Bropirimine, a class of antineoplastic agents, is known as one of the potent immunomodulators and is currently under clinical development for the treatment of cancer. However, the effect of bropirimine on the cow remains unknown as a therapeutics agent. In this experiment, the effect of bropirimine in the peripheral blood mononuclear cells (PBMCs) stimulated by lipopolysaccharide (LPS) or concanavalin-A (Con-A) was examined. Jugular venous blood was collected from Korean Hanwoo calves and PBMCs were isolated. It was used to study the effect of bropirimine upon stimulation with LPS or Con-A for 72 hours. The expression pro-inflammatory cytokines like Tumor Necrosis Factor α (TNF-α) and Interferon γ (IFN-γ) were confirmed. Bropirimine significantly inhibited LPS- or Con-A-induced TNF-α and Con-A-induced IFN-γ in dose-dependent manner. Furthermore, Bropirimine inhibited TNF-α and Con-A mRNA expression at the transcription level. These results clearly indicated that bropirimine inhibited LPS or Con-A stimulated up-regulation of proinflammatory cytokines in a dose-dependent manner without conspicuous cytotoxicity. The bropirimine has potential to protect cow from LPS or Con-A induced endotoxin shock, possibly through inhibition of the production of proinflammatory cytokines. It suggesting that bropirimine may be a novel therapeutic agent for the prevention of inflammatory diseases. This result revealed specific features of the immune responses depending on the bropirimine compound and would help to knowledge of bovine immunity.

In photobiophysics, biophoton means a kind of biological energy which enhances most metabolisms of the body. To investigate the immuno-enhancing effects of biophoton energy projector (BEP) producing light energy, pigs were irradiated with BEP for 8 weeks. Swine peripheral blood mononuclear cells (sPBMCs) were isolated from the blood of irradiated pigs. In this study, the antigen uptake and mitochondrial membrane potential of sPBMCs were measured by flow cytometric analysis. The irradiation of BEP increased the antigen uptake of sPBMCs. For functional analysis, the production of Bordetella bronchiseptica-specific IgG, measured using antigen-specific ELISA, was increased during the period of BEP irradiation. Taken together, the results suggest that the irradiation of BEP has immune-enhancing effects on sPBMCs.

Hematopoietic stem cells (HSCs) are the self‐renewing, multipotent progenitors that give rise to all types of mature blood cells. The hallmark properties of HSCs are the ability to balance self‐renewal versus differentiation cell fate decisions to provide sufficient primitive cells to sustain haematopoiesis, while generating more mature cells with specialized capacities. In the present experiment, we optimized the techniques for isolation and identification of hematopoietic stem cells from cow peripheral blood. The objective of this study was to optimize the more accurate methodology for separation of mononuclear cells (MNCs) from peripheral blood and identification of HSCs by using a specific cell surface marker i.e. CD34. A total 10 peripheral blood samples were collected from Holstein dairy cows from jugular vein. We used Ficoll 400 in different concentrations from 1 to 12% and Ficollpaque Plus (1.077 g/ml) at different centrifugation speed and time. After Giemsa staining, we found more than 98% recovery of monocytes with Ficollpaque Plus (1.077 g/ml). It was demonstrated that Ficollpaque Plus (1.077 g/ml) and centrifugation at 400xg for 30 min is the best method for separation of MNCs from bovine peripheral blood. Separated MNCs were immediately subjected for magnetic activated cell sorting (MACS) by using CD34 microbead kit. HSCs (CD34+ cells) recovery was 0.307% of peripheral blood. Peripheral blood MNCs and CD34+ cells were morphologically characterized by Giemsa staining. CD34+ cells were also confirmed by immunochemistry using FITC conjugated CD34 antibodies. HSCs were also confirmed by progenitor assay including burst forming unit‐erythroid (BFU‐E), colony forming cells‐ granulocyte (CFC‐G), colony forming cells‐ macrophage (CFC‐M), colony forming cells‐ granulocyte macrophage (CFU‐GM) and colony forming cells‐ granulocyte erythroid macrophage monocyte (CFCGEMM) on Methocult 4435.

The present study were performed to analysis the hematocrit and the red blood cells content into the blood plasma of the transgenic pigs harboring recombinent human erythropoietin gene (rhEPO). Mouse whey acidic protein (mWAP) linked to rhEPO gene was microinjected into pronuclei of porcine one-cell zygotes. After delivered of offspring, PCR analyses identified one mWAP-rhEPO transgenic founder offspring(F/sub 0/). The first generation of transgenic pig (F/sub 0/) harboring mWAP-hEPO appeared to be a male, and the second generation (F₁) pigs were made by natural mating of F/sub 0/ with domestic swine, and male and female transgenic pigs (F₁) were identified by PCR. The blood samples from transgenic and normal pigs were collected for 50 days during lactation and were counted the red blood cell (RBC) numbers and Hematocrit (HCT) content into the blood. The transgenic pigs expressing rhEPO in their blood gave rise to higher RBC numbers and HCT contents than control animals. rhEPO was secreted both in the blood and milk of genetically engineered pigs harboring rhEPO gene. Therefore, this study provides a model regarding the production of transgenic pig carrying hEPO transgene for biomedical research.

한국산 겨우살이 (Viscum album)는 면역 활성효과와 항암효과가 있다는 것으로 잘 알려져 있다. 본 연구에서는 한국산 겨우살이 추출물 M11C (비렉틴 구성물질)가 사람의 말초혈액 단구를 활성화시켜 tumor necrosis factor-alpha (TNF-α)를 생산 분비하게 하는지를 규명하기 위해 실험에 이용되었다. 단구로부터 TNF-α의 생산에 있어 M11C의 효과를 알기 위해 단구를 여러 농도 (0-2000μg/ml)의 M11C로 0.5-24시간 동안 자극한 후 배양액 MCM을 수거했다. 수거한 배양액 MCM을 TNF-α에 민감한 L929 세포에 첨가한 후 L929 세포의 독성 정도를 MTT 기법으로 검사하였는데, 배양액 MCM이 L929 독성효과를 가졌으며, 이 MCM의 L929 세포 독성효과는 TNF-α 항체에 의해 거의 완전하게 억제되었다. 이러한 결과는 배양액 MCM에 TNF-α가 존재함을 지적했으며, 또한 M11C가 단구를 자극해 TNF-α를 생산하리라고 암시했었다. 이러한 암시를 확인하기 위해 immunoblotting 기법을 사용해 배양액 MCM에 존재하는 TNF-α을 검사를 수행했다. 검사결과 배양액 MCM에 많은 량의 TNF-α가 검출되었다. 이러한 TNF-α 검출량은 단구를 자극한 M11C의 농도와 자극시간에 비례해서 증가했다. 즉, 단구로부터 TNF-α의 생산 분비량은 M11C의 농도와 자극시간에 의존적이었다. 이러한 M11C의 농도와 자극시간의 의존적 효과는 단구로부터 TNF-α mRNA 발현에도 같은 경향을 보였다. 결론적으로 한국산 겨우살이로부터 열탕 추출한 M11C가 면역활성제로 작용할 수 있음을 시사함과 동시에 한방탕제로 사용되는 한국산 겨우살이가 면역학적 효능을 가지고 있음을 말해주고 있다.