Kahraman Tamer, Yagci Fuat Cem, Ceylan Ahmet, Calik Ali, Tarman Ibrahim Oguzhan, Kiran Fadime
THORVACS Biotechnology, Bilkent Cyberpark, 06800, Ankara, Turkey.
ARBO Biotechnology, SL6 8BY, Maidenhead, United Kingdom.
Poult Sci. 2025 May;104(5):105024. doi: 10.1016/j.psj.2025.105024. Epub 2025 Mar 12.
This study aimed to develop and evaluate a CpG oligodeoxynucleotide (CpG ODN)-adjuvanted trivalent inactivated Salmonella vaccine including S. enterica subsp. Enterica serovar Typhimurium, Salmonella enterica subsp. Enterica serovar Enteritidis, and Salmonella enterica serotype Infantis, for its immunogenic efficacy in chickens. The immunomodulatory effects of various CpG ODNs were assessed based on proinflammatory cytokine secretion and the expression levels of CD80, CD86, and MHC-II in the chicken cell lines HD11 and DT40. According to the results, CpG ODNs D35 3CG PO, D35 3CG MB, 1466 Acore PO, 1466 Acore MB, and K3 which exhibited non-cytotoxicity in both HD11 and DT40 cell lines, were selected for vaccine formulation. To evaluate their effects under in vivo conditions, chicks (n = 25) were randomly assigned to fourteen groups (G1: only sterile pyrogen-free saline solution, G2: only inactivated vaccine, G3: inactivated vaccine with 150 mg/dose of ALUM, G4: commercial Salenvac T vaccine, G5-G14: various experimental vaccine formulations which included different CpG ODNs combined with inactivated bacterial strains, with or without ALUM). Immune responses were analyzed through serological assays for antigen-specific antibody titers and ex vivo splenocyte cultures for cytokine secretion. Flow cytometry was performed to assess T-cell activation and IFN-γ production. The results demonstrated that the CpG ODNs-adjuvanted vaccine formulations significantly enhanced both humoral and cellular immunity compared to the commercial vaccine. Specifically, the Vac#5+ ALUM formulation, which included the K3 CpG ODN, induced robust antibody responses against Salmonella antigens and significantly increased IFN-γ secretion, nearly two-fold higher than the commercial vaccine. This effect was primarily mediated by CD4+ helper and CD8+ cytotoxic T cells. These findings highlight the potential of CpG ODNs as effective vaccine adjuvants in poultry. To the best of our knowledge, this is the first study to investigate the use of CpG ODNs as adjuvants in inactivated Salmonella vaccine formulations. Future studies should focus on evaluating the long-term protective efficacy of this vaccine formulation and its ability to provide cross-protection against a broader spectrum of Salmonella serovars.
本研究旨在开发和评估一种含CpG寡脱氧核苷酸（CpG ODN）佐剂的三价灭活沙门氏菌疫苗，该疫苗包含肠炎沙门氏菌亚种肠道血清型鼠伤寒沙门氏菌、肠炎沙门氏菌亚种肠道血清型肠炎沙门氏菌和婴儿沙门氏菌血清型，以评估其对鸡的免疫原性效力。基于促炎细胞因子分泌以及鸡细胞系HD11和DT40中CD80、CD86和MHC-II的表达水平，评估了各种CpG ODN的免疫调节作用。根据结果，选择了在HD11和DT40细胞系中均表现出无细胞毒性的CpG ODN D35 3CG PO、D35 3CG MB、1466 Acore PO、1466 Acore MB和K3用于疫苗配方。为了评估它们在体内条件下的作用，将雏鸡（n = 25）随机分为14组（G1：仅无菌无热原生理盐水溶液；G2：仅灭活疫苗；G3：含150 mg/剂量明矾的灭活疫苗；G4：商业Salenvac T疫苗；G5 - G14：各种实验性疫苗配方，包括不同的CpG ODN与灭活细菌菌株组合，有或没有明矾）。通过血清学检测抗原特异性抗体滴度以及体外脾细胞培养检测细胞因子分泌来分析免疫反应。进行流式细胞术以评估T细胞活化和IFN - γ产生。结果表明，与商业疫苗相比，含CpG ODN佐剂的疫苗配方显著增强了体液免疫和细胞免疫。具体而言，包含K3 CpG ODN的Vac#5 + 明矾配方诱导了针对沙门氏菌抗原的强烈抗体反应，并显著增加了IFN - γ分泌，比商业疫苗高出近两倍。这种效应主要由CD4 + 辅助性T细胞和CD8 + 细胞毒性T细胞介导。这些发现突出了CpG ODN作为家禽有效疫苗佐剂的潜力。据我们所知，这是第一项研究将CpG ODN用作灭活沙门氏菌疫苗配方佐剂用途的研究。未来的研究应侧重于评估这种疫苗配方的长期保护效力及其对更广泛沙门氏菌血清型提供交叉保护的能力。
