Until now, problems related to shortage of organ for transplantation have been continuing. Pigs are the most suitable animal for xenotransplantation. Although primates are most similar to humans, they are not suitable because they have low productivity. Pigs are more productive than primates, and their organ size and physiological characteristics are similar to humans, with the exception of primates. In this study, we breeding the transgenic minipigs using natural mating to produce transgenic pigs. And, transgenic pigs has transmission rate that follow mendel’s rule. There are 20% hDAF gene, 20% US11 gene and 50% both hDAF and US11 gene in transgenic offsprings. Furthermore, transgenic pigs followed normal litter size, and piglets also has normal sex ratio. To suppress the immune function, experiments were performed using porcine ear fibroblast that transfected with hDAF and US11gene. In Cytotoxicity experiment against human complement, hDAF gene and double transgenic cell with both hDAF and US11 gene showed effect to reduce cytotoxicity rate in all of human complement condition. US11 gene and double transgenic cell were significantly reduce the cytotoxicity ratio in human NK cell. Besides, hDAF gene transgenic cell also reduce immune response in 10:1 concentration of human NK cell. In conclusion, natural mating was efficient method for breeding transgenic pigs. And, hDAF and US11 genes has effect for reduce cytotoxicity against human NK cell and human complement conditions.

Toll and IMD pathways play an important role in producing antimicrobial peptides (AMPs) through NF-κB in insects. The functions of IκB kinase (IKK) complex regulating the NF-κB signaling cascade have not yet been investigated in Tenebrio model. Here, we identified TmIKK-β (or TmIrd5) which contains 2,112 bp encoding 703 amino acid residues. Domain analysis shows that TmIKK-β contains one Serine/Threonine protein kinases catalytic domain. Developmental expression patterns indicate that TmIKK- β gene was highly expressed in early pupal (P1) and adult (A5) stages. Tissue specific profiles show that TmIKK-β was highly expressed in the integuments in last instar larvae, and fat body and hemocytes in 5 day-old adults. TmIKK-β1 transcripts were strongly induced at 3 and 12 h-post injection of E. coli, and 3 h-post injection of S. aureus or C. albicans in hemocytes. In gut, TmIKK-β transcripts were slightly induced by E. coli (at 6, 9 and 24 h) and C. albicans (at 24 h), while it was not induced by S. aureus challenge. Moreover, it was highly induced at 6 h-post injection of E. coli and then it was gradually decreased in the fat body. To understand the immunological role of TmIKK-β, gene specific RNAi and mortality assay was performed. Depletion of TmIKK-β mRNA leads to increase microbial susceptibility of larvae against E. coli, S. aureus and C. albicans. In addition, induction patterns of fourteen AMP genes in response to microbial challenge was tissue specifically investigated in TmIKK-β–silenced T. molitor larvae. The results suggest that expression of ten AMP genes out of fourteen genes were drastically decreased by TmIKK-β RNAi in fat body, suggesting that TmIKK-β plays an important role in antimicrobial innate immune responses.

IKK-γ is an essential protein to form IKK complex which regulate NF-κB. We identified TmIKK-γ (or TmKenny) gene which has 1,521 bp of nucleotides encoding 506 amino acid residues. Domain analysis of TmIKK-γ shows that there are one NF-κB essential modulator (NEMO) domain and a leucine zipper domain. Expression of TmIKK-γ gene was gradually increased from egg to 2-day-old pupal stage, dramatically decreased until 7 day-old pupal stage, and then it was gradually increased. TmIKK-γ transcripts were highly expressed in fat body and hemocytes in late instar larvae and integuments, fat body and Malpighian tubules in 5 day-old adult. TmIKK-γ was drastically induced by E. coli after 3 h challenges and by S. aureus at 3 and 12 h-post injection in hemocytes. TmIKK-γ was not induced by C. albicans although it was significantly induced by E. coli (at 3, 6 and 24 h) and S. aureus (at 9 h) in gut. In fat body, expression of TmIKK-γ was drastically induced by E. coli at 3 and 24 h-post injection while it was not significantly induced by S. aureus and C. albicans. To understand the immunological role of TmIKK-γ, gene specific RNAi and mortality assay was performed. larval mortality against microbial challenge was dramatically increased by TmIKK-γ RNAi. Furthermore, we investigate the tissue specific induction patterns of fourteen AMP genes in response TmIKK-γ dsRNA-treatment. In fat body, ten AMP genes out of fourteen was not significantly induced by microbial challenge in TmIKK-γ dsRNA-treated group. Based on these results, TmIKK-γ might play an important role in antimicrobial innate immune responses in Tenebrio molitor.

본 연구는 열 스트레스 (heat stress, HS)에 노출된 브로일러에서 혼합 생균제의 급여가 혈 액 생체지표, 면역반응, 맹장 미생물 및 생산성에 미치는 영향을 조사하였다. 총 400마리의 브로일러 수 컷 (Ross 308)을 각각 100마리씩 4그룹, C (대조군, 실온 25℃), HS (열 스트레스 33℃), HSP (HS플 러스 혼합 생균제 500, 750 mg/kg) 그룹으로 배치하였다. 브로일러의 증체량, 사료섭취, 사료요구율 및 면역기관 무게는 HSP 그룹에서 HS 그룹과 비교했을 때 증가하였다. 혈액 IgG, lymphocytes 농도는 HS 그룹에서 HSP그룹과 비교했을 때 증가하였고, heterophil과 lymphocyte(H:L)비율, 코르티코스테론 농도 및 폐사율은 낮았다. 맹장의 Lactobacillus는 HS 그룹과 비교했을 때 HSP 그룹에서 증가하였으나 Escherichia coli (E. coli), coliform bacteria, aerobic bacteria는 낮아졌다. 본 연구결과는 열 스트레스 에 노출된 브로일러에게 B. subtilis, S. galilaeus 및 Sphingobacteriaceae등 3가지 균주가 포함된 혼합 생균제를 급여해주면 면역반응 증진, 미생물 균형을 유지해줌으로써 폐사율을 낮추고 생산성을 개선할 수 있음을 시사해준다.

본 연구는 열 스트레스 (heat stress, HS)에 노출된 산란계에서 혼합 생균제의 급여가 생산성, 계란품질, 면역반응, 맹장 미생물 및 분 암모니아에 미치는 효과를 조사하였다. 총 400마리의 50주령 Hy-Line brown 산란계를 무작위로 각각 100마리씩 4 그룹, C (대조군, 실온 25℃), HS (열 스트레스 33℃), HSP (HS 플러스 혼합 생균제 500, 750 mg/kg)로 배치하였다. 산란계의 생산성, 계란품질, 비장 무게, 혈액 IgG 및 lymphocyte 농도는 HSP 그룹에서 HS 그룹과 비교했을 때 증가하였고, 코르티코스테론, heterophil과 lymphocyte의 비율 (H:L) 및 폐사율은 유의하게 낮았다. Lactobacillus는 HSP 그룹에서 HS 그룹과 비교했을 때 증가하였으나 Escherichiacoli (E. coli), coliform bacteria 및 aerobic bacteria는 유의하게 낮았다. 분에서 암모니아 발생은 HS 그룹이 HSP 그룹에 비해서 유의하게 증가하였다. 결론적으로, 이러한 혼합 생균제가 여름철 산란계의 더위 피해를 방해주고 면역반응, 맹장 미생물 균형을 경유하여 생산성, 계란품질 및 계분으로부터 악취 발생을 줄이는 데 효율적인 영양전략이 될 수 있음을 나타낸다.

We investigated the effects of two Brucella proteins expressed in a pMAL expression system, RocF and EF-Ts, as subunit vaccines on immune modulation and protective efficacy using a mouse model. Mice vaccinated with MBP-RocF and MBP-EF-Ts displayed increased production of TNF, IFN-, MCP-1, IL-10 and IL-6, and TNF and MCP-1, respectively. Furthermore, mice vaccinated with MBP-EF-Ts showed decreased induction of IFN- and Th2-related cytokines, IL-10 and IL-6. Higher proportions of CD4+ and CD8+ T cells were observed in the blood of mice vaccinated with MBP-RocF than in the PBS-vaccinated group, although the increases were not significant. Furthermore, significantly reduced Brucella proliferation in the spleens of the MBP-RocF and MBP-EF-Ts groups were observed, but inflammation of these organs was not attenuated. Overall, these results indicate that RocF and EF-Ts could be potential subunit vaccine candidates against animal brucellosis.

곤충의 면역반응에 대한 연구는 곤충 체내 침입한 미생물들과 직접 반응하는 기작들을 중심으로 연구되었다. 그러나 미생물들이 곤충 체내에 침입 한 후 발생되는 다양한 미생물 분비물질에 의한 곤충 면역반응의 시작여부 등에 대한 연구는 거의 없는 실정이다. 이를 위하여 흰점박이 꽃무지(Protaetia brevitarsis seulensis) 유충의 장내에 존재하는 공생균과 체외 병원균을 동일한 조건에서 배양 하고 다양한 분비물질들이 존재 할 거라 예상되는 배양액만을 분리, 유충에 주사하여 면역반응 여부를 조사하였다. 공생균 배양액을 주입한 유충들은 비교적 건강하고 면역반응도 발생하지 않았으나 병원균 배양액을 주입한 유충의 경우 150시간 후 60% 이상 사망하였고 주사된 자리도 짙은 갈색의 멜라닌화가 관찰 되었다. 이러한 면역반응은 과립혈구세포의 리소좀(Lysosomes) 활성화 여부로 재확인 하였다. 병원균 배양액이 주입된 유충들의 경우 12시간 후 리소좀 이 ~50% 이상 활성화 되었으나 공생균 배양액이 주입된 유충들의 경우 ~5% 미만으로 활성화 되는 것으로 나타났다. 따라서 공생균 배양액내에 는 기주면역반응을 유도하는 물질들이 없거나 량이 매우 적게 존재하는 것을 추측 할 수 있었다.

The present study investigated the effects of dietary supplementation of purified amino acid (PAA) isolated and purified from animal slaughter house blood on growth performance and immune response of weanling piglets. A total of fifteen, 28 days old, weaned pigs were randomly distributed into three treatment groups, 5 pens per treatment (one head per pen). Each experimental diet was provided to five piglets per group (T0, 0% PAA; T1, 0.1% PAA; T2, 0.5% PAA). The feed consumption was calculated daily and body weight on a weekly basis. The blood samples were analyzed for plasma concentration of biochemical parameters and cytokines using ELISA Kit assay. Pigs fed with the 0.5% PAA have greater average daily weight gain (ADG) and feed efficiency (FE) as compared to those of T0 and T1. However, ADG and FE of T0 and T1 have no remarkable differences. The plasma cytokine levels were lower in T2 as compared to T0 and T1. The blood parameters like total bilirubin, blood urea nitrogen (BUN), glutamate oxalate transaminase (GOT) and glutamate pyruvate transaminase (GPT) were within the normal range in all the treatment groups. The study indicated that supplementation of 0.5% PAA have positive effect on the growth performance of the weanling pigs. Moreover, supplementation of the amino acid isolated and purified from animal slaughter house blood has no adverse effect on palatability of the feed and health of the animals.