• Journal of Veterinary Clinics
• /
• v.14 no.2
• /
• pp.230-234
• /
• 1997

In vitro effect of cortisol on the proliferation of canine peripheral blood mononuclear cells (MNC) was examined. The MNC was isolated from peripheral blood by a gradient centrifugation with Picoll-Hypaque. The cell proliferation assayed using a noneradioactive 5-Bromo-2'-deoxy-uridine (BrdU) kit. The MNC proliferated well in response to either phrtobeRagg$]$utinin-p (PHA-P) or culture supernatant from MNC stimulated with PHA-p. However, these proliferative responses of MNC were not affected by addition of coitisol of 1 to 1,OOfl ng/ml. The addition of cortisol in MNC culture with either PHA-P or corture supernatBnt from MNC stimulated with PHA-P far 4 days wag not also influenced on the viabilities of cultured MNC. In conclusions it was able to assay the cell proliferation with BrdU instead of radioactive isotope e.g. tritiated thymidine (3H-TdR). These results suggested that cortisol does not at least influence on MNC proliferation in vitro.

• Journal of Veterinary Clinics
• /
• v.15 no.1
• /
• pp.110-115
• /
• 1998

인삼 PD saponin(GPD)으로 배양한 고양이 말초혈액 단핵구세포(afNC) 배양상충 액에서 말초혈액 다형핵백혈구(PMNC)에 대한 interleukin(IL) 8 양 유주활성에 잔하여 검토 하였다. PMNC의 유주활성은 Hoyden chamber 변법으로 측정하였다. GPD를 첨가하여 배양 한 MNC배양상층액중에는 rMNC에 대한유주활성이 인정되었다. PMNC에 대하여 GPD 로 배양한 MNC 배양상층액중에 존재하는 유주활성이 IL 8 양 물질인지를 알아보기 위해 human recombinant IL 8을 이용하여 고양이 PMNC에 대해 유주팔성을 측정한 결과, GPD 로 배양한 MNC 배양상층액의 경우와 동등한 활성이 나타났다. Human IL 8 mAb를 사용하 여 GPD로 배양한 MNC 배양상층액중의 유주활성에 대한 중화반응을 살펴본 결과, GPD로 배양잔 MNC 배양상충액 및 human IL 8에 의해 증가되었던 PMNC의 유주활성은 IL 8 cAb의 첨가농토가 증가함에 따라 활성이 완전히 억제되었다. 또한 GPD로 배양한 고양이 MNC 배양상충액중의 유주활성은 열처리(4, 30, 37, 60 및 $100{\circ}C$) 및 산(pH 3.0)과 알카리 (pH 9.0)처리에도 안정성을 보여 human IL 8의 물리화학적 성상과 매우 유사하였다. 따라서 GPD로 배양한 MNC 배양상충액중에 존재하는 고양이 PMNC에 대한 유주활성은 feline IL 8 양 물질임을 강하게 시사하였다.

• Journal of Veterinary Clinics
• /
• v.16 no.1
• /
• pp.31-36
• /
• 1999

The aim of this study is to determine the phagocytosis-promoting factor(s) for feline peripheral blood polymorphonuclear cells (PMN) by culture supernatant from mono-nuclear cells (MNC) treated with egg white derivatives (EWD). The phagocytic activity of PMN was analyzed by a flow cytometry system. The EWD did not show direct effect on the phagocytic response of PMN. The phagocytic activity of PMN was enhanced by culture supernatant from MNC but not PMN treated with EWD. Therefore, it was suggested that the enhanced phagocytic activity of feline PMN could be mediated by humoral factor(s) released from MNC treated with EWD. Thus, the phagocytosis-promoting factor(s) in supernatant fraction from MNC culture treated with EWD were isolated by reverse phase high pressure liquid chromatography. The resulting supernatant fraction on 29.02 minutes of retention time showed high phagocytic activity of PMN. The molecular weight of this supernatant fraction was 16 to 18 kDa when analyzed by capillary electrophoresis. The isoelectric point was pH 5.76 when assessed by ion-exchange chromatography. These results suggest that EWD stimulates feline MNC to elaborate a phagocytosis-promoting factor, 16 to 18 kDa of molecular weight, which could be an important mediator for the enhancement of phagocytic activity of feline peripheral blood phagocytes. Further study will be needed to elucidate this phagocytic factor.

• Korean Journal of Veterinary Research
• /
• v.40 no.2
• /
• pp.393-401
• /
• 2000

The feline chemotactic factor(s) for polymorphonuclear cells (PMN) in culture supernatant from mononuclear cells (MNC) treated with egg white derivatives (EWD) were examined. Culture supernatant from MNC treated with EWD and human recombinant (hr) IL-8 remarkably enhanced chemo-taxis of feline PMN. To investigate feline chemotactic factor(s), gel electrophoresis was performed with culture supernatant from MNC treated with EWD under denaturing (18% loading gel/5% stacking gel) and nondenaturing (12.5% loading gel/5% stacking gel) condition. Hr IL-8 and culture supernatant from MNC treated with EWD yielded a distinct band in a molecular weight, 6 to 8 kDa. Eluted solution from gel slices of 6 to 8 kDa band in denaturing condition also enhanced feline PMN chemotaxis. These chemotactic activities of feline PMN induced by culture supernatant from MNC treated with EWD, hr IL-8 and eluted solution were inhibited in a dose-dependent manner by rabbit anti-feline polyclonal IgG (RAF pIgG) and monoclonal antibody (mAb) against hr IL-8. RAF pIgG also showed a binding activity with hr IL-8, suggesting that RAF pIgG against feline IL-8-like chemotactic factor(s) had cross-reactivity with human IL-8. These results suggested that feline MNC treated with EWD might release feline IL-8-like chemotactic factor(s) with a molecular weight, 6 to 8 kDa, which induces the chemotaxis of feline PMN.

• Journal of Veterinary Clinics
• /
• v.16 no.1
• /
• pp.37-41
• /
• 1999

The immunostimulating effects of egg white derivatives (EWD) on the phagocytic response of feline peripheral blood polymorphonuclear cells (PMN) as well as mono- nuclear cells (MNC) were examined. The phagocytic activity was analyzed by a flow cytometry system. The EWD showed directly an enhanced effect on the phagocytic response of MNC but not PMN. The phagocytic activity of MNC was enhanced by culture supernatant from MNC and PMN treated with EWD, respectively. Similarly, the phagocytic activity of PMN was enhanced by culture supernatant from MNC but not PMN treated with EWD. It was, therefore, indicated that the enhanced phagocytic activity of feline PMN could be mainly mediated by humoral factor(s) released from MNC treated with EWD. These results suggested that EWD could enhance the phagocytosis of feline peripheral blood phagocytes.

• Journal of Veterinary Clinics
• /
• v.15 no.1
• /
• pp.116-123
• /
• 1998

고양이 말초혈액 탐식세포(단핵구세포(MNC) 및 다형핵백혈구(PMNC))의 탐식능 에 있어서 인삼 saponin(ginseng total saponin(G75), ginseng PT saponin(GPT) 및 ginseng PD saponin(GPD))의 면역증강 효과를 flow cytometry를 이용하여 분석하였다. 인삼 ssponins을 직접 첨가하여 배양한 MNC 및 PMNC에서는 탐식중강 효과가 나타나지 않았다. 각각의 인삼 saponin을 첨가하여 배양한 PMNC 및 MNC 배양상충액의 존재하에 PMNC 및 MNC의 탐식능을 겅토한 결과, MNC의 탐식능은 Gff 첨가 PMNC 배양상충액과 GTS 및 GPT 첨가 MNC 배양상충액의 존재하에서 약간의 탐식증강 효과를 보였다. PMNC 탐식능의 경우에는 GPD 첨가 PMNC 배양상충액에서 미약한 탐식증강 효과가 나타났으나, 각각의 인 삼 saponin 첨가 MNC 배양상충액 존재하에서는 모두 현저한 탐식중강 효과를 나타내었다. 이상의 결과로부터 고양이 말초혈액 탐식세포의 탐식증강 효과는 인삼 saponin의 직접적인 작용보다는 인삼 saponin에 의해 활성화된 단핵구세포에서 분비되는 가용성물질에 의해 단 핵구세포보다는 다형핵백혈구에서 현저하게 탐식효과가 증강되는 것으로 판단되었다.

• Journal of Veterinary Clinics
• /
• v.20 no.1
• /
• pp.1-6
• /
• 2003

Immunoenhancing effects of conjugated linoleic acid (CLA) isomers (l0t-l2c CLA, 9c-11t CLA, CLA mixture, 9c-11c CLA and 9t-11t CLA) on chemotactic activity of porcine peripheral blood polymorphonuclear cells (PMN) were examined. The chemotactic activity of PMN was evaluated by a modified Boyden chamber assay. CLA isomers at higher concentration of 50 to 200$\mu$M exhibited a low viability of cells by trypan blue exclusion. CLA isomers were used at concentration of 20uM showing no cytotoxic effect and high cell viability. CLA isomers themselves were not active or slight chemotactic for PMN. But culture supernatant from mononuclear cells (MNC) treated with 10t-12c CLA, 9c-11t CLA and CLA mixture except for 9c-11c. CLA and 9t-11t CLA enhanced remarkably chemotactic activity or porcine PMN PMN migration by culture supernatant from MNC treated with CLA mixture was found to be true chemotaxis by checkboard assay. This migration was also induced by porcine recombinant interleukin (rIL)-8. PMN chemotaxis caused both culture supernatant from MNC treated with CLA mixture and porcine rIL-8 was inhibited in a dose-dependent manner by addition of anti-porcine IL-8 polyclonal antibody. Therefore, these results strongly suggested that CLA (10t-12c CLA, 9c-11t CLA and CLA mixture) could stimulate porcine MNC to release and IL-8 like chemotactic activity.

• Journal of Veterinary Clinics
• /
• v.15 no.1
• /
• pp.104-109
• /
• 1998

고양이 말초혈액 다형핵백혈구(PMNC)의 유주성에 있어서 인삼 saponins(ginseng total ssponin(GTS), ginseng PT saponin(GfT) 및 ginseng PD saponin(GPD))의 면역증강 효과를 검토하였다. PMNC에 대한 유주성을 hoyden chamber 변법으로 측정한 결과, GTS, GPT, GPD는 PMNC에 대해 직접적인 유주활성을 나타내지 않았다. 또한 인삼 saponins을 첨가하여 배양한 PMNC 배양상충액의 경우게는 미약한 유주활성을 나타내었다. 그러나 GPf및 GPD를 첨가하여 배양한 말초혈액 단핵구세포(MNC)배양상충액에서는 PMNC의 유주활성이 현저하게 증가하였다. PMNC에 대해 유주활성이 인정된 GPI)로 배양한 MNC 배양상충액을 이용하여 checkerboard assay를 실시한 결과 배양상충액의 농도차에 의존하는 진의 유주활성임을 알 수 있었다. 이상의 결과로부터 인삼 saponine 중 GPT 및 GPD가 고양 이 말초혈액 다형핵백혈구의 유주성을 중강시키는 효과가 있으며, 이것은 이들 saponins에 의해 활성화된 단핵구세포에서 분비되는 유주성인자에 의해 다형핵백혈구의 유주활성이 증 강되는 것으로 사료되었다.

• Journal of Veterinary Clinics
• /
• v.20 no.4
• /
• pp.437-442
• /
• 2003

To examine the in vivo immunostimulating effect of conjugated linoleic acid (CLA) in pigs, the change of peripheral blood cells and the phagocytic response of phagocytes were evaluated. Spayed male pigs, 80 kg of average body weight, fed a diet containing either 0.5% 10t-12c CLA or 0.5% CLA mixture (mostly 9c-11t CLA and 10t-12c CLA) for 4 weeks. The change of blood cell values (PCV, WBC, differential count of WBC) and the phagocytic activities of phagocytes were evaluated on week 0, 2, 4, and 5, respectively. There were no change in the PCV values regardless of CLA supplement. The number of WBC, especially neutrophils, in pigs fed a diet with CLA was significantly increased (p<0.05 to 0.01) when compared with control pigs fed a diet without CLA. The phagocytosis of peripheral blood mononuclear cell (MNC) and peripheral blood polymorphonuclear cells (PMN) were analyzed by a flow cytometry system. There was no change in the phagocytic activity of MNC and monocyte-rich cells regardless of CLA supplement. However, the phagocytic activity of PMN composed by approximately 95% neutrophils was remarkably increased (p < 0.05 to 0.01) on week 2, 4, and 5 as compared wth control pigs. These results suggested that supplement of CLA into pigs induces the increase of neutrophil number and the enhancement of neutrophil phagocytosis.

• 한국생물공학회:학술대회논문집
• /
• 2003.04a
• /
• pp.95-97
• /
• 2003

Despite recent advances in the treatment of acute myocardial infarction, the ability to repair extensive myocardial damage is limited. To develop a new therapy for myocardial infarction, we examined the possibility of regenerating myocardium by implanting bone marrow-derived mononuclear cells(BM-MNC) . Histological and immunohistochemical examination showed myocardium regeneration and angiogenesis in the cell transplantation site. Isolated perfused (Langendorff) heart experiments revealed enhanced functions of heart. These results suggest that BM-MNC transplantation induce cardiac muscle regeneration and that this approach could be applied as a possible treatment for myocardial infarction.