Immune Cells In Patients Research Articles

Abstract 2635: Differential prognostic implications of programmed death-ligand 1 expression on tumor cells and tumor-infiltrating immune cells in patients with locally advanced esophageal squamous cell carcinoma treated with preoperative chemoradiotherapy

Abstract Introduction Previous studies have reported inconsistent results on the prognostic implications of program death-ligand 1 (PD-L1) expression for patients with locoregional esophageal squamous cell carcinoma (ESCC). We evaluated the differential prognostic impacts of the PD-L1 expressions on tumor cells and on tumor infiltrating immune cells in a cohort of locally advanced ESCC patients, receiving neoadjuvant chemoradiotherapy (CRT). Patients and Methods A cohort of 66 patients of locally advanced ESCC have been enrolled to a phase II clinical trial of a single-cycle chemotherapy (a combination of weekly paclitaxel, cisplatin and 24 hour infusional 5-fluorouracil, 2 week on plus one week off, followed by paclitaxel/cisplatin (TP) -based preoperative CRT and radical surgery. Primary tumor tissues were retrospectively collected for PD-L1 immunohistochemical stain by the Clone SP142 antibody (Spring Bioscience, Pleasanton, CA, USA). The expression of PD-L1 was scored as previously reported. Tumor cells (TCs) were scored according to percentage of positively-stained cells: TC3≥50%, TC2≥5% and <50%, TC1≥1% and <5%, and TC0<1%; Immune cells (ICs): IC3≥10%, IC2≥5% and <10%, IC1≥1% and <5%, and IC0<1%. Results PD-L1 expression was evaluable on tumor cells in 49 patients and on immune cells in 51 patients. TC0, TC1, TC2, and TC3 were scored in 27 (55%), 9 (18%), 11 (22%), and 2 (4%) patients, respectively; and IC0, IC1, IC2, and IC3 were in 25 (49%), 9 (18%), 15 (29%), and 2 (4%) patients, respectively. Regression analysis revealed that PD-L1 expressions on TC and IC exhibited different impacts on overall survival (OS): higher PD-L1 expression on TC trended to correlate with worse OS, whereas higher PD-L1 expression on IC trended to correlated with better OS. The median OSs of patients with TC0 and those with TC 1-3 were 52 months and 21 months, respectively, with a hazard ratio (HR) 1.5 (95% confidence interval [CI]: 0.7~3.3; P= 0.27). According to the PD-L1 expressions on TC and IC, patients with TC(+)/IC(+), TC(+)/IC(-), TC(-)/IC(+), TC(-)/IC(-) exhibited median OSs of 26, 7, 42, and 43 months, respectively. Patients with TC(+)/IC(-) PD-L1 expression had significantly worse OS than other patients (HR: 2.6, P=0.03). Among patients with TC(+) PD-L1 expression, those with IC(-) had significantly worse OS than those with IC (+), too (HR: 2.5, P=0.04). Conclusion High PD-L1 expression on TC and low PD-L1 expression on IC associate with inferior prognosis in locally advanced ESCC patients treated with neoadjuvant CRT. (The study was supported by the grant NTUH 104-M2891 and the grant MOST 105-2314-B-002-186-MY3) Citation Format: Ta-Chen Huang, Cher-Wei Liang, Chia-Chi Lin, Ya-Jhen Chen, Kuan-Ling Lin, Chih-Hung Hsu. Differential prognostic implications of programmed death-ligand 1 expression on tumor cells and tumor-infiltrating immune cells in patients with locally advanced esophageal squamous cell carcinoma treated with preoperative chemoradiotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2635.

Immunotherapy in Some Types of Tumors

Immunotherapy in Some Types of Tumors

Macrophages Represent the Major Pool of Interleukin-7 Receptor-α Expressing Immune Cells in Patients with Myocarditis

Myocarditis is an inflammatory heart disease which involves a large number of cytokine receptors. In order to identify potential key signalling cascades we screened a number of receptors by Western blot and found high expression of the interleukin-7 (IL-7) receptor-α (IL-7Rα). Since IL-7 is usually required for the development of B as well as T lymphocytes we wanted to determine the major cell type(s) expressing this receptor. Importantly, immune cells might be also a source of IL-7.

Defective formyl peptide receptor 2/3 and annexin A1 expressions associated with M2a polarization of blood immune cells in patients with chronic obstructive pulmonary disease

BackgroundControversy exists in previous studies on macrophage M1/M2 polarization in chronic obstructive pulmonary disease (COPD). We hypothesized that formyl peptide receptor (FPR), a marker of efferocytosis and mediator of M1/M2 polarization, may be involved in the development of COPD.MethodsWe examined FPR 1/2/3 expressions of blood M1/M2a monocyte, neutrophil, natural killer (NK) cell, NK T cell, T helper (Th) cell, and T cytotoxic (Tc) cell by flowcytometry method in 40 patients with cigarette smoking-related COPD and 16 healthy non-smokers. Serum levels of five FPR ligands were measured by ELISA method.ResultsThe COPD patients had lower M2a percentage and higher percentages of NK, NK T, Th, and Tc cells than the healthy non-smokers. FPR2 expressions on Th/Tc cells, FPR3 expressions of M1, M2a, NK, NK T, Th, and Tc cells, and serum annexin A1 (an endogenous FPR2 ligand) levels were all decreased in the COPD patients as compared with that in the healthy non-smokers. FPR1 expression on neutrophil was increased in the COPD patient with a high MMRC dyspnea scale, while FPR2 expression on neutrophil and annexin A1 were both decreased in the COPD patients with a history of frequent moderate exacerbation (≥ 2 events in the past 1 year). In 10 COPD patients whose blood samples were collected again after 1-year treatment, M2a percentage, FPR3 expressions of M1/NK/Th cells, FPR2 expression on Th cell, and FPR1 expression on neutrophil were all reversed to normal, in parallel with partial improvement in small airway dysfunction.ConclusionsOur findings provide evidence for defective FPR2/3 and annexin A1 expressions that, associated with decreased M2a polarization, might be involved in the development of cigarette smoking induced persistent airflow limitation in COPD.

Changes in natural killer cells and exhausted memory regulatory T Cells with corticosteroid therapy in acute autoimmune hepatitis.

Autoimmune hepatitis (AIH) is an immune‐mediated liver disease currently treated by immunosuppressive medications with significant side effects. Thus, novel mechanistic treatments are greatly needed. We performed prospective deep immunophenotyping of blood immune cells in patients with acute AIH before and after corticosteroid therapy. Blood samples from 26 patients with acute AIH (United Kingdom‐AIH Consortium) were phenotyped by flow cytometry at baseline and 4 months after starting corticosteroids. Pretreatment liver tissues were stained for forkhead box P3‐positive (FOXP3POS) regulatory T cells (Tregs), clusters of differentiation (CD)56POS natural killer (NK) cells, and chemokine (C‐X‐C motif) ligand 10. Chemokine secretion by cultured primary hepatocyte and biliary epithelial cells was measured by enzyme‐linked immunosorbent assay. Functional coculture assays with stimulated NK cells and Tregs were performed. CD161 ligand, lectin‐like transcript‐1 expression by intrahepatic immune cells was demonstrated with flow cytometry. Frequencies of NKbright cells declined with therapy (P < 0.001) and correlated with levels of alanine aminotransferase (P = 0.023). The Treg:NKbright ratio was lower pretreatment, and Tregs had an activated memory phenotype with high levels of CD39, cytotoxic T lymphocyte antigen 4, and FOXP3 but also high programmed death ligand 1, indicating exhaustion. Coculture experiments suggested the Tregs could not efficiently suppress interferon‐γ secretion by NK cells. Both Tregs and NK cells had high expression of liver infiltration and T helper 17 plasticity‐associated marker CD161 (P = 0.04). Pretreatment and CD161pos NK cells expressed high levels of perforin and granzyme B, consistent with an activated effector phenotype (P < 0.05). Lectin‐like transcript 1, a ligand for CD161, is expressed on intrahepatic B cells, monocytes, and neutrophils. Conclusion: Activated effector NK cells, which correlate with biochemical measurements of hepatitis, and exhausted memory Tregs are increased in the blood of patients with treatment‐naive AIH and decline with corticosteroid therapy. Inadequate regulation of NK cells by exhausted FOXP3pos Tregs may play a role in AIH pathogenesis and contribute to liver injury. (Hepatology Communications 2018;2:421‐436)

Overexpression of adhesion molecules and barrier molecules is associated with differential infiltration of immune cells in non-small cell lung cancer

Immunotherapy is emerging as a promising option for lung cancer treatment. Various endothelial adhesion molecules, such as integrin and selectin, as well as various cellular barrier molecules such as desmosome and tight junctions, regulate T-cell infiltration in the tumor microenvironment. However, little is known regarding how these molecules affect immune cells in patients with lung cancer. We demonstrated for the first time that overexpression of endothelial adhesion molecules and cellular barrier molecule genes was linked to differential infiltration of particular immune cells in non-small cell lung cancer. Overexpression of endothelial adhesion molecule genes is associated with significantly lower infiltration of activated CD4 and CD8 T-cells, but higher infiltration of activated B-cells and regulatory T-cells. In contrast, overexpression of desmosome genes was correlated with significantly higher infiltration of activated CD4 and CD8 T-cells, but lower infiltration of activated B-cells and regulatory T-cells in lung adenocarcinoma. This inverse relation of immune cells aligns with previous studies of tumor-infiltrating B-cells inhibiting T-cell activation. Although overexpression of endothelial adhesion molecule or cellular barrier molecule genes alone was not predictive of overall survival in our sample, these genetic signatures may serve as biomarkers of immune exclusion, or resistance to T-cell mediated immunotherapy.

Patients with idiopathic endplate inflammation displayed aberrant Th17 responses resulting in proinflammatory monocyte activation

Endplate inflammation remains a difficult task to diagnose and treat, partly due to the absence of in-depth understanding of the cellular and molecular factors driving this condition. In the current study, we investigated the circulating immune cells in patients with idiopathic endplate inflammation. Compared to healthy controls, the patients with endplate inflammation presented a significant upregulation of Th17 cells, characterized by higher frequencies of circulating IL-17+CD4+ T cells examined directly ex vivo and after PMA and ionomycin (PMA/I) stimulation. The frequency of Th17 cells in patients was not correlated with patient age, sex, or smoking status, but was significantly correlated with patient BMI. The total CD4+ T cells from patients with end plate inflammation also presented significantly higher levels of IL-17 secretion directly ex vivo and after PMA/I stimulation. The IL-17 secretion was primarily mediated by CCR4+CCR6+ CD4+ T cells, followed by CCR4-CCR6+ CD4+ T cells and was nearly absent in CCR4-CCR6- CD4+ T cells. Monocytes incubated with CCR4+CCR6+ CD4+ T cells presented significantly higher MHC class II expression, as well as higher CD80 and CD86 expression. The secretion of IL-6 and TNF was significantly higher in cell cultures containing CCR4+CCR6+ CD4+ T cells than in cell cultures containing CCR4-CCR6- CD4+ T cells. These effects were reduced when the IL-17A cytokine was neutralized. Together, we identified a systemic upregulation of Th17 responses that could contribute to proinflammatory monocyte activation in patients with endplate inflammation.

Head and Neck Carcinoma Immunotherapy: Facts and Hopes.

Cancer of the head and neck (HNC) is a heterogeneous disease of the upper aerodigestive tract, encompassing distinct histologic types, different anatomic sites, and human papillomavirus (HPV)-positive as well as HPV-negative cancers. Advanced/recurrent HNCs have poor prognosis with low survival rates. Tumor-mediated inhibition of antitumor immune responses and a high mutational burden are common features of HNCs. Both are responsible for the successful escape of these tumors from the host immune system. HNCs evolve numerous mechanisms of evasion from immune destruction. These mechanisms are linked to genetic aberrations, so that HNCs with a high mutational load are also highly immunosuppressive. The tumor microenvironment of these cancers is populated by immune cells that are dysfunctional, inhibitory cytokines, and exosomes carrying suppressive ligands. Dysfunctional immune cells in patients with recurrent/metastatic HNC can be made effective by the delivery of immunotherapies in combination with conventional treatments. With many promising immune-based strategies available, the future of immune therapies in HNC is encouraging, especially as methods for genetic profiling and mapping the immune landscape of the tumor are being integrated into a personalized approach. Efficiency of immune therapies is expected to rapidly improve with the possibility for patients' selection based on personal immunogenomic profiles. Noninvasive biomarkers of response to therapy will be emerging as a better understanding of the various molecular signals co-opted by the tumors is gained. The emerging role of immunotherapy as a potentially beneficial addition to standard treatments for recurrent/metastatic HNC offers hope to the patients for whom no other therapeutic options exist. Clin Cancer Res; 24(1); 6-13. ©2017 AACR.

Characteristics of peripheral immune cell subsets in patients with carotid atherosclerosis undergoing carotid endarterectomy

IntroductionThe role of circulating immune cells in the pathophysiology of cerebrovascular accidents is currently under debate. The aim of this study was to characterize peripheral immune cell subsets in patients undergoing carotid endarterectomy (CEA).Material and methodsWe conducted a prospective observational study in a group of 124 patients with significant carotid stenosis undergoing carotid endarterectomy, both symptomatic and asymptomatic. We compared the percentages of circulating immune cells: B lymphocytes, T lymphocytes, T helper, cytotoxic T, CD4+/CD8+ ratio, T regulatory, monocytes and NK cells before CEA and 6 h after the procedure.ResultsTotal lymphocyte count and cytotoxic T lymphocyte count decreased 6 h after CEA in both subgroups. The NK cell level decrease was statistically significant only in the symptomatic subgroup (19.41 ±9.30 before CEA and 16.52 ±9.37 after CEA; p = 0.0044), but not in the asymptomatic subgroup (17.88 ±9.14 before CEA and 15.91 ±9.51 after CEA; p = 0.0886). The T lymphocyte level showed a statistically significant increase only in symptomatic patients (69.74 ±10.16 before CEA vs. 71.45 ±9.77 after CEA; p = 0.0462), and not in the asymptomatic subgroup (70.08 ±11.19 prior to CEA and 70.21 ±12.35; p = 0.9048). B lymphocyte, helper T lymphocyte and regulatory T (Treg) lymphocyte (CD4+/CD25+) levels showed a significant increase after CEA.ConclusionsThis is the first study to compare circulating immune cells in patients undergoing carotid endarterectomy. Only the symptomatic subgroup experienced a significant decrease in the NK cell level and an increase in the T lymphocyte count after CEA. This study enriches our understanding of immune cell kinetics during carotid endarterectomy.

Unsupervised capture and profiling of rare immune cells using multi-directional magnetic ratcheting.

Immunotherapies (IT) require induction, expansion, and maintenance of specific changes to a patient's immune cell repertoire which yield a therapeutic benefit. Recently, mechanistic understanding of these changes at the cellular level has revealed that IT results in complex phenotypic transitions in target cells, and that therapeutic effectiveness may be predicted by monitoring these transitions during therapy. However, monitoring will require unique tools that enable capture, manipulation, and profiling of rare immune cell populations. In this study, we introduce a method of automated and unsupervised separation and processing of rare immune cells, using high-force and multidimensional magnetic ratcheting (MR). We demonstrate capture of target immune cells using samples with up to 1 : 10 000 target cell to background cell ratios from input volumes as small as 25 microliters (i.e. a low volume and low cell frequency sample sparing assay interface). Cell capture is shown to achieve up to 90% capture efficiency and purity, and captured cell analysis is shown using both on-chip culture/activity assays and off-chip ejection and nucleic acid analysis. These results demonstrate that multi-directional magnetic ratcheting offers a unique separation system for dealing with blood cell samples that contain either rare cells or significantly small volumes, and the "sample sparing" capability leads to an expanded spectrum of parameters that can be measured. These tools will be paramount to advancing techniques for immune monitoring under conditions in which both the sample volume and number of antigen-specific target cells are often exceedingly small, including during IT and treatment of allergy, asthma, autoimmunity, immunodeficiency, cell based therapy, transplantation, and infection.

Emerging trends in the immunotherapy of pancreatic cancer

Pancreatic cancer (PC) is the fourth leading cause of cancer-related deaths in the U.S., claiming approximately 43,000 lives every year. Much like other solid tumors, PC evades the host immune surveillance by manipulating immune cells to establish an immunosuppressive tumor microenvironment (TME). Therefore, targeting and reinstating the patient's immune system could serve as a powerful therapeutic tool. Indeed, immunotherapy has emerged in recent years as a potential adjunct treatment for solid tumors including PC. Immunotherapy modulates the host's immune response to tumor-associated antigens (TAAs), eradicates cancer cells by reducing host tolerance to TAAs and provides both short- and long-term protection against the disease. Passive immunotherapies like monoclonal antibodies or engineered T-cell based therapies directly target tumor cells by recognizing TAAs. Active immunotherapies, like cancer vaccines, on the other hand elicit a long-lasting immune response via activation of the patient's immune cells against cancer cells. Several immunotherapy strategies have been tested for anti-tumor responses alone and in combination with standard care in multiple preclinical and clinical studies. In this review, we discuss various immunotherapy strategies used currently and their efficacy in abrogating self-antigen tolerance and immunosuppression, as well as their ability to eradicate PC.

Tumour-infiltrating inflammatory and immune cells in patients with extrahepatic cholangiocarcinoma

Background:Inflammation and immune characteristics of the tumour microenvironment have therapeutic significance. The aim of this study was to investigate the clinical impact on disease progression in human extrahepatic cholangiocarcinoma (ECC).Methods:A total of 114 consecutive ECC patients with curative resection between 2000 and 2014 were enrolled. Tumour infiltrating CD66b+ neutrophils (TANs; tumour associated neutrophils), CD163+ M2 macrophages (TAMs; tumour associated macrophages), CD8+ T cells, and FOXP3+ regulatory T cells (Tregs) were assayed by immunohistochemistry, and their relationships with patient clinicopathological characteristics and prognosis were evaluated.Results:Tumour associated neutrophils were inversely correlated with CD8+ T cells (P=0.0001) and positively correlated with Tregs (P=0.001). High TANs (P=0.01), low CD8+ T cells (P=0.02), and high Tregs (P=0.04) were significantly associated with poor overall survival (OS). A high-risk signature, derived from integration of intratumoural inflammatory and immune cells, was significantly associated with poor recurrence-free survival (P=0.01) and OS (P=0.0008). A high-risk signature was correlated with postoperative distant metastases. Furthermore, a high-risk signature was related to the resistance to gemcitabine-based chemotherapy used after recurrence.Conclusions:Our data showed that tumour infiltrating inflammatory and immune cells may play a pivotal role in ECC progression and a high-risk signature predicted poor prognosis in ECC patients.

New Insight into the Pathogenesis of Erythema Nodosum Leprosum: The Role of Activated Memory T-Cells.

Memory T-cells, particularly, effector memory T cells are implicated in the pathogenesis of inflammatory diseases and may contribute to tissue injury and disease progression. Although erythema nodosum leprosum (ENL) is an inflammatory complication of leprosy, the role of memory T cell subsets has never been studied in this patient group. The aim of this study was at investigate the kinetics of memory T cell subsets in patients with ENL before and after prednisolone treatment. A case–control study design was used to recruit 35 untreated patients with ENL and 25 non-reactional lepromatous leprosy (LL) patient controls at ALERT Hospital, Ethiopia. Venous blood samples were obtained before, during, and after treatment from each patient. Peripheral blood mononuclear cells (PBMCs) were isolated and used for immunophenotyping of T cell activation and memory T-cell subsets by flow cytometry. The kinetics of these immune cells in patients with ENL before and after treatment were compared with LL patient controls as well as within ENL cases at different time points. The median percentage of CD3+, CD4+, and CD8+ T-cells expressing activated T-cells were significantly higher in the PBMCs from patients with ENL than from LL patient controls before treatment. The median percentage of central and activated memory T-cells was significantly increased in patients with ENL compared to LL patient controls before treatment. Interestingly, patients with ENL had a lower percentage of naïve T cells (27.7%) compared to LL patient controls (59.5%) (P < 0.0001) before treatment. However, after prednisolone treatment, patients with ENL had a higher median percentage of naïve T-cells (43.0%) than LL controls (33.0%) (P < 0.001). The median percentage of activated T-cells (effector memory and effector T-cells) was significantly increased in patients with ENL (59.2%) before treatment compared to after treatment with prednisolone (33.9%) (P < 0.005). This is the first work which has shown T-cell activation and the different subsets of memory T cells in untreated patients with ENL. Consequently, this study delineates the role of T-cell activation in the pathogenesis of ENL reaction and challenges the long-standing dogma of immune complex as a sole etiology of ENL reaction.

Immunohistochemical study of endometrial immune cells in patients with chronic endometritis with recurrent miscarriage

Anti-HLA class I antibodies have been suggested as a possible cause of fetal and neonatal alloimmune thrombocytopenia (FNAIT). The aim of this study was to characterize maternal anti-HLA class I alloantibodies in suspected cases of FNAIT.The study population consisted of all nationwide referrals of neonates with suspected FNAIT to the National Unit for Platelet Immunology in Tromsø, Norway, during 1998–2009 (cases), and 250 unselected pregnancies originally included in a prospective study (controls). Inclusion criterion was a positive screening for maternal anti-HLA class I antibodies. Neonates with other identifiable causes of thrombocytopenia, including maternal anti-human platelet antigens (HPA) antibodies, were excluded. Ultimately, 50 cases with suspected FNAIT were compared with 60 controls.The median neonatal platelet count nadir among cases was 24 × 109/L (range 4–98 × 109/L). Five children (10%) were reported to have intracranial hemorrhage. Maternal and neonatal HLA class I genotype was available for 33 mother/child pairs (66%). Immunization was not tied to any particular HLA class I antigen. Using epitope mapping, we could demonstrate that the maternal anti-HLA class I antibodies were specific towards mismatched paternally-inherited fetal epitopes, with little reactivity towards any third-party epitopes. Antibody reactivity patterns were similar to those found among controls, although the mean fluorescence intensities (MFI) among cases were significantly higher.This study demonstrates the value of using data on HLA epitope expression, instead of HLA antigens, to examine alloimmune responses in connection with neonatal thrombocytopenia. Our findings support the idea that maternal anti-HLA class I antibodies are involved in FNAIT.

Immunometabolism in systemic lupus erythematosus.

Systemic lupus erythematosus (SLE) is an autoimmune disease mediated by pathogenic autoantibodies directed against nucleoprotein complexes. Beyond the activation of autoreactive B cells, this process involves dysregulation in many other types of immune cells, including CD4+ T cells, dendritic cells, macrophages and neutrophils. Metabolic substrate utilization and integration of cues from energy sensors are critical checkpoints of effector functions in the immune system, with common as well as cell-specific programmes. Patients with SLE and lupus-prone mice present with activated metabolism of CD4+ T cells, and the use of metabolic inhibitors to normalize these features is associated with therapeutic effects. Far less is known about the metabolic requirements of B cells and myeloid cells in SLE. This article reviews current knowledge of the alterations in metabolism of immune cells in patients with SLE and mouse models of lupus in the context of what is known about the metabolic regulation of these cells during normal immune responses. How these alterations might contribute to lupus pathogenesis and how they can be targeted therapeutically are also discussed.

Correlation of PET/CT SUVmax with infiltration level of immune cells in patients with non-small cell lung cancer and its clinical significance

Correlation of PET/CT SUVmax with infiltration level of immune cells in patients with non-small cell lung cancer and its clinical significance

P3.03-034 Comprehensive Immunophenotyping of the Blood and Pleural Fluid from Patients with Malignant Pleural Mesothelioma by Flow Cytometry: Topic: Mesothelioma Clinical

Malignant pleural mesothelioma (MM) remains an almost universally fatal disease with limited treatment options. Immunotherapy represents a rapidly emerging therapeutic strategy for multiple malignancies including MM. However durable therapeutic responses occur in a minority (∼30%) of patients. Better understanding of the quantities and immunophenotype of circulating and intrapleural immune cells is essential to design and apply personalized immunotherapeutic strategies. We used a comprehensive flow cytometry panel (PLoS One. 2015 Mar 23;10 (3):e0121546) to prospectively characterize circulating and pleural fluid immune cells in patients with MPM (n=12) and normal volunteers (circulating cells only, n=50). Matched blood and pleural samples were available from 11 patients, including samples from 9 patients enrolled into a Phase I study investigating the intrapleural administration of the modified vaccine strain measles virus (MV-NIS), MC1023. Pre- and post-treatments samples were available from 7 patients. The immune cell counts and cell fractions were compared using a false discovery rate (FDR) of 10% and the non-parametric Mann-Whitney test and the Wilcoxon matched-pairs signed rank test (blood versus pleural fluid and different time points) (p ≤ 0.05). Cell counts and immune phenotype of circulating immune cells differ between of patients with MPM differ from normal volunteers (31 of 86 cell types). Patients with MPM had fewer B-lymphocytes, more pro-inflammatory monocytes (CD14+CD16+) and exhausted CD4 and CD8 T-lymphocytes (CD4 and CD8 PD1+TIM3+ double positive cells). As expected we observed characteristic differences between blood and pleural fluid in MPM patients. For example there are a higher numbers of antigen experienced, PD-1 expressing CD4 and CD8-memory cells within the pleural fluid compared to the peripheral blood. The intrapleural administration of a modified vaccine strain measles (MV-NIS) triggered changes in pleural and circulating immune cells. While these changes varied among patients, we observed increased CD80 and CD86 expression on CD14+ monocytes in the pleural fluid down. Comprehensive flow-cytometric immunophenotying of blood and pleural fluid is feasible in patients with MPM. This approach may help to identify patients who may respond favorably to specific immunotherapeutic interventions such as immune checkpoint inhibitors or facilitate longitudinal immune monitoring during clinical trials. Additional data is needed and we are continuing to prospectively analyze samples from patients with MPM.

Immune contexture and histological response after neoadjuvant chemotherapy predict clinical outcome of lung cancer patients

ABSTRACTThere is now growing evidence that the immune contexture influences cancer progression and clinical outcome of patients with non-small cell lung cancer (NSCLC). If chemotherapy is widely used to treat patients with advanced-stage NSCLC, it remains unclear how it could modify the immune contexture and impact its prognostic value. Here, we analyzed two retrospective cohorts, respectively composed of 122 stage III-N2 NSCLC patients treated with chemotherapy before surgery and 39 stage-matched patients treated by surgery only. In patients treated with neoadjuvant chemotherapy, the histological characteristics, the expression of PD-L1 protein, and the tumor immune microenvironment (CD8+ T cells, DC-LAMP+ mature dendritic cells, and CD68+ macrophages) were evaluated and their prognostic value assessed together with standard clinical parameters. By analyzing pre- and post-treatment specimens, we did not find any changes in the PD-L1 expression. We also found that the tumor immune contexture in patients treated with neoadjuvant chemotherapy exhibited a similar pattern that the one found in chemotherapy-naive patients, with comparable densities of tumor-infiltrating CD8+ and DC-LAMP+ cells and a similar spatial organization. The percentage of residual viable tumor cells and the immune pattern (CD8+ and DC-LAMP+ cell densities) were significantly associated with the clinical outcome and allowed the identification of short- and long-term survivors, respectively. In multivariate analysis, the immune pattern was found to be the strongest independent prognostic factor. In conclusion, this study decrypts the complex interplay between cancer and immune cells in patients undergoing chemotherapy and supports potential beneficial synergistic effect of immunotherapy and chemotherapy.

Alemtuzumab treatment alters circulating innate immune cells in multiple sclerosis.

Objective:To characterize changes in myeloid and lymphoid innate immune cells in patients with relapsing-remitting multiple sclerosis (MS) during a 6-month follow-up after alemtuzumab treatment.Methods:Circulating innate immune cells including myeloid cells and innate lymphoid cells (ILCs) were analyzed before and 6 and 12 months after onset of alemtuzumab treatment. Furthermore, a potential effect on granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)–23 production by myeloid cells and natural killer (NK) cell cytolytic activity was determined.Results:In comparison to CD4+ T lymphocytes, myeloid and lymphoid innate cell subsets of patients with MS expressed significantly lower amounts of CD52 on their cell surface. Six months after CD52 depletion, numbers of circulating plasmacytoid dendritic cells (DCs) and conventional DCs were reduced compared to baseline. GM-CSF and IL-23 production in DCs remained unchanged. Within the ILC compartment, the subset of CD56bright NK cells specifically expanded under alemtuzumab treatment, but their cytolytic activity did not change.Conclusions:Our findings demonstrate that 6 months after alemtuzumab treatment, specific DC subsets are reduced, while CD56bright NK cells expanded in patients with MS. Thus, alemtuzumab specifically restricts the DC compartment and expands the CD56bright NK cell subset with potential immunoregulatory properties in MS. We suggest that remodeling of the innate immune compartment may promote long-term efficacy of alemtuzumab and preserve immunocompetence in patients with MS.

Mammalian target of rapamycin inhibition counterbalances the inflammatory status of immune cells in patients with chronic granulomatous disease

Chronic granulomatous disease (CGD) is a primary immunodeficiency caused by defective production of reactive oxygen species in phagocytic cells that results in life-threatening infections and severe inflammatory manifestations. The treatment of inflammatory manifestations remains challenging because it can be associated with an increased risk of infections. Previous studies have shown that phagocytes from patients with CGD display a defect in autophagy and a reactive oxygen species-independent activation of the inflammasome. Because the intersections between autophagy and the inflammasome have been observed in patients with various diseases and microbial infections, we investigated the possible benefit of restoring the autophagy defect through rapamycin, a potent autophagy inducer, in the setting of CGD. We studied 15 patients given a diagnosis of CGD and followed in our institution. All patients were free of any active infection at the time of the study. We show that patients with CGD present a consistent inflammatory phenotype defined by (1) increased nonclassical and intermediate monocytes, (2) a proinflammatory state of mononuclear phagocytes with increased IL-1β and TNF-α content, (3) a TH17 bias of CD4+ Tcells, (4) and an increase in IL-17A-secreting neutrophil numbers. We document the reversion of CGD inflammatory status by the mammalian target of rapamycin inhibitor rapamycin on the different immune cell subsets. We also provide evidence for the enhancement of rapamycin's inhibitory effect on IL-1β secretion by the IL-1 receptor antagonist anakinra in phagocytes of patients with CGD. Altogether, these data open new therapeutic approaches for CGD-related inflammatory manifestations.