American Type Culture Collection

Abstract Background Human and highly pathogenic avian influenza (HPAI) viruses pose a major public health risk due to their potential for widespread illness and economic consequences. Early detection and control of outbreaks rely on effective surveillance and diagnostic testing.Figure 1:H5N1 qPCR dataFigure 1: qPCR amplification curves generated with ATCC® VR-3436SD™ (subtype H5N1) (blue) and H5N1 gRNA (pink) using (A) a Hoffmann et al., 2016 assay targeting HA, and the (B) CDC Flu SC2 Multiplex assay targeting M.Figure 2:H5N6 qPCR dataFigure 2: qPCR amplification curves generated with ATCC® VR-3439SD™ (subtype H5N6) using (A) a Hoffmann et al., 2016 assay targeting HA, and the (B) CDC Flu SC2 Multiplex assay targeting M. Methods ATCC® developed a comprehensive suite of quantitative synthetic analytical reference materials (ARMs) for HPAI virus serotypes H5N1, H5N6, H7N7, H7N9, and H9N2; human influenza A virus serotypes H1N1, H3N2, and H1N1 2009 pandemic; and Influenza B virus strains. Each synthetic ARM contains the complete sequences from segments 4, 5, 6, 7, and 8, including the HA, NP, NA, M1, M2, NS1, and NEP/NS1 genes, covering 50% of the influenza genome. These segments are key diagnostic targets for molecular tests and provide sufficient genomic context for assessing assay specificity. These ARMs are manufactured using a highly reliable synthetic biology technology, verified through next-generation sequencing, and quantitated via Droplet Digital PCR (Bio-Rad Laboratories, Inc). Further, they do not contain any viable material and can be handled in BSL-1 settings. As such, they are intended to serve as safe and reliable positive controls for molecular tests for surveillance and diagnostics.Figure 3:H9N2 qPCR dataFigure 3: qPCR amplification curves generated with ATCC® VR-3440SD™ (subtype H9N2) using (A) a Hassan et al., 2022 assay targeting HA, and the (B) CDC Flu SC2 Multiplex assay targeting M.Figure 4:pdm09 qPCR dataFigure 4: qPCR amplification curves generated with synthetic RNA ATCC® VR-3388SD™ (H1N1 pdm09) and genomic RNA samples derived from two viral cultures ATCC® VR-3441 ™ VR-1988™, which are also both of H1N1 pdm09 type, using the WHO assay for the detection of Influenza type A subtype H1pdm09. Results The synthetic ARMs were experimentally evaluated using several published quantitative PCR assays, including those from the Centers for Disease Control and Prevention, the World Health Organization, the World Organization for Animal Health, and other highly cited sources. We also conducted an in silico assessment of ARM compatibility with over 250 publicly available published assays. The synthetic products displayed equal performance to genomic RNA during all experimental tests. Conclusion The data demonstrate that all synthetic ARMs for avian and human influenza are effectively designed and suitable for developing and validating molecular-based detection and quantification assays. Furthermore, our findings indicate that the synthetic RNA ARMs are equivalent to their corresponding genomic RNA, making them a valuable BSL-1 alternative to BSL–3–derived materials. Our results suggest that these synthetic ARMs can serve as reliable and safe controls for molecular assays used in diagnostics and surveillance. Disclosures All Authors: No reported disclosures

Radiation exposure from radiological or nuclear events, medical treatments, or spaceflight poses significant health risks, yet human-specific models to investigate radiation effects on skin remain limited. This study establishes a novel in vitro platform using a full-thickness bioengineered human skin equivalent colonized with natural mixed human microbiota (coHSEs) to assess radiation-induced biological responses. We exposed coHSEs to acute doses of up to 4 Gy with x-rays and evaluated their viability, structural integrity, and molecular responses over 25 days. The coHSE model demonstrated sustained viability without dose-dependent opportunistic microbial overgrowth when procedural optimizations were applied. Radiation-induced epidermal remodeling did not compromise tissue architecture or swabbing-based sample collection. Cell proliferation analyses revealed dose- and time-dependent dynamics, with consistent dermal cell density maintained across radiation doses. Comparative multi-omic analyses, including untargeted metabolomics, targeted lipidomics, and 16 S metagenomics, revealed conserved metabolic and microbial responses to radiation in both coHSEs and skin from irradiated mice. Enriched pathways such as arachidonic acid and fatty acid metabolism, along with shifts in microbial taxa including Lachnospiraceae, support the translational relevance of the coHSE model. This system offers a scalable, ethical, and physiologically relevant platform for radiation biology, biodosimetry, and therapeutic development, advancing terrestrial health research with promising application for space research.Supplementary InformationThe online version contains supplementary material available at 10.1038/s41598-025-17153-4.

Cell models paired with curated gene expression data are vital for preclinical research. Transcriptomic data from traceable, authenticated cell lines streamline hypothesis development and experimental design while illuminating underlying biological mechanisms. In cancer research, these models have notably identified therapeutic targets and biomarkers. To advance cancer research, ATCC® sequenced the transcriptomes of 141 immune cancer cell lines, accessible through ATCC Cell Line Land, encompassing various hematopoietic lineages. This effort provided insights into gene expression patterns, revealing potential biomarkers and gene regulatory networks tailored to specific cell lines. Utilizing this data, we conducted a comparative analysis, pinpointing genes and pathways governing cellular traits. This informed our evaluation of therapeutic molecules like the Curcumin analog EF-24. Our analysis of EF-24's effects on leukemia cell lines K562 (ATCC® CCL-243™, HL-60 (ATCC® CCL-240™), THP-1 (ATCC® TIB-202™), Kasumi-1 (ATCC® CRL-2724™) demonstrated increased cell death post-treatment. EF-24 activated pathways associated with cell survival and death regulation, notably impacting STAT1-induced cell death, and enhancing innate immune system genes. This research offers a detailed understanding of EF-24's molecular actions in tumors, emphasizing the significance of STAT1 signaling in its antitumorigenic effects. The profound impact of EF-24 on inducing cell death across multiple leukemia cell lines underscores its potential as a therapeutic agent. By elucidating its specific molecular actions and highlighting the pivotal role of STAT1 signaling, this study not only deepens our comprehension of EF-24's mechanisms but also underscores its promising value in targeted antitumorigenic strategies. Citation Format: Ajeet P. Singh, Rula R. Khairi, Amy L. Reese, Jade Kirkland, James Duncan, Noah Wax, Robert Marlow, Steve King, Ana Fernandes, Jonathan L. Jacobs. Navigating next-generation pre-clinical models of drug discovery & development with RNA-seq [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 1068.

Curcumin, a polyphenolic compound derived from the plant Curcuma longa L., has demonstrated a wide range of therapeutic properties, including potential anticancer effects. However, its clinical efficacy is limited due to poor bioavailability and stability. To overcome these challenges, curcumin analogs like EF-24 have been developed with improved pharmacological properties. In this study, in order to improve our understanding of EF-24's potential mechanisms of action, we used whole-transcriptome sequencing to identify genome-wide functional impacts of EF-24 treatment in leukemia cells. These results enabled the development of a testable model system for associating druggable genes with clinical disease targets related to EF-24 treatment. To develop our model of the transcriptional response to EF-24 treatment, we used four well studied model cell lines for leukemia research, specifically the chronic myeloid leukemia (CML) cell line K-562 and acute myeloid leukemia (AML) cell lines HL-60, Kasumi-1, and THP-1. Cell viability was significantly decreased in all four of these leukemia models following EF-24 treatment as compared to untreated controls. We discovered that the genes ATF3, CLU, HSPA6, OSGIN1, ZFAND2A, and CXCL8, which are associated with reduced cell viability and proliferation, were consistently upregulated in all EF-24-treated cell lines. Further analysis of the tested cell lines revealed the activation of various signaling pathways, but notably the S100 family signaling pathway was consistently activated in all four cell lines. Our results provide critical insights into the molecular underpinnings of EF-24's antitumor efficacy against leukemia subtypes, highlighting its multifaceted impact on signaling pathways and gene networks that regulate cell survival, proliferation, and immune responses in cell line models of myeloid leukemia subtypes.

Editorial: High-level antimicrobial resistance or hypervirulence in emerging and re-emerging "super-bug" foodborne pathogens: detection, mechanism, and dissemination from omics insights

Recent studies highlight the integral role of the interferon gamma receptor (IFNγR) pathway in T cell-mediated cytotoxicity against solid but not liquid tumors. IFNγ not only directly facilitates tumor cell death by T cells but also indirectly promotes cytotoxicity via myeloid phagocytosis in the tumor microenvironment. Meanwhile, full human ex vivo immune checkpoint drug screening remains challenging. We hypothesized that an engineered gamma interferon activation site response element luciferase reporter (GAS-Luc2) can be utilized for immune checkpoint drug screening in diverse ex vivo T cell-solid tumor cell co-culture systems. We comprehensively profiled cell surface proteins in ATCC's extensive collection of human tumor and immune cell lines, identifying those with endogenously high expression of established and novel immune checkpoint molecules and binding ligands. We then engineered three GAS-Luc2 reporter tumor cell lines expressing immune checkpoints PD-L1, CD155, or B7-H3/CD276. Luciferase expression was suppressed upon relevant immune checkpoint-ligand engagement. In the presence of an immune checkpoint inhibitor, T cells released IFNγ, activating the JAK-STAT pathway in GAS-Luc2 cells, and generating a quantifiable bioluminescent signal for inhibitor evaluation. These reporter lines also detected paracrine IFNγ signaling for immune checkpoint-targeted ADCC drug screening. Further development into an artificial antigen-presenting cell line (aAPC) significantly enhanced T cell signaling for superior performance in these ex vivo immune checkpoint drug screening platforms.

Babesia microti is the primary cause of human babesiosis in North America. Despite an emergence of the disease in recent years, the pathogenesis and immune response to B. microti infection remain poorly understood. Studies in laboratory mice have shown a critical role for macrophages in the elimination of parasites and infected red blood cells (iRBCs). Importantly, the underlying mechanisms that activate macrophages are still unknown. Recent evidence identified the release of extracellular vesicles (EVs) from Babesia iRBCs. EVs are spherical particles released from cell membranes under natural or pathological conditions that have been suggested to play roles in host-pathogen interactions among diseases caused by protozoan parasites. The present study examined whether EVs released from cultured Babesia iRBCs activated macrophages resulting in changes in the secretion of cytokines. An analysis of vesicle size in EV fractions from Babesia iRBCs showed diverse populations in the <100 nm size range compared to EVs from uninfected RBCs. Uptake of EVs released from B. microti-iRBCs was observed in macrophages in vitro. In addition, incubation of macrophages with EVs isolated from Babesia iRBC culture supernatants resulted in the activation of NF-kappa B and modulation of pro-inflammatory cytokines. These results support a role for EVs in the activation of macrophage functions and provide new insights into the mechanisms involved in the induction of the innate immune response during babesiosis.

Functional human brown and white adipose tissue (BAT and WAT) are vital for thermoregulation and nutritional homeostasis, while obesity and other stressors lead, respectively, to cold intolerance and metabolic disease. Understanding BAT and WAT physiology and dysfunction necessitates clinical trials complemented by mechanistic experiments at the cellular level. These require standardized in vitro models, currently lacking, that establish references for gene expression and function. We generated and characterized a pair of immortalized, clonal human brown (hBA) and white (hWA) preadipocytes derived from the perirenal and subcutaneous depots, respectively, of a 40-year-old male individual. Cells were immortalized with hTERT and confirmed to be of a mesenchymal, nonhematopoietic lineage based on fluorescence-activated cell sorting and DNA barcoding. Functional assessments showed that the hWA and hBA phenocopied primary adipocytes in terms of adrenergic signaling, lipolysis, and thermogenesis. Compared to hWA, hBA were metabolically distinct, with higher rates of glucose uptake and lactate metabolism, and greater basal, maximal, and nonmitochondrial respiration, providing a mechanistic explanation for the association between obesity and BAT dysfunction. The hBA also responded to the stress of maximal respiration by using both endogenous and exogenous fatty acids. In contrast to certain mouse models, hBA adrenergic thermogenesis was mediated by several mechanisms, not principally via uncoupling protein 1 (UCP1). Transcriptomics via RNA-seq were consistent with the functional studies and established a molecular signature for each cell type before and after differentiation. These standardized cells are anticipated to become a common resource for future physiological, pharmacological, and genetic studies of human adipocytes.

African swine fever is a lethal disease of domestic pigs, geographically expanding as a pandemic, that is affecting countries across Eurasia and severely damaging their swine production industry. After more than 40 years of being absent in the Western hemisphere, in 2020 ASF reappeared in the Dominican Republic and Haiti. The recent outbreak strain in the Dominican Republic has been identified as a genotype II ASFV a derivative of the ASF strain circulating in Asia and Europe. However, to date no full-length genome sequence from either the 1978–1980 Here we report the complete genome sequence of an African swine fever virus (ASFV) (DR-1980) that was previously isolated from blood collected in 1980 from the Dominican Republic at the end of the last outbreak, before culling of all swine on the island of Hispaniola and stored in the Plum Island Animal Disease Center ASFV repository. A contig representing the full-length genome (183,687 base pairs) was de novo assembled into a single contig using both Nanopore and Illumina sequences. DR-1980 was determined to belong to genotype I and, as determined by full genome comparison, a close relative to the sequenced Sardinia viruses that were causing outbreaks at this time.

The Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial is a prospective cohort study of nearly 155,000 U.S. volunteers aged 55–74 at enrollment in 1993–2001. We developed the PLCO Atlas Project, a large resource for multi-trait genome-wide association studies (GWAS), by genotyping participants with available DNA and genomic consent. Genotyping on high-density arrays and imputation was performed, and GWAS were conducted using a custom semi-automated pipeline. Association summary statistics were generated from a total of 110,562 participants of European, African and Asian ancestry. Application programming interfaces (APIs) and open-source software development kits (SKDs) enable exploring, visualizing and open data access through the PLCO Atlas GWAS Explorer website, promoting Findable, Accessible, Interoperable, and Re-usable (FAIR) principles. Currently the GWAS Explorer hosts association data for 90 traits and >78,000,000 genomic markers, focusing on cancer and cancer-related phenotypes. New traits will be posted as association data becomes available. The PLCO Atlas is a FAIR resource of high-quality genetic and phenotypic data with many potential reuse opportunities for cancer research and genetic epidemiology.