• 대한수의학회지
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• 제36권2호
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• pp.337-348
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• 1996

The proportion of leukocyte subpopulation in the host is of a great importance in understanding their functions and disease progress. Many methods have been developed to seperate leukocytes and to measure their activities. Characterization of immune cell subpopulations in Korean native cattle was performed using a set of monoclonal antibodies specific which are specific to bovine leukocyte differentiation antigen. Peripheral blood leukocytes from fifty Korean native and ten Holstein cattle were collected and analyzed for the investigation of leukocyte subpopulation by using monoclonal antibodies and flow cytometry. The result indicated that Korean native cattle have significantly higher proportion of leukocyte subpopulations expressing MHC class II molecules and BoCD4 than Holstein cattle.

• 대한수의학회지
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• 제34권2호
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• pp.275-286
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• 1994

Plasma protein which has been known as one of nonspecific immunostimulators was added to feedstuff to examine its effect on the enhancement of cellular immune response in porcine immune system. A total of 40 piglets, 20 male and 20 female each, were fed for 30 days with or without plasma protein. The peripheral blood were collected and analyzed for the investigation of leukocyte subpopulations and their activities by using a panel of monoclonal antibodies specific to porcine leukocyte differentiation antigens and flow cytometry. The results obtained as follows. 1. Total weight gain, daily feed intake and feed conversion rate for 10 days were significantly improved to 56%, 20% and 22% in the piglets fed plasma protein, respectively. 2. A significant increase in N (null or non T/non B) cells was also noticed. Leukocyte proportion from piglets fed plasma protein was 20.2-24.7%, otherwise that from piglets fed without plasma protein was 12.3-13.4%, respectively. 3. A significant increase in the proportion of B cells and cells expressing poCD1 was not found in piglets fed plasma protein. 4. Reaction with monoclonal antibodies specific to adhesion molecules, poCD11a, poCD11b, poCD44 and poCD45A and poCD45B, has shown that leukocyte subpopulation from piglets fed plasma protein did not significantly higher than that from piglets fed without plasma protein. 5. Total proportion of granulocytes and monocytes was about 50% in both group and the proportion after treated with Hypaque/Ficoll was 2.7% and 5.8% in each group, respectively.

• 한국동물위생학회지
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• 제30권1호
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• pp.51-66
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• 2007

The purpose of this study was to evaluate the effect of a subsequent infection of porcine reproductive and respiratory syndrome(PRRS) virus to pigs with A pleuropneumonia. Twenty three 7-week-old commercial pigs were infected intratracheally with PRRS virus and/or A pleuropneumoniae serotype 5. Serum antibody titers were examined by an enzyme-linked immunosorbent assay(ELISA) and proportion of porcine leukocyte subpopulations in peripheral blood was examined by flow cytometry. In this experiment, antibodies against PRRS virus and A pleuropneumoniae were detected at 2 weeks and 1 week postinfection and the number of antibody positive pigs were gradually increased. And in proportion to leukocyte subpopulations in peripheral blood of pigs infected with A pleuropneumoniae compared with pigs administrated with saline, the proportion of PoCD4 and N cells were increased(P<0.1). Furthermore, in proportion to leukocyte subpopulations in peripheral blood of pigs infected with PRRS virus followed by A pleuropneumoniae compared with pigs administrated with saline, the proportion of MHC class II, PoCD4 and B cells were significantly increased(P<0.1). The results indicated that dual infection with PRRS virus and A pleuropneumoniae induced the stronger immune responses associated with macrophages and Th cells in pigs than single infection with PRRS virus or A pleuropneumoniae.

• 대한수의학회지
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• 제34권2호
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• pp.287-299
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• 1994

Plasma protein which has been known as one of nonspecific immunostimulators was added to feedstuff to examine its effect on the enhancement of cellular immune response in porcine immune system. A total of 40 piglets, 20 male and 20 female each, were fed for 30 days with or without plasma protein. The peripheral blood were collected and analyzed for the investigation of leukocyte subpopulations and their activities by using a panel of monoclonal antibodies specific to porcine leukocyte differentiation antigens and flow cytometry. The results obtained as follows. 1. Subpopulations expressing major histocompatibility complex(MHC) class I antigen were $96.2{\pm}3.1%$ and $86.6{\pm}3.8%$ in piglets fed with plasma protein and in piglets fed without plasma protein, respectively. 2. Proportion of leukocyte subpopulation expressing MHC class II antigens were significantly higher in the piglets fed with plasma protein than ones without plasma protein. The proportion was $27.6{\pm}3.6%$ and $16.6{\pm}2.2%$ in MHC class II DQ antigen, and $28.1{\pm}2.0%$ and $20.0{\pm}0.3%$ in MHC class II DR antigen, respectively. 3. A significant increase in the proportion of cells expressing poCD2 was not found in piglets fed plasma protein. 4. Proportion of subpopulation expressed porcine(Po) CD4 antigens which specific to helper T lymphocytes were not increased (18.3-19.1% vs. 25.6-28.8%), rather slightly decreased, in plasma protein-treated group. 5. The most important increase of proportion in plasma protein-treated group was the leukocyte subpopulation specific to $poCD8^+$ T cytotoxic/suppressor lymphocytes. The expression level was significantly higher up to 45.9-47.1% in plasma protein-treated group in comparing with 29.7-33.0% in non-plasma protein-treated group. 6. Lymphoblastogenetic responses using different concentrations of Con A mitogen and plasma protein has found that the responses of lymphocyte from piglets fed plasma protein was significantly activated (p<0.01). The activities measured by 3[H]-thymidine incorporation showed 3-6 times stronger in plasma protein-treated group than those in non-plasma protein-treated group. The study has concluded that plasma protein, which has known as a nonspecific immunostimulator, may have an immunoenhancing activities in porcine lymphoid system by increase the activated cell proportions and their blastogenetic properties which is critical to host immune responses.

• 대한수의학회지
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• 제38권1호
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• pp.91-99
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• 1998

The immunogenetic analysis was performed to characterize the Korean native chickens (KNC) determined by monoclonal antibodies specific to chicken leukocyte differentiation antigens and flow cytometry. A total of 174 chickens including 58 KNC (black, brown and darkbrown colored), 77 foreign breed (Nagoya, White Reghorn, Rhode Island and Cornish) and 39 mixed breed (19 KNC with Nagoya and 20 KNC with Rhode Island) separately growing at Animal Science and Technology Institute were examined. The proportion of cells expressing MHC class II molecule (B-L in chicken) was significantly high in KNC. Proportion of CD4+ T helper cells was also higher in KNC and two mixed breed than that in foreign breed. However, proportion of CD8+ cells and TCR1 + (${\gamma}^{\delta}$ T cell receptor) cells was the lowest among the breed examined. Otherwise, those proportions were significantly high in White leghorn and two mixed breeds with two exclusive subpopulations. The two subpopulations were also typically shown in MHC class $II^+$ cells in KNC and one mixed breed, black-colored KNC with Nagoya. Although genotypic analysis was not pursued to characterize the immunogenetic properties of KNC, difference of phenotypic expression based on leukocyte differentiation molecules could be elucidated in KNC in this study.

• 한국동물위생학회지
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• 제29권3호
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• pp.365-376
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• 2006

To identify immune response of leukocytes in peripheral blood of cattle vaccinated with an attenuated live Akabane virus vaccine, leukocytes were reacted with monoclonal antibodies which are specific to bovine lymphocyte surface antigens and assayed by the flow cytometry. Serum neutralizing (SN) test was used to measure antibody titers after vaccination, SN antibody was appeared to 7 days post-vaccination (PV) and 2-8 antibody titers were observed in 14 days PV. Proportion of $CD8^-$ MHC $class II^+$ expressing cells were rapidly increased at 3 days PV. $CD8^+$ MHC $class II^-$ cells were increased at 7 days PV. $CD4^+CD8^-,\;WC^+CD4^-,\;CD4^+CD8^+,\;WC1^-CD4^+, \;WC1^-CD8^+$, and $CD4^-CD8^+$ cells were highly increased at 3, 3, 7, 7, 14, 14 days PV, respectively.

• 대한수의학회지
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• 제45권4호
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• pp.501-505
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• 2005

Immu-Forte composed of chitosan, ${\beta}-glucan$, manno-oligosaccharide and pangamic acid was evaluated for its effectiveness as a nonspecific immunostimulator in mice. The effects of Immu-Forte were determined by analysis of cytokines using ELISA and phenotype of leukocyte subpopulations using monoclonal antibodies specific to mouse leukocyte differentiation antigens and flow cytometry. All T cells, all B cells, CD4 T cells, CD8 T cells, macrophages, IL-2, IL-4, IL-12 and IFN-r in Immu-Forte A-treated group increased in 1 months posttreatment and were significantly higher (p < 0.05) than that of control at 1 months posttreatment. All T cells, all B cells, CD4 T cells, CD8 T cells, macrophages and IL-2 in Immu-Forte EX-treated low and middle dose groups increased in 1 months posttreatment and were significantly higher (p < 0.05) than that of control at 1 months posttreatment. In the Immu-Forte soybean-treated group, NK cells and IL-4 were significantly higher in middle dose-treated group, and IL-2, IL-4 and IFN-r were significantly higher in low dose-treated group. In the Immu-Forte F-treated group, all T cells, all B cells, CD4 T cells, CD8 T cells, macrophages, NK cells, IL-2, IL-4, IL-12 and IFN-r in high dose-treated group and all T cells, all B cells, CD4 T cells, CD8 T cells, macrophages, IL-2, IL-4, IL-12 and IFN-r in middle dose-treated group and NK cells, IL-2, IL-4, IL-12 and IFN-r in low dose-treated group were significantly higher (p < 0.05) than that of control at 1 months posttreatment. In conclusion, this study has demonstrated that Immu-Forte had an immunostimulatory effect on mice through proliferation and activation of mouse immune cells.

• 대한의생명과학회지
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• 제1권1호
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• pp.19-25
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• 1995

소는 인간에게 고기와 우유를 제공하는 영양자원으로서 뿐만 아니라 노동력을 제공하는 중요한 가축 동물 중의 하나이다. 또한 사람의 질병을 규명하고 예방 및 치료의 실험적 도구로서도 많이 이용되고 있으나, 대동물이라는 점과 경제적인 이유로서 일반적인 응용과 실험적 적용에 어려움이 되어왔다. 저자들은 이와 같은 사정을 감안하여 실험적 기초 자료를 얻기 위하여, 한우를 대상으로 말초혈액상과 특히 림프아군의 특성을 알아본 결과 다음과 같이 일부 결과를 얻었다. 즉, 한우 22마리(암컷 12마리, 숫컷 10마리)의 말초혈액을 채혈하여 림프구 표면항원의 특성을 단클론항체와 반응시키고 flow cytometry로 측정한 결과 CD2는 숫컷에서 53%, 암컷에서 54%의 반응을 보였고, CD4는 숫컷에서 30% 암컷에서 32%의 반응을 보였으며, CD8에서는 숫컷에서 13%, 암컷에서 13%의 반응을 보였다. 그리고 말초혈액상은 RBC가 숫컷 7.20$\times10^6{mm}^3$, 암컷 6.35$\times10^6{mm}^3$이었고 WBC는 숫컷 8.09$\times10^3{mm}^3$, 암컷 7.09$\times10^3{mm}^3$었으며 leukocyte differential counts 에서는 lymphocytes, Neutrophils, Eosinophils의 순으로 높은 성적을 보였다.

• 대한수의학회지
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• 제36권2호
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• pp.359-369
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• 1996

An effective candidate subunit vaccine was prepared by using the immunostimulating complexs(ISCOMs) with Quil A and recombinant protein(gp50, gIII and inactive $\alpha$-ADV) Aujeszky's disease virus(ADV). The weaned pigs were twice immunized with a ADV-ISCOMs, and followed by intramuscular challenge with $1{\times}10^4$ $TCID_{50}$ ADV(strain Yangsan). The unvaccinated pigs were also challenged with same dose of ADV. At 5 days after challenge, the control pigs have developed ADV clinical signs. Whereas, the vaccinated pigs protected them from ADV-induced acute symptoms and death. Also, to identify the lymphocyte subpopulation in peripheral blood with pigs from ADV-ISCOMs vaccinated and control group, lymphocyte reacted with a panel of monoclonal antibodies which are specific to swine leukocyte surface antigens and assayed by the flow cytometry. MHC class I, CD2, CD8, N cells, CD11a, and CD45 antigen positive cells were decreased after inoculating virulent ADV Yangsan strain in control group. The data indicated that ISCOMs technique was useful in ADV subunit vaccine preparation and demonstrated the importance of gp50, gIII as a component of ADV vaccine.

• Asian-Australasian Journal of Animal Sciences
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• 제22권3호
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• pp.396-403
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• 2009

Male broiler chicks were fed graded levels of organic zinc (zinc-methionine) supplementation to investigate the effects of partial or complete substitution of the organic zinc source for inorganic ones on the development of lymphoid organs and immunological responses. A total of 450 day-old male broilers were distributed into groups of 10 chicks and randomly assigned to nine experimental diets during a 42-day feeding trial. Dietary treatments consisted of two basal diets supplemented with 40 mg/kg added zinc as feed-grade Zn sulfate or Zn oxide in which, Zn was replaced with that provided from zinc-methionine (ZnMet) complex at the levels of 25, 50, 75 or 100%. Two randomly-selected birds from each pen replicate were bled and then slaughtered by cervical cutting on the final day of the trial to measure leukocyte subpopulations and relative weights of lymphoid organs. Among lymphoid organs, only thymus weight was affected (p<0.05) by dietary treatments. The sulfate-supplemented birds were heavier (p<0.01) in relative weight of thymus than oxide-supplemented birds. The 10 days of age-assessed cutaneous hypersensivity reaction was stronger in chicks fed ZnMet-containing diets. Dietary ZnMet supplementation caused (p<0.05) an increase in proportion of lymphocytes and consequently a decrease in heterophil to lymphocyte ratio. Diet fortification by zinc-methionine complex increased (p<0.01) Newcastle antibody titer at 19 days of age. Also, a similar response was observed in antibody titers at 6 and 12 d after infectious bronchitis vaccine administration. There was no significant effect of replacement of dietary zinc on antibody titer against infectious bursal disease virus (IBDV) at the 6th d post-vaccine inoculation; however, at d 12 after vaccination, ZnMet-fortified diets improved antibody titer against IBDV. Although dietary inclusion of ZnMet had no marked effect on primary antibody titer against sheep erythrocytes, effective responses were observed during secondary reaction from the viewpoint of both total antibody and immunoglobulin Y (IgY) titers. From the present findings, it can be concluded that dietary supplementation with organic zinc improves both cellular and humoral immune responses. It is necessary to replace 75% of supplemental inorganic zinc with organic ZnMet complex to achieve the optimum immunological responses in broiler chicks.