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The moderate pathogenicity coupled with high host susceptibility of Eimeria maxima has precipitated substantial economic losses in the poultry industry. Addressing challenges such as emerging drug resistance underscores the imperative for innovative vaccine strategies. This study developed a novel DNA vaccine to solve this challenge by fusing E. maxima elongation factor-1α (EmEF1α) with chicken chemokine XCL1 (ChXCL1) in the pVAX1 vector. The recombinant plasmid, designated pVAX1-ChXCL1-EmEF1α, was successfully constructed and confirmed to express the ChXCL1-EmEF1α fusion protein in vitro. Immunization of chickens with this DNA vaccine elicited a robust and balanced immune response, characterized by significantly increased proportions of CD4⁺ (11.76%) and CD8⁺ (5.58%) T lymphocytes, elevated levels of Th1-associated cytokines (IFN-γ and IL-12), and strong antigen-specific IgG and IgA antibody responses. Following experimental challenge with E. maxima, vaccinated birds exhibited substantial protection: a 66.4% reduction in oocyst shedding, a 71.7% improvement in relative weight gain, marked attenuation of intestinal lesions, and an anticoccidial index (ACI) of 170. These findings demonstrate that the ChXCL1-EmEF1α DNA vaccine effectively enhances both cellular and humoral immunity. Collectively, this study validates ChXCL1 as a potent molecular adjuvant and establishes the “antigen–adjuvant” fusion DNA platform as a promising strategy for developing next-generation vaccines against avian coccidiosis. Full article

Coccidiosis is a major parasitic disease that suppresses poultry productivity and causes significant global economic losses. Currently, controlling Eimeria parasites relies primarily on the use of anticoccidial drugs or live vaccines. However, these conventional control strategies face the dual constraints of escalating drug resistance and unsustainable economic expenditures. In this study, the efficacy of a chimeric subunit vaccine comprising Eimeria maxima Elongation Factor-1α (EmEF1α) and chicken chemokine Ligand-1 (ChXCL1) was assessed for protection against experimental Eimeria maxima infection. The synthetic gene fragment ChXCL1-EmEF1α was ligated to the pET28a vector and expressed in vitro. Western blot analysis confirmed the successful expression of the recombinant ChXCL1-EmEF1α protein. Chickens immunized with the ChXCL1-EmEF1α exhibited a significantly stronger IgY response and higher secretion of IL-2 and IL-17 compared to those vaccinated with recombinant ChXCL1 alone or challenged solely with E. maxima. Furthermore, the ChXCL1-EmEF1α group demonstrated enhanced anticoccidial effects, including reduced intestinal lesions, higher body weight gain, and lower oocyst shedding compared to control groups. Following E. maxima challenge, the EmEF1α and ChXCL1-EmEF1α groups demonstrated robust protective efficacy, achieving high ACI values of 182 and 178, respectively. In contrast, the ChXCL1 and UC groups exhibited significantly lower ACI values (150 and 149, respectively), indicating minimal protection. This improvement was also reflected in the immune response, with significantly elevated levels of CD4⁺ and CD8⁺ T cells in the ChXCL1-EmEF1α-treated chickens. Moreover, ChXCL1 acts as an effective adjuvant when fused with EmEF1α, enhancing the vaccine’s anticoccidial efficacy. These results suggest that the ChXCL1-EmEF1α chimeric immunogen is a promising candidate for developing subunit vaccines against E. maxima infections. Full article