3c7b89d50434dac0dc4bc58c122ef3eb 327607.pdf 8168b3921a2fc1f6fefd35d26d44e33213253270 327607.pdf 1066582dc709937fa23091117e363d89c897adf106ab55b964751a1d441d90cb 327607.pdf Title: Premature Neuroimmune and Redox-Inflammatory Breakdown at the Prodromal Stage in Male and Female Triple-Transgenic Alzheimer’s Disease Mice Subject: Background/Objectives: Homeostatic (nervous, immune and endocrine) systems and their communications network are crucial for health and aging rate. We previously reported behavioral and peritoneal leukocyte function alterations and oxidative-inflammatory stress in young female triple-transgenic (3xTg) mice for Alzheimer’s disease (AD). Here, the deterioration of the homeostatic systems and their interplay was investigated, in an integrated way, at prodromal stages and in both sexes of 3xTg-AD mice. Methods: An integrative analysis of the behavioral profile, peripheral immune splenic and thymic leukocyte functions, splenic oxidative-inflammatory state, and plasmatic corticosterone in both sexes of 3xTg-AD mice at 4 months of age was compared to that of age- and sex-matched NTg counterparts. Results: The prodromal stage of 3xTg-AD, characterized by anxiety-like behaviors and disrupted exploration, was aligned with reduced chemotaxis, natural killer activity, and lymphoproliferation—especially in the spleen. In addition, 3xTg-AD mice exhibited lower anti-inflammatory (IL-10) and higher pro-inflammatory (IL-2, IL-1, and TNF-) cytokine concentrations and oxidative stress (higher oxidants and lower antioxidants). Several of these alterations displayed sex-dependent differences (worse in males). However, no differences in corticosterone were found. Conclusions: These findings suggest that neuroimmune and redox-inflammatory dysfunctions, indicative of premature aging, emerge at the prodromal stage of AD, preceding corticosterone changes, unveiling a time lag in the neuroimmunoendocrine alterations in these animals. They may act as early indicators of premature aging in AD pathology and provide potential targets for sex-specific prodromal intervention. Keywords: Alzheimer’s disease; 3xTg-AD mice; prodromal stage; behavior; immune functions; oxidative-inflammatory stress; corticosterone; neuroimmunoendocrine communications; spleen; thymus; males and females; premature aging Author: Lydia Giménez-Llort, Carmen Vida, Judith Félix, Silvia Quer-Palomas, Rashed Manassra and Monica De la Fuente Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25; modified using OpenPDF 1.4.2 CreationDate: Wed Feb 11 05:40:48 2026 CET ModDate: Wed Feb 11 05:46:08 2026 CET Custom Metadata: yes Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 28 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 1354859 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- VVSYYB+EURM10 Type 1 Builtin yes yes yes 199 0 OEUGJS+URWPalladioL-Roma Type 1 Custom yes yes yes 200 0 EDLHRM+URWPalladioL-Ital Type 1 Custom yes yes yes 201 0 QKZKVH+VnURWPalladioL Type 1 Custom yes yes yes 202 0 XGGTRI+URWPalladioL-Bold Type 1 Custom yes yes yes 203 0 YFFKLH+CMSY10 Type 1 Builtin yes yes yes 333 0 URYLRZ+URWPalladioL-BoldItal Type 1 Custom yes yes yes 402 0 NLDHEE+PalatinoLinotype TrueType WinAnsi yes yes no 410 0 NLDHEE+PalatinoLinotype TrueType WinAnsi yes yes no 448 0 NLDHEE+PalatinoLinotype TrueType WinAnsi yes yes no 455 0 NLDHEE+PalatinoLinotype TrueType WinAnsi yes yes no 489 0 NLDHEE+PalatinoLinotype TrueType WinAnsi yes yes no 516 0 NLDHEG+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 573 0 NLDHEE+PalatinoLinotype TrueType WinAnsi yes yes no 574 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2026-04-11 04:18:55 CEST RepresentationInformation: 327607.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2026-04-10 09:49:29 CEST Size: 1354859 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 899 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: ViewerPreferences: HideToolbar: false HideMenubar: false HideWindowUI: false FitWindow: true CenterWindow: false DisplayDocTitle: false NonFullScreenPageMode: UseNone Direction: L2R ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Item: Title: Materials and Methods Children: Item: Title: Animals Item: Title: Experimental Design Item: Title: Prodromal Behavioral Profile Item: Title: Organometrics Item: Title: Immune Functions Children: Item: Title: Collection of Lymphocyte Suspensions Item: Title: Chemotaxis Assay Item: Title: Natural Killer (NK) Cytotoxicity Assay Item: Title: Lymphoproliferation Assay Item: Title: Cytokine Concentrations in Cultures of Spleen Leukocytes Item: Title: Redox-Inflammatory State Children: Item: Title: Glutathione Peroxidase (GPx) Activity Item: Title: Glutathione Reductase (GR) Activity Item: Title: Total Glutathione (GSH) Content Item: Title: Xanthine Oxidase (XO) Activity Item: Title: Plasma Corticosterone Item: Title: Statistical Analysis Item: Title: Results Children: Item: Title: Prodromal Behavioral State of 4-Month-Old Male and Female 3xTg-AD Mice Item: Title: Changes in Physical Condition, Organometrics, and the Endocrine System Are Sex-Dependent and Observable at 4 Months of Age in Male and Female 3xTg-AD Mice Item: Title: An Impairment of the Immune Functions Is Presented in 4-Month-Old Female and Male 3xTg-AD Mice Item: Title: Oxidative Stress Is Present in the Spleen of 4-Month-Old Female and Male 3xTg-AD Mice Item: Title: Discussion Children: Item: Title: Prodromal Behavioral Profile Children: Item: Title: Characteristics of Early BPSD-like Profile Item: Title: Specific Sex-Per Genotype-Effects Item: Title: Organometrics and Plasma Corticosterone Item: Title: Immune Functions Children: Item: Title: Early Innate and Adaptive Immunity Impairment Item: Title: Unbalanced Anti-Inflammatory/Proinflammatory Cytokines Item: Title: Redox-Inflammatory State Item: Title: Conclusions Item: Title: References Info: Title: Premature Neuroimmune and Redox-Inflammatory Breakdown at the Prodromal Stage in Male and Female Triple-Transgenic Alzheimer’s Disease Mice Author: Lydia Giménez-Llort, Carmen Vida, Judith Félix, Silvia Quer-Palomas, Rashed Manassra and Monica De la Fuente Subject: Background/Objectives: Homeostatic (nervous, immune and endocrine) systems and their communications network are crucial for health and aging rate. We previously reported behavioral and peritoneal leukocyte function alterations and oxidative-inflammatory stress in young female triple-transgenic (3xTg) mice for Alzheimer’s disease (AD). Here, the deterioration of the homeostatic systems and their interplay was investigated, in an integrated way, at prodromal stages and in both sexes of 3xTg-AD mice. Methods: An integrative analysis of the behavioral profile, peripheral immune splenic and thymic leukocyte functions, splenic oxidative-inflammatory state, and plasmatic corticosterone in both sexes of 3xTg-AD mice at 4 months of age was compared to that of age- and sex-matched NTg counterparts. Results: The prodromal stage of 3xTg-AD, characterized by anxiety-like behaviors and disrupted exploration, was aligned with reduced chemotaxis, natural killer activity, and lymphoproliferation—especially in the spleen. In addition, 3xTg-AD mice exhibited lower anti-inflammatory (IL-10) and higher pro-inflammatory (IL-2, IL-1, and TNF-) cytokine concentrations and oxidative stress (higher oxidants and lower antioxidants). Several of these alterations displayed sex-dependent differences (worse in males). However, no differences in corticosterone were found. Conclusions: These findings suggest that neuroimmune and redox-inflammatory dysfunctions, indicative of premature aging, emerge at the prodromal stage of AD, preceding corticosterone changes, unveiling a time lag in the neuroimmunoendocrine alterations in these animals. They may act as early indicators of premature aging in AD pathology and provide potential targets for sex-specific prodromal intervention. 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