mouse igg antigen elisa kit  (Innovative Research Inc)

Journal: Alzheimer's & Dementia

Article Title: Female‐biased astrocytic priming shapes early locus coeruleus vulnerability in an Aβ oligomer milieu

doi: 10.1002/alz.71168

Figure Lengend Snippet: Astrocyte‐linked C3 elevation and lactate–MCT2–OXPHOS axis engagement in the pons of female APP/PS1 mice. Astrocyte‐enriched pons fraction shows increased C3 expression in 2‐ to 3‐month‐old female APP/PS1 mice. A, Schematic illustrating MACS‐based isolation of pontine astrocytes. Schematic created using Biorender. B, Assessment of C3 expression by qPCR followed by 1% agarose gel electrophoresis–gel image showing C3 bands at 172 bp in astrocyte‐enriched fractions (lane 1–4) and astrocyte‐depleted fractions (lane 5–8). C, D, Bar graphs demonstrating elevated C3 levels in the astrocyte‐enriched fraction of female APP/PS1 mice compared to other groups and to their corresponding negative fraction. Each bar represents pooled pons samples from 8 mice per group. Lactate–MCT2 axis and OXPHOS pathway engagement in female APP/PS1 mice. Female APP/PS1 mice showed higher (E) lactate levels in the pons measured by colorimetric assay and increased (F) MCT2 expression assessed by qPCR ( n = 6 per group; two‐way ANOVA with Tukey post hoc test; data shown as mean ± SEM; * p < 0.05, ** p < 0.01). G, H, mRNA expression profiling of mitochondrial markers by qPCR in the pons. Female APP/PS1 mice showed significantly elevated expression of the mitochondrial complex I subunit Ndufs8 and complex V subunit Atp5a1 , indicating adaptive mitochondrial activation in the pons ( n = 5‐6; two‐way ANOVA with Tukey post hoc test; data shown as mean ± SEM; * p < 0.05, ** p < 0.01). I, Graphical summary. Elevated Aβ oligomers in female APP/PS1 mice shift astrocytes from a resting to an active GFAP +ve state, with associated C3 and NF‐κB2 upregulation and engagement of lactate–OXPHOS axis, supporting increased energetic demands. Graphical summary created using Biorender. Aβ, amyloid beta; ACSA‐1, astrocyte cell surface antigen 1; ANOVA, analysis of variance; Atp5a1, ATP synthase F1 subunit α; C3, complement component 3; GLUT1, glucose transporter 1; MACS, magnetic‐activated cell sorting; MCT2, monocarboxylate transporter 2; Ndufs8, NADH dehydrogenase iron‐sulfur protein 8; NF‐κB2, nuclear factor‐kappa‐B p100/p52 subunit; OXPHOS, oxidative phosphorylation; qPCR, quantitative polymerase chain reaction; SEM, standard error of mean; WT, wild type.

Article Snippet: Cells were incubated with FcR blocking reagent (Miltenyi Biotec, 130‐092‐575) at a 1:9 dilution for 10 minutes to prevent non‐specific antibody binding, followed by incubation with Anti‐GLAST or Astrocyte Cell Surface Antigen‐1 (ACSA‐1)‐Biotin (Miltenyi Biotec, 130‐095‐826) for 10 minutes.

Techniques: Expressing, Isolation, Agarose Gel Electrophoresis, Colorimetric Assay, Activation Assay, FACS, Phospho-proteomics, Real-time Polymerase Chain Reaction

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: HBVZ10, an AAV8 vector-based new HBV therapy candidate for cccDNA elimination

doi: 10.1016/j.omtm.2025.101646

Figure Lengend Snippet: Progressive reduction of both serum HBeAg and HBsAg upon blocking of cccDNA replenishment (A) Experimental procedures and time course of HBV infection and treatment evaluation using uPA/SCID chimeric mice model. (B1) The expressed anti-HBs antibody levels were >100,000 mIU/mL and sustained at steady levels for 200 days after a single injection of HBVZ10 in 8 chimeric mice (T1 and CT1-CT7) infected with HBV. A pretreatment serum sample from each replicate was used as a negative control, and all pretreatment anti-HBs antibody levels were plotted as 1 mIU/mL for consistency. (B2) Average anti-HBs antibody levels per group. HBVZ10 was administered on day 11 with a dose of 2.5E10 copies for mice CT1-CT4 on day 22 with 1.8E11 copies for mice T1 and CT5-CT6, or on day 44pi with 7E11 copies of HBV10 for mouse CT7. All 7 mice (CT1-CT7) also received 12-week ETV 10 days after HBVZ10 injection. T. HBVZ10 monotherapy. CT: Combination of HBVZ10 with ETV. (C1) Serum HBeAg kinetics among 7 mock-treated mice (MT1-MT7 blue) and 8 treated mice (T1 and CT1-CT7 green). Serum HBeAg levels are expressed as the OD ratio of the tested sample to the mean OD of three negative controls. A ratio greater than 2.1 is considered positive. (C2) Average serum HBeAg levels per group between mock-treated (blue) and treated (green) from B1. (D1) Kinetic serum HBsAg levels (IU/mL) among mock-treated 7 mice (blue) and 8 treated mice (green) from B1. (D2) Average serum HBsAg levels per group between mock-treated (blue) and treated group (green) from C1. (E) Comparison of average cccDNA levels between mock-treated and combination treated groups (cccDNA was undetectable in mice CT2 and CT4 and not plotted). (F) Comparison of average rcDNA levels between mock-treated and combination-treated groups. Statistical significance was determined using Student’s t test; p < 0.05 was considered significant. Error bars: standard deviation (SD).

Article Snippet: Blood was collected triweekly for quantification of serum HBV DNA (qPCR), HBeAg (CSB-E13557h, CUSABIO), HBsAg (GS HBsAg EIA 32591, Bio-Rad), anti-HBs antibody (MONOLISA anti-HBs EIA 25200, Bio-Rad) with calibrators (MONOLISA anti-HBs 20-Calibrator kit 25219, Bio-Rad), and human albumin (human albumin ELISA kit E−80AL, Immunology Consultants Laboratory) levels by ELISA per instructions.

Techniques: Blocking Assay, Infection, Injection, Negative Control, Comparison, Standard Deviation

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: HBVZ10, an AAV8 vector-based new HBV therapy candidate for cccDNA elimination

doi: 10.1016/j.omtm.2025.101646

Figure Lengend Snippet: Add-on of HBVZ10/anti-HBs antibodies to 9-week ETV treatment prevented HBV relapse, leading to progressive reduction of both serum HBeAg and HBsAg levels (A) Experimental procedures and time course of HBV infection and treatment evaluation using uPA/SCID chimeric mice model. (B) Serum HBV DNA (copies/mL log10) kinetics in mock-treated (MT1-MT8 blue) and ETV monotherapy (ET1-ET4 orange) group. (C) Serum HBV DNA kinetics in 8 ETV-treated mice (CT1-CT8) with add-on with a single dose of 1E10 copies of HBVZ10 on day 70pi, then anti-HBs levels were further boosted with administration of mouse anti-HBs at a dose of 250 μg/injection triweekly started on day 99 or later. (D) Average serum HBV DNA levels (copies/mL log10) among 3 groups. (E1) Serum kinetic anti-HBs antibody levels (mIU/mL) expressed by HBVZ10 and boosted by infusions of mouse anti-HBs antibody among the 8 mice (CT1-CT8). Dashed line: 100,000 mIU/mL. Note: Anti-HBs antibody levels exhibited significant fluctuations in some mice receiving triweekly infusions of mouse anti-HBs antibody, contrasting the steady and consistent antibody levels expressed by sufficient HBVZ10 doses ( A). A pretreatment serum sample from each replicate was used as a negative control, and all pretreatment anti-HBs antibody levels were plotted as 1 mIU/mL for consistency. (E2) Average anti-HBs antibody levels (mIU/mL) per group. (F1) Kinetic serum HBsAg levels (IU/mL) among 2 mice with mock-treated (MT1 and MT2 blue), 2 mice with 9-week of ETV therapy (ET1 and ET2 yellow), and 8 mice with 9-week of ETV and anti-HBs antibodies add-on (CT1-CT8 green). (F2) Average serum HBsAg levels (IU/mL) per group. (G1) Kinetic serum HBeAg levels among 2 mice with mock-treated (MT1 and MT2 blue), 2 mice with 9-week of ETV therapy (ET1 and ET2 yellow), and 8 mice with 9-week of ETV and anti-HBs antibodies add-on (CT1-CT8 green). Note. Mouse CT5 had relatively higher baseline HBeAg level and HBeAg remained detectable on termination day (day 218 PI) despite progressive reduction. Serum HBeAg levels are expressed as the OD ratio of the tested sample to the mean OD of three negative controls. A ratio greater than 2.1 is considered positive. (G2) Average serum HBeAg levels per group. (H and I) Comparison of intracellular cccDNA and rcDNA levels among 2 livers with 9-week of ETV therapy (ET1 and ET2 yellow), 2 livers with mock-treated (MT1 and MT2 blue), and 6 livers with 9-week of ETV and anti-HBs antibodies add-on, 5 (CT1-CT3, CT7 and CT8) of them achieved progressive reduction of both serum HBeAg and HBsAg to undetectable levels and the remaining one with detectable serum HBeAg on the termination day (CT5). The differences in cccDNA and rcDNA levels were analyzed by Student t test, p < 0.05 considered significant. Error bars: standard deviations (SD). The limit of detection for HBV DNA is 100 copies/mL and 0.05 IU/mL for serum HBsAg.MT: mock-treated. ET: ETV Monotherapy. CT: 9-week of ETV and anti-HBs antibodies add-on.

Article Snippet: Blood was collected triweekly for quantification of serum HBV DNA (qPCR), HBeAg (CSB-E13557h, CUSABIO), HBsAg (GS HBsAg EIA 32591, Bio-Rad), anti-HBs antibody (MONOLISA anti-HBs EIA 25200, Bio-Rad) with calibrators (MONOLISA anti-HBs 20-Calibrator kit 25219, Bio-Rad), and human albumin (human albumin ELISA kit E−80AL, Immunology Consultants Laboratory) levels by ELISA per instructions.

Techniques: Infection, Injection, Negative Control, Comparison