumors (or present in ascites) is deleterious. Several markers that characterize Tregs include glucocorticoid-induced tumor necrosis factor receptor, cytotoxic T lymphocyte antigen-4, and forkhead box protein 3 (Foxp3). Research has shown that Tregs can infiltrate cancerous tissue and contribute to tumor growth by secreting immunosuppressive cytokines such as transforming growth factor beta and interleukin (IL)-10. Importantly, these cells might hamper the efficacy of immunotherapeutic approaches, thus strategies involving depletion or regulation of this population have been proposed and tested in experimental models. In this Minireview, we will discuss the relevance of Tregs in ovarian cancer and the experimental approaches destined to impair their immunosuppressive effects.     

Regulatory T Cells and Cancer: A Two-Sided Story

ABSTRACT

Regulatory T cells (Tregs) play pivotal roles in limiting the duration and magnitude of immune response against infectious agents and self-antigens. This is accomplished through contact-dependent and -independent mechanisms that involve crosstalk between Treg cells and other immune and tissue-specific cell types. The same machinery is employed by Tregs to regulate immune responses to cancer, limiting both pro-tumor inflammation and anti-tumor immunity. Factors produced by Treg cells also act directly on transformed epithelial cells and exert opposing effects during different stages of cancer development. Therefore, the immune regulatory cell population serves as a double-edged sword for the development, progression, and treatment of cancers. In this review, we summarize current knowledge on the roles of Treg lymphocytes during cancer development, as well as the underlying cellular and molecular mechanism.     

Regulatory T Cells in Lupus

Naturally occurring, CD4⁺CD25⁺ regulatory T cells that are exported from the thymus early in life play an important role in controlling organ-specific autoimmune diseases, but they may not be critical for suppressing systemic autoimmunity in lupus. On the other hand, lupus-prone subjects appear to be deficient in generation of adaptive T-regulatory cells that can be induced by various means. We review autoantigen-specific therapeutic approaches that induce such regulatory T cells. Of particular interest are TGF-β producing CD4⁺CD25⁺ and CD8⁺ regulatory T cells that are induced by low dose tolerance therapy of lupus-prone mice with nucleosomal histone peptide epitopes, administered subcutaneously in subnanomolar doses. These regulatory T cells are not only efficient in suppressing autoantigen recognition and autoantibody production, but they also inhibit migration/accumulation of pathogenic autoimmune cells in the target organ, such as the kidneys of mice prone to develop lupus nephritis. We discuss why and under what conditions such therapeutic approaches would be beneficial in lupus patients and lupus-prone subjects.     

Clinical Implications of Microsatellite Instability in T1 Colorectal Cancer

Purpose

The estimation of regional lymph node metastasis (LNM) risk in T1 colorectal cancer is based on histologic examination and imaging of the primary tumor. High-frequency microsatellite instability (MSI-H) is likely to decrease the possibility of metastasis to either regional lymph nodes or distant organs in colorectal cancers. This study evaluated the clinical implications of MSI in T1 colorectal cancer with emphasis on the usefulness of MSI as a predictive factor for regional LNM.

Materials and Methods

A total of 133 patients who underwent radical resection for T1 colorectal cancer were included. Genomic DNA was extracted from normal and tumor tissues and amplified by polymerase chain reaction (PCR). Five microsatellite markers, BAT-25, BAT-26, D2S123, D5S346, and D17S250, were used. MSI and clinicopathological parameters were evaluated as potential predictors of LNM using univariate and multivariate analyses.

Results

Among 133 T1 colorectal cancer patients, MSI-H, low-frequency microsatellite instability (MSI-L), and microsatellite stable (MSS) colorectal cancers accounted for 7.5%, 6%, and 86.5%, respectively. MSI-H tumors showed a female predominance, a proximal location and more retrieved lymph nodes. Twenty-two patients (16.5%) had regional LNM. Lymphovascular invasion and depth of invasion were significantly associated with LNM. There was no LNM in 10 MSI-H patients; however, MSI status was not significantly correlated with LNM. Disease-free survival did not differ between patients with MSI-H and those with MSI-L/MSS.

Conclusion

MSI status could serve as a negative predictive factor in estimating LNM in T1 colorectal cancer, given that LNM was not detected in MSI-H patients. However, validation of our result in a different cohort is necessary.     

Regulatory T Cells in Autoimmune Diseases: Anti-Ergotypic T Cells

T regulatory cells play an important role in regulating T-cell responses to self-antigens and control autoimmunity and autoimmune disease. Anti-ergotypic T cells are a subset of such regulatory T cells that respond to activation markers, ergotopes, expressed on other activated T cells. Anti-ergotypic T cells do not respond to nonactivated T cells. Ergotopes include the α-chain of the IL-2 receptor (CD25). Anti-ergotypic T cells were found to downregulate experimental diseases such as experimental autoimmune encephalomyelitis (EAE) and adjuvant arthritis (AA). Anti-ergotypic T cells are present in humans and are activated after T-cell vaccination. Here we review anti-ergotypic T cells in animal models and in humans and contrast anti-ergotypic T cells with other regulatory T-cell subsets.     

T Regulatory Cells in Allergen-Specific Immunotherapy

A dramatic increase in the prevalence of allergy and asthma has occurred during the past few decades. Although the symptoms of many allergic disorders can be suppressed quite effectively by pharmacological interventions, these do not provide a curative solution and therefore involve lifelong use of medication. Allergen-specific immunotherapy (SIT) on the other hand provides a long-lasting effect on the immune response to common environmental antigens, therefore allowing cessation of the therapy after several years. The changes in the immune response brought about by allergen-SIT are slowly being unveiled and explained. Mechanisms underlying allergen-SIT and in particular the role of regulatory T cells will be discussed in this review, based on recent findings and current concepts.     

Multiple Sclerosis and Regulatory T Cells

Introduction  Multiple sclerosis (MS) is a complex genetic disease characterized by chronic inflammation of the central nervous system (CNS). The pathology of MS is largely attributed to autoreactive effector T cells that penetrate the blood–brain barrier and become activated within the CNS. As autoreactive T cells are present in the blood of both patients with MS and healthy individuals, other regulatory mechanisms exist to prevent autoreactive T cells from causing immune disorders. Active suppression by regulatory T (Treg) cells plays a key role in the control of self-antigen-reactive T cells and the induction of peripheral tolerance in vivo. In particular, the importance of antigen-specific Treg cells in conferring genetic resistance to organ-specific autoimmunity and in limiting autoimmune tissue damage has been documented in many disease models including MS. Results  We have found that the frequency of Tregs in MS patients is unchanged from controls, but their function measured in vitro may be diminished, correlating with impaired inhibitory activity in vivo. This review discusses the immunopathology of MS with particular focus given to regulatory T cells and their potential for the development of new therapies to treat this disease.     

调节性T细胞与自身免疫性疾病

自身免疫性疾病是由于机体正常免疫耐受功能受损导致免疫系统对自身组织结构和功能的破坏,并出现一定临床表现的一类疾病.调节性T细胞作为一类具有负向免疫调节功能的淋巴细胞亚群在免疫自稳和免疫耐受中起关键作用,既能抑制不恰当的免疫反应,又能限制免疫应答的范围、程度及作用时间,对效应性T细胞的增殖及免疫活性的发挥产生抑制,因此在许多自身免疫性疾病的发病中扮演重要角色.近年来的研究表明调节性T细胞可以通过细胞接触、分泌细胞因子、基因调控等多种途径发挥作用,在不同的疾病,不同的内环境因素作用下可以表现出不同的特点,转录因子Foxp3作为调节性T细胞的特异性标志是其分化成熟及功能维持的根本.     

Type 1 Regulatory T Cells and Regulatory B Cells Induced by Tolerogenic Dendritic Cells

Dendritic cells (DC) are professional antigen‐presenting cells that are capable of both activating immune responses and inducing tolerance. Several studies have revealed efficiency of therapeutic vaccination with tolerogenic DC (tolDC) in inhibition of experimental autoimmunity. The purpose of this study was to compare four different protocols for generation of tolDC – the antidiabetic drug troglitazone (TGZ DC), NF‐κB inhibitor BAY 11‐7082 (BAY DC), prostaglandin D2 metabolite 15d‐PGJ₂ (PGJ DC) and a combination of dexamethasone and 1α,25‐dihydroxyvitamin D3 (DexVD3 DC) regarding phenotype, cytokine production and T cell stimulatory capacity. TGZ DC and BAY DC had a phenotype comparable to immature DC, while DexVD3 DC were more macrophage like. Analysis of cytokine production using cell culture supernatants from all DC populations revealed that DexVD3 DC were efficient producers of IL‐10 and produced less pro‐inflammatory cytokines. T cells primed with DexVD3 DC showed reduced proliferation, and further analyses of these T cells revealed that functionally effective type 1 regulatory T cells (Tr1) but not FoxP3⁺ Treg were induced. Furthermore, DexVD3 DC promoted the induction of regulatory B cells (Breg). Together, these results indicate that DexVD3 DC have the best potential to be used in a tolerogenic antigen‐presenting cell‐based immunotherapy setting.     

Translating the Concept of Suppressor/Regulatory T Cells to Clinical Applications

The in vivo expansion of suppressor/regulatory T cells (Tregs) is a desirable event in autoimmunity and transplantation. Here we summarize the general rules involved in antigen recognition by T cells and describe Tregs and their requirements, discussing different levels of immune intervention.     

Putative Precursor Cancer Cells in Human Colorectal Cancer Tissue

Multistage carcinogenesis is an important concept in cancer biology. Each new stage is triggered by the acquisition of an additional genetic aberration, leading to clonal expansion of the cancer cell. The resulting tumor mass consists of cancer cells with all genetic aberrations, but may include precursor cells at some point of carcinogenesis. We analyzed six colorectal cancer tissues with APC, K-ras, and p53 mutations. From each sample, 40–50 areas (100×100×40μm) consisting only of cancer cells were microdissected, and genomic DNA was purified. Ratios of mutated and normal alleles were quantitated by the SNaPshot assay, a primer extension assay. In five tumor tissues, we identified cancer cell subpopulations corresponding to putative precursors, i.e., cells with mutations in one or two of the three genes. All samples were likely to be of monoclonal origin, and temporal sequences of the mutations could be deduced from the mutation patterns of putative precursors. The orders of mutation events were variable. However, the two carcinoma tissues accompanying adenoma regions started with the APC mutation, not contradicting the previous studies. The analysis also revealed considerable heterogeneity in allele ratios of one or two of the chromosomes. The current findings are promising to uncover the process of carcinogenesis directly from the tumor tissue of the patient.     

晚期肺癌患者调节性T细胞和IL-10表达的变化

探讨晚期肺癌患者的CD4＋CD25＋调节性T细胞（Treg）、IL-10及其他T细胞亚群表达的临床意义。检测晚期肺癌患者92例和正常对照者64名的IL-10（ELISA法）、CD4＋CD25＋调节性T细胞及其他T细胞亚群（流式细胞仪法）。结果显示：肺癌组血清IL-10明显大于对照组（278±34ng/L vs 122±65ng/L,P〈0.01）、CD4＋CD25＋Treg细胞明显大于对照组[（17.8±5.2）%vs（7.1±0.4）%,P〈0.01]、CD3＋、CD4＋、CD8＋CD28＋和NK细胞明显小于对照组[（58.4±7.8）%vs（78.2±6.4）%,P≤0.05;（34.4±7.6）%vs（44.9±8.4）%,P〈0.01;（9.4±3.6）%vs（16.5±2.7）%,P〈0.01;（9.4±3.6）%vs（18.5±7.2）%,P〈0.01]。CD8＋T细胞明显高于对照组[（37.8±6.5）%vs（31.8±5.1）%,P〈0.01]。CD4＋CD25＋Treg细胞、IL-10与CD8＋CD28＋细胞、NK细胞呈明显负相关。这些结果表明,CD4＋CD25＋Treg细胞与IL-10增多为晚期肺癌患者免疫功能受损的表现。     

Circulating Regulatory T Cells in Endometrial Cancer: A Role for Age and Menopausal Status

Regulatory T cells (Treg) are a sub-population of T cells that suppress self-reactivity and are implicated in immune tolerance towards malignant cells. Circulating Treg cells are increased in several cancers. In endometrial cancer Treg cells have been investigated only in tumour tissues and, in contrast to some other tumours, fewer Treg cells were reported in endometrial cancer compared with benign controls. Flow cytometry was used to determine the frequency of circulating Treg cells in women undergoing hysterectomy for either endometrial cancer (n = 24) or non- cancer-related conditions (n = 21). Circulating Treg cells were more abundant in women with cancer compared to those without (4.68% vs. 3.66%, p = 0.05, Mann-Whitney test). This relationship disappeared, however, when only data from post-menopausal women were included in the analysis. Mean Treg cell frequency was 4.65% in postmenopausal women with cancer (n = 23) and 4.73% in postmenopausal controls (n = 5) (p = 0.9). In women without cancer we found that mean Treg cell frequency was higher in postmenopausal women (4.73%, n = 5) in comparison to premenopausal controls (3.33%, n = 16) (p = 0.02). These results suggest that the increased proportion of Treg cells seen in endometrial cancer patients might be, at least in part, attributed to their postmenopausal status or age.