Veterinary immunology and immunopathology最新文献

Veterinary vaccine quality assurance has traditionally relied on animal-based potency, safety, and efficacy tests, including challenge protection and in vivo titration assays. Although historically regarded as regulatory gold standards, these methods are increasingly constrained by ethical considerations, high costs, limited throughput, and prolonged timelines. In response, strong scientific and regulatory momentum now supports alternative approaches that reduce or replace animal use while maintaining confidence in vaccine performance. This review critically examines the current landscape of non-animal and reduced-animal methodologies for veterinary vaccine evaluation within the 3Rs framework (Replacement, Reduction, and Refinement). We synthesize advances across five methodological domains: molecular and genetic quantification (including qPCR, digital PCR, and DIVA strategies); cell-based and in vitro functional potency assays; immunochemical and point-of-care platforms; physicochemical and analytical characterization; and systems-level immunological and multi-omics approaches. For each category, we discuss scientific principles, analytical performance, validation requirements, and regulatory acceptance, with emphasis on alignment with WOAH and VICH guidance. Practical challenges related to assay standardization, reference materials, infrastructure, and inter-laboratory reproducibility are addressed. Species-specific case studies spanning livestock, companion animal, and poultry vaccines including foot-and-mouth disease, lumpy skin disease, rabies, and avian influenza illustrate how integrated, weight-of-evidence strategies support batch release, consistency testing, and post-licensure surveillance. Collectively, the evidence supports a paradigm shift toward data-rich, mechanism-informed quality-control frameworks that enhance efficiency, ethical compliance, and regulatory confidence in veterinary vaccine evaluation.

To investigate the variances in inhibitory effects and potential immunological mechanisms of transfer factors (TFs) from different origins against avian reticuloendotheliosis virus (REV) infection in chickens, this study firstly compared the protein expression profiles of spleens from REV-infected chickens, Newcastle disease (ND) vaccine-immunized chickens, and specific pathogen-free (SPF) chickens. TFs were subsequently extracted from the spleens of these groups, and the alterations in splenic proteomics were monitored post-administration of these TFs to chickens. Furthermore, the CD4⁺/CD8⁺ T cell subsets and the levels of cytokines such as IFN-γ and IL-2 were determined, followed by REV challenge experiments. The results demonstrated that TFs sourced from REV-infected spleens, which enriched virus-specific immune-related proteins such as STAT1 and OASL, significantly boosted cellular immune responses by activating the NOD-like receptor signaling pathway and establishing an efficient STAT1-OASL-PARP9-IRF7 protein-protein interaction network, consequently inhibiting REV replication efficiently. TFs obtained from vaccine-immunized spleens only amplified humoral immunity with restricted anti-REV effects, while TFs derived from SPF spleens had no substantial immunomodulatory activity. Notably, TFs extracted from REV-infected spleens exhibited superior anti-REV efficacy. This study furnishes crucial evidence for the selection of sources and molecular mechanism analysis of high-efficiency TF preparations.

In cattle, passive immunity relies entirely on the intake and intestinal absorption of colostral immunoglobulins after birth; however, breed-specific differences in passive transfer efficiency remain incompletely characterized. This study aimed to compare passive immunity status among Holstein, Jersey, and Montbéliarde calves under standardized colostrum management conditions, thereby minimizing colostrum-related variability and evaluating potential breed-associated differences in immunoglobulin G (IgG) absorption. The study was conducted on a single commercial dairy farm under uniform management conditions. A total of 42 newborn calves were enrolled, including Holstein (n = 15), Jersey (n = 15), and Montbéliarde (n = 12) calves. All calves received colostrum within the first six hours postpartum at a standardized volume corresponding to 10-12% of body weight. Colostrum quality was evaluated using Brix refractometry, and ELISA determined colostral IgG concentrations. Blood samples were collected from calves at approximately 36 h after birth to assess serum IgG concentrations. No significant differences were observed among breeds in colostrum Brix values, colostral IgG concentrations, serum Brix values, or serum total protein concentrations (p > 0.05). In contrast, serum IgG concentrations differed significantly among breeds (p < 0.05), with Holstein calves exhibiting the highest values (20.90 ± 3.61 g/L), followed by Jersey (17.52 ± 6.95 g/L) and Montbéliarde calves (12.70 ± 8.55 g/L). Breed-specific correlation patterns between colostrum and serum parameters were identified, particularly in Montbéliarde calves. In conclusion, despite comparable colostrum quality and standardized colostrum intake, Montbéliarde calves demonstrated significantly lower serum IgG concentrations than Holstein and Jersey calves. These findings suggest that passive transfer efficiency may be influenced by breed-specific physiological or genetic factors independent of colostrum IgG content.

Streptococcosis caused by Streptococcus agalactiae is widespread in the world, especially in south Asia and southeast Asia. In this study, an S. agalactiae isolate collected from diseased fish in India was molecularly confirmed through GAPDH-targeted PCR, and further validated by sequencing. Genotyping revealed distinct genomic relationships among regional isolates; pulsed-field gel electrophoresis identified three pulsotypes with the Indian isolate clustering closely with two Taiwanese isolates under both ApaI and SmaI digestion, while ERIC-PCR grouped the Indian strain with a Taiwanese strain at > 92% similarity. Antibiotic susceptibility test showed that the Indian isolate was resistant to tetracycline, in contrast to the control strain. Pathogenicity assays determined an LD₅₀ of 1.7 × 10⁵ CFU/ml, and infected fish displayed typical clinical signs consistent with streptococcosis. Pro-inflammatory cytokines (IL-1β, IL-8, IFN-γ) were significantly upregulated in head kidney and spleen tissues at 48 h post infection, although IL-8 and IL-1β showed earlier expression in the spleen. These results characterize the molecular identity, virulence, and immunopathological profile of an Indian S. agalactiae strain, providing critical insights for disease surveillance in tilapia aquaculture.

Research related to the investigation of the activation and regulation of the immune system of domestic buffaloes is still very limited. The main factor linked to this limitation is the scanty availability of commercial monoclonal antibodies (mAb) against buffalo immune markers that allow the use of multiparametric techniques, such as flow cytometry, to study the modulation of the immune system of these animals. The evaluation of the cross-reactivity of commercially available mAbs against other species for cytokines and components of the immune system of buffaloes, is the fastest and least expensive way to expand the tools to assess immune response in those animals. Therefore, sixteen commercial mAbs anti-cell markers (CD4, CD8, CD14, CD21, CD25, CD28, CD45RB, CD45RO, WC1, and CD80) and anti-cytokines or other effector molecules of the immune system (TNF-α, IFN-γ, IL-4, IL-17A, granzyme B and perforin) that recognize molecules from bovine and human species were evaluated in whole blood samples from buffaloes and cattle. Two experiments were performed: Experiment 1, with six buffaloes, evaluated the specificity of mAb binding, and experiment 2, with twelve buffaloes and twelve cattle, evaluated the cross-reactivity of the mAbs. Samples were processed and analyzed by flow cytometry. Of the mAbs analyzed 15 clones were validated for flow cytometry of buffalo cells; only the CD4 clone did not show cross-reactivity to buffaloes amplifying the spectrum analyses of immune response in this animal species. This validated panel enables standardized flow cytometric immunophenotyping in buffaloes for research on vaccination, infectious disease, and immune regulation.