IgA potentiates NETosis in response to viral infection

IgA is the second most abundant antibody present in circulation and is enriched at mucosal surfaces. As such, IgA plays a key role in protection against a variety of mucosal pathogens including viruses. In addition to neutralizing viruses directly, IgA can also stimulate Fc-dependent effector functions via engagement of Fc alpha receptors (Fc-αRI) expressed on the surface of certain immune effector cells. Neutrophils are the most abundant leukocyte, express Fc-αRI, and are often the first to respond to sites of injury and infection. Here, we describe a function for IgA–virus immune complexes (ICs) during viral infections. We show that IgA–virus ICs potentiate NETosis—the programmed cell-death pathway through which neutrophils release neutrophil extracellular traps (NETs). Mechanistically, IgA–virus ICs potentiated a suicidal NETosis pathway via engagement of Fc-αRI on neutrophils through a toll-like receptor–independent, NADPH oxidase complex–dependent pathway. NETs also were capable of trapping and inactivating viruses, consistent with an antiviral function.

Design and validation of the Gingkolide estimation using RP-HPLC analytical tool

Gingkolide is an antiseizure medicine used as an adjuvant of partial seizures and GAD to relieve neuropathic pain. It binds to the very high affinity alpha delta site in the CNS. Although the drug's mechanism remains unclear, in genetically engineered mice and other anticonvulsive models, findings showed that it binds to alpha receptors. A rapid rise in the number of drugs added to each class of drugs has been noted. Whether in a single or multi-drug delivery form, these medications are developed into newer formulations. These newest formulations put on the market need a new investigation to estimate the medication in the formulations. In the scientific literature, the current analytical procedures for such drugs are available, but not all approaches are stable and economical to use. Few other techniques are often time-consuming. The goal of this work was to develop an RP-HPLC analytical tool for Gingkolide estimation. The drug's RP-PLC study meets the drug's optimum integrity, suitability, regeneration. The drug's LOQ and LOD were reached with elevated sensitivity. Overall, the results show that the recommended analytical approach in the formulation should be used to evaluate the drug. For regular study of the medication in its dosage form, this approach may be recommended.

IgA Potentiates NETosis in Response to Viral Infection

ABSTRACTIgA is the second most abundant antibody present in circulation and is enriched at mucosal surfaces. As such, IgA plays a key role in protection against a variety of mucosal pathogens, including viruses. In addition to neutralizing viruses directly, IgA can also stimulate Fc-dependent effector functions via engagement of Fc alpha receptors (FcαRI) expressed on the surface of certain immune effector cells. Neutrophils are the most abundant leukocyte, express FcαRI, and are often the first to respond to sites of injury and infection. Here, we describe a novel function for IgA:virus immune complexes (ICs) during viral infections. We show that IgA:virus ICs potentiate NETosis – the programmed cell death pathway through which neutrophils release neutrophil extracellular traps (NETs). Mechanistically, IgA:virus ICs potentiated a suicidal NETosis pathway via engagement of FcαRI on neutrophils through a toll-like receptor (TLR)-independent, NADPH oxidase complex-dependent pathway. NETs also were capable of trapping and inactivating viruses, consistent with an antiviral function.

S32. ARE SELECTIVE ESTROGEN RECEPTOR BETA AGONISTS POTENTIAL THERAPEUTICS FOR SCHIZOPHRENIA?

Background Estrogen therapies, such as estradiol, have shown promise as therapeutics for schizophrenia; however, safety and tolerability concerns, including feminization effects in men and cancer and stroke risk in pre-menopausal women, may limit their broader use. Estradiol binds to both the estrogen alpha (ERA) and beta (ERB) receptors. ERB receptors appear not to mediate many of the concerning side effects of estrogen therapies. In addition, beta receptors have unique localization in cortical regions (i.e., hippocampus), and improve social behaviors and cognition in some animal models, which has led to interests in these compounds for testing in schizophrenia. To our knowledge, there have been no previous clinical trials of selective ERB agonists in schizophrenia. LY500307 is a highly selective agent for beta receptors without effects on estrogen alpha receptors when doses are constrained. Doses that are too high may engage alpha receptors but the alpha engaging threshold dose has not been fully determined in patient groups. The purpose of this dose-response study was to determine: ERB selectivity doses of LY500307 (i.e., without engaging alpha receptors); safety and tolerability; brain target engagement; and effects on cognition and symptoms. Methods A two-staged, double-blind, 8-week, adjunctive to APDs, adaptive phase 1b/2a trial design was conducted in men with schizophrenia (women were not included because of the lack of toxicology, safety, phase 1 and clinical data supporting use in this population). Three LY500307 doses and placebo were evaluated: 25 mg/day, 75 mg/day, and 150 mg/day. The primary markers for estrogen beta receptor selectivity was lack of effects on total testosterone levels (TT) and no feminization signs. Target engagement was assessed with an N-back working memory fMRI task and the electrophysiology measure mismatch negativity (MMN). Cognitive effects were assessed by the MCCB Composite score. Negative and total symptoms were assessed by the NSA-16 and PANSS, respectively. The primary analyses included all subjects and compared the slope from the three LY500307 dosing arms to the placebo slope in order to evaluate the dose responses. The linear mixed model with random intercept was employed and secondary analyses assessed differences between mean changes of the two higher dose arms combined (75 mg and 150 mg) versus placebo. Results Ninety-four patients were randomized across the placebo and three LY500307 dosing arms. There were no effects on plasma TT levels and no evidence of feminization, suggesting all doses were selective for the beta receptor. No significant adverse events were observed. There were no significant differences between the slopes of the three drug doses versus placebo on the brain target engagement variables (fMRI/N-back: F=0.24, p=0.868; MMN (Duration): F=1.08, p=0.358; MMN (Frequency): F=0.89, p=0.446) or on the cognitive/symptom measures (MCCB composite: F=0.87, p=0.458; NSA-16: F=1.79, p=0.148; and PANSS Total: F=0.69, p=0.558.) Secondary analyses also failed to show any significant effects of LY500307 versus placebo on any of the study variables. Discussion Conclusions: This study indicates that the ERB agonist LY500307 was selective, safe, and well tolerated in patients with schizophrenia. This selective ERB agonist, however, failed to demonstrate any significant effects on brain targets, cognition, negative and total symptoms. Potential issues related to dosing and characteristics of the patient population will be discussed. These data suggest that estrogen alpha receptor activation may be necessary to yield positive results in this patient population. Future studies are needed to confirm these findings.

Control of cancer progression by extracorporeal removal of soluble TNF-alpha receptors.

20 Background: Tumor necrosis factor (TNF)-alpha is a potent antineoplastic cytokine which exerts tumor destruction in part through vascular disruption. Local administration of TNF-alpha exerts profound anti-cancer effects as observed in studies using isolated limb perfusion. However, systemic administration of this cytokine is associated with vascular leakage and limits clinical use. Methods: One method of stimulating localized effects of TNF-alpha is through removal of its soluble receptors, sTNF-R1 and sTNF-R2 using extracorporeal apheresis; ie, Immunopheresis. In this comparative oncology study, we utilized the proprietary LW-02 device to remove sTNF-Rs in pet dogs with varying types of solid malignancies (Part A; n = 12) or with oral malignant melanoma (Part B; n = 8). Canine patients received between 12-24 apheresis treatments over the course of 4 to 8 weeks. Results: In the over 300 immunopheresis treatments, no device associated adverse effects were reported. Consistent reduction of sTNF-Rs was observed, as measured by decreased plasma levels during treatment as well as elution of sTNF-Rs from the cartridge. Of 17 evaluable dogs who completed treatment, 7 had disease stabilization or regression as confirmed by cRECIST criteria. Conclusions: These data strongly support the clinical translation of the LW-02 device as a potential therapeutic approach for treating solid malignancies/melanoma, either as mono- or as adjuvant therapy.

CLONING AND EXPRESSION OF VACCINIA VIRUS PROTEIN (B18) IN HETEROLOGOUS HOST

B18 is an immunoglobulin superfamily glycoprotein produced by vaccinia virus which resembles to interferon alpha receptors. It acts both in solution and also when associated with the cell surface. This B18 protein was expressed in heterologous host and purified based on affinity column chromatography. The purified B18 protein has been evaluated for biological activity in Pig Kidney- 15 cells (PK-15) which has been reported to have α –interferon receptors and signaling mechanism. The CPE was more than 90% in the presence of protein and around 50% in the absence. B-18 can enhance the infectivity of FMDV to cells having α –interferon receptors. This provides the basis for developing new strategies for improving attenuated vaccines and its capacity to act as an enhancer protein.