Immunological Effector Cells Research Articles

Macrophages reprogramming improves immunotherapy of IL-33 in peritoneal metastasis of gastric cancer

Peritoneal metastasis (PM) has a suppressive tumor immune microenvironment (TIME) that limits the effects of immunotherapy. This study aimed to investigate the immunomodulatory effects of intraperitoneal administration of IL-33, a cytokine that is reported to potentiate antitumor immunity and inhibit metastasis. We found survival was significantly prolonged in patients with high IL-33 mRNA expression. In immunocompetent mice, intraperitoneal administration of IL-33 could induce a celiac inflammatory environment, activate immunologic effector cells, and reverse the immunosuppressive tumor microenvironment, which effectively delayed tumor progression and PM of gastric cancer. Mechanistically, IL-33 could induce M2 polarization by activating p38-GATA-binding protein 3 signaling. IL-33 combined with anti-CSF1R or p38 inhibitor to regulate tumor-associated macrophages (TAMs) had a synergistic antitumor effect. Inducing a local inflammatory milieu by IL-33 administration provided a novel approach for treating peritoneal metastasis, which, when combined with TAM reprogramming to reshape TIME, can achieve better treatment efficacy.

NNV024, a Humanized Anti-CD37 Antibody with Enhanced ADCC and Extended Plasma Half-Life for the Treatment of B-Cell Malignancies

Background: There is a significant unmet clinical need for new therapeutic approaches for patients with B-cell non-Hodgkin lymphomas (B-NHL) who relapse or are refractory to anti-CD20 therapies. Moreover, certain B-cell malignancies do not express CD20. High expression of the CD37 antigen on malignant B-cells associated with limited or no expression on other hematopoietic cells makes CD37 a highly attractive alternative target for B-cell depleting therapies. We have therefore developed a humanized anti-CD37 monoclonal antibody (NNV024) engineered to exhibit a strong antibody-dependent cellular cytotoxicity (ADCC). Methods: Generation of NNV024 was performed by combining in silico CDR-grafting and structure-based back mutation. Candidates were then selected for their affinity to CD37 expressing cell line, Ramos. In silico immunogenicity prediction tools were integrated in the selection process to support the generation of a candidate with low immunogenicity potential (NNV023). Last, to increase ADCC, the selected candidate was afucosylated at the N-linked bi-antennary glycan at N297 of the Fc region (NNV024). The NNV024 was characterized in vitro and in vivo. Binding to human Fc gamma receptors (FcγRs), neonatal Fc receptor (FcRn), and complement component 1q (C1q) was confirmed by ELISA and SPR. ADCC and antibody-dependent cellular phagocytosis (ADCP) induction were measured using cell-based reporter assays with eleven cell lines derived from B-NHL subtypes such as Burkitt's lymphoma, diffuse large B-Cell lymphoma (DLBCL), and mantel cell lymphoma (MCL). All cell lines express both CD20 and CD37 on their cell surface. In addition, B cell chronic lymphocytic leukaemia (B-CLL) cells isolated from patient samples were used to test induction of ADCC, ADCP and complement-dependent cytotoxicity (CDC) in vitro. NNV024 plasma pharmacokinetics were assessed in Tg32 transgenic mice expressing human FcRn, which is considered a key regulator of IgG plasma half-life. A therapy study comparing NNV024 (n=10) and obinutuzumab (n=10) was performed using a disseminated tumor (Daudi) model in CB17-SCID mice (107 cells/mouse, i.v.). The anti-CD20 Abs rituximab and obinutuzumab as well as the anti-CD37 Ab, Duohexabody-CD37, were used for benchmarking. Results: As expected, the glyco-engineering increased affinity of NNV024 to FcγRIIIa (KD before afucosylation: 460.8 ng/mL; KD after: 104.1 ng/mL). Binding of Abs to FcγRIIIa activates immunological effector cells such as Natural Killer cells, macrophages, monocytes, and eosinophils. NNV024 demonstrated ADCC superior to that observed with obinutuzumab, rituximab, and Duohexabody-CD37. NNV024 exhibited a 2-fold prolonged plasma half-life in mice (8.9±0.8 days) compared to obinutuzumab (4.4±1.3 days) and Duohexabody-CD37 (4.1±1.2 days). Mice survival was enhanced up to ~ 40% when treated with NNV024 (10 µg/mouse - 70.5 days; 50 µg/mouse - 84 days) compared to obinutuzumab at a dose of 10 µg (70.5 days vs. 53 days) or 50 µg (84 days vs. 62 days). Conclusion: These encouraging preclinical results support further development of NNV024 as a potential therapeutic mAb candidate for the treatment of B-cell malignancies and B-cell driven autoimmune disorders.

A Low Dose of Pure Cannabidiol Is Sufficient to Stimulate the Cytotoxic Function of CIK Cells without Exerting the Downstream Mediators in Pancreatic Cancer Cells.

Despite numerous studies conducted over the past decade, the exact role of the cannabinoid system in cancer development remains unclear. Though research has focused on two cannabinoid receptors (CB1, CB2) activated by most cannabinoids, CB2 holds greater attention due to its expression in cells of the immune system. In particular, cytokine-induced killer cells (CIKs), which are pivotal cytotoxic immunological effector cells, express a high-level of CB2 receptors. Herein, we sought to investigate whether inducing CIK cells with cannabidiol can enhance their cytotoxicity and if there are any possible counter effects in its downstream cascade of phosphorylated p38 and CREB using a pancreatic ductal adenocarcinoma cell line (PANC-1). Our results showed that IL-2 modulates primarily the expression of the CB2 receptor on CIK cells used during ex vivo CIK expansion. The autophagosomal-associated scaffold protein p62 was found to co-localize with CB2 receptors in CIK cells and the PANC-1 cell line. CIK cells showed a low level of intracellular phospho-p38 and, when stimulated with cannabidiol (CBD), a donor specific variability in phospho-CREB. CBD significantly decreases the viability of PANC-1 cells presumably by increasing the cytotoxicity of CIK cells. Taken together, in our preclinical in vitro study, we propose that a low effective dose of CBD is sufficient to stimulate the cytotoxic function of CIK without exerting any associated mediator. Thus, the combinatorial approach of non-psychoactive CBD and CIK cells appears to be safe and can be considered for a clinical perspective in pancreatic cancer.

Assessment of FcgRIIIA single nucleotide polymorphisms (SNPs) on the efficacy of IgG1 monoclonal antibodies (mAbs) in PANGEA study patients (pts) with advanced gastroesophageal adenocarcinoma (aGEA).

4526 Background: Targeted therapies (Ttx) have had limited efficacy in aGEA. The phase IIa PANGEA trial assessed the outcomes of pts treated with IgG1 mAbs targeting receptor tyrosine kinases (RTKs) or PD-1 based on predefined molecular groups. The fragment C (Fc) portion of mAbs binds to IgG receptors (FcgR) of immunologic effector cells such as natural killer (NK) cells, leading to antibody-dependent cell-mediated cytotoxicity (ADCC). The FcgR subclass, FcgRIIIA, has genetic variants with different Fc binding affinities. A single nucleotide polymorphism (SNP) in FcgRIIIA substitutes phenylalanine (F) with valine (V) at amino acid position 158, enhancing FcgR’s affinity for the IgG1 Fc domain. Pts with V/V or V/F FcgRIIIA allotypes have enhanced NK cell binding affinity compared to the homozygous F/F allotype. We evaluated the association of FcgRIIIA SNPs on Ttx outcomes amongst PANGEA pts and another cohort of aGEA pts treated with IgG1 mAbs. Methods: Whole-blood samples were identified from aGEA pts (N = 104), including 70/80 available PANGEA pts, who were treated with an IgG1 mAb (trastuzumab 24, anti-EGFR 21, anti-PD1 30, ramucirumab 48) in at least 1 Ttx line. After lymphocyte DNA extraction, FcgRIIIA genotyping was performed. The Cox proportional hazard model and log-rank tests, adjusted for pt age, were used to assess for an association of genotype with overall survival (OS). Results: Of 104 genotyped pts, the F/F, F/V & V/V genotypes were observed at a frequency of 32%, 51% and 17% respectively. There was no significant difference in median OS (mOS) between the F/F, F/V or V/V or comparing F/F vs V/F+V/V overall, nor in the PANGEA-only cohort. A trend of increased mOS was seen in 20 non-PANGEA pts harboring F/V or V/V compared to 14 F/F pts (mOS 43.4 vs 23.1 months, HR 0.41 [0.15-1.14] p = 0.09). However, 3-year OS rates trended higher in V/F+V/V pts (22%, 16/71) compared to F/F pts (7%, 2/33) (p = 0.09). At 3 years, 50% of V/V+V/F non-PANGEA pts were alive versus 13% of F/F pts (p = 0.04), while 13% of V/V+V/F PANGEA pts were alive versus 0% of F/F pts (p = 0.32). Conclusions: Amongst pts receiving IgG1 mAbs, the high affinity V FcgRIIIA SNP enriched for a subgroup of ‘extreme responders’ alive 3 years from diagnosis. Multivariate analyses accounting for baseline characteristics in a larger number of pts are ongoing to further elucidate the role of FcgRIIIA SNPs as predictive biomarkers. These findings may have implications on IgG1 mAb ADCC optimization.

Clinical Trials with Combination of Cytokine-Induced Killer Cells and Dendritic Cells for Cancer Therapy.

Adoptive cellular immunotherapy (ACI) is a promising treatment for a number of cancers. Cytokine-induced killer cells (CIKs) are considered to be major cytotoxic immunologic effector cells. Usually cancer cells are able to suppress antitumor responses by secreting immunosuppressive factors. CIKs have significant antitumor activity and are capable of eradicating tumors with few side effects. They are a very encouraging cell population used against hematological and solid tumors, with an inexpensive expansion protocol which could yield to superior clinical outcome in clinical trials employing adoptive cellular therapy combination. In the last decade, clinical protocols have been modified by enriching lymphocytes with CIK cells. They are a subpopulation of lymphocytes characterized by the expression of CD3+ and CD56+ wich are surface markers common to T lymphocytes and natural killer NK cells. CIK cells are mainly used in two diseases: in hematological patients who suffer relapse after allogeneic transplantation and in patients with hepatic carcinoma after surgical ablation to eliminate residual tumor cells. Dendritic cells DCs could play a pivotal role in enhancing the antitumor efficacy of CIKs.

Novel TLR7 agonist stimulates activity of CIK/NK immunological effector cells to enhance antitumor cytotoxicity.

Toll-like receptor (TLR) 7/8 agonists have been applied in combination with chemo-, radio- or immunotherapy for lymphoma, and used as topical drugs for the treatment of viral skin lesions and skin tumors. In the present study, the role of an adenine analog, 9-(4-carboxyphenyl)-8-hydroxy-2-(2-methoxyethoxy)-adenine [termed Gao Dong (GD)], a novel TLR7 agonist, in the activation of cytokine-induced killer/natural killer (CIK/NK) cells was determined. The results of the present study indicated that GD was able to activate CIK/NK cells. The proportion of GD-induced CD3+CD56+ CIK and CD3-CD56+ NK cells was ~4% higher respectively compared with the control. Notably, combination therapy with CIK/NK cells stimulated by GD, markedly suppressed the proliferation of the chronic myelogenous leukemia K562 cell line. Following GD treatment, the cytotoxicity improved by ~25 and 21% when the effector/target ratio was 20:1 and 10:1, respectively. The results of the present study suggested a novel protocol for CIK/NK cell proliferation and revealed that GD may serve as a potent innate and adaptive immunomodulator in immunocyte culture.

Blood components – so much more than clots and oxygen delivery!

Blood, and the blood components, packed red blood cells (pRBC), platelet concentrate (PC), plasma and cryoprecipitate, have traditionally been transfused to maintain or improve oxygen delivery, blood volume and/or haemostatic capacity. There is, however, emerging evidence that blood components may support other alternative functions beyond oxygen delivery, blood volume and haemostasis. Thus, PC may through their functions as innate immunological effector cells improve immune competence. Plasma may through its endothelial protective and repairing functions improve vascular integrity and hence outcome in critical illness. Erythrocytes and thus pRBC may be critical regulators of the vascular, haemostatic and immune systems through their haemostatic function, their chemokine scavenging capacity and their endothelial crosstalk functions. Finally, the blood cell derived vesicles present in all blood components seem to influence the vascular, haemostatic and immune systems in a way that goes far beyond that of passive by‐products. The present review reveals, and discusses, some emerging new functions of blood and blood components and thus alternative indications for transfusion.

Immunotherapy and gene therapy as novel treatments for cancer.

The immune system interacts closely with tumors during the disease development and progression to metastasis. The complex communication between the immune system and the tumor cells can prevent or promote tumor growth. New therapeutic approaches harnessing protective immunological mechanisms have recently shown very promising results. This is performed by blocking inhibitory signals or by activating immunological effector cells directly. Immune checkpoint blockade with monoclonal antibodies directed against the inhibitory immune receptors CTLA-4 and PD-1 has emerged as a successful treatment approach for patients with advanced melanoma. Ipilimumab is an anti-CTLA-4 antibody which demonstrated good results when administered to patients with melanoma. Gene therapy has also shown promising results in clinical trials. Particularly, Herpes simplex virus (HSV)-mediated delivery of the HSV thymidine kinase (TK) gene to tumor cells in combination with ganciclovir (GCV) may provide an effective suicide gene therapy for destruction of glioblastomas, prostate tumors and other neoplasias by recruiting tumor-infiltrating lymphocytes into the tumor. The development of new treatment strategies or combination of available innovative therapies to improve cell cytotoxic T lymphocytes trafficking into the tumor mass and the production of inhibitory molecules blocking tumor tissue immune-tolerance are crucial to improve the efficacy of cancer therapy.

Novel vaccines for glioblastoma: clinical update and perspective.

Glioblastoma is the most common primary brain cancer. Aggressive treatment with surgery, radiation therapy and chemotherapy provides limited overall survival benefit. Glioblastomas have a formidable tumor microenvironment that is hostile to immunological effector cells and these cancers produce profound systemic immunosuppression. However, surgical resection of these tumors creates conditions that favor the use of immunotherapeutic strategies. Therefore, extensive surgical resection, when feasible, will remain part of the equation to provide an environment in which active specific immunotherapy has the greatest chance of working. Toward that end, a number of vaccination protocols are under investigation. Vaccines studied to date have produced cellular and humoral antitumor responses, but unequivocal clinical efficacy has yet to be demonstrated. In addition, focus is shifting toward the prospect of therapies involving vaccines in combination with immune checkpoint inhibitors and other immunomodulatory agents so that effector cells remain active against their targets systemically and within the tumor microenvironment.

Reduction of Minimal Residual Disease in Pediatric B-lineage Acute Lymphoblastic Leukemia by an Fc-optimized CD19 Antibody.

Prognosis of primary refractory and relapsed pediatric B-lineage acute lymphoblastic leukemia (ALL) is very poor. Relapse rates significantly correlate with persistent minimal residual disease (MRD). In MRD, favorable effector-target ratios prevail and thus this situation might be optimally suited for immunotherapy with antibodies recruiting immunological effector cells. We here report on the generation, preclinical characterization and first clinical application in B-lineage ALL of an Fc-optimized CD19 antibody. This third-generation antibody (4G7SDIE) mediated enhanced antibody-dependent cellular cytotoxicity (ADCC) against leukemic blasts with effector cells from healthy volunteers and B-lineage ALL patients. The antibody was produced in a university-owned production unit and was applied on a compassionate use basis to 14 pediatric patients with refractory and relapsed B-lineage ALL at the stage of MRD. In 10/14 patients, MRD was reduced by ≥ 1 log or below the patient-individual detection limit, and 5/14 patients have achieved ongoing complete molecular remission with a median leukemia-free survival of 428 days. Two additional patients died in complete molecular remission due to complications not related to antibody therapy. Besides profound in vivo B-cell depletion, side effects were negligible. A clinical phase 1/2 study to further assess the therapeutic activity of 4G7SDIE is in preparation.

Mycobacterium tuberculosis-Induced Polarization of Human Macrophage Orchestrates the Formation and Development of Tuberculous Granulomas In Vitro.

The tuberculous granuloma is an elaborately organized structure and one of the main histological hallmarks of tuberculosis. Macrophages, which are important immunologic effector and antigen-presenting cells, are the main cell type found in the tuberculous granuloma and have high plasticity. Macrophage polarization during bacterial infection has been elucidated in numerous recent studies; however, macrophage polarization during tuberculous granuloma formation and development has rarely been reported. It remains to be clarified whether differences in the activation status of macrophages affect granuloma formation. In this study, the variation in macrophage polarization during the formation and development of tuberculous granulomas was investigated in both sections of lung tissues from tuberculosis patients and an in vitro tuberculous granuloma model. The roles of macrophage polarization in this process were also investigated. Mycobacterium tuberculosis (M. tuberculosis) infection was found to induce monocyte-derived macrophage polarization. In the in vitro tuberculous granuloma model, macrophage transformation from M1 to M2 was observed over time following M. tuberculosis infection. M2 macrophages were found to predominate in both necrotic and non-necrotic granulomas from tuberculosis patients, while both M1 and M2 polarized macrophages were found in the non-granulomatous lung tissues. Furthermore, it was found that M1 macrophages promote granuloma formation and macrophage bactericidal activity in vitro, while M2 macrophages inhibit these effects. The findings of this study provide insights into the mechanism by which M. tuberculosis circumvents the host immune system as well as a theoretical foundation for the development of novel tuberculosis therapies based on reprogramming macrophage polarization.

P0241 : CD40/CD40Ligand interaction between tumor lysate-pulsed dendritic cells induces a Th1-environment and enhances the cytotoxic activity of immunologic effector cells towards human hepatocellular carcinoma and cholangiocellular carcinoma cells

P0241 : CD40/CD40Ligand interaction between tumor lysate-pulsed dendritic cells induces a Th1-environment and enhances the cytotoxic activity of immunologic effector cells towards human hepatocellular carcinoma and cholangiocellular carcinoma cells

Immunotherapy of Ovarian Cancer: The Role of Checkpoint Inhibitors.

Ovarian cancer is the most important cause of gynecological cancer-related mortality, with the majority of women presenting with advanced disease. Although surgery and chemotherapy can improve survival rates, it is necessary to integrate alternative strategies to improve the outcomes. Advances in understanding the role of immune system in the pathogenesis of cancer have led to the rapid evolvement of immunotherapy, which might establish a sustained immune system response against recurring cancer cells. Recently, it has emerged that powerful immunologic effector cells may be blocked by inhibitory regulatory pathways controlled by specific molecules often called “immune checkpoints,” which turn off the immune system. Similarly, cancer cells are able to use these checkpoints to avoid immune control and rejection. Inhibition of these inhibitory pathways represents a potent strategy in the fight against cancer and is currently under investigation with encouraging results in some cancers, such as melanoma. In ovarian cancer researches are still in an early phase, but with promising results. In this review we will explore the rationale of immunotherapy in ovarian cancer with a special focus on these emerging molecules.

Advances in Bispecific Antibodies Engineering: Novel Concepts for Immunotherapies

Bispecific antibodies are emerging as novel approach for immunotherapies by combining two antigen-recognizing elements into a single construct that is able to simultaneously bind to two distinct targets. Bispecific antibodies can be applied to recruit immunological effector cells for killing of tumor cells or to simultaneously block two signaling pathways or cytokines. Subsequently, they have prompted significant interest for a number of therapeutic applications, both in cancer and in other indications. The growing interest in therapeutic antibodies along with the rapid progress in antibody engineering have yield a multitude of bispecific antibodies formats and derived molecules that differ in size, shape and function. Presently, two major classes of bispecific antibodies are most widely studied: immunoglobulin-like and small single chain Fv (scFv)-based bispecific antibodies. This review summarizes selected key methods used to generate bispecific antibodies, reports new concepts developed for immunotherapy and discusses their potential development and therapeutic benefits.

Inflammatory response to subcutaneous allergen-specific immunotherapy in patients with bronchial asthma and allergic rhinitis

Background: Bronchial asthma (BA) and allergic rhinitis (AR) are chronic inflammatory disorders of the airways. The allergic response is driven by the production of different immunological effector cells cytokines like interleukin-5 (IL-5) and IL-6 among others. Subcutaneous allergen-specific immunotherapy (SCIT) modifies basic immunological mechanisms, reducing IL-5 production. The effect of SCIT on levels of IL-6 is undetermined. Objective: The aim is to study the changes in immunological parameters that follow SCIT in patients suffering from BA and/or AR. Materials and Methods: Twenty-nine patients (18-48 years, mean 25.5 years) diagnosed with BA and/or AR were evaluated for allergic sensitivity using skin prick test (SPT). The patients were started on standardized treatment for BA and AR as per global initiative for asthma and AR and its impact on asthma guidelines, respectively. SCIT was initiated as per the standard Indian guidelines. IL-5 and IL-6 levels were obtained at 0, 3 and 6 months during the course of SCIT and the response was evaluated using Friedman test. Results: Twenty-nine patients; 16 males and 13 females were evaluated and initiated on SCIT. The decreasing order of antigen sensitivity on SPT was mosquito (65.5%), housefly (58.6%), female cockroach (58.6%), male cockroach (48.2%), moth (34.4%) and house dust mite (17.2%). The IL-5 and IL-6 levels, for 0, 3 and 6 months were compared, and it was noted that with an increase in duration of treatment, the levels of inflammatory markers significantly decreases (P = 0.003). On comparison, the inflammatory response between male and female, duration of symptoms and number of positive antigens was not statistically significant. Conclusion: Immunologic changes associated with immunotherapy are complex and allergic patients suffering from asthma, and/or rhinitis showed a significant reduction in levels of inflammatory markers.

Immune checkpoint inhibitors in clinical trials.

Immunology-based therapy is rapidly developing into an effective treatment option for a surprising range of cancers. We have learned over the last decade that powerful immunologic effector cells may be blocked by inhibitory regulatory pathways controlled by specific molecules often called “immune checkpoints.” These checkpoints serve to control or turn off the immune response when it is no longer needed to prevent tissue injury and autoimmunity. Cancer cells have learned or evolved to use these mechanisms to evade immune control and elimination. The development of a new therapeutic class of drugs that inhibit these inhibitory pathways has recently emerged as a potent strategy in oncology. Three sets of agents have emerged in clinical trials exploiting this strategy. These agents are antibody-based therapies targeting cytotoxic T-lymphocyte antigen 4 (CTLA4), programmed cell death 1 (PD-1), and programmed cell death ligand 1 (PD-L1). These inhibitors of immune inhibition have demonstrated extensive activity as single agents and in combinations. Clinical responses have been seen in melanoma, renal cell carcinoma, non-small cell lung cancer, and several other tumor types. Despite the autoimmune or inflammatory immune-mediated adverse effects which have been seen, the responses and overall survival benefits exhibited thus far warrant further clinical development.

Cytotoxic T Lymphocytes: The Future of Cancer Stem Cell Eradication?

Cancer stem-like cells (CSCs)/cancer-initiating cells (CICs) are a hot topic in cancer research since they are highly tumorigenic and are resistant to standard cancer therapies. Recent studies have revealed that cancer immunotherapy is a possible and promising candidate to target CSCs/CICs. Among the various immunological effector cells, cytotoxic T lymphocytes (CTLs) are a good candidate for CSC/CIC-targeted immunotherapy as CTLs are antigen-specific effector cells. In this article, we summarize advances in studies on CTLs and discuss the future of CSC/CIC-targeted therapy. Since the first identification of leukemia stem cells from an acute myeloid leukemia sample, studies of CSCs/CICs have made great advances [1]. CSCs/CICs are a small population of cancer cells that have tumor-initiating, self-renewal and differentiation abilities. Recent studies have revealed that CSCs/CICs are resistant to cancer therapies because they are in a quiescent cellcycle state, they express high levels of transporters and apoptosis inhibitors, and they express low levels of reactive oxygen species. CSCs are therefore regarded as major causes of cancer recurrence, distant metastasis and treatment resistance; studies of CSCs/CICs have been focusing on how CSCs/CICs can be targeted efficiently.

The differential immunological activities of Ganoderma lucidum on human pre-cancerous uroepithelial cells

Ganoderma lucidum is active to stimulate immunological effector cells, but the effects on uroepithelial cells have never been explored. The present study compared the expression of major cytokines induced by the water (GLw) and ethanol (GLe) extracts of G. lucidum. The pre-cancerous human uroepithelial cell (HUC-PC) line was employed. A total of 15 cytokines, including major Th1/Th2 cytokines and chemokines, were measured in the complete media after 24h incubation with GLw and GLe. Additionally, the following assays were performed: cytotoxicity, apoptosis, migration of neutrophils, and nuclear factor-kappaB (NF-κB) DNA binding activity. GLe inhibited the growth of HUC-PC cells through apoptosis. Interleukins IL-2, IL-6, and IL-8 were significantly up-regulated by GLe in dose-dependent manners, but not by GLw. However, MCP-1 level was significantly increased by GLw but was oppositely reduced by GLe. Furthermore, the elevation of cytokine expression was correlated with the enhancement of p50/p65 NF-κB activity induced by GLe. The elevated IL-8 levels in GLe-treated cells were also correlated with the migration of neutrophils. GLe and GLw exhibited different immunological activities on the HUC-PC cells. In particular, the activities of GLe may favor the clearance of high risk urothelial cells, suggesting potent chemopreventive ingredients are extractable by ethanol from G. lucidum.

Multimodality treatment of malignant pleural mesothelioma with or without immunotherapy: does it change anything?☆

The purpose of this study was to investigate immunological effector cells and angiogenesis in malignant pleural mesothelioma (MPM) patients, who underwent multimodality treatments. Clinical and pathological characteristics of 57 patients, with International Mesothelioma Interest Group stage II-III MPM, who underwent two different multimodality treatments (with and without immunotherapy) between 1999 and 2008 were analyzed. CD8+, CD4+ and Foxp3+ tumor-infiltrating lymphocytes, tryptase and chymase mast cells (MCs), CD34, number of microvessels and vascular endothelial growth factor were determined by immunohistochemistry. The histology was 51 epitheliomorf and 6 biphasic. The stage was III in 41 cases and II in 16 cases. With an average follow-up of 69 months (range 9-115) 14 patients are still alive and the overall median actuarial survival is 21.4 months. Tryptase MCs, CD8+ and Foxp3+ lymphocytes had significantly increased in the interleukin 2 (IL-2) treated group. Moreover, the number of microvessels was significantly lower in IL-2 treated patients. This study indicates that immunotherapy leads to an increase in cytotoxic CD8+ lymphocytes and tryptase MCs and to a decrease of the tumoral neoangiogenesis. Changes in MPM microenvironment induced by immunotherapy may play a major role in the local control of this disease and need further investigations.

Treatment with interleukin-2 in malignant pleural mesothelioma: immunological and angiogenetic assessment and prognostic impact

Background:Administration of interleukin-2 (IL-2) has shown some effects on malignant pleural mesothelioma (MPM) tumour regression. The purpose of this study was to investigate the ability of IL-2 to modify immunological effector cells and angiogenesis in MPM patients and their prognostic value.Methods:Tumour-infiltrating lymphocytes (CD4, CD8, Foxp3), mast cells (MCs) (tryptase and chymase), microvessel count (MVC) and VEGF were determined by immunohistochemistry in two series of MPM patients: 60 patients treated with intra-pleural preoperative IL-2 and 33 patients untreated.Results:Tryptase MCs, and CD8 and Foxp3 lymphocytes were significantly increased in the IL-2-treated group, whereas MVC was significantly lower in the same group. Moreover, in the IL-2-treated group, greater tryptase+MCs and greater Foxp3 lymphocytes were associated with improved and poorer clinical outcomes, respectively. Notably, when these two immunological parameters were combined, they predicted outcomes more effectively.Conclusions:This study showed that IL-2 treatment leads to a significant increase of immunological parameters, concomitantly with a reduction in vasculature, providing new insight into the cancer mechanisms mediated by IL-2. Moreover, these results suggest that tryptase-positive MCs and Foxp3+ lymphocytes predict clinical outcomes in IL-2-treated patients, highlighting the critical role of the inflammatory response in mesothelioma cancer progression.