Cellular Metabolism in B Cells in Type 1 Diabetes

Background/Objective: Type 1 diabetes (T1D) is an immune-mediated disease that results in the destruction of pancreatic beta cells. While beta cell destruction is classically considered T cell mediated, autoreactive B cells play important roles in disease progression. B cell depletion prevents disease in non-obese diabetic (NOD) mice, and B cell depletion temporarily slows disease progression in individuals with new-onset T1D. However, mechanisms of autoreactive B cell function in T1D are not fully known. Cellular metabolism has been shown to drive autoimmune B cell development in other mouse models. We hypothesize that metabolic characteristics of B cells from NOD mice are distinct from metabolic characteristics of B cells from non-autoimmune C57BL/6J (B6) mice; therefore, making cellular metabolic pathways viable targets for therapeutic intervention. Methods: Lymphocytes from spleen, pancreas, pancreatic lymph nodes, and mesenteric/lumbar lymph nodes were processed into single-cell suspensions. Glucose uptake was measured using fluorescent glucose analog 2-NBDG. Mitochondrial polarity was measured using fluorescent probes for mass and membrane potential. Cells were stained for surface markers and analyzed on an Attune Nxt flow cytometer. Results: No statistically significant differences in glucose uptake or mitochondrial polarity for lymphocyte subsets in the spleen or PLNs of NOD and B6 mice were identified. In NOD mice, polarity was significantly higher in B cells in the pancreas compared to the spleen and PLNs. Polarity was also higher in B cells in PLNs compared to non-specific lymph nodes in NOD mice. Conclusions/Impact: While no differences in glucose uptake or polarity in lymphocytes from NOD and B6 mice ex vivo were identified, future studies are needed to determine whether their activation drives metabolic alterations. Differences in polarity in the pancreas in NOD mice suggest that cellular metabolism is influenced by the islet microenvironment and has the potential to influence their function at the site of autoimmune attack

Neurofibromin Deficient Myeloid Cells are Critical Mediators of Aneurysm Formation In Vivo

Background Neurofibromatosis Type 1 (NF1) is a genetic disorder resulting from mutations in the NF1 tumor suppressor gene. Neurofibromin, the protein product of NF1, functions as a negative regulator of Ras activity in circulating hematopoietic and vascular wall cells, which are critical for maintaining vessel wall homeostasis. NF1 patients have evidence of chronic inflammation resulting in development of premature cardiovascular disease, including arterial aneurysms, which may manifest as sudden death. However, the molecular pathogenesis of NF1 aneurysm formation is unknown. Method and Results Utilizing an angiotensin II-induced aneurysm model, we demonstrate that heterozygous inactivation of Nf1 (Nf1+/−) enhanced aneurysm formation with myeloid cell infiltration and increased oxidative stress in the vessel wall. Using lineage-restricted transgenic mice, we show loss of a single Nf1 allele in myeloid cells is sufficient to recapitulate the Nf1+/− aneurysm phenotype in vivo. Finally, oral administration of simvastatin or the antioxidant apocynin, reduced aneurysm formation in Nf1+/− mice. Conclusion These data provide genetic and pharmacologic evidence that Nf1+/− myeloid cells are the cellular triggers for aneurysm formation in a novel model of NF1 vasculopathy and provide a potential therapeutic target

Classical and controlled auditory mismatch responses to multiple physical deviances in anaesthetised and conscious mice

Human mismatch negativity (MMN) is modelled in rodents and other non-human species to examine its underlying neurological mechanisms, primarily described in terms of deviance-detection and adaptation. Using the mouse model, we aim to elucidate subtle dependencies between the mismatch response (MMR) and different physical properties of sound. Epidural field potentials were recorded from urethane anaesthetised and conscious mice during oddball and many-standards control paradigms with stimuli varying in duration, frequency, intensity, and inter-stimulus interval. Resulting auditory evoked potentials, classical MMR (oddball – standard), and controlled MMR (oddball – control) waveforms were analysed. Stimulus duration correlated with stimulus-off response peak latency, whereas frequency, intensity, and inter-stimulus interval correlated with stimulus-on N1 and P1 (conscious only) peak amplitudes. These relationships were instrumental in shaping classical MMR morphology in both anaesthetised and conscious animals, suggesting these waveforms reflect modification of normal auditory processing by different physical properties of sound. Controlled MMR waveforms appeared to exhibit habituation to auditory stimulation over time, which was equally observed in response to oddball and standard stimuli. These findings are inconsistent with the mechanisms thought to underlie human MMN, which currently do not address differences due to specific physical features of sound. Thus, no evidence was found to objectively support the deviance-detection or adaptation hypotheses of MMN in relation to anaesthetised or conscious mice. These findings highlight the potential risk of mischaracterising difference waveform components that are principally influenced by physical sensitivities and habituation of the auditory system

Serology describes a profile of declining malaria transmission in Farafenni, The Gambia

BACKGROUND: Malaria morbidity and mortality has declined in recent years in a number of settings. The ability to describe changes in malaria transmission associated with these declines is important in terms of assessing the potential effects of control interventions, and for monitoring and evaluation purposes. METHODS: Data from five cross-sectional surveys conducted in Farafenni and surrounding villages on the north bank of River Gambia between 1988 and 2011 were compiled. Antibody responses to MSP-119 were measured in samples from all surveys, data were normalized and expressed as seroprevalence and seroconversion rates (SCR) using different mathematical models. RESULTS: Results showed declines in serological metrics with seroprevalence in children aged one to 5 years dropping from 19 % (95 % CI 15-23 %) in 1988 to 1 % (0-2 %) in 2011 (p value for trend in proportions < 0.001) and the SCR dropping from 0.069 year(-1) (0.059-0.080) to 0.022 year(-1) (0.017-0.028; p = 0.004). The serological data were consistent with previously described drops in both parasite prevalence in children aged 1-5 years (62 %, 57-66 %, in 1988 to 2 %, 0-4 %, in 2011; p < 0.001), and all-cause under five mortality rates (37 per 1000 person-years, 34-41, in 1990 to 17, 15-19, in 2006; p = 0.059). CONCLUSIONS: This analysis shows accurate reconstruction of historical malaria transmission patterns in the Farafenni area using anti-malarial antibody responses. Demonstrating congruence between serological measures, and conventional clinical and parasitological measures suggests broader utility for serology in monitoring and evaluation of malaria transmission

B Quiet: Autoantigen-Specific Strategies to Silence Raucous B Lymphocytes and Halt Cross-Talk with T Cells in Type 1 Diabetes

Islet autoantibodies are the primary biomarkers used to predict type 1 diabetes (T1D) disease risk. They signal immune tolerance breach by islet autoantigen-specific B lymphocytes. T-B lymphocyte interactions that lead to expansion of pathogenic T cells underlie T1D development. Promising strategies to broadly prevent this T-B crosstalk include T cell elimination (anti-CD3, teplizumab), B cell elimination (anti-CD20, rituximab), and disruption of T cell costimulation/activation (CTLA-4/Fc fusion, abatacept). However, global disruption or depletion of immune cell subsets is associated with significant risk, particularly in children. Therefore, antigen-specific therapy is an area of active investigation for T1D prevention. We provide an overview of strategies to eliminate antigen-specific B lymphocytes as a means to limit pathogenic T cell expansion to prevent beta cell attack in T1D. Such approaches could be used to prevent T1D in at-risk individuals. Patients with established T1D would also benefit from such targeted therapies if endogenous beta cell function can be recovered or islet transplant becomes clinically feasible for T1D treatment