Acidosis-mediated increase in IFN-γ-induced PD-L1 expression on cancer cells as an immune escape mechanism in solid tumors.

Immune checkpoint inhibitors have revolutionized cancer therapy, yet the efficacy of these treatments is often limited by the heterogeneous and hypoxic tumor microenvironment (TME) of solid tumors. In the TME, programmed death-ligand 1 (PD-L1) expression on cancer cells is mainly regulated by Interferon-gamma (IFN-γ), which induces T cell exhaustion and enables tumor immune evasion. In this study, we demonstrate that acidosis, a common characteristic of solid tumors, significantly increases IFN-γ-induced PD-L1 expression on aggressive cancer cells, thus promoting immune escape. Using preclinical models, we found that acidosis enhances the genomic expression and phosphorylation of signal transducer and activator of transcription 1 (STAT1), and the translation of STAT1 mRNA by eukaryotic initiation factor 4F (elF4F), resulting in an increased PD-L1 expression. We observed this effect in murine and human anti-PD-L1-responsive tumor cell lines, but not in anti-PD-L1-nonresponsive tumor cell lines. In vivo studies fully validated our in vitro findings and revealed that neutralizing the acidic extracellular tumor pH by sodium bicarbonate treatment suppresses IFN-γ-induced PD-L1 expression and promotes immune cell infiltration in responsive tumors and thus reduces tumor growth. However, this effect was not observed in anti-PD-L1-nonresponsive tumors. In vivo experiments in tumor-bearing IFN-γ-/- mice validated the dependency on immune cell-derived IFN-γ for acidosis-mediated cancer cell PD-L1 induction and tumor immune escape. Thus, acidosis and IFN-γ-induced elevation of PD-L1 expression on cancer cells represent a previously unknown immune escape mechanism that may serve as a novel biomarker for anti-PD-L1/PD-1 treatment response. These findings have important implications for the development of new strategies to enhance the efficacy of immunotherapy in cancer patients.

Immunological Responses to Cancer Therapy.

The use of immunotherapy for cancer has taken flight in the last two decades, from experimental therapy envisioned mainly by laboratory scientists to everyday treatment used by physicians to treat many patients [...].

Consuming media, consuming food: investigating concurrent TV viewing and eating using a 7-d time use diary survey.

OBJECTIVE: One explanation for the relationship between TV viewing and obesity is that people may (over)eat while watching TV. The current study investigated associations between TV viewing and the time spent on (concurrent) eating in a naturalistic setting among a general population sample. DESIGN: Preregistered secondary data analyses were performed of a diary survey in which respondents reported their time use in 10-min blocks for 7 d. SETTING: Concurrent TV viewing and eating was operationalised as all blocks in which TV viewing and eating occurred simultaneously. Furthermore, the TV content respondents watched was coded as food-related (i.e. culinary content) or non-food related. PARTICIPANTS: The sample composed of 2292 adults (58·9 % female) in the Netherlands, aged ≥ 20 years, from all educational levels (18·1 % low, 29·8 % middle and 51·4 % high). RESULTS: More than half of the respondents (51·3 %) reported concurrent TV viewing and eating at least once during the 7-d diary period. The average eating occasion was longer in duration while watching TV (v. without media use), and the total time spent on eating was longer on days of concurrent TV viewing and eating (v. days of eating without media use). The percentage of TV viewing time spent on concurrent eating did not differ between food-related and non-food-related TV content. CONCLUSIONS: Eating while watching TV was related to an increased time spent on eating. Even though energy intake was not assessed, these findings from a naturalistic setting provide further evidence that concurrent TV viewing and eating may contribute to overeating.

Immune Checkpoint Therapy: Tumor Draining Lymph Nodes in the Spotlights.

Tumor-draining lymph nodes play a paradoxical role in cancer. Surgeons often resect these sentinel lymph nodes to determine metastatic spread, thereby enabling prognosis and treatment. However, lymph nodes are vital organs for the orchestration of immune responses, due to the close encounters of dedicated immune cells. In view of the success of immunotherapy, the removal of tumor-draining lymph nodes needs to be re-evaluated and viewed in a different light. Recently, an important role for tumor-draining lymph nodes has been proposed in the immunotherapy of cancer. This new insight can change the use of immune checkpoint therapy, particularly with respect to the use in neoadjuvant settings in which lymph nodes are still operational.

Ipilimumab plus nivolumab and chemoradiotherapy followed by surgery in patients with resectable and borderline resectable T3-4N0-1 non-small cell lung cancer: the INCREASE trial.

BACKGROUND: The likelihood of a tumor recurrence in patients with T3-4N0-1 non-small cell lung cancer following multimodality treatment remains substantial, mainly due distant metastases. As pathological complete responses (pCR) in resected specimens are seen in only a minority (28-38%) of patients following chemoradiotherapy, we designed the INCREASE trial (EudraCT-Number: 2019-003454-83; Netherlands Trial Register number: NL8435) to assess if pCR rates could be further improved by adding short course immunotherapy to induction chemoradiotherapy. Translational studies will correlate changes in loco-regional and systemic immune status with patterns of recurrence. METHODS/DESIGN: This single-arm, prospective phase II trial will enroll 29 patients with either resectable, or borderline resectable, T3-4N0-1 NSCLC. The protocol was approved by the institutional ethics committee. Study enrollment commenced in February 2020. On day 1 of guideline-recommended concurrent chemoradiotherapy (CRT), ipilimumab (IPI, 1 mg/kg IV) and nivolumab (NIVO, 360 mg flat dose IV) will be administered, followed by nivolumab (360 mg flat dose IV) after 3 weeks. Radiotherapy consists of once-daily doses of 2 Gy to a total of 50 Gy, and chemotherapy will consist of a platinum-doublet. An anatomical pulmonary resection is planned 6 weeks after the last day of radiotherapy. The primary study objective is to establish the safety of adding IPI/NIVO to pre-operative CRT, and its impact on pathological tumor response. Secondary objectives are to assess the impact of adding IPI/NIVO to CRT on disease free and overall survival. Exploratory objectives are to characterize tumor inflammation and the immune contexture in the tumor and tumor-draining lymph nodes (TDLN), and to explore the effects of IPI/NIVO and CRT and surgery on distribution and phenotype of peripheral blood immune subsets. DISCUSSION: The INCREASE trial will evaluate the safety and local efficacy of a combination of 4 modalities in patients with resectable, T3-4N0-1 NSCLC. Translational research will investigate the mechanisms of action and drug related adverse events. TRIAL REGISTRATION: Netherlands Trial Registration (NTR): NL8435 , Registered 03 March 2020.

High FcγR Expression on Intratumoral Macrophages Enhances Tumor-Targeting Antibody Therapy.

Therapy with tumor-specific Abs is common in the clinic but has limited success against solid malignancies. We aimed at improving the efficacy of this therapy by combining a tumor-specific Ab with immune-activating compounds. In this study, we demonstrate in the aggressive B16F10 mouse melanoma model that concomitant application of the anti-TRP1 Ab (clone TA99) with TLR3-7/8 or -9 ligands, and IL-2 strongly enhanced tumor control in a therapeutic setting. Depletion of NK cells, macrophages, or CD8+ T cells all mitigated the therapeutic response, showing a coordinated immune rejection by innate and adaptive immune cells. FcγRs were essential for the therapeutic effect, with a dominant role for FcγRI and a minor role for FcγRIII and FcγRIV. FcγR expression on NK cells and granulocytes was dispensable, indicating that other tumoricidal functions of NK cells were involved and implicating that FcγRI, -III, and -IV exerted their activity on macrophages. Indeed, F4/80+Ly-6C+ inflammatory macrophages in the tumor microenvironment displayed high levels of these receptors. Whereas administration of the anti-TRP1 Ab alone reduced the frequency of these macrophages, the combination with a TLR agonist retained these cells in the tumor microenvironment. Thus, the addition of innate stimulatory compounds, such as TLR ligands, to tumor-specific Ab therapy could greatly enhance its efficacy in solid cancers via optimal exploitation of FcγRs.

A Restricted Role for FcγR in the Regulation of Adaptive Immunity.

By their interaction with IgG immune complexes, FcγR and complement link innate and adaptive immunity, showing functional redundancy. In complement-deficient mice, IgG downstream effector functions are often impaired, as well as adaptive immunity. Based on a variety of model systems using FcγR-knockout mice, it has been concluded that FcγRs are also key regulators of innate and adaptive immunity; however, several of the model systems underpinning these conclusions suffer from flawed experimental design. To address this issue, we generated a novel mouse model deficient for all FcγRs (FcγRI/II/III/IV-/- mice). These mice displayed normal development and lymphoid and myeloid ontogeny. Although IgG effector pathways were impaired, adaptive immune responses to a variety of challenges, including bacterial infection and IgG immune complexes, were not. Like FcγRIIb-deficient mice, FcγRI/II/III/IV-/- mice developed higher Ab titers but no autoantibodies. These observations indicate a redundant role for activating FcγRs in the modulation of the adaptive immune response in vivo. We conclude that FcγRs are downstream IgG effector molecules with a restricted role in the ontogeny and maintenance of the immune system, as well as the regulation of adaptive immunity.

Food at first sight: Visual attention to palatable food cues on TV and subsequent unhealthy food intake in unsuccessful restrained eaters.

BACKGROUND: This study investigated whether unsuccessful dieters show heightened visual attention to food cues in TV content and how visual attention influences subsequent unhealthy food intake. This study adds to prior literature by investigating the influence of visual attention to food cues on food intake with actual media content (i.e., instead of isolated food cues such as pictures or words) and by differentiating between chronic dieters (i.e., restrained eaters) who vary in dieting success (i.e., perceived self-regulatory success [PSRS]). To get a more detailed insight into different processes of visual attention, two measures of attention (i.e., initial orientation and attention duration) were examined. METHODS: Unrestrained (n = 34) and restrained eaters (n = 28) varying in PSRS watched a talk show containing subtly depicted, palatable food cues. While watching, their visual attention to the food cues was measured with an eye-tracker. Unhealthy food intake was assessed afterwards in a taste test. RESULTS: A two-way interaction between eating restraint and PSRS on initial visual orientation was found: unsuccessful restrained eaters' initial orientation to food cues was faster compared to that of successful restrained eaters. There were no significant findings on attention duration. Furthermore, visual attention did not predict unhealthy food intake. DISCUSSION: Unsuccessful restrained eaters' fast initial orientation, but no longer attention duration, suggests that self-regulation may be important at early stages of visual attention. Future research on this topic should continue to differentiate between initial orientation and attention duration, as well as between more and less successful restrained eaters. The lack of findings on unhealthy food intake suggest that food cues embedded in actual media content might have less influence on eating behavior compared to isolated food cues.

FcγR interaction is not required for effective anti-PD-L1 immunotherapy but can add additional benefit depending on the tumor model.

Immunomodulatory antibodies blocking interactions of coinhibitory receptors to their ligands such as CTLA-4, PD1 and PD-L1 on immune cells have shown impressive therapeutic efficacy in clinical studies. The therapeutic effect of these antibodies is mainly mediated by reactivating antitumor T cell immune responses. Detailed analysis of anti-CTLA4 antibody therapy revealed that an optimal therapeutic efficacy also requires binding to Fc receptors for IgG, FcγR, mediating depletion of intratumoral regulatory T cells. Here, we investigated the role of Fc binding in anti-PD-L1 antibody therapy in the MC38 C57BL/6 and CT26 BALB/c colon adenocarcinoma tumor models. In the MC38 tumor model, all IgG subclasses anti-PD-L1 showed similar therapeutic efficacy when compared to each other in either wild-type mice or in mice deficient for all FcγR. In contrast, in the CT26 tumor model, anti-PD-L1 mIgG2a, the IgG subclass with the highest affinity for activating FcγR, showed stronger therapeutic efficacy than other IgG subclasses. This was associated with a reduction of a myeloid cell subset with high expression of PD-L1 in the tumor microenvironment. This subclass preference for mIgG2a was lost in C57BL/6 × BALB/c F1 mice, indicating that the genetic background of the host may determine the additional clinical benefit of the high affinity antibody subclasses. Based on these data, we conclude that FcγR are not crucial for anti-PD-L1 antibody therapy but might play a role in some tumor models.

Watch what you watch: The effect of exposure to food-related television content on the accessibility of a hedonic eating goal.

BACKGROUND: This study examined whether seeing food-related TV content affected the accessibility of a hedonic eating goal differently for people scoring relatively high or low on chronic dieting (i.e., eating restraint) and perceived self-regulatory success (i.e., PSRS). METHODS: Three between-subjects experiments were conducted in which participants were exposed to food-related or non-food related TV content. In Experiment 1 (student sample, N = 111) and Experiment 2 (community sample, N = 69) participants watched TV commercials for food or non-food products and in Experiment 3 (student sample, N = 102) a cooking show or a non-food TV show. Hedonic eating goal accessibility was assessed by means of a lexical decision task (LDT). Eating restraint and PSRS were measured afterwards. RESULTS: The expected three-way interaction between TV content, eating restraint, and PSRS on hedonic eating goal accessibility was not found in Experiments 1 and 2. In Experiment 3, a three-way interaction was found although effects were short-lived. As expected, watching food-related versus non-food related TV content resulted in more hedonic eating goal accessibility among people relatively high in eating restraint but low in PSRS (i.e., unsuccessful restrained eaters), but in less accessibility among participants relatively high in both eating restraint and PSRS (i.e., successful restrained eaters). DISCUSSION: As effects were found after watching a cooking show (Experiment 3) but not after watching TV commercials (Experiments 1 and 2), future research should explore whether the type of TV content might play a role in the effects of food-related TV content on hedonic eating goal accessibility, as well as whether the effects found on goal accessibility translate into actual food choices.

"I was Right about Vaccination": Confirmation Bias and Health Literacy in Online Health Information Seeking.

When looking for health information, many people turn to the Internet. Searching for online health information (OHI), however, also involves the risk of confirmation bias by means of selective exposure to information that confirms one's existing beliefs and a biased evaluation of this information. This study tests whether biased selection and biased evaluation of OHI occur in the context of early-childhood vaccination and whether people's health literacy (HL) level either prevents or facilitates these processes. Vaccination beliefs were measured for 480 parents of young children (aged 0-4 years) using an online survey, after which they were exposed to a list of ten vaccine-related message headers. People were asked to select those headers that interested them most. They also had to evaluate two texts which discussed vaccination positively and negatively for credibility, usefulness, and convincingness. The results showed that people select more belief-consistent information compared to belief-inconsistent information and perceived belief-confirming information as being more credible, useful, and convincing. Biased selection and biased perceptions of message convincingness were more prevalent among people with higher HL, and health communication professionals should be aware of this finding in their practice.

Self-Assembling Peptide Epitopes as Novel Platform for Anticancer Vaccination.

The aim of the present study was to improve the immunogenicity of peptide epitope vaccines using novel nanocarriers based on self-assembling materials. Several studies demonstrated that peptide antigens in nanoparticulate form induce stronger immune responses than their soluble forms. However, several issues such as poor loading and risk of inducing T cell anergy due to premature release of antigenic epitopes have challenged the clinical success of such systems. In the present study, we developed two vaccine delivery systems by appending a self-assembling peptide (Ac-AAVVLLLW-COOH) or a thermosensitive polymer poly(N-isopropylacrylamide (pNIPAm) to the N-terminus of different peptide antigens (OVA250-264, HPV-E743-57) to generate self-assembling peptide epitopes (SAPEs). The obtained results showed that the SAPEs were able to form nanostructures with a diameter from 20 to 200 nm. The SAPEs adjuvanted with CpG induced and expanded antigen-specific CD8+ T cells in mice. Furthermore, tumor-bearing mice vaccinated with SAPEs harboring the HPV E743-57 peptide showed a delayed tumor growth and an increased survival compared to sham-treated mice. In conclusion, self-assembling peptide based systems increase the immunogenicity of peptide epitope vaccines and therefore warrants further development toward clinical use.

Signalling product healthiness through symbolic package cues: Effects of package shape and goal congruence on consumer behaviour.

Three studies show that product packaging shape serves as a cue that communicates healthiness of food products. Inspired by embodiment accounts, we show that packaging that simulates a slim body shape acts as a symbolic cue for product healthiness (e.g., low in calories), as opposed to packaging that simulates a wide body shape. Furthermore, we show that the effect of slim package shape on consumer behaviour is goal dependent. Whereas simulation of a slim (vs. wide) body shape increases choice likelihood and product attitude when consumers have a health-relevant shopping goal, packaging shape does not affect these outcomes when consumers have a hedonic shopping goal. In Study 3, we adopt a realistic shopping paradigm using a shelf with authentic products, and find that a slim (as opposed to wide) package shape increases on-shelf product recognition and increases product attitude for healthy products. We discuss results and implications regarding product positioning and the packaging design process.

Framing in Entertainment-Education: Effects on Processes of Narrative Persuasion.

Nowadays, entertainment-education (E-E) is often used as a persuasive strategy to stimulate prosocial behavior. Although E-E is mostly regarded as a persuasive strategy in itself, in an increasing number of E-E programs several persuasive strategies are used to communicate the educational message to the audience. This study investigates the effects of a strategy widely used in health communication, but not previously studied in the field of E-E: framing. To this means we examined the effect of two different ways an E-E message can be framed: by emphasizing either the losses of not performing the behavior in question or the gains of performing this behavior. A serial multiple mediation model showed that framing affected intention to refrain from drunk cycling via counterarguing and attitude toward drunk cycling; the use of a gain frame decreased counterarguing, which decreased the attitude toward drunk cycling. This subsequently resulted in a higher intention to refrain from this behavior. Implications of these results are discussed.

IgG-mediated anaphylaxis to a synthetic long peptide vaccine containing a B cell epitope can be avoided by slow-release formulation.

Synthetic long peptides (SLP) are a promising vaccine modality to induce therapeutic T cell responses in patients with chronic infections and tumors. We studied different vaccine formulations in mice using SLP derived from carcinoembryonic Ag. We discovered that one of the SLP contains a linear Ab epitope in combination with a CD4 epitope. Repeated vaccination with this carcinoembryonic Ag SLP in mice shows improved T cell responses and simultaneously induced high titers of peptide-specific Abs. These Abs resulted in unexpected anaphylaxis after a third or subsequent vaccinations with the SLP when formulated in saline. Administration of low SLP doses in the slow-release vehicle IFA prevented the anaphylaxis after repeated vaccination. This study underscores both the immunogenicity of SLP vaccination, for inducing T cell as well as B cell responses, and the necessity of safe administration routes.

Effects of Disclosing Sponsored Content in Blogs: How the Use of Resistance Strategies Mediates Effects on Persuasion.

This article presents two studies examining the effects of disclosing online native advertising (i.e., sponsored content in blogs) on people's brand attitude and purchase intentions. To investigate the mechanisms underlying these effects, we integrated resistance theories with the persuasion knowledge model. We theorize that disclosures activate people's persuasion knowledge, which in turn evokes resistance strategies that people use to cope with the persuasion attempt made in the blog. We tested our predications with two experiments (N = 118 and N = 134). We found that participants indeed activated persuasion knowledge in response to disclosures, after which they used both cognitive (counterarguing) and affective (negative affect) resistance strategies to decrease persuasion. The obtained insights do not only advance our theoretical understanding of how disclosures of sponsored blogs affect persuasion but also provide valuable insights for legislators, advertisers, and bloggers.

Educational storylines in entertainment television: audience reactions toward persuasive strategies in medical dramas.

Medical television drama series provide an important source of health information. This form of entertainment-education (E-E) can be used to influence knowledge, attitudes, and behaviors toward health-related issues. In the literature, E-E is generally regarded as a persuasive strategy in itself, whereas in an increasing number of E-E programs, several different persuasive strategies are used. An important question is how the audience ethically evaluates these strategies. The aim of the present study is to examine viewers' ethical judgments toward the use of three persuasive strategies in E-E: product placement, framing, and persuasion toward a controversial position. A survey among 525 viewers of 5 popular medical dramas demonstrates that viewers evaluate the use of the currently investigated attitudinal statements about potential persuasive strategies in E-E as being immoral and that viewers prefer neutral storylines. Adopting a strategy that viewers find inappropriate may interfere with the intended prosocial effects of E-E. A broader understanding of the appropriate and inappropriate uses of persuasive strategies in E-E is indispensable for effective E-E productions.

Vaccine-induced effector-memory CD8+ T cell responses predict therapeutic efficacy against tumors.

CD8(+) T cells have the potential to attack and eradicate cancer cells. The efficacy of therapeutic vaccines against cancer, however, lacks defined immune correlates of tumor eradication after (therapeutic) vaccination based on features of Ag-specific T cell responses. In this study, we examined CD8(+) T cell responses elicited by various peptide and TLR agonist-based vaccine formulations in nontumor settings and show that the formation of CD62L(-)KLRG1(+) effector-memory CD8(+) T cells producing the effector cytokines IFN-γ and TNF predicts the degree of therapeutic efficacy of these vaccines against established s.c. tumors. Thus, characteristics of vaccine-induced CD8(+) T cell responses instill a predictive determinant for the efficacy of vaccines during tumor therapy.

Multi-modal cell-free DNA genomic and fragmentomic patterns enhance cancer survival and recurrence analysis.

The structure of cell-free DNA (cfDNA) is altered in the blood of patients with cancer. From whole-genome sequencing, we retrieve the cfDNA fragment-end composition using a new software (FrEIA [fragment end integrated analysis]), as well as the cfDNA size and tumor fraction in three independent cohorts (n = 925 cancer from >10 types and 321 control samples). At 95% specificity, we detect 72% cancer samples using at least one cfDNA measure, including 64% early-stage cancer (n = 220). cfDNA detection correlates with a shorter overall (p = 0.0086) and recurrence-free (p = 0.017) survival in patients with resectable esophageal adenocarcinoma. Integrating cfDNA measures with machine learning in an independent test set (n = 396 cancer, 90 controls) achieve a detection accuracy of 82% and area under the receiver operating characteristic curve of 0.96. In conclusion, harnessing the biological features of cfDNA can improve, at no extra cost, the diagnostic performance of liquid biopsies.

Single-arm trial of neoadjuvant ipilimumab plus nivolumab with chemoradiotherapy in patients with resectable and borderline resectable lung cancer: the INCREASE study.

BACKGROUND: In non-small cell lung cancer (NSCLC), chemoradiotherapy (CRT) yields pathological complete response (pCR) rates of approximately 30%. We investigated using ipilimumab plus nivolumab (IPI-NIVO) with neoadjuvant CRT in resectable, and borderline resectable NSCLC. METHODS: This single-arm, phase-II trial enrolled operable T3-4N0-2 patients with NSCLC without oncogenic drivers. Primary study endpoints were safety, major pathological response (MPR) and pCR. Treatment encompassed platinum-doublet concurrent CRT, IPI 1 mg/kg intravenous and NIVO 360 mg intravenous on day-1, followed by chemotherapy plus NIVO 360 mg 3 weeks later. Thoracic radiotherapy was 50 or 60 Gy, in once-daily doses of 2 Gy. Resections were 6 weeks post-radiotherapy. RESULTS: In a total of 30 patients in the intention-to-treat (ITT) population, grades 3-4 treatment-related adverse events (TRAEs) occurred in 70%, one TRAE grade 5 late-onset pneumonitis on day 96 post-surgery (1/30, 3.3%) occurred, and one non-TRAE COVID-19 death (1/30, 3.3%). pCR and MPR were achieved in 50% (15/30) and 63% (19/30) of the ITT; and in 58% (15/26) and 73% (19/26) of the 26 patients who underwent surgery, respectively. Postoperative melanoma was seen in one non-pCR patient. The R0 rate was 100% (26/26), and no patient failed surgery due to TRAEs. In peripheral blood, proliferative CD8+ T cells were increased, while proliferative regulatory T cells (Tregs) were not. On-treatment, pCR-positives had higher CD8+CD39+ T cells and lower HLA-DR+ Tregs. CONCLUSIONS: Neoadjuvant IPI-NIVO-CRT in T3-4N0-2 NSCLC showed acceptable safety with pCR and MPR in 58% and 73% of operated patients, respectively. No patient failed surgery due to TRAEs. TRIAL REGISTRATION NUMBER: NCT04245514.