Biology subjects

Burleigh, K.

Publications and source records attributed to Burleigh, K..

4 recordsLinked to original sources

bioRxiv2026

Coordinated expansion of CD163⁺ monocytes and immature CD177⁺ neutrophils marks severe neurotoxicity after CD19 CAR T cell therapy

Immune effector cell-associated neurotoxicity syndrome (ICANS) is a major complication after CAR T cell therapy, but its underlying mechanisms remain poorly understood. We performed longitudinal immune profiling of paired whole blood and serum samples from patients with relapsed or refractory diffuse large B cell lymphoma (DLBCL) treated with CD19 CAR T cells. At peak neurotoxicity, high-dimensional mass cytometry and serum proteomics identified the expansion of CD163 monocytes and immature CD10lowCD101low neutrophils correlated with elevated serum ST2 and IL-2RA concentrations. Integrative immune module analysis identified these features among the strongest predictors of ICANS severity. Independent single-cell transcriptomic profiling validated the emergence of immunoregulatory CD163 monocytes and identified CD177 as a biomarker of ICANS-associated immature neutrophils. Together, these findings reveal a coordinated myeloid inflammatory network associated with ICANS and nominate candidate biomarkers and therapeutic targets for improving the safety of CAR T cell therapy. SignificanceWe demonstrate that immunoregulatory CD163+ monocytes and immature, activated CD177hiCD10lowCD101low neutrophils emerge in patients with moderate to severe ICANS at peak toxicity following CD19 CAR T cell therapy. These findings identify an uncharacterized myeloid network potentially contributing towards ICANS pathogenesis.

immunology↗

bioRxiv2025

Endothelial-leukocyte interaction in CAR T cell neurotoxicity

CAR-T cells treat cancer, but also cause systemic cytokine release and immune effector cell associated neurotoxicity syndrome (ICANS). In an immunocompetent mouse model, we show by in vivo two-photon imaging that CD19-CAR T treatment causes brain capillary plugging by circulating CAR-T cells and other CD45+ leukocytes, as well as cortical hypoxia. This is accompanied by increased endothelial ICAM-1 and VCAM-1 expression in the brain capillary-venule transition zone, where most of the capillary stalls occur. In the mouse model, circulating CAR-T cells strongly upregulate integrin 4{beta}1 affinity to VCAM-1, but not affinity of integrin L{beta}2 to ICAM-1. Blockade of integrin 4 but not integrin L improves locomotion behavior. In vitro, human brain microendothelial cells upregulate ICAM-1 more than VCAM-1 in response to TNF, IFN-{gamma}, and IL-1{beta}. In a 3D brain human microvessel model, treatment with TNF and IFN-{gamma} is sufficient to induce adhesion of CAR T cells under flow conditions, which is blocked synergistically by antibodies against integrins 4 and L. Finally, patients with the highest levels of TNF and IFN-{gamma} also have the highest blood levels of soluble ICAM-1 and VCAM-1, which in turn correlate with ICANS. Integrin 4 but not L increases in CAR-T cells after they are infused into patients. Combined data from patients, mouse models and in vitro microvessels indicate differential regulation of interactions of ICAM-1 and VCAM-1 with their respective leukocyte integrins. Overall, our study supports the hypothesis that cytokine-driven upregulation of endothelial-leukocyte adhesion is sufficient to induce acute, reversible neurotoxicity. One Sentence SummaryDuring CAR T cell therapy, cytokine release induces white blood cell stalling in brain capillaries by upregulating ICAM-1/VCAM-1-integrin interactions.

immunology↗

bioRxiv2020

{Phi}X174 Attenuation by Whole Genome Codon Deoptimization

Natural selection acting on synonymous mutations in protein-coding genes influences genome composition and evolution. In viruses, introducing synonymous mutations in genes encoding structural proteins can drastically reduce viral growth, providing a means to generate potent, live attenuated vaccine candidates. However, an improved understanding of what compositional features are under selection and how combinations of synonymous mutations affect viral growth is needed to predictably attenuate viruses and make them resistant to reversion. We systematically recoded all non-overlapping genes of the bacteriophage {Phi}X174 with codons rarely used in its E. coli host. The fitness of recombinant viruses decreases as additional deoptimizing mutations are made to the genome, although not always linearly, and not consistently across genes. Combining deoptimizing mutations may reduce viral fitness more or less than expected from the effect size of the constituent mutations and we point out difficulties in untangling correlated compositional features. We test our model by optimizing the same genes and find that the relationship between codon usage and fitness does not hold for optimization, suggesting that wild-type {Phi}X174 is at a fitness optimum. This work highlights the need to better understand how selection acts on patterns of synonymous codon usage across the genome and provides a convenient system to investigate the genetic determinants of virulence.

genomics↗

bioRxiv2019

Human DNA-PK activates a STING-independent DNA sensing pathway

Detection of intracellular DNA by the cGAS-STING pathway activates a type I interferon-mediated innate immune response that protects from virus infection and can be harnessed to promote anti-tumor immunity. Whether there are additional DNA sensing pathways, and how such pathways might function, remains controversial. We show here that humans - but not mice - have a second, potent, STING-independent DNA sensing pathway that is blocked by the E1A viral oncogene of human adenovirus 5. We identify human DNA-PK as the sensor of this pathway and demonstrate that DNA-PK kinase activity drives a robust and broad antiviral response. We discover that the heat shock protein HSPA8/HSC70 is a unique target of DNA-PK. Finally, we demonstrate that detection of foreign DNA and DNA damage trigger distinct modalities of DNA-PK activity. These findings reveal the existence, sensor, unique target, and viral antagonists of a STING-independent DNA sensing pathway (SIDSP) in human cells.

immunology↗