ABSTRACT
Triple negative breast cancer holds a dismal clinical outcome and as such, patients routinely undergo aggressive, highly toxic treatment regimens. Clinical trials for TNBC employing immune checkpoint blockade in combination with chemotherapy show modest prognostic benefit, but the percentage of patients that respond to treatment is low, and patients often succumb to relapsed disease. Here, we show that a combination immunotherapy platform utilizing low dose chemotherapy (FEC) combined with oncolytic virotherapy (oHSV-1) increases tumor-infiltrating lymphocytes, in otherwise immune-bare tumors, allowing 60% of mice to achieve durable tumor regression when treated with immune checkpoint blockade. Whole-tumor RNA sequencing of mice treated with FEC + oHSV-1 shows an upregulation of B cell receptor signaling pathways and depletion of B cells prior to the start of treatment in mice results in complete loss of therapeutic efficacy and expansion of myeloid-derived suppressor cells. Additionally, RNA sequencing data shows that FEC + oHSV-1 suppresses genes associated with myeloid-derived suppressor cells, a key population of cells that drive immune escape and mediate therapeutic resistance. These findings highlight the importance of tumor-infiltrating B cells as drivers of antitumor immunity and their potential role in the regulation of myeloid-derived suppressor cells.
Subject(s)
B-Lymphocytes/drug effects , Immune Checkpoint Inhibitors/therapeutic use , Immunotherapy/methods , Lymphocytes, Tumor-Infiltrating/drug effects , Myeloid-Derived Suppressor Cells/drug effects , Triple Negative Breast Neoplasms/therapy , Animals , Antineoplastic Combined Chemotherapy Protocols/therapeutic use , B-Lymphocytes/immunology , B-Lymphocytes/metabolism , Cell Line, Tumor , Chlorocebus aethiops , Combined Modality Therapy , Female , Humans , Immune Checkpoint Inhibitors/immunology , Kaplan-Meier Estimate , Lymphocytes, Tumor-Infiltrating/immunology , Lymphocytes, Tumor-Infiltrating/metabolism , Mice, Inbred C57BL , Myeloid-Derived Suppressor Cells/immunology , Myeloid-Derived Suppressor Cells/metabolism , Oncolytic Virotherapy/methods , Triple Negative Breast Neoplasms/immunology , Triple Negative Breast Neoplasms/pathology , Vero CellsABSTRACT
CLEC16A has been shown to play a role in autophagy/mitophagy processes. Additionally, genetic variants in CLEC16A have been implicated in multiple autoimmune diseases. We generated an inducible whole-body knockout, Clec16aΔUBC mice, to investigate the loss of function of CLEC16A. The mice exhibited a neuronal phenotype including tremors and impaired gait that rapidly progressed to dystonic postures. Nerve conduction studies and pathological analysis revealed loss of sensory axons that are associated with this phenotype. Activated microglia and astrocytes were found in regions of the CNS. Several mitochondrial-related proteins were up- or down-regulated. Upregulation of interferon stimulated gene 15 (IGS15) were observed in neuronal tissues. CLEC16A expression inversely related to IGS15 expression. ISG15 may be the link between CLEC16A and downstream autoimmune, inflammatory processes. Our results demonstrate that a whole-body, inducible knockout of Clec16a in mice results in an inflammatory neurodegenerative phenotype resembling spinocerebellar ataxia.
Subject(s)
Lectins, C-Type/physiology , Monosaccharide Transport Proteins/physiology , Nervous System Autoimmune Disease, Experimental , Spinocerebellar Ataxias , Animals , Cytokines/metabolism , Female , Gene Knockout Techniques , Male , Mice, Knockout , Neurons/ultrastructure , Ubiquitins/metabolismABSTRACT
OBJECTIVE: To compare the use of two ophthalmic sponges for tear collection in dogs and cats. ANIMALS STUDIED: Ten healthy dogs and 10 healthy cats. PROCEDURES: A strip (4 × 10 mm) of either cellulose or polyvinyl acetal (PVA) sponge was inserted into the ventral fornix of each eye for either 15, 30, or 60 s. The wetted strip was placed into a 0.2-mL tube that was first punctured at its bottom. Tears were eluted through the drainage hole into a 1.5-mL tube via centrifugation. Tear volume absorbed (VA) and tear volume recovered (VR) were calculated as the difference of the post- and precollection weight of the 0.2-mL tube and 1.5-mL tube, respectively. Recovery ratio (RR) was determined as the ratio between VR and VA. RESULTS: Ophthalmic sponges were well tolerated by all subjects. In dogs and cats, median (95% range) VA, VR, and RR were as follows: 44 µL (11-106 µL) and 16 µL (2-43 µL); 27 µL (1-84 µL) and 6 µL (0-29 µL); 64% (7-91%) and 35% (0-86%), respectively. PVA sponges achieved significantly greater VR in cats and RR in both species. All parameters were significantly greater with a collection time of 60 vs. 30 and 15 s. Body weight was associated with VA and VR in dogs but not cats. CONCLUSIONS: Polyvinyl acetal is better than cellulose for tear collection given its superior recovery. Ophthalmic sponges could facilitate routine analysis of tear fluid in dogs and cats, although further studies are needed to evaluate the quality of tears obtained with this method.
Subject(s)
Cats , Dogs , Porifera , Specimen Handling/veterinary , Tears , Animals , Cellulose , Female , Male , Polyvinyls , Specimen Handling/instrumentation , Specimen Handling/methodsABSTRACT
When analytes containing color are irradiated with a pulsed UV laser in the ion source of a mass spectrometer, molecules such as dyes or pigments absorb energy, resulting in their desorption and ionization. This method, laser desorption mass spectrometry (LDMS), has been used successfully to analyze colorants of forensic interest in a wide variety of materials. Here, we present and interpret the most complex of such spectra obtained to date from a sample of fingernail polish. Interpretation of the spectrum provides a unique opportunity to characterize the laser desorption mass spectra of some unexpected inorganic materials found in cosmetics, such as "broken glass", cyanide compounds, and heavy metals. Also, the possibility of a useful forensic database of LDMS spectra of fingernail polishes is considered.
