African swine fever virus (ASFV) remains a serious global threat to the swine industry due to its high mortality rate and the lack of a widely effective vaccine. In this study, we developed an attenuated Salmonella Typhimurium-based delivery platform, JOL2500, to deliver constructs encoding selected ASFV antigens, including the structural proteins p54 and p72, and virulence- or immune-associated proteins 9GL, HA/CD2v, and C-type lectin. The expression of these recombinant antigens was confirmed in vitro using SDS-PAGE, RT-PCR, and indirect immunofluorescence assay (IFA) in RAW 264.7 cells. The immunogenicity of the recombinant constructs was then evaluated in BALB/c mice following intramuscular immunization. Compared with the vector control group, vaccinated mice showed increased antigen-specific systemic antibody responses, including IgM, IgG1, and IgG2a, as well as enhanced mucosal secretory IgA responses. In addition, splenocytes from immunized mice showed increased proliferation and metabolic activity after antigen simulation. Cytokine analysis revealed a pro-inflammatory and Th1-associated cytokine profile, marked by elevated levels of IFN-γ, TNF-α, and IL-17. Flow cytometry further demonstrated immunization-associated shifts in splenocyte immune-cell populations. Overall, these findings demonstrate that Salmonellamediated delivery of multiple ASFV antigens can induce broad humoral, mucosal, and cellular immune responses in mice. Although protective efficacy against ASFV infection still needs to be confirmed in future studies, this work provides a promising proof of concept for developing safe, practical, and cost-effective ASFV vaccine candidates.
The gut microbiota of companion animals is recognized as a key factor involved not only in digestion and nutrient metabolism, but also in immune regulation and disease susceptibility. Dietary composition plays a crucial role in regulating this gut microbial environment. Insect protein, recently attracting attention as a sustainable protein source, is being proposed as an alternative feed ingredient that can meet the nutritional needs of companion dogs and cats. Rather than drastically altering the overall diversity of the gut microbiota, insect protein-based diets tend to induce gradual changes in specific microbial communities and microbial-derived metabolites. These changes suggest the possibility of interactions with the host immune response. In particular, gut microbial metabolites are known to influence immune homeostasis through association with TLR, NF-κB, and MAPK signaling pathways. However, studies directly elucidating the molecular mechanisms by which insect protein-based diets affect the gut microbiota-immune axis in companion animal models are limited. In the future, systematic, mechanism-focused research will be needed to clarify whether a diet utilizing insect proteins can contribute to the improvement of immune homeostasis and health in companion animals through regulation of intestinal microflora.
Lactobacillus johnsonii JERA01-supplemented feed additive (Lj-A) was produced by fermenting dried porcine blood with Lactobacillus johnsonii (Lj). Lj-A has highly digestible nutrients, bioactive peptides, and probiotic effects. To assess the immunomodulatory potential of Lj-A, it was tested on splenocytes of C57BL/6 mouse. Lj-A was treated on splenocytes in a range of concentration, 0-100 μg/ml. The metabolic activity of splenocytes was enhanced by Lj-A, as shown in MTT assay. Also, some splenocyte clusters were observed under a bright-field microscope on the wells treated with Lj-A. The splenocyte clusters indicated that the cells were activated and proliferating in response to Lj-A. These findings suggest that Lj-A stimulates splenocytes to promote immune cell activation, as evidenced by increased tumor necrosis factor-alpha and interleukin-12 production, thereby enhancing immunological defense functions. In vitro treatment of splenocytes with Lj-A increased the proportions of T cells, B cells, and CD25+ cells. In vivo, immune cell activity was evaluated in C57BL/6 mice orally administered with Lj-A at a dose of 100 mg/day. The proportion of dendritic cells in peritoneal cells was increased along with elevated CD54+ expression. Additionally, the proportions of B cells, CD25+ cells in Peyer’s patch cells increased as well. These results suggest that Lj-A may contribute to the enhancement of immune function and the maintenance of long-term health in animals.
The importance of the immune system in the pathogenesis of periodontitis is being emphasized. To investigate the effects of different immune statuses on the progression of periodontitis, we induced periodontitis in neutrophil-deficient, lymphocytedeficient, and dual neutrophil- and lymphocyte-deficient mice. The groups included BALB/c mice (Con-lym+/neu+), periodontitisinduced BALB/c and SCID mice (Peri-lym+/neu+ and Peri-lym–/neu+), and periodontitis-induced BALB/c and SCID mice treated with anti-Ly6G antibody (Peri-lym+/neu– and Peri-lym–/neu–). Periodontitis was induced by ligation of the mandibular first molar. Mice were sacrificed on days 0 (D0), 3 (D3), and 7 (D7) after ligation, and blood and periodontal tissue were analyzed. On D3, the AB area in the Peri-lym–/neu+ group was larger than in the Peri-lym+/neu+ group. The osteoclast on D3 was lower in the Peri-lym+/neu– group compared to the Peri-lym+/neu+ group. On D7, Ly6G+ cells were lower in both blood and periodontal tissue in the Peri-lym+/neu– group compared to the Peri-lym+/neu+ group, whereas CD3+ cell counts were higher only in periodontal tissue. In the Peri-lym–/neu– group, Ly6G+ cells were lower in both blood and periodontal tissue than in the Peri-lym–/ neu+ group, whereas CD3+ cells were higher in blood only. These results suggest that lymphocyte depletion delays AB loss and neutrophil depletion influences osteoclast formation at the early stage of periodontitis. In the late stage of periodontitis, lymphocyte depletion does not increase lymphocyte infiltration into periodontal tissue, even with neutrophil depletion. Collectively, different immune states may influence distinct stages of periodontitis progression.
국내 양돈 산업의 대형화와 산업화에 따라 축산 악취에 대한 민원 발생이 지속적으로 증가하고 있다. 돼지 분뇨 에서 주로 발생하는 악취는 후각적 불쾌감을 유발할 뿐 아니라, 돼지의 폐사율 증가, 생산성 저하, 농장 근로자의 작업 환경 악화 등 다양한 부정적 영향을 미친다. 따라서 전국적인 양돈장 환경 개선과 효과적인 악취 저감 대책의 마련이 필요하다. 본 연구에서는 천연 추출물 복합체인 바 이오매직®의 양돈장 악취 저감 효과와 그에 따른 부수적 인 생리학적 개선 효과를 평가하였다. 본 실험에 사용된 바이오매직®은 오렌지, 파파야, 라임을 발효하여 제조한 것으로, 우선 돼지 농장에서 분리된 메티실린 내성 황색 포도상구균(MRSA)을 대상으로 항균 활성을 평가하였다. CLSI 기준에 따른 최소억제농도(MIC) 및 최소살균농도 (MBC) 시험 결과, 바이오매직®은 15개 MRSA 균주에 대 해 낮은 MIC와 MBC 값을 나타내어 우수한 항균 활성을 보였다. 양돈장 내 주요 악취 유발 물질인 암모니아(NH3), 황화수소(H2S), 이산화탄소(CO2)에 대한 저감 효과를 조사 한 결과, 바이오매직®을 살포한 후 NH3와 C O2 농도가 유 의적으로 감소하였으나(P<0.05), H2S는 연구 전기간에 걸 쳐 측정되지 않았다. 또한, 돼지의 스트레스 수준은 혈중 코르티솔 농도를 효소면역측정법(ELISA)으로 측정하여 평 가하였다. 그 결과, 실험군의 코르티솔 농도 감소율이 대조 군에 비해 유의적으로 높게 나타났다(P<0.05). 스트레스 감 소에 따른 면역력 향상 여부를 평가하기 위해 유세포 분석 (flow cytometry)을 실시한 결과, 항원제시세포의 비율이 다 소 증가하였고, CD4-T cell 비율이 유의적으로 증가하였다 (P<0.05). 이상의 결과를 종합하면, 본 실험에서 사용한 바 이오매직®은 양돈장에서 악취 저감뿐만 아니라 돼지의 스 트레스 완화 및 면역 기능 향상에도 긍정적인 효과를 나타 내었다. 이러한 개선 효과는 폐사율 감소 및 생산성 향상, 나아가 동물 복지 증진에 기여할 수 있을 것으로 판단된다.
Fenbendazole (FBZ) is one of benzimidazole drugs, which is well known for its broad spectrum of anthelmintics. During recent research on FBZ, it is also expected to have anti-inflammatory characteristics. However, the related research on FBZ with its anti-inflammatory effects is still lacking. This study focuses on the effects of FBZ on macrophage cell line of mouse, RAW264.7, and also on the inflammatory condition induced by lipopolysaccharide (LPS). FBZ alone treatment on RAW264.7 cells reduced the metabolic activity on the range of 1-5 μM, but the metabolic activity in the presence of LPS was higher than that of absence of LPS. Given that LPS has the ability to enhance glucose uptake in macrophage and rapid ATP production, there is a possibility that LPS increases the metabolic activity. Whereas TNF-alpha was hardly produced at FBZ 1-5 μM in the presence of LPS, IL-10 was still being produced, suggesting on the possibility of anti-inflammatory effect. Interestingly, despite the decreased survival of cells treated with FBZ, MHC class II and CD86 expression in surviving cells was markedly increased. Further investigation of FBZ is required through in vivo experiments to validate these findings.
현대 사회의 산업화와 서구화된 생활습관으로 인해 만성 염증성 질환이 급증하고 있으며, 염증성 사이토카인의 과도한 분비가 다양한 질환의 근본 원인으로 주목받고 있다. 본 연구는 국내외 학술 데이터베이스를 활용하여 최근 10년간 발표된 염증성 사이토카인 관련 연구와 약용식물의 항염 효과에 관한 논문을 체계적으로 수집·분석 하고, 전통 본초 문헌에 기록된 약재의 효능과 현대 생명과학·약리학적 연구 결과를 비교 검토하였다. 연구 결과, IL-6 억제 효과를 나타내는 한약재로는 상황, 방기, 신 이, 적소두가 확인되었으며, TNF-α 억제 효과를 보이는 한약재로는 모과, 황금, 인 동등, 자초가 규명되었다. 이들은 RAW 264.7 대식세포와 다양한 염증 모델에서 염 증성 사이토카인 발현을 유의하게 억제하였으며, NF-κB 및 MAPK 신호 경로 차 단을 통한 분자적 기전이 입증되었다. 전통적으로 해열, 해독, 활혈, 소종의 효능으 로 기록된 이들 약재의 항염 효능이 현대 분자생물학적 기전을 통해 과학적으로 입 증되었으며, 이는 전통 지식이 과학적 근거를 내포하고 있음을 시사한다. 따라서 해 당 약재들은 항염증 보조치료제, 기능성 식품, 천연물 신약 개발의 유망한 후보군이 될 수 있으며, 향후 통합적 염증 치료 전략 수립에 기여할 것으로 기대된다.