Significant Population Differences Research Articles

42 EXPLORING THE IMPACT OF AGE AND FRAILTY STATUS ON THE IMMUNE RESPONSE TO COVID-19 ILLNESS USING DETAILED IMMUNO-PHENOTYPING

BackgroundWhilst age and frailty are independently associated with mortality in COVID-19 illness, the underlying immunological mechanisms explaining this remain unexplored. We aimed to explore the impact of age and frailty on the acute immune response to COVID-19 illness.MethodsWe recruited older (aged 65+; n = 33) and younger (aged 20–50; n = 43) participants with acute COVID-19 illness for clinical assessment and detailed immunophenotyping for T-cell, neutrophil, monocyte and inflammatory markers using flow cytometry. We additionally assessed circulating levels of several important pro-inflammatory cytokines. Wilxocon rank-sum, chi-square tests and linear regression were used to examine the impact of age and frailty (Clinical Frailty Score 3–9) on the immune response in COVID-19 illness. Results were compared to age-matched pre-pandemic controls (n = 40).ResultsCOVID-19 illness was associated with a marked pro-inflammatory response (raised CRP, IL-6, IL-2R), lymphopenia and emergency myelopoesis (an expansion of non-classical/intermediate monocytes and immature neutrophils) in both age groups (all P < 0.001). In comparison to younger participants (35.7 ± 8.6 years), older adults (76.7 ± 7.8 years) had a more marked reduction in naïve CD4+ and CD8+ cells and a more marked expansion of activated CD4+, CD8+ and effector CD8+ T-cells (all P < 0.001). These findings were independent of illness severity (all P < 0.001). In comparison to their younger counterparts, older adults had a reduced number of CD10+ neutrophils/non-classical monocytes and greater CRP, IL-6 and IL-2R (P < 0.05), although results were attenuated on adjusting for COVID-19 severity. Frailty was not associated with any significant difference in immune cell population or pro-inflammatory response in acute COVID-19 illness.ConclusionIncreasing age, but not frailty status, was associated with reduced naïve T-cells and a more marked expansion of activated/effector T-cells in acute COVID-19 illness. These findings have important implications for understanding impact of age on anti-viral and pro-inflammatory immune responses.

Immune Profiling of Responses to Influenza Vaccination in Patients with Myeloproliferative Neoplasms

▪Myeloproliferative neoplasms (MPNs) are clonal stem cell neoplasms that include polycythemia vera (PV), essential thrombocythemia (ET), and myelofibrosis (MF). MPN patients are immunocompromised due to immune dysregulation from clonal hematopoiesis, heightened inflammation, and immunosuppressive treatments. Infection is a leading cause of morbidity and mortality in MPNs, and as a result annual influenza vaccination is a critical component of care. However, efficacy of the influenza vaccine in MPN patients is unclear. This study sought to identify differences between MPN patients and healthy donors in their immune response to influenza vaccination and subsequent in vitro stimulation.The Massachusetts General Hospital (MGH) maintains a tissue bank of peripheral blood and bone marrow samples collected in consented patients with hematologic malignancy, including MPNs. The tissue bank was interrogated for MPN patients with peripheral blood samples banked 1-6 months after yearly trivalent/quadrivalent influenza vaccination. A total of 13 MPN patients were included, with a median age of 70 (32-86). Five patients had MF, 3 patients had PV, 4 patients had ET, and 1 patient had pre-fibrotic MF. Five patients were on treatment with ruxolitinib, 4 patients were on hydroxyurea (HU), and 4 patients were without treatment. Peripheral blood mononuclear cells (PBMCs) from 11 healthy donors 3-6 months post-vaccination, with a median age of 50 (28-56), were sourced commercially as controls.Cryopreserved samples were thawed and PBMC samples divided equally prior to incubation for 18 hours with peptides derived from Influenza A or sham. Subsequently samples were labeled with a 41-antibody panel and analyzed by mass cytometry. Manual gating together with FlowSOM clustering followed by EdgeR using T-test and FDR to correct for multiple comparisons were used to identify differentially abundant immune cell clusters between cohorts and treatment conditions.Without stimulation, MPN patients were decreased in B cell and memory CD4+ T cell populations as compared to healthy donors (p<0.05), in agreement with a previous report. MPN patients also exhibited an increase in a subset of mature B cells (p<0.05) and lineage negative CD123+ cells (p<0.05) as compared to controls. Stimulation included in this study aimed to expand on these distinctions and identified that baseline differences were retained with additional increases in CD4+ and CD8+ T cell responses in healthy donors as compared to MPN patients. Conversely, stimulation increased the frequency of inflammatory monocytic cells in MPN patients compared to healthy controls (p<0.05).In healthy donors stimulation induced several alterations in, most notably an increase in CD4+ effector memory cells (p<0.03), as compared to matched unstimulated controls. In contrast, comparison of pre- and post-stimulation within the MPN cohort revealed no significant differences in immune cell populations apart from a decrease in naïve CD4+ T cells (p<0.05). Patients receiving HU were enriched for naïve CD4+ and effector CD8+ T cells as compared to patients receiving ruxolitinib (p<0.05); however, these differences were diminished following stimulation. Within MPN patients, pre-stimulation samples in MF patients were characterized by increased abundance of activated monocyte/macrophage clusters and a lineage-negative CD123+ population; there were no significant differences in post-stimulation samples between MF and ET/PV/pre-fibrotic MF patients.We evaluated immunologic responses to influenza vaccination, with and without peptide stimulation, in MPN patients and compared to healthy controls. Overall, MPN patients demonstrated less robust B- and T-cell responses and a muted response to stimulation as compared to healthy controls, which may indicate reduced cellular responses following influenza vaccination. Ongoing analysis will interrogate specific immune populations and subsets in detail to refine current observations and will evaluate associations between immunologic cell clusters with serologic responses and additional patient characteristics. The results of our study highlight important alterations in the immune response to influenza vaccination and muted response to stimulation in MPN patients and will inform a future prospective study of vaccine responsiveness and immune dysfunction in patients with MPNs. [Display omitted] DisclosuresReeves: PR Fludigm: Honoraria; Immunoscape: Consultancy. Hobbs: Merck: Research Funding; Incyte Corporation: Research Funding; AbbVie.: Consultancy; Novartis: Consultancy; Celgene/Bristol Myers Squibb: Consultancy; Constellation Pharmaceuticals: Consultancy, Research Funding; Bayer: Research Funding.

Identifikasi dan Keragaman Genetik Longtail Tuna (Thunnus tonggol) Yang Didaratkan di PPI Kedonganan dan PPP Muncar Menggunakan Marka D-loop Mitokondria

Longtail tuna (Thunnus tonggol) is one of the oceanodromus neritic species and the migration pattern follows the water currents. Currently, this species has not been widely studied in Indonesian waters, so it is nesessary to study the identification of morphology and genetic diversity. The molecular approach employed in this study is DNA barcoding using mitochondrial D – loop locus. This study can explain the importance of species genetic information in stability and resilience. This study aims to determine the identification of morphology, phylogenetic and genetic diversity of longtail tuna at two locations in PPI Kedonganan, Bali dan PPP Muncar, Banyuwangi. The molecular analysis was done in several stages, i.e DNA extraction, Polymerase Chain Reaction, electrophoresis and sequencing. Based on the result of sequencing and analysis, 33 samples of longtail tuna was found. The result of phylogenetic tree reconstruction from two locations showed one clade with genetic distance value among longtail tuna species ranging from 0.000 – 0.042 for all close kinship samples. The haplotype diversity (Hd) value of longtail tuna was 0.9905 and nucleotide diversity (?) was 0.020. The high value of genetic diversity indicated that two longtail tuna populations have a high survival ability to adapt on environmental changes. Index fixation analysis (Fst) has a value of 0.0299, p - value &gt; 0.05. The index fixation value indicated no significant population difference. The result of this study can be use as basic data in planning genetic conservation strategies with sustainable fisheries management efforts.&#x0D; Keywords: Tuna; Thunnus tonggol; Genetic Marker

Information access and preventive measures during COVID-19 among migrants and general population

BackgroundAccess to information and adherence to preventive measures are crucial for containing the COVID-19 outbreak. This study examines access to information and adherence to preventive measures during the COVID-19 pandemic among migrant origin persons and the general population.MethodsData from the Finnish MigCOVID Survey conducted 10/2020 to 2/2021 among persons aged 20 to 66 years was used (n = 3 668). Participants in Health 2017 follow-up survey were the reference group (n = 3 490). We examined age and sex-adjusted access to information by language skills and adherence to preventive measures (staying home if fluish; keeping a safety distance; wearing a face mask; good hand and coughing hygiene; avoiding traveling abroad; avoiding shaking hands with others; downloading the Koronavilkku contact tracing app).ResultsPersons with excellent Finnish/Swedish language skills (96.8%; 95% CI 95.0-97.9) reported receiving sufficient information more often those with intermediate (92.3; 95% CI 88.8-94.8) or basic (91.0%; 95% CI 88.0-93.3) language skills. Overall high adherence to preventive measures was observed both among the migrant origin and the general population. However, significant differences in the migrant origin population by region of origin were observed. More pronounced differences between migrant origin persons and general population persons were observed for avoiding traveling abroad (83.7%; 95% CI 81.1-86.0 vs. 96.6%; 95% CI 95.6-97.3) and downloading the Koronavilkku app (42.8%; 95% CI 39.8-45.9 vs. 64.3%; 95% CI 61.7-66.8).ConclusionsPoorer perceived access to information by language skills points to the continuous need for use of simple language and multilingual communication materials. Self-reported adherence to key preventive measures campaigned by health authorities was generally high both among persons of migrant origin and the general population, although concerns were noted in adherence to these recommendation in some regional groups.Key messages Multilingual and multichannel communication is central in crisis situations to ensure adequate access to information among different population groups.Higher incidence of COVID-19 among migrant origin persons is likely related not only to information access and adherence to preventive measures, but also to various social and structural factors.

Frequencies and mechanisms of pesticide resistance in Tetranychus urticae field populations in China.

The two-spotted spider mite Tetranychus urticate is an important agricultural pest worldwide. It is extremely polyphagous and has developed resistance to many pesticides. Here, we assessed the pesticide resistance of seven field populations of T. urticae in China, their target site mutations and the activities of their detoxification enzymes. The results showed that abamectin and the traditional pesticides pyridaben, profenofos and bifenthrin had higher resistance or lower toxicity than more recently developed pesticides including chlorfenapyr, spinetoram, cyflumetofen, cyenopyrafen, bifenazate and B-azolemiteacrylic. The frequency of point mutations related to abamectin resistance, G314D in the glutamate-gated chloride channel 1 (GluCl1) and G326E in GluCl3, ranged 47%-70% and 0%-97%, respectively. The frequency of point mutations in A1215D and F1538I of the voltage-gated sodium channel gene (VGSC), which may increase resistance to pyrethroids, ranged 88%-100% and 10%-100%, respectively. For target sites related to organophosphate resistance, mutation frequencies ranged 25%-92% for G119S and 0%-23% for A201S in the acetycholinesterase gene (Ace). Mutation G126S in the bifenazate resistance-related cytochrome b gene (Cytb) was observed in three of the seven T. urticae populations. Higher activities of detoxification enzymes (P450, GST, CarEs and UGTs) were observed in two T. urticae populations, with significant difference in the XY-SX population. These results provide useful information on the status of pesticide resistance of T. urticae in China and suggest that T. urticae field populations may have multiple resistance mechanisms.

Steady Habits in the Constitution State: Connecticut's Inequitable System of Representation, 1639–1965

Steady Habits in the Constitution State: Connecticut's Inequitable System of Representation, 1639–1965

The Influence of Hospital Urbanicity on Mortality in Patients With Acute Respiratory Failure: A National Cohort Retrospective Analysis.

The primary objective of this study was to employ a national database to evaluate the association of hospital urbanicity, urban versus rural, on mortality and length of hospital stay in patients hospitalized with acute respiratory failure. We used the 2014 National Inpatient Sample database to evaluate the association of hospital urbanicity with (1) mortality and (2) prolonged hospital stay, defined as ≥ 75th percentile of the study population. We conducted a mixed-effects logistic regression analysis adjusting for sociodemographic variables and medical comorbidities. The random effect was hospital identification number (a unique value assigned in the NIS database for a specific institution). The odds ratio (OR), 95% CI, and P values were reported for each independent variable. The odds of inpatient mortality were significantly higher among urban teaching (OR 1.39, 95% CI 1.39-1.66, P < .001) and urban nonteaching hospitals (OR = 1.39, 95% CI 1.26-1.52, P < .001) compared to rural hospitals. The odds of prolonged hospital stay were significantly higher among urban teaching (OR = 1.82, 95% CI 1.66-2.0, P < .001) and urban nonteaching compared to rural hospitals (OR = 1.50, 95% CI 1.36-1.65, P < .001). This study supports the current body of literature that there are significant differences in patient populations among hospital type. Differences in health outcomes among different types of hospitals should be considered when designing policies to address health equity as these are unique populations with specific needs.

Observation of ionization trends in a laboratory photoionized plasma experiment at Z.

We report experimental and modeling results for the charge state distribution of laboratory photoionized neon plasmas in the first systematic study over nearly an order of magnitude range of ionization parameter ξ∝F/N_{e}. The range of ξ is achieved by flexibility in the experimental platform to adjust either the x-ray drive flux F at the sample or the electron number density N_{e} or both. Experimental measurements of photoionized plasma conditions over such a range of parameters enable a stringent test of atomic kinetics models used within codes that are applied to photoionized plasmas in the laboratory and astrophysics. From experimental transmission data, ion areal densities are extracted by spectroscopic analysis that is independent of atomic kinetics modeling. The measurements reveal the net result of the competition between photon-driven ionization and electron-driven recombination atomic processes as a function of ξ as it affects the charge state distribution. Results from radiation-hydrodynamics modeling calculations with detailed inline atomic kinetics modeling are compared with the experimental results. There is good agreement in the mean charge and overall qualitative similarities in the trends observed with ξ but significant quantitative differences in the fractional populations of individual ions.

Characterization of fecal microbiome in biopsy positive prostate cancer patients

ObjectivesTo characterize the fecal microbiome in newly diagnosed prostate cancer patients.Patients and methodsForty‐nine consecutive patients who were referred for trans rectal prostate biopsy were tested. Patients who received antibiotics 3 months prior to the biopsy, patients with history of pelvic irradiation, prostate or colon cancer, inflammatory bowel disease and urinary tract infection were excluded. A rectal swab was obtained just prior to the biopsy, immediately placed in a sterile tube and kept in −80°C. Sequencing was performed for the 16S rRNA 515F + 806R gene fragment using the QIIME2 software. Analytic tests included Beta diversity (Weighted Unifrac, Unweighted Unifrac, Bray‐Curtis), Alpha diversity (Faith, Evenness), Taxa bar plots and PCoA plots.ResultsForty‐five samples were suitable for analysis with at least 8000 readings per sample. All patients were Caucasian. Twenty patients had prostate cancer and 25 had benign prostates (BPH). Among prostate cancer patients, Gleason Score was 3 + 3 in 11 patients, 3 + 4 in 5, 4 + 3 in 3, and 4 + 4 in 2. There was no statistical difference in demographic parameters between the groups. We identified over 1000 bacterial species, typical for the colonic microbiome. No significant differences in bacterial populations were found between prostate cancer versus benign prostate patients nor between age groups or between subgroups of Gleason or International Society of Uro‐pathology (ISUP) scores.ConclusionsAlthough the microbiome has previously been shown to have an impact on the human microenvironment and cancer risk, we could not demonstrate a significant difference between the flora diversity of newly diagnosed prostate cancer patients and BPH patients. Further research into distinct bacterial metabolic pathways may reveal unique risk factors for prostate cancer.

Incidence of Fungal Aerosols from Selected Crowded Places in Port Harcourt, Nigeria

The role of aerosols in the spread of infectious diseases cannot be overemphasized in the face of increased environmental pollution from different sources. This study aimed at determining the distribution of fungi occurring in the air around human dwellings. The fungal aerosols were randomly collected from five crowded environments (market, church, school, motor park and crowded home) in Port Harcourt, Rivers State, Nigeria and examined. The sedimentation method was used for the microbiological sampling of air and fungi were identified based on macroscopic and microscopic method. The mean fungal load of the different crowded environments were as follows; market (3.19±0.43 log10 SFU cm−2 min−1), church (2.96±0.33 log10 SFU cm−2 min−1), school (3.22±0.29 log10 SFU cm−2 min−1), motor park (3.25±0.35 log10 SFU cm−2 min−1) and home (3.04±0.46 log10 SFU cm−2 min−1), with motor park having the highest and church having the lowest. A total of 16 fungal isolates belonging to twelve (12) genera were identified. They included Aspergillus spp. (31.25%) and Paecilomyces sp., Mucor sp., Fusarium sp., Aspergillus sp., Rhizopus sp., Colletotrichum sp., Cryptococcus sp., Alternaria sp., Cladosporium sp., Chrysosporium sp. and Lophophyton sp. each having a percentage occurrence of 6.25%. Although there was no significant difference (p≥0.05) in the fungal population in the different sampled locations, Motor Park was observed to have the highest percentage occurrence (34.50%) and church the least (9.35%). The percentage occurrence from all sampled sites was in the order motor park>school>market>home>church. The presence of some potential pathogenic fungi such as Aspergillus flavus pose serious public health risks.

MP48-02 SINGLE CELL RNA SEQUENCING OF UTUC IS FEASIBLE AND REVEALS HETEROGENEITY OF THE TUMOR AND IMMUNE MICROENVIRONMENT

You have accessJournal of UrologyBladder Cancer: Upper Tract Transitional Cell Carcinoma II (MP48)1 Sep 2021MP48-02 SINGLE CELL RNA SEQUENCING OF UTUC IS FEASIBLE AND REVEALS HETEROGENEITY OF THE TUMOR AND IMMUNE MICROENVIRONMENT Peter Reisz, Andrew Tracey, Fengshen Kuo, Jasmine Thomas, Quinlan Demac, Tim Clinton, Hong Truong, Andrew Lenis, Lucas Nogueira, Karan Nagar, Taha Merghoub, Bernard Bochner, Eugene Pietzak, David Solit, Kwanghee Kim, and Jonathan Coleman Peter ReiszPeter Reisz More articles by this author , Andrew TraceyAndrew Tracey More articles by this author , Fengshen KuoFengshen Kuo More articles by this author , Jasmine ThomasJasmine Thomas More articles by this author , Quinlan DemacQuinlan Demac More articles by this author , Tim ClintonTim Clinton More articles by this author , Hong TruongHong Truong More articles by this author , Andrew LenisAndrew Lenis More articles by this author , Lucas NogueiraLucas Nogueira More articles by this author , Karan NagarKaran Nagar More articles by this author , Taha MerghoubTaha Merghoub More articles by this author , Bernard BochnerBernard Bochner More articles by this author , Eugene PietzakEugene Pietzak More articles by this author , David SolitDavid Solit More articles by this author , Kwanghee KimKwanghee Kim More articles by this author , and Jonathan ColemanJonathan Coleman More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002074.02AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Upper tract urothelial carcinoma (UTUC) has unique clinical and molecular characteristics compared to bladder UC. Prior investigations have used bulk profiling of tumor tissue to identify molecular subtypes, classifying the majority of UTUC as luminal and T-cell depleted. Single-cell RNA sequencing (scRNA-seq) allows examination of intra-tumoral heterogeneity and the immune tumor microenvironment (TME). We applied scRNA-seq to better characterize UTUC. METHODS: scRNA-seq was performed on 16 UTUC tissue specimens using an established institutional process and the 10X Genomics platform. Sequencing reads were aligned/captured by 10X Cellranger, R/Seurat package. We assessed the composition of each specimen with known marker genes for molecular subtypes. We then assessed the composition of immune cells in each specimen using known marker genes. We compared high- (HG) and low-grade (LG) specimens by subtype composition and immune cell infiltrates. RESULTS: Table 1 describes cohort characteristics and sequencing metrics. Lineage density analyses demonstrate intra-tumoral heterogeneity with luminal subtype predominance across all samples. CD45+ cell analysis revealed significant heterogeneity of immune cell infiltrates (Figure 1). There were no significant differences in immune cell populations in HG vs. LG. Increased interferon gamma (IFNg) expression in tumor cells is associated with significantly increased immune cell infiltration (Spearman’s rank correlation r = 0.71, p=0.009). CONCLUSIONS: Our scRNA-seq data confirmed luminal subtype predominance and described the heterogeneous TME in UTUC. The statistically significant relationship of tumor cell IFNg with increased overall immune cell infiltration is notable and may have implications for systemic therapy response. Further high-resolution studies are needed to expand our knowledge of the molecular biology and TME of UTUC with implications for treatment selection and disease surveillance. Source of Funding: Thompson Family Foundation © 2021 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 206Issue Supplement 3September 2021Page: e863-e863 Advertisement Copyright & Permissions© 2021 by American Urological Association Education and Research, Inc.MetricsAuthor Information Peter Reisz More articles by this author Andrew Tracey More articles by this author Fengshen Kuo More articles by this author Jasmine Thomas More articles by this author Quinlan Demac More articles by this author Tim Clinton More articles by this author Hong Truong More articles by this author Andrew Lenis More articles by this author Lucas Nogueira More articles by this author Karan Nagar More articles by this author Taha Merghoub More articles by this author Bernard Bochner More articles by this author Eugene Pietzak More articles by this author David Solit More articles by this author Kwanghee Kim More articles by this author Jonathan Coleman More articles by this author Expand All Advertisement Loading ...

Prognostic value of derived neutrophil-to-lymphocyte ratio (dNLR) in patients with non-small cell lung cancer receiving immune checkpoint inhibitors: a meta-analysis

ObjectivesDerived neutrophil-to-lymphocytes ratio (dNLR) has recently been reported as a novel potential biomarker associated with prognosis of non-small cell lung cancer (NSCLC). However, evidence for the prognostic utility of dNLR...

Adaptive plasticity in plant traits increases time to hydraulic failure under drought in a foundation tree.

The viability of forest trees, in response to climate change-associated drought, will depend on their capacity to survive through genetic adaptation and phenotypic plasticity in drought tolerance traits. Genotypes with enhanced plasticity for drought tolerance (adaptive plasticity) will have a greater ability to persist and delay the onset of hydraulic failure. By examining populations from different climate-origins grown under contrasting soil water availability, we tested for genotype (G), environment (E) and genotype-by-environment (G×E) effects on traits that determine the time it takes for saplings to desiccate from stomatal closure to 88% loss of stem hydraulic conductance (time to hydraulic failure, THF). Specifically, we hypothesized that: (i) THF is dependent on a G×E interaction, with longer THF for warm, dry climate populations in response to chronic water deficit treatment compared with cool, wet populations, and (ii) hydraulic and allometric traits explain the observed patterns in THF. Corymbia calophylla saplings from two populations originating from contrasting climates (warm-dry or cool-wet) were grown under well-watered and chronic soil water deficit treatments in large containers. Hydraulic and allometric traits were measured and then saplings were dried-down to critical levels of drought stress to estimate THF. Significant plasticity was detected in the warm-dry population in response to water-deficit, with enhanced drought tolerance compared with the cool-wet population. Projected leaf area and total plant water storage showed treatment variation, and minimum conductance showed significant population differences driving longer THF in trees from warm-dry origins grown in water-limited conditions. Our findings contribute information on intraspecific variation in key drought traits, including hydraulic and allometric determinants of THF. It highlights the need to quantify adaptive capacity in populations of forest trees in climate change-type drought to improve predictions of forest die-back.

Effects of rhizosphere fungi on the chemical composition of fruits of the medicinal plant Cinnamomum migao endemic to southwestern China

BackgroundThis study examined how rhizosphere fungi influence the accumulation of chemical components in fruits of a small population species of Cinnamomum migao.ResultsAscomycota and Basidiomycota were dominant in the rhizosphere fungal community of C. migao. Pestalotiopsis and Gibellulopsis were associated with α-Terpineol and sabinene content, and Gibellulopsis was associated with crude fat and carbohydrate content. There were significant differences in rhizosphere fungal populations between watersheds, and there was no obvious change between fruiting periods. Gibberella, Ilyonectria, Micropsalliota, and Geminibasidium promoted sabinene accumulation, and Clitocybula promoted α-Terpineol accumulation.ConclusionThe climate-related differentiation of rhizosphere fungal communities in watershed areas is the main driver of the chemical composition of C. migao fruit. The control of the production of biologically active compounds by the rhizosphere fungal community provides new opportunities to increase the industrial and medicinal value of the fruit of C. migao.

Abstract 1606: Potentiating effect of olaparib on anti-PD-1 treatment in colorectal cancer

Abstract Immune checkpoint inhibitors (ICIs) are approved for microsatellite unstable colorectal cancer (CRC), however, microsatellite stable (MSS) CRC that make up the majority of tumors in clinical practice have not seen any benefit with ICI. This is in part due to the lack of sufficient tumor infiltrating immune cells, particularly CD8+ T cells, in the milieu. Immunogenic cell death (ICD) is a particular form of cell death that elicits robust cross-priming of CD8 T cells against antigens expressed by dying cells in the absence of exogenous adjuvants. ICD-inducing drugs such as poly (ADP-ribose) polymerase inhibitors (PARPis) are poised to enhance cancer cell killing, boost tumor immunogenicity, increase in vivo immune infiltration, and thereby accelerate a tumor response to ICI treatment. Here, we tested the combinatorial therapeutic efficacy of a PARPi, olaparib (OLA), and an ICI, anti-PD-1, in in vitro and in vivo conditions. Combined treatment of OLA and anti-mouse PD-1 reduced tumor growth and increased survival in an immunocompetent, syngeneic animal model of BALB/c carrying allograft CT26 tumor that is MSS. Flow cytometry analysis revealed that the combo treatment significantly reduced percentages of anti-inflammatory macrophages and increased inflammatory monocytes. There was no significant difference in dendritic cell populations. We observed increased anti-tumor lymphocyte response via activation of CD4 and CD8 T cell populations upon combo treatment with OLA and anti-mouse PD-1. In addition, we noticed an increased association of expression of PD-L1 and its receptor PD-1 with OLA treatment in CRC cell lines and CT26 tumors. Our preliminary data suggests altered expressions of the CD8+ T cell exhaustion marker TIM-3 and its ligands galectin-9 and high mobility group protein 1 (HMGB1), an indicator of ICD. The pilot studies also revealed increased levels of STING-dependent type I interferon signaling initiator cGAS upon treatment with OLA in vivo. Mechanistically, OLA might induce DNA damage among tumor cells and trigger innate immune signaling including NF-κB activation and STING-dependent type I interferon signaling, both of which contribute to the maturation of myeloid antigen-presenting cells and subsequent activation of T cells. In conclusion, OLA potentiated the antitumor efficacy of anti-PD-1 in CT26 induced syngeneic CRC mouse model. Our findings indicate that OLA might trigger tumor immunogenicity, and enable a potential targeted-immunotherapeutic angle for exercising ICD and activating cytotoxic T lymphocytes in CT26 syngeneic CRC mouse model; however, further studies are needed to elucidate the mechanism. Despite ICD markers' identification as immuno-stimulating characteristics of dead cells, whether ICD-associated molecules can be used as a prognostic biomarker or therapeutic target, and the rational for such markers on tumor cells in eliciting antitumor immunity are still under investigation. Citation Format: Chintal Bhavsar, Peter John, Magnolia Abreu, Laura Bencosme, Radhashree Maitra, Sanjay Goel, Titto Augustine. Potentiating effect of olaparib on anti-PD-1 treatment in colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1606.

Abstract LB134: The rexinoid MSU-42011 enhances dendritic-T cell cross talk in a HER2 positive model of breast cancer

Abstract Breast cancer (BC) was considered an immunologically silent tumor, however recent findings suggest that immune cells present in the tumor microenvironment play important roles in tumor growth. Retinoid X receptors (RXRs) are a subclass of nuclear receptors that act as ligand-dependent transcription factors that regulate a variety of cellular processes including proliferation and differentiation. Rexinoids are synthetic molecules that bind and activate RXRs. Bexarotene (Targretin®) is the only rexinoid approved by the FDA and is used for the treatment of refractory cutaneous T-cell lymphoma. Other more potent rexinoids have been synthesized, such as LG100268 and LG101506, which will not be tested in clinical trials because of intellectual property issues. Because recent studies have suggested that RXRs are involved in immune regulation, we synthesized novel rexinoids and selected MSU-42011 as a lead compound. MMTV-neu mice with tumors approximately 32 mm3 in volume were treated with control diet or MSU-42011 (100 mg/kg diet) for 10 days. Rexinoid treatment significantly (p=0.0069) reduced tumor fold change normalized to initial volume from 8.33 in controls to 4.80 on treatment. Analysis of the tumor by flow cytometry showed no significant differences in the major immune populations, such as T cells, macrophages and dendritic cells. However, expression of PDL1 was increased in CD11c+, MHC-II+ dendritic cells (5484 vs 7140 MFI, p=0.0456), suggesting engaged dendritic cells in activating cytotoxic T cells. To further elucidate the cross talk between dendritic and T cells, cytokines associated with T cell recruitment and activation were analyzed by RT-PCR. Tumor treated with MSU42011 had a 38.5 fold increase in IL12a expression (p=0.011, n=4), and a 20.8 fold increase in CCL9 (p=ns, n=4), suggesting that dendritic cells are actively recruiting and activating T cells. The activation of cytotoxic T cells was confirmed by a 75.5 fold increase in interferon gamma (p=0.0026, n=4) and a decrease of FOXP3 expression (p=ns, n=4). To test if the effects seen in tumor reduction were dependent on the presence of T cells, CD4 and CD8 were depleted. Depletion of CD4 alone, but not CD8, reversed the tumor reduction induced by MSU-42011. These results show that RXR agonists can be used to modulate immune cell populations in the tumor microenvironment, specifically by recruiting and activating dendritic cells that then activate cytotoxic T cells. We thank the Breast Cancer Research Foundation for support of these studies. Citation Format: Ana Sofia Leal, Sarah Carapellucci, Karen T. Liby. The rexinoid MSU-42011 enhances dendritic-T cell cross talk in a HER2 positive model of breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr LB134.

C-C chemokine hepatocellular carcinoma motif ligand 5-deficiency promotes hepatocellular carcinoma progression by affecting B cell recruitment.

To evaluate the expression of C-C motif chemokine ligand 5 (CCL5) in hepatocellular carcinoma (HCC) and to explore its role in regulating the immune microenvironment and the related mechanism in tumor immunity. The mRNA expression level of CCL5 in HCC and adjacent non-cancerous tissues was measured by quantitative polymerase chain reaction and the protein expression was examined by immunohistochemistry. Serum CCL5 expression was measured by an enzyme-linked immunosorbent assay (ELISA). C57BL/6 wild-type (WT) and Ccl5-knockout (Ccl5-/- ) mice were utilized to conduct the diethylnitrosamine-induced HCC model. The immune cell population was determined by flow cytometry, and peripheral serum immunoglobulin M (IgM) level was quantified by ELISA. CCL5 expression was low in HCC tissue and peripheral blood compared with adjacent non-cancerous tissues or controls, and its expression was correlated with the overall survival, cancer recurrence and distant metastasis. In the HCC mouse model, liver-to-body weight ratio was of the Ccl5-/- group were higher than that of the WT group. Moreover, compared with the WT mice, the number of B cells in the tumor tissue of the Ccl5-/- mice was lower, while there were no significant differences in the other immune cell populations. Furthermore, serum IgM level of the Ccl5-/- mice was significantly lower than that of the WT mice. CCL5 expression is decreased in HCC tissues. CCL5 deficiency reduces B cell recruitment and decreases IgM secretion in HCC, potentially leading to tumor progression.

FIELD COMPARISON OF AUTOCIDAL GRAVID OVITRAPS AND IN2CARE TRAPS AGAINST AEDES AEGYPTI IN DOWNTOWN SAINT AUGUSTINE, NORTHEASTERN FLORIDA

Mosquito Control programs are utilizing cost-effective long term autocidal gravid traps because they minimize labor needs while targeting the gravid population of container-breeding mosquitoes. This field study compared the efficacy of the In2Care Mosquito Trap and the Centers for Disease Control and Prevention autocidal gravid ovitrap (CDC-AGO). The study consisted of two control and two treatment sites, and each treatment site had either 100 In2Care Mosquito Traps or 100 CDC-AGOs. Aedes aegypti populations in each site were monitored using Biogent (BG) Sentinel 2 mosquito traps and ovitraps. Analysis of pre- and post-treatment data indicated no significant difference in adult mosquito populations detected by BG traps from either the In2Care or CDC-AGO sites. However, the mean number of eggs collected by ovitraps showed significant reduction in both trap type treated areas posttreatment, compared to pre-treatment. Furthermore, the mean number of egg collections from the In2Care mosquito trap treated area was much less than the collection from the CDC-AGO trap treated area post-treatment.

Syncytiotrophoblast Extracellular Vesicles From Late-Onset Preeclampsia Placentae Suppress Pro-Inflammatory Immune Response in THP-1 Macrophages

Syncytiotrophoblast derived Extracellular Vesicles (STBEV) from normal pregnancy (NP) have previously been shown to interact with circulating monocytes and B cells and induce pro-inflammatory cytokine release. Early-onset preeclampsia (EOPE) is associated with an exacerbated inflammatory response, yet there is little data regarding late-onset PE (LOPE) and immune function. Here, using a macrophage/monocyte cell line THP-1, we investigated the inflammatory potential of STBEV, comprising medium/large-STBEV (>200nm) and small-STBEV (<200nm), isolated from LOPE (n=6) and normal (NP) (n=6) placentae via dual-lobe ex-vivo placental perfusion and differential centrifugation. THP-1 cells bound and internalised STBEV isolated from NP and LOPE placentae, as revealed by flow cytometry, confocal microscopy, and ELISA. STBEV-treated THP-1 cells were examined for cytokine gene expression by RT-qPCR and the cell culture media examined for secreted cytokines/chemokines. As expected, NP medium/large-STBEV significantly upregulated the transcriptional expression of TNF-α, IL-10, IL-6, IL-12, IL-8 and TGF-β compared to PE medium/large-STBEV. However, there was no significant difference in the small STBEV population between the two groups, although in general, NP small STBEVs slightly upregulated the same cytokines. In contrast, LOPE STBEV (medium and large) did not induce pro-inflammatory responses by differentiated THP-1 macrophages. This decreased effect of LOPE STBEV was echoed in cytokine/chemokine release. Our results appear to suggest that STBEV from LOPE placentae do not have a major immune-modulatory effect on macrophages. In contrast, NP STBEV caused THP-1 cells to release pro-inflammatory cytokines. Thus, syncytiotrophoblast extracellular vesicles from LOPE dampen immune functions of THP-1 macrophages, suggesting an alternative mechanism leading to the pro-inflammatory environment observed in LOPE.

Yeast and lactic acid population relationship in palm wine and their effects on the production of ethanol in Enugu Ezike south eastern Nigeria

Palm wine produced traditionally and consumed by many people in the South eastern part of Nigeria is a whitish liquid formed by naturally fermenting the sugary sap from various palm plants. In this study, we evaluated the population of yeast and lactic acid bacteria in palm wine with regards to its ethanol concentration from three communities in Enugu Ezike, Enugu State. We monitored the isolation of microbial population as well as biochemical characteristics of fermenting palm wine samples using culture-dependent and culture-independent methods. The fermentation step was performed within time interval and completed after 48hrs. Its acidity increasedprogressively with the production of lactic and acetic acids by bacteria.During the fermentation, palm wine microbial numbers (log10 CFU/ml) varied between 1.6 x104 to 7.6 x107 for yeast and 4.5 x 104 to 7.8 x 107 for lactic acid bacteria respectively. Similarly, pH and alcohol level of palm wine samples during the fermentation varied between 3.0 to 5.8 and 0.2 to 5.3 respectively. Statistical analysis revealed that there was no significant difference in both yeast and lactic acid bacteria population at 24hrs. At this time, there was an increased metabolic activity, leading to an increase in ethanol production. At 36-48hrs, there was a slight decrease in both the lactic acid bacteria and the yeast, indicating a reduction in source of carbon.The analysis of the yeast and the lactic acid bacterial population revealed different yeast and bacterial population. Bacteria compete with yeast for nutrients during ethanol production process, potentially causing economic losses. Therefore, it is important to have a better understanding of the abundance and the change in population of lactic acid bacteria and yeast throughout the bio-ethanol process in order to achieve design a more efficient relationship in lactic acid, yeast and ethanol production processes.&#x0D; Keywords: Bacteria, ethanol, lactic acid, palm wine, yeast