IFN-I Release Research Articles

Molecular Function of cGAS-STING in SARS-CoV-2: A Novel Approach to COVID-19 Treatment.

Coronavirus illness 2019 is a significant worldwide health danger that began with severe acute respiratory syndrome coronavirus two infections. It is the largest pandemic of our lifetime to date, affecting millions of people and crippling economies globally. There is currently no viable therapy for this devastating condition. The fast spread of SARS-CoV-2 underlines the critical need for favorable treatments to prevent SARS-CoV-2 infection and dissemination. Regulating the upstream cytokine release might be a possible method for COVID-19 therapy. We propose that more consideration be paid to the dysregulated IFN-I release in COVID-19 and that cGAS and STING be considered therapeutic targets for avoiding cytokine storms and as critical components in host antiviral defense mechanisms.

A Phase 1 Study of TAK-676, a Novel STING Agonist, Plus Pembrolizumab Following Radiation Therapy in Patients with Advanced Non–Small-Cell Lung Cancer (NSCLC), Triple-Negative Breast Cancer (TNBC), or Squamous-Cell Carcinoma of the Head and Neck (SCCHN)

<h3>Purpose/Objective(s)</h3> Radiation therapy-induced cell death produces cytosolic DNA that activates the cyclic GMP-AMP synthase (cGAS)–STimulator of INterferon Genes (STING) pathway, crucial for the induction of Type I interferons (IFN-I). Checkpoint inhibitor (CPI) resistance mechanisms have been linked to impaired IFN signaling. Preclinical data have shown STING agonists to reverse CPI resistance in tumors with prior exposure, particularly when used with anti-PD-1/PD-L1 therapies. TAK-676 (a synthetic STING agonist) potently modulates the innate immune system, leading to cytokine release, adaptive immune activation, and antitumor responses in preclinical studies (Appleman et al., AACR 2022). TAK-676 is being investigated as a single agent, and in combination with pembrolizumab, for advanced solid tumors, in a first-in-human phase 1 study (NCT04420884). TAK-676 is optimally designed for intravenous (IV) delivery, with a prolonged half-life in serum and enhanced tissue permeability, allowing access to tumor sites and lymphatic tissue. Following radiation therapy, TAK-676 has the potential to stimulate T-cell mediated antitumor immunity via STING-mediated IFN-I release, particularly when used with anti-PD-1/PD-L1 therapies. Here, we present a second phase 1 study to investigate the safety and preliminary antitumor activity of TAK-676 in combination with pembrolizumab following radiation therapy, in patients with advanced or metastatic NSCLC, TNBC, or SCCHN who have progressed on CPIs (NCT04879849). <h3>Materials/Methods</h3> Patients aged ≥18 years who progressed on CPIs and have ≥2 lesions, of which one is targetable with radiation, are being enrolled. Patients receive 8 Gy x 3 followed by (after a minimum of 40 hours) escalating doses of IV TAK-676 on days 1, 8 and 15 of a 21-day cycle, and 200 mg of IV pembrolizumab on day 1 of each cycle until disease progression, intolerance, or withdrawal of consent. Dose escalation of TAK-676 will be guided by the Bayesian Optimal Interval design. At screening and between days 15–21 of cycle 1, patients with a safely accessible lesion outside of the radiation field will have paired biopsies collected once the pharmacologically active dose levels of TAK-676 have been observed. The primary objective is to determine the safety and tolerability of TAK-676 in combination with pembrolizumab following radiation therapy. Secondary objectives are to determine the recommended phase 2 dose of TAK-676 in combination with pembrolizumab following radiation therapy, and to assess the local (within the radiation field) and systemic (non-radiated lesions) preliminary antitumor activity. As of February 2022, ∼10% of the planned patients have been enrolled. <h3>Results</h3> TBD <h3>Conclusion</h3> TBD

Monocyte-derived macrophages aggravate pulmonary vasculitis via cGAS/STING/IFN-mediated nucleic acid sensing.

Autoimmune vasculitis is a group of life-threatening diseases, whose underlying pathogenic mechanisms are incompletely understood, hampering development of targeted therapies. Here, we demonstrate that patients suffering from anti-neutrophil cytoplasmic antibodies (ANCA)-associated vasculitis (AAV) showed increased levels of cGAMP and enhanced IFN-I signature. To identify disease mechanisms and potential therapeutic targets, we developed a mouse model for pulmonary AAV that mimics severe disease in patients. Immunogenic DNA accumulated during disease onset, triggering cGAS/STING/IRF3-dependent IFN-I release that promoted endothelial damage, pulmonary hemorrhages, and lung dysfunction. Macrophage subsets played dichotomic roles in disease. While recruited monocyte-derived macrophages were major disease drivers by producing most IFN-β, resident alveolar macrophages contributed to tissue homeostasis by clearing red blood cells and limiting infiltration of IFN-β-producing macrophages. Moreover, pharmacological inhibition of STING, IFNAR-I, or its downstream JAK/STAT signaling reduced disease severity and accelerated recovery. Our study unveils the importance of STING/IFN-I axis in promoting pulmonary AAV progression and identifies cellular and molecular targets to ameliorate disease outcomes.

Abstract 2102: NFAT2 determines susceptibility of chronic lymphocytic leukemia cells to constitutive and Rituximab-induced NK cell killing

Abstract Genetic loss of the transcription factor NFAT2 (NFATc1), known as important player in lymphocyte development, causes an aggressive course of CLL, as it enhances BCR signaling and leads to highly malignant and proliferative disease (Märklin et al. Nat Commun 2017). To investigate whether and how NFAT2 affects treatment resistance in CLL, we generated a CRISPR/Cas9 based NFAT2 knockout (KO) in MEC-1 CLL cells to model aggressive CLL. In immunodeficient NSG-mice, the NFAT2 KO cells showed a ten-fold higher engraftment in the bone marrow after three weeks compared to scrambled (SCR) controls. Cytotoxicity assays revealed a profoundly higher resistance of NFAT2 KO cells to NK cell killing without detectable differences at the level of recognition as revealed by analyses of NK cell activation, degranulation and IFNy release, which excluded resistance to NK cells was due to altered immunogenicity. As CLL patients are routinely treated with anti-CD20 antibody Rituximab, which mediates its therapeutic efficacy in large part by inducing antibody dependent cellular cytotoxicity (ADCC) of NK cells via secretion of perforin and granzymes. When we exposed the target cells to the cytotoxic content isolated out of NK cells, MEC-1 NFAT2 KO cells showed higher resistance for membrane permeabilization compared to SCR controls. In line, perforin blockade equalized susceptibility to membrane permeabilization of both, NFAT2 KO cells and controls, confirming the involvement of NFAT2 in maintaining membrane integrity. In addition, caspase 3/7 activation was reduced in NFAT2 KO cells upon exposure to NK cell granule content as well as upon treatment with staurosporine. This shows an important role of NFAT2 for resistance of the leukemic cells at early stages of apoptosis. The results on the role of NFAT2 in resistance to NK effector function were confirmed with primary CLL patient cells, which were grouped according to high and low NFAT2 expression followed by treatment with NK cell content. Our results demonstrate that loss of NFAT2 allows CLLs cells to evade NK cell effector function. This holds true for both, constitutive cytotoxicity and therapeutically induced ADCC and is due to lowered susceptibility of the leukemic cells to perforin mediated membrane permeabilization and intrinsic resistance to granzyme induced apoptosis. Thus, NFAT2 loss, which correlates with aggressive clinical course in patients, facilitates resistance of CLL cells to CD20 antibody-based treatment. Citation Format: Samuel J. Holzmayer, Sarah M. Greiner, Kuebra Kaban, Jonas S. Heitmann, Helmut R. Salih, Melanie Maerklin. NFAT2 determines susceptibility of chronic lymphocytic leukemia cells to constitutive and Rituximab-induced NK cell killing [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2102.

Abstract CT243: Phase 1 study of TAK-676 + pembrolizumab following radiation therapy in patients with advanced non-small-cell lung cancer (NSCLC), triple-negative breast cancer (TNBC), or squamous-cell carcinoma of the head and neck (SCCHN)

Abstract Background: Radiation therapy has immune-modulating effects resulting from apoptosis of tumor cells and DNA damage. The downstream generation of cytosolic DNA activates the cyclic GMP-AMP Synthase (cGAS)-STimulator of INterferon Gene (STING) signaling axis in both tumor and nearby immune cells, leading to increased induction of type I interferon (IFN-I) and other immune stimulating molecules. TAK-676 is a novel synthetic STING agonist being investigated (+/- pembrolizumab) in an ongoing first-in-human phase 1 study (NCT04420884). TAK-676 is a potent modulator of the innate immune system and leads to downstream activation of the adaptive immune system to produce antitumor responses in preclinical studies. In contrast to intratumorally injected STING agonists, TAK-676 is optimally designed for reduced serum degradation and enhanced permeability, allowing systemic IV delivery and access to tumor sites and lymphatics. The addition of TAK-676 following radiation therapy may enhance the immune response by increasing the STING-mediated IFN-I release and further stimulate T cell-mediated antitumor immunity, particularly in combination with anti-PD-1/PD-L1 therapies. Impaired IFN signaling has been linked to checkpoint inhibitor (CPI) resistance. Preclinical studies show that addition of a STING agonist may reverse the mechanisms of resistance in tumors with prior exposure to CPIs. This phase 1 trial was designed to investigate the safety and preliminary antitumor activity of TAK-676 + pembrolizumab following radiation therapy (NCT04879849). Methods: Patients aged ≥18 years with advanced NSCLC, TNBC, or SCCHN who have progressed on CPIs with ≥2 lesions, one of which can be targeted with radiation, are being enrolled. Patients receive 8 Gy x 3 fractions of image-guided radiation therapy followed by (after a minimum of 40 hours) IV pembrolizumab 200 mg on day 1 plus escalating doses of IV TAK-676 on days 1, 8, and 15 of a 21-day cycle. TAK-676 dose escalation is guided by the Bayesian Optimal Interval design. Patients receive TAK-676 + pembrolizumab until disease progression, intolerance, or withdrawal of consent. Once pharmacologically active dose levels of TAK-676 have been observed, paired biopsies will be collected in patients with a safely accessible lesion outside the radiation field at screening and between days 15 and 21 of cycle 1. The primary objective is to determine the safety and tolerability of TAK-676 + pembrolizumab following radiation therapy; secondary objectives are to determine the recommended phase 2 dose of TAK-676 + pembrolizumab following radiation therapy, and to assess preliminary antitumor activity both locally (in the radiation field) and systemically (non-radiated lesions). As of January 2022, nearly 10% of the planned patients have been enrolled. Citation Format: Benjamin Cooper, Steven J. Chmura, Jason J. Luke, Stephen L. Shiao, Reva K. Basho, Wade T. Iams, David B. Page, Cong Li, Richard C. Gregory, Michael H. Shaw, Kristin H. Horn, John P. Gibbs, Vicky A. Appleman, Allison J. Berger, Adnan O. Abu-Yousif, Neil B. Lineberry, Kate F. Stumpo, Aymen Elfiky, Naamit K. Gerber. Phase 1 study of TAK-676 + pembrolizumab following radiation therapy in patients with advanced non-small-cell lung cancer (NSCLC), triple-negative breast cancer (TNBC), or squamous-cell carcinoma of the head and neck (SCCHN) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr CT243.

TAK-676 in combination with pembrolizumab after radiation therapy in patients (pts) with advanced non–small cell lung cancer (NSCLC), triple-negative breast cancer (TNBC), or squamous-cell carcinoma of the head and neck (SCCHN): Phase 1 study design.

TPS2698 Background: The cyclic GMP-AMP Synthase (cGAS)–STimulator of INterferon Genes (STING) pathway is an important modulator of the innate immune system via induction of type I interferon (IFN-I). Cytosolic DNA generated as a result of tumor cell death following radiation therapy has been demonstrated to activate the cGAS-STING signaling axis resulting in antitumor immunogenicity. TAK-676 is a novel, synthetic STING agonist and it has been shown in preclinical studies to potently modulate the innate immune system and subsequently activate the adaptive immune system to produce antitumor responses. TAK-676 is designed for prolonged half-life in serum and enhanced tissue permeability compared with other STING agonists designed for intratumoral injection, allowing for systemic IV delivery with access to tumor sites and lymphatic tissue. IFN signaling impairment has been linked to checkpoint inhibitor (CPI) resistance in tumors. Treatment with TAK-676 after radiation therapy has the potential to stimulate T cell-mediated antitumor immunity via STING-mediated IFN-I release, particularly when used with anti-PD-1/PD-L1 therapies. Preclinical data support the addition of STING agonists to reverse resistance in tumors with prior exposure to CPIs. TAK-676 is being investigated (+/- pembrolizumab) in an ongoing first-in-human phase 1 study (NCT04420884). Here, we describe another phase 1 trial to investigate the safety and preliminary antitumor activity of TAK-676 plus pembrolizumab following radiation therapy in pts with advanced or metastatic NSCLC, TNBC, or SCCHN (NCT04879849). Methods: Adult pts with progressive disease (PD) following CPI treatment and who have ≥2 lesions, 1 of which can be targeted with radiation, are being enrolled. Pts receive 8 Gy x 3 fractions of image-guided radiation followed (after ≥40 hours) by IV pembrolizumab 200 mg on day 1 plus escalating doses of IV TAK-676 on days 1, 8, and 15 of a 21-day cycle. TAK-676 dose escalation is guided by the Bayesian optimal interval design. Pts receive TAK-676 plus pembrolizumab until PD, intolerance to treatment, or withdrawal. Pts enrolled at TAK-676 dose levels shown to have pharmacodynamic activity, and who have a safely accessible lesion outside the radiation field, will have paired biopsies collected at screening and between days 15 and 21 of cycle 1. The primary objective is to determine the safety and tolerability of TAK-676 plus pembrolizumab following radiation therapy; secondary objectives are to establish the recommended phase 2 dose of TAK-676 plus pembrolizumab following radiation therapy, and to assess preliminary antitumor activity both locally (within the radiation field) and systemically (non-radiated lesions). As of February 2022, we have enrolled ̃10% of the planned pts. Clinical trial information: NCT04879849.

Natural Killer Cells and Host Defense Against Human Rhinoviruses Is Partially Dependent on Type I IFN Signaling.

Rhinovirus (RV), the causative agent of the common cold, causes only mild upper respiratory tract infections in healthy individuals, but can cause longer lasting and more severe pulmonary infections in people with chronic lung diseases and in the setting of immune suppression or immune deficiency. RV-infected lung structural cells release type I interferon (IFN-I), initiating the immune response, leading to protection against viruses in conjunction with migratory immune cells. However, IFN-I release is deficient in some people with asthma. Innate immune cells, such as natural killer (NK) cells, are proposed to play major roles in the control of viral infections, and may contribute to exacerbations of chronic lung diseases, such as asthma. In this study, we characterized the NK cell response to RV infection using an in vitro model of infection in healthy individuals, and determined the extent to which IFN-I signaling mediates this response. The results indicate that RV stimulation in vitro induces NK cell activation in healthy donors, leading to degranulation and the release of cytotoxic mediators and cytokines. IFN-I signaling was partly responsible for NK cell activation and functional responses to RV. Overall, our findings suggest the involvement of NK cells in the control of RV infection in healthy individuals. Further understanding of NK cell regulation may deepen our understanding of the mechanisms that contribute to susceptibility to RV infections in asthma and other chronic lung diseases.

Detection of native interferon-γ in nyala (Tragelaphus angasii): Towards diagnosing tuberculosis.

Mycobacterium bovis is the main cause of tuberculosis in wildlife. In South Africa, African buffaloes (Syncerus caffer) are a wildlife maintenance host while a number of other species are considered spillover hosts. Nyala (Tragelaphus angasii), a large antelope species from Southern Africa, is frequently traded and can be infected with M. bovis. Interferon gamma (IFN-γ) release assays that detect cell-mediated immune (CMI) responses to M. bovis infection have shown promise in elephants, rhinoceroses and buffaloes. The BOVIGAM® assay is a commercial IFN-γ release assay designed to detect tuberculosis in cattle and has been validated in buffaloes. We tested the suitability of the BOVIGAM® assay to detect native IFN-γ release in nyala. Blood samples collected from 17 nyalas were stimulated with different mitogens and IFN-γ release measured. We found that incubating whole blood with phorbol 12-myristate 13-acetate and calcium ionophore (PMA/CaI) resulted in the highest levels of IFN-y release. Samples stimulated with tuberculin purified protein derivatives of M. bovis (PPDb) and M. avium (PPDa) did not show significant IFN-γ production. An intradermal tuberculin test (IDT) and culture of tissues from 15 of the 17 culled nyala were also performed, which supported the findings of the BOVIGAM® assay, suggesting the potential value of this assay for the diagnosis of tuberculosis in nyala.

Metabolic stress in cancer cells induces immune escape through a PI3K-dependent blockade of IFN\u03b3 receptor signaling

BackgroundT-cell mediated immunotherapy brought clinical success for many cancer patients. Nonetheless, downregulation of MHC class I antigen presentation, frequently occurring in solid cancers, limits the efficacy of these therapies. Unraveling the mechanisms underlying this type of immune escape is therefore of great importance. We here investigated the immunological effects of metabolic stress in cancer cells as a result of nutrient deprivation.MethodsTC1 and B16F10 tumor cell lines were cultured under oxygen- and glucose-deprivation conditions that mimicked the tumor microenvironment of solid tumors. Presentation of peptide antigens by MHC class I molecules was measured by flow cytometry and via activation of tumor-specific CD8 T cell clones. The proficiency of the IFNy-STAT1 pathway was investigated by Western blots on phosphorylated proteins, transfection of constitutive active STAT1 constructs and qPCR of downstream targets. Kinase inhibitors for PI3K were used to examine its role in IFNy receptor signal transduction.ResultsCombination of oxygen- and glucose-deprivation resulted in decreased presentation of MHC class I antigens on cancer cells, even in the presence of the stimulatory cytokine IFNy. This unresponsiveness to IFNy was the result of failure to phosphorylate the signal transducer STAT1. Forced expression of constitutive active STAT1 fully rescued the MHC class I presentation. Furthermore, oxygen- and glucose-deprivation increased PI3K activity in tumor cells. Pharmacological inhibition of this pathway not only restored signal transduction through IFNy-STAT1 but also improved MHC class I presentation. Importantly, PI3K inhibitors also rendered tumor cells sensitive for recognition by CD8 T cells in culture conditions of metabolic stress.ConclusionsThese data revealed a strong impact of metabolic stress on the presentation of tumor antigens by MHC class I and suggest that this type of tumor escape takes place at hypoxic areas even during times of active T cell immunity and IFNy release.

Abstract B57: Effect of ADAM17 inhibition on the release of IFN-y during natural killer cell-mediated antibody dependent cellular cytotoxicity in cancer

Abstract Introduction: Antibody-dependent cell cytotoxicity (ADCC) by NK cells is a key mechanism in the anti-cancer effects of therapeutic antibodies. CD16a is an IgG Fc receptor (FcRIII) present on the surface of NK cells and plays essential role in recognizing tumor-bound antibodies to initiate ADCC. NK cells are triggered to release cytokines like IFN-y during ADCC which has been reported to have a number of antitumor activities. A disintegrin and metalloproteinase 17 (ADAM17) plays key role in downregulating CD16a in the tumor microenvironment and facilitate tumor induced anergy through the inactivation of cell cytotoxic function. Downregulation of CD16a on NK cells and decreased levels of IFN-y have been reported in ovarian cancer patients. We hypothesize that by blocking the function of ADAM17 we can enhance IFN-y release during ADCC by maintaining higher CD16a levels on the NK cells. To test this hypothesis, we used ADAM17 function blocking antibody MEDI3622 (MedImmune), HER2 receptor positive ovarian (SKOV-3 and MA148) and CD20 receptor positive lymphoblastoid (Raji) cancer cell lines. To further study the role of CD16a in this process we used monoclonal function block anti-CD16 antibody (clone: 3G8) and cleavage resistant CD16a NK92 cells (CD16a/S197P) where we substituted serine with proline at the location of 197 in the membrane proximal region. Description: NK cells derived from peripheral blood (PBNK) were isolated through negative selection EasySepTM (StemCell, Cat: 19055). These PBNK cells were incubated with target cells in the presence of a therapeutic antibody to initiate ADCC with or without blocking ADAM17 by MEDI3622. The samples were incubated at 37°C in CO2 incubator and the supernatant were collected after 4 hrs of incubation by briefly spinning them at 1500 rpm for 5 minutes. IFN-y was measured through CBA (BD Bioscience, Cat: 560111). Summary: The blocking of ADAM17 by MEDI3622 in the presence of herceptin showed an increase in the IFN-y release in comparison to herceptin alone group during ADCC with two ovarian cancer cell lines i.e. SKOV-3 (p&amp;lt;0.0001) and MA148 (p&amp;lt;0.0001). Furthermore, this increase was consistent when we performed the ADCC assay with a different lymphoblastoid related cancer cell line (Raji) with a dissimilar target (CD20 receptor) and therapeutic antibody (rituximab) where we again found a significant increase in IFN-y release in MEDI3622 with rituximab group in comparison to rituximab alone group (p&amp;lt;0.0001). Next we investigated the role of CD16a receptor in the release of IFN-y. We blocked the CD16a receptor with a monoclonal antibody (clone: 3G8) during herceptin mediated ADCC by PBNK cells in SKOV-3 cancer cell line with or without MEDI3622 and found a substantial decrease in the IFN-y release (P&amp;lt;0.0001) in both the groups (herceptin+MEDI3622 and herceptin alone groups). In another experiment, where we expressed a cleavage resistant CD16a (CD16a/S197P) in human NK cells line (NK92), we compared the IFN-y release by CD16a/S197P to CD16a/WT NK92 cells during herceptin mediated ADCC in SKOV-3 tumor. In this experiment, NK92a/S197P showed higher release of IFN-y in comparison to NK92aWT (p&amp;lt;0.0001). Upon blocking ADAM17 on CD16a/WT there was a significant increase in the IFN-y (p&amp;lt;0.0003) release. However, NK92a/S197P did not show any further increase in the presence of MEDI3622 (p=ns) signifying the key role of CD16a in this process. Conclusion: Our results indicate that the inhibition of ADAM17 enhances the IFN-y release by NK cells during ADCC by blocking the shedding of CD16a. Based on these results, ADAM17 may be considered a checkpoint target to improve tumor killing and its utility can be exploited in cancer immunotherapy. Citation Format: Hemant K. Mishra, Wu Jianming, Nabendu Pore, Bruce Walcheck. Effect of ADAM17 inhibition on the release of IFN-y during natural killer cell-mediated antibody dependent cellular cytotoxicity in cancer [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2017 Oct 1-4; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2018;6(9 Suppl):Abstract nr B57.

PD1-CD28 Fusion Protein Enables CD4+ T Cell Help for Adoptive T Cell Therapy in Models of Pancreatic Cancer and Non-hodgkin Lymphoma.

Background: Interaction of the programmed death receptor 1 (PD-1) and its ligand, PD-L1, suppresses T cell activity and permits tumors to evade T cell-mediated immune surveillance. We have recently demonstrated that antigen-specific CD8+ T cells transduced with a PD1-CD28 fusion protein are protected from PD-1-mediated inhibition. We have now investigated the potential of PD1-CD28 fusion protein-transduced CD4+ T cells alone or in combination with CD8+ T cells for immunotherapy of pancreatic cancer and non-Hodgkin lymphoma.Methods: OVA-specific CD4+ and CD8+ were retrovirally transduced with the PD1-CD28 fusion protein. Cytokine release, proliferation, cytotoxic activity, and phenotype of transduced T cells were assessed in the context of Panc02-OVA (murine pancreatic cancer model) and E.G7-PD-L1 (murine T cell lymphoma model) cells.Results: Stimulation of PD1-CD28 fusion protein-transduced CD4+ T cells with anti-CD3 and recombinant PD-L1 induced specific T cell activation, as measured by IFN-y release and T cell proliferation. Coculture with Panc02-OVA or E.G7-PD-L1 tumor cells also led to specific activation of CD4+ T cells. Cytokine release and T cell proliferation was most effective when tumor cells simultaneously encountered genetically engineered CD4+ and CD8+ T cells. Synergy between both cell populations was also observed for specific tumor cell lysis. T cell cytotoxicity was mediated via granzyme B release and mediated enhanced tumor control in vivo. Transduced CD4+ and CD8+ T cells in co-culture with tumor cells developed a predominant central memory phenotype over time. Different ratios of CD4+ and CD8+ transduced T cells led to a significant increase of IFN-y and IL-2 secretion positively correlating with CD4+ T cell numbers used. Mechanistically, IL-2 and MHC-I were central to the synergistic activity of CD4+ and CD8+ T cells, since neutralization of IL-2 prevented the crosstalk between these cell populations.Conclusion: PD1-CD28 fusion protein-transduced CD4+ T cells significantly improved anti-tumoral effect of fusion protein-transduced CD8+ T cells. Thus, our results indicate that PD1-CD28 fusion protein-transduced CD4+ T cells have the potential to overcome the PD-1-PD-L1 immunosuppressive axis in pancreatic cancer and non-Hodgkin lymphoma.

Validation of PASD1 as an immunotherapeutic target in colorectal cancer (CRC)

Event Abstract Back to Event Validation of PASD1 as an immunotherapeutic target in colorectal cancer (CRC) Joanne E. Soh1, Nadiah Abu1, Hui-min Neoh1, Luqman Mazlan2, Azyani Yahaya3 and Rahman Jamal1* 1 UKM Medical Molecular Biology Institute (UMBI), Malaysia 2 Department of Surgery, Faculty of medicine, University of Malaya, Malaysia 3 Department of Pathology, Faculty of Medicine, National University of Malaysia, Malaysia Background Colorectal cancer (CRC) is the third commonest malignancy in Asia including Malaysia. Despite the advances in screening, surgery and conventional therapies, a significant proportion of CRC patients failed to achieve long term remission or experienced toxic side effects of treatment. The identification of PAS (Per ARNT Sim) domain containing 1 (PASD1), a promising cancer testis antigen (CTA) as an immunotherapeutic target, represents one approach to improve treatment options for CRC. The aim of the present study was to investigate the expression of PASD1 and CD8 T-cells pulsed with PASD1 immune responses in CRC patients. Methods PASD1 mRNA expression was determined via RT-PCR on paired tumors and normal tissue samples from 25 CRC patients. Tissue expression of PASD1 was assessed via immunochemistry (IHC) in another 18 CRC samples. Four immunogenic PASD1 peptides predicted to bind to Major Histocompatibility Complex (MHC) Class 1 allele were identified using web-based algorithms. Peripheral blood mononuclear cells from CRC patients were used to investigate the immunogenicity of these HLA-A*2402 restricted peptides in vitro using IFN-y release ELISpot assay. The effects of effector CD8+ T-cells pulsed with PASD1 peptides against different cancer cells were detected in cytolytic and granzyme-B release ELISpot cytotoxicity assays. Results Gene expression of PASD1 was detected in 20% (5/25) CRC samples but not its protein expression. One of the four PASD1 peptides, PASD1(4), was shown to be immunogenic in CRC patients studied via the IFN-y release ELISpot assay. CD8-positive cytotoxic T cells from three patients raised against PASD1(4) were able to lyse HLA-A*2402 positive-SW480 cell line expressing endogenous PASD1 protein in all three effector/target ratios. Conclusion This is the first report of CD8-positive CTL response to PASD1 protein in CRC patients. Preliminary results show that PASD1 peptide is a probable immunogenic antigen and represents a potential target for peptide-based immunotherapy in CRC Keywords: PASD1, colorectal cancer, HLA-A*2402, cancer immunotherapy, Cytotoxic T-cell Conference: International Conference on Drug Discovery and Translational Medicine 2018 (ICDDTM '18) “Seizing Opportunities and Addressing Challenges of Precision Medicine”, Putrajaya, Malaysia, 3 Dec - 5 Feb, 2019. Presentation Type: Poster Presentation Topic: Cancer Citation: Soh JE, Abu N, Neoh H, Mazlan L, Yahaya A and Jamal R (2019). Validation of PASD1 as an immunotherapeutic target in colorectal cancer (CRC). Front. Pharmacol. Conference Abstract: International Conference on Drug Discovery and Translational Medicine 2018 (ICDDTM '18) “Seizing Opportunities and Addressing Challenges of Precision Medicine”. doi: 10.3389/conf.fphar.2018.63.00047 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 30 Sep 2018; Published Online: 17 Jan 2019. * Correspondence: Prof. Rahman Jamal, UKM Medical Molecular Biology Institute (UMBI), Kuala Lumpur, Malaysia, rahmanj@ppukm.ukm.edu.my Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Joanne E Soh Nadiah Abu Hui-min Neoh Luqman Mazlan Azyani Yahaya Rahman Jamal Google Joanne E Soh Nadiah Abu Hui-min Neoh Luqman Mazlan Azyani Yahaya Rahman Jamal Google Scholar Joanne E Soh Nadiah Abu Hui-min Neoh Luqman Mazlan Azyani Yahaya Rahman Jamal PubMed Joanne E Soh Nadiah Abu Hui-min Neoh Luqman Mazlan Azyani Yahaya Rahman Jamal Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Interferon type I responses in primary and secondary infections

The mammalian host responds to a microbial infection with a rapid innate immune reaction that is dominated by type I interferon (IFN-I) release. Most cells of vertebrates can respond to microbial attack with IFN-I production, but the cell type responsible for most of the systemic IFN-I release is thought to be plasmacytoid dendritic cells (pDCs). Besides its anti-microbial and especially anti-viral properties IFN-I also exerts a regulatory role on many facets of the sequential adaptive immune response. One of these is being the recently described partial, systemic activation of the vast majority of B and T lymphocytes in mice, irrespective of antigen reactivity. The biological significance of this partial activation of lymphocytes is at present speculative. Secondary infections occurring within a short time span of a primary infection fail to elicit a similar lymphocyte activation response due to a refractory period in systemic IFN-I production. This period of exhaustion in IFN-I responses is associated with an increased susceptibility of the host to secondary infections. The latter correlates with well-established clinical observations of heightened susceptibility of patients to secondary microbial infections after viral episodes.

Type I Interferons Trigger Systemic, Partial Lymphocyte Activation in Response to Viral Infection

The vast majority of both T and B cells in mice were found to up-regulate cell surface expression of the early activation markers CD69 and CD86, but not CD25, within 24 h of infection with Semliki Forest virus. Kinetics and magnitude of activation marker expression was dependent on live virus, dose, and correlated with strain virulence. Activation marker expression declined to baseline levels over the next 96 h. This very early "activation" of such a high percentage of lymphocytes required the presence of type I IFN receptor genes, was inducible with poly(I:C), and correlated with IFN-I levels in serum. We conclude that virus-induced IFN-I release systemically affects most of the hosts T and B cells by triggering them rapidly and independently of Ag-reactivity into a semiactivated state.

The suppression of Crk inhibits the metastatic profile of breast cancer cells lines

The suppression of Crk inhibits the metastatic profile of breast cancer cells lines