is the most common oncogenic driver in lung adenocarcinoma (LUAC). We previously reported that (KL) or (KP) comutations define distinct subgroups of -mutant LUAC. Here, we examine the efficacy of PD-1 inhibitors in these subgroups. Objective response rates to PD-1 blockade differed significantly among KL (7.4%), KP (35.7%), and K-only (28.6%) subgroups ( < 0.001) in the Stand Up To Cancer (SU2C) cohort (174 patients) with -mutant LUAC and in patients treated with nivolumab in the CheckMate-057 phase III trial (0% vs. 57.1% vs. 18.2%; = 0.047). In the SU2C cohort, KL LUAC exhibited shorter progression-free ( < 0.001) and overall ( = 0.0015) survival compared with ; LUAC. Among 924 LUACs, alterations were the only marker significantly associated with PD-L1 negativity in TMB LUAC. The impact of alterations on clinical outcomes with PD-1/PD-L1 inhibitors extended to PD-L1-positive non-small cell lung cancer. In-mutant murine LUAC models, loss promoted PD-1/PD-L1 inhibitor resistance, suggesting a causal role. Our results identify alterations as a major driver of primary resistance to PD-1 blockade in -mutant LUAC. This work identifies alterations as the most prevalent genomic driver of primary resistance to PD-1 axis inhibitors in-mutant lung adenocarcinoma. Genomic profiling may enhance the predictive utility of PD-L1 expression and tumor mutation burden and facilitate establishment of personalized combination immunotherapy approaches for genomically defined LUAC subsets. .

Interleukin (IL)-10 and glucocorticoids (GCs) inhibit the ability of antigen-presenting dendritic cells (DCs) to stimulate T lymphocytes. We show that induction of GILZ (GC-induced leucine zipper) is involved in this phenomenon. IL-10, dexamethasone (DEX), and transforming growth factor (TGF)beta stimulate GILZ production in human immature DCs derived from monocytes and from CD34+ cells. GILZ is necessary and sufficient for DEX, IL-10, and TGFbeta modulation of CD80, CD83, CD86, immunoglobulin-like transcript (ILT)-3, and B7-H1 expression by DCs, and alteration of DC functions. GILZ stimulates the production of IL-10 by immature DCs and prevents the production of inflammatory chemokines by CD40L-activated DCs. In contrast, GILZ does not prevent CD40 ligand-mediated inhibition of phagocytosis, indicating that it affects some but not all aspects of DC maturation. GILZ prevents DCs from activating antigen-specific T lymphocyte responses. Administration of GCs to patients stimulates GILZ expression in their circulating antigen-presenting cells, and this contributes to the weak lymphocyte responses of GC-treated patients. Thus, regulation of GILZ expression is an important factor determining the decision of DCs whether or not to stimulate T lymphocytes, and IL-10, GCs, and TGFbeta share this mechanism for influencing DC functions and the balance between immune response and tolerance.

IntroductionRegulatory T lymphocytes (Tregs) play a key role in controlling immune responses. Initially, 2 types of Tregs were identified. Natural (or constitutive) Tregs are generated in the thymus and they spontaneously express the transcription factor FOXP3 and high levels of CD25. Inducible (or adaptive) Tregs are generated by peripheral activation, particularly in the presence of IL-10 or TGF␤. However, the relationship between these 2 subpopulations has not been completely elucidated. Tregs inhibit immune responses by a combination of effects requiring direct contact with their targets and the production of inhibitory cytokines, including IL-10 and TGF␤. In addition to the coexpression of CD25 hi and FOXP3 and the production of IL-10 and TGF␤, several markers potentially characteristic of Tregs have been identified, but none of them has been unequivocally associated with the suppressive function of Tregs or with their classification into natural or inducible Tregs. [1][2][3][4] Antigen presentation by dendritic cells (DCs) leads to either immune stimulation or tolerance. The mechanisms underlying the decision of DCs to orientate the immune response toward one of these 2 opposite outcomes are not fully understood. Toleranceinducing DCs are also called regulatory DCs. One method used by regulatory DCs to prevent immune activation is to trigger T-lymphocyte anergy or apoptosis during antigen presentation. This involves production of the enzyme indoleamine 2,3-dioxygenase (IDO) and of nitric oxide by DCs. 5,6 The other method is to generate regulatory cells, including Tregs. Although IDO production by DCs may play a role in the induction of Tregs, 7 additional mechanisms are presumably involved. The state of maturation and activation of DCs is critical to Treg development: DCs activated and maturing in response to inflammatory stimuli trigger immune responses, but immature or "semimature" DCs, in contrast, induce tolerance, [8][9][10][11] and this is in part mediated by the generation of Tregs. [12][13][14][15] Phenotypically mature DCs can also be tolerogenic, and certain environmental signals can induce maturation of DCs in a tolerogenic mode. 10 IL-10, TGF␤, glucocorticoids (GCs), vasoactive intestinal peptide, vitamin D3, and antioxidative vitamins, used alone or in combination, orientate DC maturation to induce tolerance, [16][17][18][19][20][21][22][23][24] and Treg development has been demonstrated for several of these agents. 18,19,22,24 The molecular mechanisms involved in this switch of DC maturation toward regulatory DCs are largely unknown. Several tolerogenic agents down-regulate the NF-kB and p38 MAPK transduction pathways in DCs, which contrasts with the potency of inflammatory agents to activate them. 24,25 In addition, DCs from RelB-deficient mice induce immune tolerance and antigen-specific Tregs. 26 This suggests that modulation of NF-kB and p38 MAPK function contributes to the decision of DCs to differentiate into regulatory DCs.An intracellular factor called glucocorticoid-induced leucine ...

Introduction: Latent tuberculosis infection (LTBI) is detected with the tuberculin skin test (TST) before anti-TNF therapy. We aimed to investigate in vitro blood assays with TB-specific antigens (CFP-10, ESAT-6), in immune-mediated inflammatory diseases (IMID) for LTBI screening. Patients and methods: Sixty-eight IMID patients with (n = 35) or without (n = 33) LTBI according to clinicoradiographic findings or TST results (10 mm cutoff value) underwent cell proliferation assessed by thymidine incorporation and PKH-26 dilution assays, and IFNc-release enzyme-linked immunosorbent spot (ELISPOT) assays with TB-specific antigens. Results: In vitro blood assays gave higher positive results in patients with LTBI than without (p,0.05), with some variations between tests. Among the 13 patients with LTBI diagnosed independently of TST results, 5 had a negative TST (38.5%) and only 2 a negative blood assays result (15.4%). The 5 LTBI patients with negative TST results all had positive blood assays results. Ten patients without LTBI but with intermediate TST results (6-10 mm) had no different result than patients with TST result (5 mm (p.0.3) and lower results than those with LTBI (p,0.05) on CFP-10+ESAT-6 ELISPOT and CFP-10 proliferation assays. Conclusion: Anti-TB blood assays are beneficial for LTBI diagnosis in IMID. Compared with TST, they show a better sensitivity, as seen by positive results in 5 patients with certain LTBI and negative TST, and better specificity, as seen by negative results in most patients with intermediate TST as the only criteria of LTBI. In the absence of clinico-radiographic findings for LTBI, blood assays could replace TST for antibiotherapy decision before anti-TNF.

Glucocorticoid-induced leucine zipper (GILZ), a recently identified protein induced by glucocorticoids (GCs), inhibits the nuclear factor B pathway and the activation of monocytes/macrophages by lipopolysaccharides (LPS). This study aimed to elucidate the contribution of GILZ to the pathogenesis of alcoholic hepatitis (AH): we (1) assessed GILZ expression in the livers of patients with AH and (2) treated patients with severe AH with GCs (prednisolone 40 mg/day) and studied the effect of GILZ modulation on circulating monocyte function. We quantified GILZ expression in the livers of 42 consecutive alcoholic patients (21 with and 21 without AH). GILZ messenger RNA (mRNA) levels were lower in the livers of patients with AH versus those without AH (P < 0.05). We collected circulating monocytes from patients with severe AH before and 48 hours after GC treatment to quantify GILZ expression and cytokine secretion. GC treatment induced significantly higher levels of GILZ mRNA than that observed before treatment and impaired LPS-induced tumor necrosis factor-␣ (TNF-␣) and regulated upon activation, normal T cell-expressed secretion (RANTES) by these monocytes. We transfected circulating monocytes with GILZ small interfering RNA (siRNA), specifically blocking GILZ expression, to demonstrate the role of GILZ in mediating GC effect. GILZ siRNA abrogated the effect of GC treatment on LPS-induced TNF-␣ and RANTES secretion. Conclusion: Low expression of GILZ may contribute to liver inflammation in AH. GCs enhance GILZ expression, abrogating macrophage sensitivity to LPS and proinflammatory cytokine secretion. These findings may explain the beneficial effect of GC treatment in patients with severe AH. (HEPATOLOGY

Introduction: Blockade of immune checkpoints has improved clinical outcomes for patients with metastatic non-small cell lung cancer (NSCLC), but its role in the perioperative setting for early-stage disease is unclear. We generated preclinical models of resectable NSCLC expressing an antigen that permits quantitative assessment of the immune response and compared survival, tumor recurrence, and immune response after neoadjuvant or adjuvant immunotherapy. Experimental Procedures: We transfected murine 344SQ NSCLC cells with an ovalbumin-expression plasmid to generate OVA+ cells that can be identified with an antibody against the peptide SIINFEKL bound to H-2Kb of MHC-I. We implanted 344SQ-OVA+ cells in the flank of syngeneic mice and then randomized mice with established tumors to either 3 doses of neoadjuvant IgG, anti-PD-1, anti-CTLA-4, anti-PD-1+anti-CTLA-4, or to observation. Primary tumors were resected in all mice 2 days after mice in the neoadjuvant group had received their last dose of therapy. Two days post-surgery, mice in the observation group were treated with 3 doses of adjuvant IgG, anti-PD-1, anti-CTLA-4, anti-PD-1+anti-CTLA-4. Mice were euthanized 4 weeks after injection or when moribund and their survival recorded and lung metastases counted. We determined the composition of CD3+CD8+ tumor-infiltrating lymphocytes (TILs) with flow cytometry using tetramers against SIINFEKL-specific T cell receptors and immunohistochemical staining of tumors. Results: Single agent and combined immunotherapy significantly prolonged survival compared to controls in both neoadjuvant and adjuvant groups (p&lt;0.05). Combined therapy was superior to either monotherapy in the neoadjuvant setting (combo vs. anti-PD-1 and vs. anti-CTLA-4 p&lt;0.05), whereas there were no significant differences between combined treatment and monotherapies in the adjuvant setting. Moreover, combined therapy in the neoadjuvant setting significantly prolonged survival compared to adjuvant combined treatment (p&lt;0.05). Neoadjuvant combined therapy produced the most profound reduction in lung metastases when compared to monotherapy (combo vs. IgG, vs. anti-PD-1 and vs. anti-CTLA-4 p&lt;0.01), or to adjuvant combined treatment. Neoadjuvant combined therapy was associated with the greatest amounts of SIINFEKL-tetramer+ TILs in resected primary tumors and also with increased CD8+ TIL density in both resected primary tumors (combo vs. IgG and vs. anti-PD-1 p&lt;0.05) and secondary lung metastases (combo vs. IgG, vs. anti-PD1 and vs. anti-CTLA-4 p&lt;0.05). Functional studies investigating the immune response of tumors treated with perioperative immunotherapies are ongoing. Conclusions: Neoadjuvant combined immune checkpoint blockade is superior to adjuvant immunotherapy at prolonging survival, reducing distal recurrence and inducing anti-tumor immunity in preclinical models of resectable NSCLC. Citation Format: Tina Cascone, Haifa Hamdi, Fahao Zhang, Alissa Poteete, Lerong Li, Courtney W. Hudgens, Leila J. Williams, Qiuyu Wu, Jayanthi Gudikote, Weiyi Peng, Patrick Hwu, Jing Wang, Michael Tetzlaff, William N. William, John V. Heymach. Superior efficacy of neoadjuvant compared to adjuvant immune checkpoint blockade in non-small cell lung cancer [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 1719.

T cells chronically stimulated by a persistent antigen often become dysfunctional and lose effector functions and proliferative capacity. To identify the importance of micro-RNA-155 (miR-155) in this phenomenon, we analyzed mouse miR-155-deficient CD4 + T cells in a model where the chronic exposure to a systemic antigen led to T-cell functional unresponsiveness. We found that miR-155 was required for restoring function of T cells after programmed death receptor 1 blockade. Heme oxygenase 1 (HO-1) was identified as a specific target of miR-155 and inhibition of HO-1 activity restored the expansion and tissue migration capacity of miR-155 −/− CD4 + T cells. Moreover, miR-155-mediated control of HO-1 expression in CD4 + T cells was shown to sustain in vivo antigen-specific expansion and IL-2 production. Thus, our data identify HO-1 regulation as a mechanism by which miR-155 promotes T-cell-driven inflammation.Keywords: Adaptive tolerance r Heme oxygenase 1 (HO-1) r Immunoregulation r Micro-RNA r Th1Additional supporting information may be found in the online version of this article at the publisher's web-site IntroductionT lymphocytes are a crucial part of the immune system and are a major cell type involved in the adaptive immune response. Usually, T cells expand rapidly in response to antigenic stimulation, and then, as a result of antigen clearance, they die by apoptosis. However, in situations of chronic antigenic exposure, such as those observed in cancer, chronic infections, or autoimmunity, antigen-specific T cells may persist for extended periods of time and these conditions are often associated with T-cell Correspondence: Dr. Michel Y. Braun e-mail: mbraun@ulb.ac.be dysfunction, such as decreased cytokine expression and effector function. Recent experimental evidence indicates that this unresponsive state is maintained by the inhibition of signaling events, particularly downstream proximal signals of the T-cell receptor, that in turn promotes T-cell dysfunction [1][2][3][4]. Since the attenuation of T-cell effector functions by the programmed death receptor 1 (PD-1) pathway has been demonstrated in various chronic infectious diseases and cancer and in the control of autoimmune T cells, the modulation of this inhibitory pathway is considered a promising means to control the function of T cells or enhance immune responses in patients.Micro-RNAs (miRNAs) are small genome-encoded RNAs that regulate gene expression by targeting complementary messenger RNAs (mRNAs) and block their translation [5,6]. The first C 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim www.eji-journal.eu 830Jinyu Zhang et al. Eur. J. Immunol. 2015. 45: 829-842 evidence for the involvement of miRNAs in regulating T-cell differentiation and function came from the observation that the specific deletion of the endoribonuclease DICER, a RNaseIII-like enzyme required for generating miRNAs, in the CD4 + T-cell lineage resulted in impaired T-cell development and aberrant Th1-cell differentiation and cytokine production [7]. Among miRNAs the mo...

Reactivation of latent Mycobacterium tuberculosis (Mtb) infection is a major complication of anti-tumour necrosis factor (TNF)-α treatment, but its mechanism is not fully understood. We evaluated the effect of the TNF antagonists infliximab (Ifx), adalimumab (Ada) and etanercept (Eta) on anti-mycobacterial immune responses in two conditions: with ex vivo studies from patients treated with TNF antagonists and with the in vitro addition of TNF antagonists to cells stimulated with mycobacterial antigens. In both cases, we analysed the response of CD4 + T lymphocytes to purified protein derivative (PPD) and to culture filtrate protein (CFP)-10, an antigen restricted to Mtb. The tests performed were lymphoproliferation and immediate production of interferon (IFN)-γ. In the 68 patients with inflammatory diseases (rheumatoid arthritis, spondylarthropathy or Crohn's disease), including 31 patients with a previous or latent tuberculosis (TB), 14 weeks of anti-TNF-α treatment had no effect on the proliferation of CD4 + T lymphocytes. In contrast, the number of IFN-γ-releasing CD4 + T lymphocytes decreased for PPD (p < 0.005) and CFP-10 (p < 0.01) in patients with previous TB and for PPD (p < 0.05) in Ada = adalimumab; BCG = Bacille de Calmette Guérin; CD = Crohn's disease; Cd = Candida; CFP-10 culture filtrate protein-10; CMV = cytomegalovirus; EC 50 = median effective concentration; ELISPOT = enzyme-linked immunosorbent spot; ESAT-6 = early secretory antigen target-6; Eta = etanercept; IFN = interferon; Ifx = infliximab; IL = interleukin; IMID = immune-mediated inflammatory disease; mAb = monoclonal antibody; Mtb = Mycobacterium tuberculosis; mTNF = membrane-bound tumour necrosis factor; PBMC = peripheral blood mononuclear cell; PPD = purified protein derivative (or tuberculin); RA = rheumatoid arthritis; SA = sponlylarthropathy; TB = tuberculosis; TNF = tumour necrosis factor; TNFR = tumour necrosis factor receptor; Toxo = Toxoplasma gondii; TT = Tetanus toxoid. Arthritis Research & Therapy Vol 8 No 4 Hamdi et al. Page 2 of 15 (page number not for citation purposes)other patients (all vaccinated with Bacille Calmette-Guérin). Treatments with Ifx and with Eta affected IFN-γ release to a similar extent. In vitro addition of TNF antagonists to CD4 + T lymphocytes stimulated with mycobacterial antigens inhibited their proliferation and their expression of membrane-bound TNF (mTNF). These effects occurred late in cultures, suggesting a direct effect of TNF antagonists on activated mTNF + CD4 + T lymphocytes, and Ifx and Ada were more efficient than Eta. Therefore, TNF antagonists have a dual action on antimycobacterial CD4 + T lymphocytes. Administered in vivo, they decrease the frequency of the subpopulation of memory CD4 + T lymphocytes rapidly releasing IFN-γ upon challenge with mycobacterial antigens. Added in vitro, they inhibit the activation of CD4 + T lymphocytes by mycobacterial antigens. Such a dual effect may explain the increased incidence of TB in patients treated with TNF antagonists as well as possible...