Abstract
Type 1 diabetes (T1D) is an autoimmune disease characterized by inflammatory lesions within islets (insulitis) and β cell loss which is associated T cell and B cell responses specific for ...pancreatic islet β cell proteins. Data show that treatment with poly(lactide-co-glycolide) (PLGA) nanoparticles (i.e. TIMP/COUR’s CNP) containing a single diabetogenic peptide blocks T1D induced by transfer of CD4+ BDC2.5 or CD8+ NY8.3 TCR transgenic T cells to NOD-scid mice. In contrast, Ag-specific tolerance approaches targeting single β-islet cell protein epitopes, e.g. GAD65 or Insulin, have failed to achieve efficacy in both wildtype NOD mice and clinical trials. The possibility exists that T1D initiation and progression is due to activation of T cell populations specific for multiple diabetogenic epitopes. To test this hypothesis, recombinant Chormogranin A, GAD65, and Insulin proteins were encapsulated within CNPs to assess Treg/Tr1 cell induction, inhibition of Ag-specific T cell responses, and blockade of T1D in NOD mice. While treatment of NOD mice with CNPs containing a single protein inhibited the corresponding Ag-specific T cell response, the inhibition of T1D development only occurred if all three diabetogenic proteins were included within the CNPs (CNP-T1D). CNP-T1D-induced blockade of T1D was characterized by Treg/Tr1 cell induction and a significant decrease in both peri-insulitis and immune cell infiltration into pancreatic islets. We have recently published that CNP treatment is both safe and induces Ag-specific tolerance in Phase I/IIa celiac disease clinical trials. Therefore, utilization of this technology including multiple diabetogenic proteins is a promising Ag-specific tolerance approach to T1D treatment.
Abstract
Older people exhibit dysregulated innate immunity to respiratory viral infections, including influenza and SARS-CoV-2, to increase morbidity and mortality. Nanoparticles are a potential ...practical therapeutic that could divert exaggerated innate immune responses away from the lungs during viral infection. However, such therapeutics have not been examined for effectiveness during respiratory viral infection, particular in aged hosts. Here, we employed a highly lethal model of influenza viral infection in vulnerable aged mice to examine the ability of biodegradable, cargo-free nanoparticles, designated ONP-302, to resolve innate immune dysfunction and improve outcomes during infection. We administered ONP-302 via intravenous injection to aged mice at day 3 post-infection when the hyperinflammatory innate immune response is already established. We found that ONP-302 treatment diverted tissue-damaging inflammatory monocytes from the lungs to the spleen without impacting viral clearance or reducing chemo-attractive signals in the infected lung. Importantly, cargo-free nanoparticles reduced lung damage and histological lung inflammation and improved gas exchange and, ultimately, the clinical outcomes in influenza-infected aged mice. ONP-302 improves outcomes in influenza-infected, vulnerable, aged mice. Thus, our study provides vital fundamental information concerning a practical therapeutic which, if translated clinically, could improve disease outcomes for vulnerable older patients suffering from respiratory viral infections.
Supported by COUR Pharmaceuticals
Mass spectrometry (MS) is widely used in the characterization of biomolecules including peptide and protein therapeutics. These biotechnology products have seen rapid growth over the past few decades ...and continue to dominate the global pharmaceutical market. Advances in MS instrumentation and techniques have enhanced protein characterization capabilities and supported an increased development of biopharmaceutical products. Areas covered: This review describes recent developments in MS-based biotherapeutic analysis including sequence determination, post-translational modifications (PTMs) and higher order structure (HOS) analysis along with improvements in ionization and dissociation methods. An outlook of emerging applications of MS in the lifecycle of product development such as comparability, biosimilarity and quality control practices is also presented. Expert commentary: MS-based methods have established their utility in the analysis of new biotechnology products and their lifecycle appropriate implementation. In the future, MS will likely continue to grow as one of the leading protein identification and characterization techniques in the biopharmaceutical industry landscape.
Weight gain in adulthood is now common in many populations, ranging from modest gains in developing countries to a substantial percentage of body weight in some Western societies. To examine the rate ...of change across the spectrum of low to high-income countries we compared rates of weight change in samples drawn from three countries, Nigeria, Jamaica and the United States.
Population samples from Nigeria (n = 1,242), Jamaica (n = 1,409), and the US (n = 809) were selected during the period 1995-1999 in adults over the age of 19; participation rates in the original survey were 96%, 60%, and 60%, respectively. Weight in (kg) was measured on 3 different occasions, ending in 2005. Multi-level regression models were used to estimate weight change over time and pattern-mixture models were applied to assess the potential effect of missing data on estimates of the model parameters.
The unadjusted weight gain rate (standard error) was 0.34(0.06), 1.26(0.12), 0.34(0.19) kg/year among men and 0.43(0.06), 1.28(0.10), 0.40(0.15) kg/year among women in Nigeria, Jamaica, US, respectively. Regression-adjusted weight change rates were significantly different across country, sex, and baseline BMI. Adjusted weight gain in Nigeria, Jamaica and US was 0.31(0.05), 1.37(.04), and 0.52(0.05) kg/year respectively. Women in Nigeria and the US had higher weight gains than men, with the converse observed among Jamaicans. The obese experienced weight loss across all three samples, whereas the normal weight (BMI < 25) had significant weight gains. Missing data patterns had an effect on the rates of weight change.
Weight change in sample cohorts from a middle-income country was greater than in cohorts from either of the low- or high-income countries. The steep trajectory of weight gain in Jamaica, relative to Nigeria and the US, is most likely attributable to the accelerating effects of the cultural and behavioral shifts which have come to bear on transitional societies.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Summary Hyperactivity of the hypothalamic–pituitary–adrenal axis (HPAA) resulting from fetal programming may play a role in the development of high blood pressure (BP) in black people. We assessed ...the diurnal salivary cortisol profile in children with and without increased BP and evaluated their mother's HPAA. In a cross-sectional study, 20 Afro-Caribbean children (mean age 9.6 years) with higher blood pressures and 20 children with lower blood pressures were chosen from a prospective study of 569 mothers and children in Jamaica. Daytime salivary cortisol profiles were collected in the children and their mothers. The mothers were also assessed for features of the metabolic syndrome. Children with higher BP had higher mean morning salivary cortisol concentrations than those with lower BP (7.9 S.D. 1.9 vs. 4.5 S.D. 2.4 nmol/l; p = 0.03). Their mothers also had increased morning salivary cortisol concentrations (9.9 S.D. 1.8 vs. 5.5 S.D. 2.5 nmol/l; p = 0.02), but no changes in fasting glucose, insulin, lipids, BP or adiposity. Maternal and offspring cortisol concentrations correlated significantly ( r = 0.465, p = 0.004). Maternal cortisol concentrations were significantly associated with the child's BP. We conclude that Afro-Caribbean children with higher BP have higher morning salivary cortisol concentrations. The children's cortisol concentrations correlate significantly with the mother's cortisol concentrations. These findings suggest that the HPAA may play a role in the development of raised BP in Afro-Caribbean people.
While the International Diabetes Federation (IDF) has ethnic specific waist circumference (WC) cut-points for the metabolic syndrome for Asian populations it is not known whether the cut-points for ...black populations should differ from those for European populations. We examined the validity of IDF WC cut points for identifying insulin resistance (IR), the underlying cause of the metabolic syndrome, in predominantly black, young Jamaican adults.
Participants from a 1986 birth cohort were evaluated between 2005 and 2007 when they were 18-20 years old. Trained observers took anthropometric measurements and collected a fasting blood sample. IR was assessed using the homeostasis model assessment computer programme (HOMA-IR). Sex specific quartiles for IR were generated using HOMA-IR values and participants in the highest quartile were classified as "insulin resistant". Receiver operator characteristic (ROC) curves were used to estimate the best WC to identify insulin resistance. The sensitivity and specificity of these values were compared with the IDF recommended WC cut-points.
Data from 707 participants (315 males; 392females) were analysed. In both sexes those with IR were more obese, had higher mean systolic blood pressure, glucose and triglycerides and lower mean HDL cholesterol. The WC was a good predictor of IR with an ROC area under the curve (95% CI) of 0.71(0.64,0.79) for men and 0.72(0.65,0.79) for women. Using the Youden Index (J) the best WC cut point for identifying IR in male participants was 82 cm (sensitivity 45%, specificity 93%, J 0.38) while the standard cut point of 94 cm had a sensitivity of 14% and specificity of 98% (J 0.12). In the female participants 82 cm was also a good cut point for identifying IR (sensitivity 52%, specificity 87%, J 0.39) and was similar to the standard IDF 80 cm cut point (sensitivity 53%, specificity 82%, J 0.35).
The WC that identified IR in young black men is lower than the IDF recommended WC cut point. Sex differences in WC cut points for identifying IR were less marked in this population than in other ethnic groups.
In celiac disease (CeD), gluten induces immune activation, leading to enteropathy. TAK-101, gluten protein (gliadin) encapsulated in negatively charged poly(dl-lactide-co-glycolic acid) ...nanoparticles, is designed to induce gluten-specific tolerance.
TAK-101 was evaluated in phase 1 dose escalation safety and phase 2a double-blind, randomized, placebo-controlled studies. Primary endpoints included pharmacokinetics, safety, and tolerability of TAK-101 (phase 1) and change from baseline in circulating gliadin-specific interferon-γ–producing cells at day 6 of gluten challenge, in patients with CeD (phase 2a). Secondary endpoints in the phase 2a study included changes from baseline in enteropathy (villus height to crypt depth ratio Vh:Cd), and frequency of intestinal intraepithelial lymphocytes and peripheral gut-homing T cells.
In phase 2a, 33 randomized patients completed the 14-day gluten challenge. TAK-101 induced an 88% reduction in change from baseline in interferon-γ spot-forming units vs placebo (2.01 vs 17.58, P = .006). Vh:Cd deteriorated in the placebo group (−0.63, P = .002), but not in the TAK-101 group (−0.18, P = .110), although the intergroup change from baseline was not significant (P = .08). Intraepithelial lymphocyte numbers remained equal. TAK-101 reduced changes in circulating α4β7+CD4+ (0.26 vs 1.05, P = .032), αEβ7+CD8+ (0.69 vs 3.64, P = .003), and γδ (0.15 vs 1.59, P = .010) effector memory T cells. TAK-101 (up to 8 mg/kg) induced no clinically meaningful changes in vital signs or routine clinical laboratory evaluations. No serious adverse events occurred.
TAK-101 was well tolerated and prevented gluten-induced immune activation in CeD. The findings from the present clinical trial suggest that antigen-specific tolerance was induced and represent a novel approach translatable to other immune-mediated diseases.
ClinicalTrials.gov identifiers: NCT03486990 and NCT03738475.
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TAK-101 is a novel therapy that reduced immune activation and intestinal damage induced by oral gluten exposure in patients with celiac disease.
Abstract
There are many autoimmune diseases where pathologic self T cells are the underlying cause of disease symptoms and progression. T cells are found at the site of tissue destruction, resulting ...not only in the exacerbation of disease, but also the release of self-epitopes in the context of inflammation resulting in the activation of additional T cell populations perpetuating disease. Published data have shown treatment with antigen-containing biodegradablepoly(lactide-co-glycolide) (PLGA) nanoparticles, i.e. tolerogenic immune-modifying particles (TIMP), is both safe and induces antigen-specific tolerance in mouse models of autoimmunity and allergy, as well as in a celiac disease Phase I/IIa clinical trial. This study further investigated the role and mechanism of TIMP-induced tolerance. The present data show that TIMP treatment modulated T cells specific for spread epitopes associated with disease progression, but not encapsulated within the TIMP, i.e. tissue-specific bystander suppression. The PLP139–151 TCR transgenic model systems was utilized to determine the cellular and molecular mechanisms driving antigen specific tolerance mediated by tolerogenic nanoparticle treatment. These data show antigen-specific TIMP treatment induced both FoxP3+ iTregs and IL-10+ Tr1 regulatory phenotypes within the antigen-specific T cell populations in both naïve mice and mice pre-primed with antigen/CFA. Additionally, both functional Tregs and IL-10 are required for TIMP-induced tolerance. Treatment thus activates various antigen-specific Treg subsets capable of regulating responses to disease-relevant autoepitopes not encapsulated within the TIMP via release of immunoregulatory cytokines within the inflammatory site.
Supported by funding provided by Cour Pharmaceuticals Development, Inc.
Abstract
Antigen (Ag)-specific T cells are the underlying cause of many autoimmune diseases. T cells found at the site of tissue destruction not only in the exacerbation of disease, but contribute to ...the release of additiotnal self-epitopes resulting in the activation of spread eptitope-specific T cell populations perpetuating disease. Treatment with Ag-containingbiodegradable poly(lactide-co-glycolide) (PLGA) nanoparticles, i.e. tolerogenic immune-modifying particles (TIMP), has been shown to be both safe and induces Ag-specific tolerance in mouse models of autoimmunity and allergy, as well as in a celiac disease Phase I/IIa clinical trial. We are in the process of assessing if the mechanism of action between Ag-specific TIMP treatment and Ag-coupled cell treatment, to thereby identify self-tolerance mechanisms in common between the two highly efficacious therapies. The PLP 139-151TCR transgenic model system was utilized to determine the cellular and molecular mechanisms driving Ag-specific tolerance mediated by tolerogenic nanoparticle treatment. These data show Ag-specific TIMP treatment induced both FoxP3 +iTregs and IL-10 +Tr1 regulatory in both naïve mice and mice pre-primed with Ag/CFA. Additionally, the expression of STING is required for the induction of tolerance. The involvement of the STING pathway for both TIMP and Ag-coupled cell treatment was confirmed in MOG 35-55EAE. The data also show that Ag-specific tolerance induction requires both Type I IFNs and PD-1/PD-L1 expression, which are downstream of STING activation and signaling. Treatment thus activates various Ag-specific Treg subsets capable of regulating responses to disease-relevant autoepitopes via a STING/IFNAR/PD-1/PD-L1 pathway.
Abstract
Patients with locally advanced and metastatic urothelial carcinoma have a low survival rate (median 15.7 months, 13.1–17.8), with a 23% response rate to monotherapy treatment with anti-PD-L1 ...immunotherapy. We recently reported the potent effects of i.v. infusions of immune modifying poly(lactic-co-glycolic acid) (PLGA) nanoparticles (IMPs; named ONP-302) to both decrease B16.F10, MC-38, LLC, and 4T1 tumor growth and allow for B16.F10 tumor responsiveness to anti-PD-1 treatment. We next sought to determine if ONP-302 treatment could significantly decrease the more clinically relevant BBN-carcinogen induced bladder cancer. The present data show that ONP-302 treatment significantly decreases bladder cancer stage and overall bladder weight. While minor epithelial hyperplasia remained, ONP-302 treatment inhibited widespread carcinoma in situ and muscle invasive tumor development, as compared to saline treated mice. Mechanistically, ONP-302 infusion decreased tumor growth via the activation of the cGAS/STING pathway within myeloid cells, and subsequently increased CD8+ T cell and NK cell activation via IL-15. In subsequent studies to assess the cellular mechanisms involved in ONP-302 function, we found that in vitro treatment with ONP-302 rapidly induced oxidized DNA within the cells that took up PLGA nanoparticles and oxidized DNA was present within the supernatant as well. Furthermore, co-treatment of ONP-302 with DNase inhibited the STING mediated cytokine production. These findings indicate that ONP-302 allows for tumor control via reprogramming myeloid cells and inducing a STING pathway activation, increases anti-PD-1 response rates, and significantly decreases BBN-carcinogen induced bladder cancer.
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