Viral Glycoproteins Induce NLRP3 Inflammasome Activation and Pyroptosis in Macrophages.

Infections with viral pathogens are widespread and can cause a variety of different diseases. In-depth knowledge about viral triggers initiating an immune response is necessary to decipher viral pathogenesis. Inflammasomes, as part of the innate immune system, can be activated by viral pathogens. However, viral structural components responsible for inflammasome activation remain largely unknown. Here we analyzed glycoproteins derived from SARS-CoV-1/2, HCMV and HCV, required for viral entry and fusion, as potential triggers of NLRP3 inflammasome activation and pyroptosis in THP-1 macrophages. All tested glycoproteins were able to potently induce NLRP3 inflammasome activation, indicated by ASC-SPECK formation and secretion of cleaved IL-1ß. Lytic cell death via gasdermin D (GSDMD), pore formation, and pyroptosis are required for IL-1ß release. As a hallmark of pyroptosis, we were able to detect cleavage of GSDMD and, correspondingly, cell death in THP-1 macrophages. CRISPR-Cas9 knockout of NLRP3 and GSDMD in THP-1 macrophages confirmed and strongly support the evidence that viral glycoproteins can act as innate immunity triggers. With our study, we decipher key mechanisms of viral pathogenesis by showing that viral glycoproteins potently induce innate immune responses. These insights could be beneficial in vaccine development and provide new impulses for the investigation of vaccine-induced innate immunity.

Gasdermin D mediates host cell death but not interleukin-1ß secretion in Mycobacterium tuberculosis-infected macrophages.

Necrotic cell death represents a major pathogenic mechanism of Mycobacterium tuberculosis (Mtb) infection. It is increasingly evident that Mtb induces several types of regulated necrosis but how these are interconnected and linked to the release of pro-inflammatory cytokines remains unknown. Exploiting a clinical cohort of tuberculosis patients, we show here that the number and size of necrotic lesions correlates with IL-1ß plasma levels as a strong indicator of inflammasome activation. Our mechanistic studies reveal that Mtb triggers mitochondrial permeability transition (mPT) and subsequently extensive macrophage necrosis, which requires activation of the NLRP3 inflammasome. NLRP3-driven mitochondrial damage is dependent on proteolytic activation of the pore-forming effector protein gasdermin D (GSDMD), which links two distinct cell death machineries. Intriguingly, GSDMD, but not the membranolytic mycobacterial ESX-1 secretion system, is dispensable for IL-1ß secretion from Mtb-infected macrophages. Thus, our study dissects a novel mechanism of pathogen-induced regulated necrosis by identifying mitochondria as central regulatory hubs capable of delineating cytokine secretion and lytic cell death.

Long-lived macrophage reprogramming drives spike protein-mediated inflammasome activation in COVID-19.

Innate immunity triggers responsible for viral control or hyperinflammation in COVID-19 are largely unknown. Here we show that the SARS-CoV-2 spike protein (S-protein) primes inflammasome formation and release of mature interleukin-1ß (IL-1ß) in macrophages derived from COVID-19 patients but not in macrophages from healthy SARS-CoV-2 naïve individuals. Furthermore, longitudinal analyses reveal robust S-protein-driven inflammasome activation in macrophages isolated from convalescent COVID-19 patients, which correlates with distinct epigenetic and gene expression signatures suggesting innate immune memory after recovery from COVID-19. Importantly, we show that S-protein-driven IL-1ß secretion from patient-derived macrophages requires non-specific monocyte pre-activation in vivo to trigger NLRP3-inflammasome signaling. Our findings reveal that SARS-CoV-2 infection causes profound and long-lived reprogramming of macrophages resulting in augmented immunogenicity of the SARS-CoV-2 S-protein, a major vaccine antigen and potent driver of adaptive and innate immune signaling.

Importance of and Satisfaction with Information about Their Disease in Cancer Patients.

To learn more about information needs and satisfaction with provided information among cancer patients and whether dissatisfaction with information has any association with how therapy decisions are made. An online survey was conducted during March 2015 and January 2016 by the German non-profit patient organization "Das Lebenshaus e.V." among their members with rare solid tumors. A total of 338 records was analyzed. The majority found information on their disease important and was satisfied with the provided information. The participants were less satisfied with the information concerning management of side effects than with other aspects of information (p < .001). Support groups, lectures, and the oncologist were rated as the most helpful sources of information followed by a second opinion and media. Participants who were dissatisfied with the information more often made the decision on the treatment alone by themselves (p < .001). Our results show a high satisfaction with disease-related information among our study participants. Improvements could be made by offering more information on the management of side effects and by giving more information about support groups, reliable websites, and other helpful media.

Not all cancer patients with an interest in CAM are the same. Differences between patients with a CAM interest prior to the cancer diagnosis and those with first-time interest since diagnosis.

OBJECTIVES: The aim was to assess differences in age, gender, education and personality (Big Five) between cancer patients already interested in CAM prior to the diagnosis and cancer patients only interested in CAM since their diagnosis. DESIGN: 323 members of the support and information platform "Lebenshaus e.V." with gastrointestinal stromal tumours (GIST), renal cell carcinomas (RCC) and sarcomas participated in an online survey. MAIN OUTCOME MEASURES: Interest in CAM prior to cancer diagnosis, CAM interest since diagnosis, CAM use and disclosure of CAM use to doctors. RESULTS: 39% were already interested in CAM before the diagnosis and 40.6% were first interested in CAM after their diagnosis. 44.9% stated a current/past CAM use. Female gender and high education were significant predictors for interest in CAM before the diagnosis and general CAM use. The Big Five did not become significant predictors. Predictors were different for patients interested in CAM before the diagnosis and patients only interested since the diagnosis. Sarcoma patients were significantly more likely to be interested in CAM after the diagnosis than patients with GIST. Disclosure of CAM use could not be predicted by sociodemographic variables and personality. CONCLUSION: Known predictors for interest in CAM such as female gender, higher education and younger age do not seem to apply in cancer patients with no prior CAM affinity. This result together with a high CAM prevalence raises the importance of addressing CAM in all cancer patients for a need-oriented treatment and to avoid side effects and interactions with conventional treatment.

Information needs and usage of complementary and alternative medicine in members of a German self-help group for gastrointestinal stroma tumours, sarcoma, and renal cancer.

OBJECTIVE: 40-50% of German cancer patients use some method of complementary and alternative medicine (CAM) and both patients and doctors often feel insufficiently informed. We examined the information-seeking behaviour and satisfaction with information on patients' interest in CAM and the therapy decision. DESIGN AND SETTING: An anonymous, voluntary online survey was conducted among the members of "Das Lebenshaus e.V." (House of Life), a decentralized support group for patients with gastrointestinal stroma tumours (GIST), sarcoma, and renal cancer. Data was collected from March 2015 until January 2016 using closed questions with multiple choice if appropriate and in case of ranking, a 5-point Likert scale. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Correlations between CAM interest, usage, information needs, sources of information and therapy decision were calculated using chi square tests for univariate analyses. RESULTS: Overall, 431 patients took part in our survey, thus return rate was 19.6%. 43.9% (n = 189) of the participants were female, 37.1% (n = 160) were male, 19.0% (n = 82) did not respond. Mean age was 59.8 years. The most common tumours were GIST (346%, n = 149), renal cancer (22.3%, n = 96) and sarcoma (20.0%, n = 86). 55.2% (n = 138) of the respondents were patients undergoing treatment, 19.7% (n = 85) were after treatment, 2.6% (n = 11) were relatives and 4.4% (n = 19) others while 18.1% (n = 78) did not respond. A total of 81.8% (n = 337) of the participants were interested in CAM, but only 44.7% (n = 152) used one of the methods. Women were more commonly interested in CAM (87.2%, n = 163) and used it more often: 53.0% (n = 97) vs. 36.2% (n = 55). Information about CAM was considered important by 85.5% (n = 360) and the Internet was the most commonly used source for information about CAM (77.9%, n = 205). However, 61.4% (n = 233) were not satisfied with the information received about CAM, especially from doctors and hospitals. Patients unsatisfied with the information they had formally received about the course of their disease significantly more often used CAM (p = 0.029). Users would also make the therapy decision by themselves more often (p = 0.036). Nearly a fifth did not disclose their use to a doctor. CONCLUSIONS: Dissatisfaction with received information reveals a strong need for scientific information to be available to both patients and doctors. Physicians should get special training about CAM. As the Internet is an important source, high-quality and scientific information should be portrayed on webpages easily accessible to patients.

The use of complementary and alternative medicine by patients in routine care and the risk of interactions.

BACKGROUND: Patients suffering from cancer often make use of complementary and alternative medicine (CAM). Only few data exist on the prevalence and clinical significance of interactions of a biological CAM method and conventional drugs. METHODS: From February 2014 to March 2016, consecutive patients from five oncological practices in Germany were asked to fulfill a standardized questionnaire regarding use of CAM. Data on diagnosis, date of first diagnosis, ECOG and the past and current treatment were derived from the patients' files. Interactions were evaluated by systematically using a database on potential interactions. RESULTS: From 1000 patients asked to participate, we received a total of 720 questionnaires of which 711 were completed and eligible for evaluation. 29% of the patients reported any CAM usage. Women showed a significantly higher use of CAM with 35.6 versus 23.6% of men. For 54.9% of CAM users (15.9% of all patients), we found a combination of conventional drugs and biological based CAM methods with a risk for interactions. Vitamins A, C and E were the most frequently used CAM substances in these cases (39.3%), followed by herbs with 17.5%. CONCLUSION: There was a risk of interactions between a biological CAM method and conventional drugs in 54.9% of the patients using CAM. To raise knowledge on interactions a better training for doctors with respect to CAM is strongly needed. Furthermore, patients' awareness should also be raised and communication between physician and patient on the topic improved.