2023806d53d92b616e1ef8317e8fc0c9 VAC_a2025v13p1076.pdf 87649f65b62e4338efb470d62871f9444574e20f VAC_a2025v13p1076.pdf 809e2e01058ed4aed4d1be376d4d375f6d506519333c3f560c864acd76de3ff3 VAC_a2025v13p1076.pdf Title: Efficacy of a Novel PCV2d and Mycoplasma hyopneumoniae Combined Vaccine in Piglets with High and Low Levels of PCV2 Maternally Derived Antibodies at Vaccination Subject: Background/Objectives: Maternally derived antibody (MDA) levels of porcine circovirus 2 (PCV2) may eventually interfere with humoral response and vaccination efficacy. This study aimed to evaluate the efficacy of a ready-to-use PCV2d and Mycoplasma hyopneumoniae combined vaccine in piglets with different PCV2 MDA levels at vaccination in an experimental inoculation with a heterologous viral genotype. Methods: Forty-eight piglets were allocated into vaccinated (V) and non-vaccinated (NV) groups with high (H) and low (L) PCV2 MDA subgroups (H-V, H-NV, L-V, L-NV). At 3 weeks of age, the piglets received either one dose of vaccine or placebo. Five weeks later, all animals were intranasally challenged with a PCV2b inoculum. Body weight was registered at different time points. Blood samples, peripheral blood mononuclear cells and tracheobronchial lymph nodes (TBLN) were collected and used to assess viraemia, viral load, humoral and cellular responses and histological lesions. Results: The V group showed higher PCV2 antibody levels from challenge onwards, along with a lower percentage of viraemic pigs and reduced viral load in serum at 2 and 3 weeks post-challenge (wpc) and in TBLN tissues compared to the NV group. The H-V group had the highest antibody levels post-challenge, showed no detectable viraemia and had a lower overall amount of virus in tissues. The NV group (especially H-NV) exhibited increased levels of IFN-, IFN- and TNF- post-challenge. Conclusions: The tested vaccine elicited humoral and cellular immune responses and reduced viral presence in serum and tissues, demonstrating efficacy in a PCV2 subclinical infection model despite high MDA levels at the time of vaccination. Understanding both humoral and cellular immune responses according to different MDA levels can help design more effective vaccination strategies against PCV2. Keywords: porcine circovirus 2; maternally derived antibodies; vaccine; cytokines; quantitative PCR; in situ hybridisation Author: Mònica Sagrera, Laura Garza-Moreno, Àlex Cobos, Anna Maria Llorens, Eva Huerta, Mónica Pérez, Diego Pérez, David Espigares, Joaquim Segalés and Marina Sibila Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25; modified using OpenPDF 1.4.2 CreationDate: Wed Oct 22 10:19:40 2025 CEST ModDate: Wed Oct 22 11:00:52 2025 CEST Custom Metadata: yes Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 22 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 9019027 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- NQOWZC+URWPalladioL-BoldItal Type 1 Custom yes yes yes 168 0 WMCBHI+URWPalladioL-Roma Type 1 Custom yes yes yes 169 0 WTPVWN+URWPalladioL-Ital Type 1 Custom yes yes yes 170 0 XKWFDB+VnURWPalladioL Type 1 Custom yes yes yes 171 0 LEVXPH+EURM10 Type 1 Builtin yes yes yes 172 0 GTWTJX+URWPalladioL-Bold Type 1 Custom yes yes yes 173 0 ONJBTP+VnURWPalladioL-Bold Type 1 Custom yes yes yes 174 0 YFFKLH+CMSY10 Type 1 Builtin yes yes yes 264 0 GPKIAL+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 256 0 GPKIAJ+PalatinoLinotype TrueType WinAnsi yes yes no 257 0 GPKIAJ+PalatinoLinotype TrueType WinAnsi yes yes no 305 0 GPKIAJ+PalatinoLinotype TrueType WinAnsi yes yes no 453 0 GPKIAH+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 454 0 GPKIAJ+PalatinoLinotype TrueType WinAnsi yes yes no 470 0 GPLDAO+TimesNewRoman TrueType WinAnsi yes yes no 471 0 GPKIAJ+PalatinoLinotype TrueType WinAnsi yes yes no 480 0 GPKIAJ+PalatinoLinotype TrueType WinAnsi yes yes no 501 0 GPKIAH+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 502 0 GPKIAJ+PalatinoLinotype TrueType WinAnsi yes yes no 734 0 GPKIAH+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 735 0 GPKIAJ+PalatinoLinotype TrueType WinAnsi yes yes no 1065 0 GPKIAH+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 1066 0 GPKIAL+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 1125 0 GPKIAJ+PalatinoLinotype TrueType WinAnsi yes yes no 1126 0 HBEKOI+Arial TrueType WinAnsi yes yes no 1127 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2025-11-25 04:11:05 CET RepresentationInformation: VAC_a2025v13p1076.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2025-11-24 11:48:53 CET Size: 9019027 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 1368 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: Animal Selection and Housing Item: Title: Experimental Study Design and Sample Collection Item: Title: DNA Extraction and Detection of PCV2 by qPCR Item: Title: PCV2 Antibody Levels Measured by PCV2 IgG ELISA Item: Title: Histopathology and In Situ Hybridisation Item: Title: Peripheral Blood Mononuclear Cells (PBMCs) Isolation and Stimulation Item: Title: Multiplex Immunoassay for the Quantification of Cytokines Item: Title: Statistical Analyses Item: Title: Results Children: Item: Title: Clinical Assessment Item: Title: Comparison Between V and NV Groups Children: Item: Title: BW and ADWG Item: Title: Dynamics of PCV2 IgG Antibody Levels Item: Title: PCV2 Infection Dynamics Item: Title: Lesion Assessment and PCV2 Antigen Detection in TBLN Item: Title: Cytokine Concentration Item: Title: Comparison Among H-V, H-NV, L-V and L-NV Groups Children: Item: Title: BW and ADWG Item: Title: Dynamics of PCV2 IgG Antibody Levels Item: Title: PCV2 Infection Dynamics Item: Title: Lesion Assessment and PCV2 Antigen Detection in TBLN Samples Item: Title: Cytokine Concentration Item: Title: Discussion Item: Title: Conclusions Item: Title: Appendix A Item: Title: References Info: Title: Efficacy of a Novel PCV2d and Mycoplasma hyopneumoniae Combined Vaccine in Piglets with High and Low Levels of PCV2 Maternally Derived Antibodies at Vaccination Author: Mònica Sagrera, Laura Garza-Moreno, Àlex Cobos, Anna Maria Llorens, Eva Huerta, Mónica Pérez, Diego Pérez, David Espigares, Joaquim Segalés and Marina Sibila Subject: Background/Objectives: Maternally derived antibody (MDA) levels of porcine circovirus 2 (PCV2) may eventually interfere with humoral response and vaccination efficacy. This study aimed to evaluate the efficacy of a ready-to-use PCV2d and Mycoplasma hyopneumoniae combined vaccine in piglets with different PCV2 MDA levels at vaccination in an experimental inoculation with a heterologous viral genotype. Methods: Forty-eight piglets were allocated into vaccinated (V) and non-vaccinated (NV) groups with high (H) and low (L) PCV2 MDA subgroups (H-V, H-NV, L-V, L-NV). At 3 weeks of age, the piglets received either one dose of vaccine or placebo. Five weeks later, all animals were intranasally challenged with a PCV2b inoculum. Body weight was registered at different time points. Blood samples, peripheral blood mononuclear cells and tracheobronchial lymph nodes (TBLN) were collected and used to assess viraemia, viral load, humoral and cellular responses and histological lesions. Results: The V group showed higher PCV2 antibody levels from challenge onwards, along with a lower percentage of viraemic pigs and reduced viral load in serum at 2 and 3 weeks post-challenge (wpc) and in TBLN tissues compared to the NV group. The H-V group had the highest antibody levels post-challenge, showed no detectable viraemia and had a lower overall amount of virus in tissues. The NV group (especially H-NV) exhibited increased levels of IFN-, IFN- and TNF- post-challenge. Conclusions: The tested vaccine elicited humoral and cellular immune responses and reduced viral presence in serum and tissues, demonstrating efficacy in a PCV2 subclinical infection model despite high MDA levels at the time of vaccination. Understanding both humoral and cellular immune responses according to different MDA levels can help design more effective vaccination strategies against PCV2. 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