ObjectiveAs the current therapeutic strategies for human hepatocellular carcinoma (HCC) have been proven to have limited effectiveness, immunotherapy becomes a compelling way to tackle the disease. We aim to provide humanised mouse (humice) models for the understanding of the interaction between human cancer and immune system, particularly for human-specific drug testing.DesignPatient-derived xenograft tumours are established with type I human leucocyte antigen matched human immune system in NOD-scid Il2rg−/− (NSG) mice. The longitudinal changes of the tumour and immune responses as well as the efficacy of immune checkpoint inhibitors are investigated.ResultsSimilar to the clinical outcomes, the human immune system in our model is educated by the tumour and exhibits exhaustion phenotypes such as a significant declination of leucocyte numbers, upregulation of exhaustion markers and decreased the production of human proinflammatory cytokines. Notably, cytotoxic immune cells decreased more rapidly compared with other cell types. Tumour infiltrated T cells have much higher expression of exhaustion markers and lower cytokine production compared with peripheral T cells. In addition, tumour-associated macrophages and myeloid-derived suppressor cells are found to be highly enriched in the tumour microenvironment. Interestingly, the tumour also changes gene expression profiles in response to immune responses by upregulating immune checkpoint ligands. Most importantly, in contrast to the NSG model, our model demonstrates both therapeutic and side effects of immune checkpoint inhibitors pembrolizumab and ipilimumab.ConclusionsOur work provides a model for immune-oncology study and a useful parallel-to-human platform for anti-HCC drug testing, especially immunotherapy.

Successful translation of laboratory-based surface-enhanced Raman scattering (SERS) platforms to clinical applications requires multiplex and ultratrace detection of small metabolites from a complex biofluid. However, these metabolites exhibit low Raman scattering cross-sections and do not possess specific affinity to plasmonic nanoparticle surfaces, significantly increasing the challenge of detecting them at low concentrations. Herein, a 'confine-and-capture' approach is demonstrated for multiplex detection of two families of urine metabolites correlated with miscarriage risks, 5β-pregnane-3α,20α-diol-3α-glucuronide and tetrahydrocortisone. To enhance SERS signals by 10 12 -fold, specific nanoscale surface chemistry is used for targeted metabolite capture from a complex urine matrix prior to confining them on a superhydrophobic SERS platform. Applying chemometrics, including principal component analysis and partial least square regression, enables conversion of molecular fingerprint information into quantifiable readouts. The whole screening procedure requires only 30 minutes, including urine pretreatment, sample drying on the SPHB-mirror platform, SERS measurements and chemometric analyses. These readouts correlate well with the pregnancy outcomes in a case-control study of 40 patients presenting threatened miscarriage symptoms.Keywords. surface-enhanced Raman spectroscopy (SERS), superhydrophobic SERS platform, chemometrics, metabolomics, urine-based diagnostic test 3 Achieving ultratrace detection of small molecules with low Raman scattering cross-sections and without specific affinity to plasmonic nanoparticle surfaces remains challenging in surfaceenhanced Raman scattering (SERS) spectroscopy. [1][2][3] This difficulty is further compounded by the need to perform multiplex and quantitative molecular detection from a complex matrix.Successfully addressing these issues is instrumental towards the translating laboratory-based SERS platforms into practical sensing devices. 4 SERS offers multiple advantages over conventional analytical platforms such as fluorescence-based techniques. 5 SERS platforms can be tailored to generate intense electromagnetic field enhancements and dense plasmonic hotspots, in turn enhancing molecule-specific Raman vibrational fingerprint intensities by >10 9 -fold. 6,7 These fingerprints exhibit substantially narrower peak widths as compared to the broad fluorescence emission bands (full-width half-maximum of ~ 2 nm versus 30 nm respectively), further enabling SERS to achieve label-free multiplex analysis with ease. 8 SERS measurements also require significantly shorter time as compared to conventional chromatography-or mass spectrometrybased analytical approaches, whereby SERS analyses can be completed within an hour. 9 More importantly, the fingerprint specificity of SERS readouts enables differentiation of isomeric structures which cannot be easily achieved using other techniques. [10][11][12] However, majority of current SERS research focus predominantly on platform design, using s...

Immunodeficient mouse-human chimeras provide a powerful approach to study host-specific pathogens, such as Plasmodium falciparum that causes human malaria. Supplementation of immunodeficient mice with human RBCs supports infection by human Plasmodium parasites, but these mice lack the human immune system. By combining human RBC supplementation and humanized mice that are optimized for human immune cell reconstitution, we have developed RBC-supplemented, immune cell-optimized humanized (RICH) mice that support multiple cycles of P. falciparum infection. Depletion of human natural killer (NK) cells, but not macrophages, in RICH mice results in a significant increase in parasitemia. Further studies in vitro show that NK cells preferentially interact with infected RBCs (iRBCs), resulting in the activation of NK cells and the elimination of iRBCs in a contact-dependent manner. We show that the adhesion molecule lymphocyte-associated antigen 1 is required for NK cell interaction with and elimination of iRBCs. Development of RICH mice and validation of P. falciparum infection should facilitate the dissection of human immune responses to malaria parasite infection and the evaluation of therapeutics and vaccines. malaria infection | humanized mouse model | LFA-1 | NK killing

ObjectiveHCV infection affects millions of people worldwide, and many patients develop chronic infection leading to liver cancers. For decades, the lack of a small animal model that can recapitulate HCV infection, its immunopathogenesis and disease progression has impeded the development of an effective vaccine and therapeutics. We aim to provide a humanised mouse model for the understanding of HCV-specific human immune responses and HCV-associated disease pathologies.DesignRecently, we have established human liver cells with a matched human immune system in NOD-scid Il2rg−/− (NSG) mice (HIL mice). These mice are infected with HCV by intravenous injection, and the pathologies are investigated.ResultsIn this study, we demonstrate that HIL mouse is capable of supporting HCV infection and can present some of the clinical symptoms found in HCV-infected patients including hepatitis, robust virus-specific human immune cell and cytokine responses as well as liver fibrosis and cirrhosis. Similar to results obtained from the analysis of patient samples, the human immune cells, particularly T cells and macrophages, play critical roles during the HCV-associated liver disease development in the HIL mice. Furthermore, our model is demonstrated to be able to reproduce the therapeutic effects of human interferon alpha 2a antiviral treatment.ConclusionsThe HIL mouse provides a model for the understanding of HCV-specific human immune responses and HCV-associated disease pathologies. It could also serve as a platform for antifibrosis and immune-modulatory drug testing.