Biology subjects

Le, X.

Publications and source records attributed to Le, X..

5 recordsLinked to original sources

bioRxiv2026

Maternal antiviral history synergizes with pregnancy and lactation to transfer intergenerational systemic immunity through IgG in milk

Maternal immune transfer is essential for early-life health, yet whether immune experiences before pregnancy shape maternal physiology to optimize immunity in subsequent offspring is unclear. Here, we show that respiratory viral infection before pregnancy confers robust protection against lethal neonatal influenza through antibodies transferred postpartum in milk. Despite the predominance of IgA in milk, antiviral IgG is indispensable for protection. Pregnancy amplifies pre-existing antiviral B cell responses, while prior infection durably reprograms the mammary gland to promote transfer of circulating antiviral IgG into milk. These antibodies retain their epitope specificity, are enriched for broadly protective influenza epitopes, remain functional after passage through the neonatal intestine, and enter offspring circulation through FcRn to provide protection beyond weaning. Natural transmission of virus from infected offspring to mothers establishes maternal immunity that protects future offspring, revealing a coordinated adaptive program that links maternal immune history, pregnancy, and lactation to optimize intergenerational immunity. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=178 SRC="FIGDIR/small/744935v1_ufig1.gif" ALT="Figure 1"> View larger version (66K): org.highwire.dtl.DTLVardef@1f82cbeorg.highwire.dtl.DTLVardef@41ac81org.highwire.dtl.DTLVardef@1a4538aorg.highwire.dtl.DTLVardef@168bc21_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOGRAPHICAL ABSTRACTC_FLOATNO C_FIG HIGHLIGHTSO_LIPreconceptual maternal intranasal influenza infection confers complete neonatal B cell mediated protection against lethal neonatal influenza infection that persists beyond weaning into early adulthood. C_LIO_LIProtection can be transmitted postnatally through milk and is fully dependent on maternal IgG. C_LIO_LIPregnancy enhances rather than suppresses antiviral B cell programs in the mother. C_LIO_LIMilk IgG targets a restricted set of conserved influenza Hemagglutinin epitopes, suggesting selective transfer of broadly protective antibody populations. C_LIO_LIProtective IgG in milk derives from maternal circulation, not from local B cell mammary gland production. C_LIO_LIRespiratory infection before pregnancy induces long-lived vascular, stromal, and epithelial transcriptional remodeling of the mammary gland. C_LIO_LINeonatal Fc Receptor (FcRn) mediated transport of milk IgG into circulation is required for protection. C_LIO_LIInfected neonates transmit virus back to mothers, extending protection to subsequent litters for multi-generation protection. C_LI

immunology↗

bioRxiv2026

HER2 mutation-derived neoantigens in NSCLC as actionable targets for TCR therapy

HER2 mutations are oncogenic drivers in 1-6% of non-small cell lung cancers (NSCLC), but therapeutic resistance limits the durability of current HER2-targeted treatments. Here, we identify T-cell receptors (TCRs) targeting recurrent HER2 hotspot mutations as a potential immunotherapeutic strategy for HER2-mutant NSCLC. Using neoepitope prediction and antigen-specific T-cell enrichment, we isolated HLA-A*02:01restricted TCRs recognizing HER2 A775insYVMA, S310F, and G776delinsVC mutations, collectively covering approximately 60% of HER2-mutant NSCLC. These TCRs selectively recognized mutant HER2 epitopes without detectable wild-type reactivity and some displayed cross-recognition of related hotspot variants, expanding the spectrum of targetable tumors. The G776delinsVC-specific TCR also exhibited co-receptorindependent activity showcased by its ability to activate CD4+ T cells. Importantly, timelapse single-cell flow cytometry analyses demonstrated that TCR-engineered T cells repeatedly reacquired activated polyfunctional states following serial antigen stimulation, while serial tumor rechallenge assays confirmed sustained cytotoxic activity across multiple rounds of tumor killing. These findings identify recurrent HER2 mutations as shared immunotherapeutic targets and provide a foundation for the development of TCR-based therapies for HER2-mutant NSCLC.

immunology↗

bioRxiv2025

Mapping the phenotypic landscape of a transcriptional repressor using Deep Mutational Scanning and Growth-based Quantitative Sequencing

CymR is a TetR-family transcriptional repressor that recognizes a well-defined operator sequence in the promoter PcymRC. The native ligand cumate and several structurally related aromatic acids bind at an allosteric site and induce a conformational change in CymR, resulting in release from the DNA operator and de-repression of the promoter. The amino acid residues that contribute to these core functions have not been mapped, nor has the protein been subjected to extensive mutagenesis to modify its function. Here, for the first time, we integrate Deep Mutational Scanning (DMS) with Growth-based Quantitative Sequencing (GROQ-Seq) to evaluate a comprehensive phenotypic landscape of CymR variants, including single amino acid insertions and deletions. We measure this library across a concentration gradient of small molecule inducers to construct an induction curve for all library members. From this analysis, we identify amino acids throughout the protein that are essential for repressor function and discover several mutations that improve the sensitivity of CymR to the ligand perillic acid. In addition, rarely investigated insertion mutants are revealed to be a key driver of novel phenotypes, including several regions of CymR where insertions result in an inverted phenotype and the isolation of variants exhibiting an unusual band-stop phenotype. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=81 SRC="FIGDIR/small/693801v1_ufig1.gif" ALT="Figure 1"> View larger version (30K): org.highwire.dtl.DTLVardef@121b16org.highwire.dtl.DTLVardef@b0478aorg.highwire.dtl.DTLVardef@128f2ecorg.highwire.dtl.DTLVardef@164a426_HPS_FORMAT_FIGEXP M_FIG C_FIG

synthetic biology↗

bioRxiv2021

Metabolic evidence for distinct pyruvate pools inside plant mitochondria

The majority of the pyruvate inside plant mitochondria is either transported into the matrix from the cytosol via the mitochondria pyruvate carrier (MPC) or synthesised in the matrix by alanine aminotransferase (AlaAT) or NAD-malic enzyme (NAD-ME). Pyruvate from these origins could mix into a single pool in the matrix and contribute indistinguishably to respiration, or they could maintain a degree of independence in metabolic regulation. Here, we demonstrated that feeding isolated mitochondria with U-13C-pyruvate and unlabelled malate enables the assessment of pyruvate contribution from different sources to TCA cycle intermediate production. Imported pyruvate is the preferred source for citrate production even when the synthesis of NAD-ME-derived pyruvate was optimised. Genetic or pharmacological elimination of MPC activity removed this preference and allowed an equivalent amount of citrate to be generated from the pyruvate produced by NAD-ME. Increasing mitochondrial pyruvate pool size by exogenous addition only affected metabolites from pyruvate transported by MPC whereas depleting pyruvate pool size by transamination to alanine only affected metabolic products derived from NAD-ME. Together, these data reveal respiratory substrate supply in plants involves distinct pyruvate pools inside the matrix that can be flexibly mixed based on the rate of pyruvate transport from the cytosol. These pools are independently regulated and contribute differentially to organic acids export from plant mitochondria. Significance statementPyruvate is the primary respiratory substrate for energy production to support plant growth and development. However, it is also the starting material of many other pathways. Prioritisation of respiratory use over other competing pathways would enable a level of control when pyruvate is delivered to mitochondria via the mitochondrial pyruvate transporter. We demonstrated the existence of two distinct pyruvate pools in plant mitochondria suggesting inner mitochondrial organisation allows metabolic heterogeneity, hence metabolic specialisation. This explains why NAD-ME flux into plant respiration is low and confirms the prominent link between imported pyruvate and energy production. This compartmentation also reveals how NAD-ME supplies substrate to the mitochondrial pyruvate exporter in plants, especially during C4 metabolism.

plant biology↗

bioRxiv2021

The mitochondrial pyruvate carrier (MPC) complex is one of three pyruvate-supplying pathways that sustain Arabidopsis respiratory metabolism

Malate oxidation by plant mitochondria enables the generation of both oxaloacetate (OAA) and pyruvate for tricarboxylic acid (TCA) cycle function, potentially eliminating the need for pyruvate transport into mitochondria in plants. Here we show that the absence of the mitochondrial pyruvate carrier 1 (MPC1) causes the co-commitment loss of its orthologs, MPC3/MPC4, and eliminates pyruvate transport into Arabidopsis mitochondria, proving it is essential for MPC complex function. While the loss of either MPC or mitochondrial pyruvate-generating NAD-malic enzyme (NAD-ME) did not cause vegetative phenotypes, the lack of both reduced plant growth and caused an increase in cellular pyruvate levels, indicating a block in respiratory metabolism, and elevated the levels of branched-chain amino acids at night, a sign of alterative substrate provision for respiration. 13C-pyruvate feeding of leaves lacking MPC showed metabolic homeostasis were largely maintained except for alanine and glutamate, indicating that transamination contributes to restoration of the metabolic network to an operating equilibrium by delivering pyruvate independently of MPC into the matrix. Inhibition of alanine aminotransferases (AlaAT) when MPC1 is absent resulted in extremely retarded phenotypes in Arabidopsis, suggesting all pyruvate-supplying enzymes work synergistically to support the TCA cycle for sustained plant growth.

plant biology↗