• Korean Journal of Veterinary Research
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• v.34 no.2
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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.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.8
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• pp.1189-1195
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• 2014

Natural resistance-associated macrophage protein 1 encoding gene (NRAMP1) plays an important role in immune response against intracellular pathogens. To evaluate the effects of NRAMP1 gene on immune capacity in pigs, tissue expression of NRAMP1 mRNA was observed by real time quantitative polymerase chain reaction (PCR), and the results revealed NRAMP1 expressed widely in nine tissues. One single nucleotide polymorphism (SNP) (ENSSSCG00000025058: g.130 C>T) in exon1 and one SNP (ENSSSCG00000025058: g.657 A>G) in intron1 region of porcine NRAMP1 gene were demonstrated by DNA sequencing and PCR-RFLP analysis. A further analysis of SNP genotypes associated with immune traits including contain of white blood cell (WBC), granulocyte, lymphocyte, monocyte (MO), rate of cytotoxin in monocyte (MC) and $CD4^-CD8^+$ T lymphocyte subpopulations in blood was carried out in four pig populations including Large White and three Chinese indigenous breeds (Wannan Black, Huai pig and Wei pig). The results showed that the SNP (ENSSSCG00000025058: g.130 C>T) was significantly associated with level of WBC % (p = 0.031), MO% (p = 0.024), MC% (p = 0.013) and $CD4^-CD8^+$ T lymphocyte (p = 0.023). The other SNP (ENSSSCG00000025058: g.657 A>G) was significantly associated with the level of MO% (p = 0.012), MC% (p = 0.019) and $CD4^-CD8^+$ T lymphocyte (p = 0.037). These results indicate that the NRAMP1 gene can be regarded as a molecular marker for genetic selection of disease susceptibility in pig breeding.

• Korean Journal of Veterinary Pathology
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• v.1 no.1
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• pp.1-6
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• 1997

The cellularity and composition of the spleen lymph node thymus and peripheral blood and tempo of regeneration were studied at various time points after syngeneic bone marrow transplantation(BMT) in C3H/Hen mice. Significant depression of absolute lymphocyte count was noted on week 1 after lethal whole-body irradiation and BMT. In comparison to the lymph node thymus and spleen had an rapid regeneration of cellularity. The distinct cell populations($CD4^+,\;CD8^+,\;CD28^+,\;B220^+) have determined in the lymphoid tissue of mice subjected to irradiation. The relative representation of these subpopulations was significantly different from that in nonirradiated control. $CD4^+\;and\;CD8^+$ cells were present in very low numbers whereas the $B220^+$ cells reached more than normal range at 2 weeks after BMT. The number of $CD4^+$ cells returned to normal relatively soon than $CD8^+$ cell. At week 4 after BMT, the cellularity and composition of spleen lymph node and peripheral blood lymphocyte reached about 50% of the normal range therefore we can choose this time point for the other tests of immune function after BMT.

• Korean Journal of Veterinary Research
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• v.35 no.4
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• pp.769-776
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• 1995

This study was undertaken to develop the monoclonal antibody(MAb) for lymphocytes of Korean native cattle by the cell hybridization of myeloma P3/NS-1/ 1-Ag-4-1 and spleen cells from BALB/c mice hyperimmunized with nylon wool column eluted peripheral T lymphocytes of Korean native cattle. The isotype of MAb KCT-14 against T lymphocyte was mouse $IgG_1$. KCT-14 positivity of mononuclear cells(MNC) from peripheral blood lymphocytes, nylon wool nonadherent and adherent-lymphocytes was 41.7%, 58.4% and 22.6%, respectively. And that of mesenteric lymph node-, spleen and thymus-MNC was 43.3%, 40.2% and 33.6%, respectively. Immunoperoxidase staining of frozen tissue sections showed that the MAb positive cells were located in the medulla of the thymus and in the paracortical area and the mantle zone of the germinal center in the lymph nodes. These results indicated that KCT-14 was one of the MAb for investigate of T lymphocyte subpopulations in the Korean native cattle.

• Korean Journal of Veterinary Research
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• v.46 no.2
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• pp.149-158
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• 2006

To determine the effects of ${\beta}$-carotene on the control of mastitis in dairy cows during the dry period, 38 dairy cows (18 mastitic cows and 20 healthy cows) were administered with 5 ml of ${\beta}$-carotene (30 mg/ml) intramuscularly twice (4 week intervals). Blood samples were taken from the cows before the injection and two weeks after the second injection, respectively, and were measured for the proportion of lymphocyte subpopulations and lymphocyte proliferation responses. With ${\beta}$-carotene injection, the proportion of cells expressing BoCD2, BoCD4 and B and N cells increased in healthy cows. In the presence of mitogens, lymphocytes from healthy cows showed significantly higher proliferation responses after ${\beta}$-carotene injection than before (p < 0.05), The somatic cell counts in ${\beta}$-carotene injected group decreased from 1,001,00 cells/ml at dry off to 647,000 cell/ml and 447 cells/ml at the stage of first and second weeks after calving, respectively (p < 0,05), This study indicated that ${\beta}$-carotene as a nonspecific immunostimulator could have a definite role for the prevention of intramammary infections in dairy cows at dry period.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.8
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• pp.1141-1149
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• 2018

Objective: Cluster of differentiation 4 protein (CD4) gene is an important immune related gene which plays a significant role in T cell development and host resistance during viral infection. Methods: In order to unravel the relationship of CpG island methylation level of CD4 gene with its gene expression and T lymphocyte subpopulation traits, we used one typical Chinese indigenous breed (Dapulian, DP) and one commercial breed (Landrace), then predicted the CpG island of CD4 gene, determined the methylation status of CpG sites by bisulfite sequencing polymerase chain reaction (BSP), and carried out the correlation analyses of methylation frequencies of CpG sites with mRNA expression and T lymphocyte subpopulation traits. Results: There was one CpG island predicted in the upstream -2 kb region and exon one of porcine CD4 gene, which located 333 bp upstream from the start site of gene and contained nine CpG sites. The correlation analysis results indicated that the methylation frequency of CpG_2 significantly correlated with CD4 mRNA expression in the DP and Landrace combined population, though it did not reach significance level in DP and Landrace separately. Additionally, 15 potential binding transcription factors (TFs) were predicted within the CpG island, and one of them (Jumonji) contained CpG_2 site, suggesting that it may influence the CD4 gene expression through the potential binding TFs. We also found methylation frequency of CpG_2 negatively correlated with T lymphocyte subpopulation traits CD4+CD8-CD3-, CD4-CD8+CD3- and CD4+/CD8+, and positively correlated with CD4-CD8+CD3+ and CD4+CD8+CD3+ (for all correlation, p<0.01) in DP and Landrace combined population. Thus, the CpG_2 was a critical methylation site for porcine CD4 gene expression and T lymphocyte subpopulation traits. Conclusion: We speculated that increased methylation frequency of CpG_2 may lead to the decreased expression of CD4, which may have some kind of influence on T lymphocyte subpopulation traits and the immunity of DP population.

• Biomedical Science Letters
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• v.2 no.1
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• pp.13-20
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• 1996

A Recent discovery of surface antigens in cells has led to the success of quantitative measurement of T-cell subpopulations, and this has especially opened the way for an epoch-making development in the understanding and classification of cellular immune mechanisms. It is known that phenotypes of T-cell subpopulations exist in many forms according to the variation of species or animal experimental models. In Korea, Clonorchis sinensis still gives rise to public concern as it infects more than eighty million people and threatens the public by causing cirrhosis of the liver, or liver cancer when liver infection becomes prolonged and chronic. Up until now there has been much progress in research and improvement in the classification system of Clonorchis sinensis in the area of humoral immunity, but as for research in the area of cellular immune mechanisms, there is almost none. Knowing all these circumstances, the authors delved for the characterization of Iymphocyte subpopulations with mice as Clonorchis sinensis in the area of cellular immunity, and obtained the following results. That is, we injected Clonorchis sinensis antigens mixed in Freund's ajuvant solution intraperitoneally in mice and measured the T-cell subpopulation characterization of spleen lymphocytes with flow cytometry. The results of these measurements showed that CD2, CD5 and CD8 decreased early following injections but then in-creased again seven weeks after the injections. CD4, however, showed a slight increase shortly after the injection but then a fair increase seven weeks after the injection.

• Korean Journal of Veterinary Research
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• v.36 no.2
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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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• v.24 no.5
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• pp.696-706
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• 2011

The objective of this study was to characterize serum immunoglobulins and lymphocytes subpopulations in the peripheral blood mononuclear cells (PBMCs) of Holstein calves in response to lipopolysaccharide (LPS) challenge from Escherichia coli. Fourteen calves received subcutaneous injections of E. coli LPS at 10 weeks of age, and six calves were injected with saline as a control. The concentrations of total serum IgG and the relative amount of LPS-specific IgG in calves challenged with LPS were significantly higher (p<0.05) compared to control animals and LPS challenge significantly increased (p<0.05) the percentage of $CD5^+$ and $CD21^+$ T cells in PBMCs. Meanwhile, LPS challenge significantly increased (p<0.05, p<0.01) the percentage of $CD8^+$ and $CD25^+$ T cells in peripheral blood mononuclear cells (PBMC) at 7 and 14 Day-post LPS challenge (DPLC), respectively. The composition of $CD4^+CD25^+$ T cells and $CD8^+CD25^+$ T cells from calves challenged with LPS was also higher (p<0.05 and p = 0.562, respectively) than those of control calves at 14 DPLC. In conclusion, LPS challenge not only induces production of IgG with expression of B-cell immune response related cell surface molecules, but also stimulates activation of T-lymphocytes in PBMC. Our results suggest that LPS challenge in calves is a good model to elucidate cellular immune response against Gram-negative bacterial infections.

• Biomedical Science Letters
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• v.1 no.1
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• pp.19-25
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• 1995

Characterization of immune cell subpopulations in the bovine was performed using a direct immunofluorescence technique adaptable for routine and repeated monitoring. This whole blood procedure is faster and requires less volume than conventional density gradient isolation methods. Low inter-and intra-animal variations were seen in hematology parameters and in CD4, CD8 and CD2 lymphocyte subtypes. CD4 values were 30% of lymphocytes in male and 32% in females. Thriteen percent were CD8 in males and 13% in females. CD4: CD8 ratios were approximately two in both sexes. Fiffty three percent were CD2 in males and 54% in females. The mean RBC counts of peripheral blood were 7.20$\times10^6/{mm}^3$ for male cattle and 6.36$\times10^6/{mm}^3$ for females. The mean WBC counts were 8.09$\times10^3/{mm}^3$ for males and 7.09$\times10^3/{mm}^3$ for females. The percent of lymphocytes(63-65%) was higher than the percent of neutrophils(17-18%), the percent of eosinophils(11-15%), te percent of monocytes(4-5%), and percent of basophiles(1%).