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.
We studied the T lymphocyte and its subpopulation percentage change in 40 patients immediately after the radiation therapy. Study population consisted of 12 patients treated at the site of head and neck region,14 patients treated at the site of thoracic region, and 14 patients treated at the site of pelvic region. Twenty two patients received radiotherapy as radical modality, and remaining 18 patients received radiotherapy as postoperative modality. Immediately after radiotherapy, total T lymphocyte (T1) percentage was decreased from $56.4\%$ to $55.2\%$, helper T cell (T4) percentage was decreased from $36.4\%$ to $34.1\%$, but suppressor T lymphocyte (T8) percentage was increased from $23.5\%$ to $25.4\%$. As a result, T4/T8 ratio was decreased from 1.57 to 1.39. This study suggested that immediate change after radiotherapy of the T lymphocyte and its subpopulation percentage was not related to the treatment volume and the degree of helper T cell decrement was not pronounced by the radiation dose increment. Long-term follow-up study En larger scale is needed to determine long term changing pattern in T lymphocyte subpopulation and its relationship to the prognosis of patients.
The purpose of this research was to investigate effects of Bambusae Caulis in Liquamen(BCL) on T-lymphocytes and peritoneal macrophages in mice. The apoptosis and subpopulation of T-lymphocytes were tested using a flow cytometer. The phagocytic activity of mouse peritoneal macrophage was tested using a luminometer. Nitric oxide production was tested using a Griess reagents. BCL induced T-lymphocytes apoptosis. BCL increased $T_H$ cells population and decreased $T_C$ cells population of T-lymphocyte, but did not affect splenocytes subpopulation. BCL increased nitric oxide production and phagocytic activity of peritoneal macrophage in mice. These results suggest that BCL regulates the immune system in consequence of an increase in helper T cell population and macrophages activation.
This study was designed 1) to compare the distributions of periapical inflammatory cells and 2) to identify lymphocytes and compare the lymphocyte distribution with T lymphocyte subpopulation and then 3) to examine the distribution of cycling cell in human dental periapical lesions. From each of the twenty-five human dental periapical lesions observed one small portion was fixed, embeded in paraffin, sectioned serially and stained with HE. The periapical inflammatory cells were counted to obtain the relative concentration of lymphocyte, plasma cell, macrophage and neutrophil. The large part of each lesion was analysed using Flow cytometer and monoclonal antibodies to obtain the relative concentration of T lymphocyte, B lymphocyte, T'helper cell and T suppressor/cytotoxic cell. In addition to that, seven human dental periapical lesions were examined with DNA analysis to observe the distribution of cycling cell. Following results were obtained: 1. 24 cases of the 32 periapical lesions examined were diagnosed as periapical granuloma and the remaining 8 cases as periapical cyst. Lymphocytes comprised 42.1% of total inflammatory cells in periapical granuloma and 41.8% in periapical cyst. Corresponding percentages for macrophages were 33.8% and 30.3%; for plasma cells, 15.9% and 19.0%; for neutrophils, 8.2% and 8.8%. 2. All of the periapical lesions examined had T lymphocyte, B lymphocyte, T helper cell, T suppressor/cytotoxic cell. And in all cases, T lymphocytes were observed predominantly more than B lymphocytes. 3. In 2 cases of the control group only T lymphocytes were found, and in the remaining 2 cases T lymphocytes were observed predominantly. 4. T helper cells were observed predominantly more than T suppressor/cytotoxic cells in all cases of perapical granulomas. 5. T suppressor/cytotoxic cells were observed predominantly more than T helper cells in 4 cases of periapical cysts (total 5 cases were examined) and only in one case T helper cells were more than T suppressor/cytotoxic cells. 6. In control group, T helper cells were predominant in 2 cases and T helper cells were equivalent to T suppressor/cytotoxic cells in one case. In remaining one case T suppressor/cytotoxic cells were predominant. 7. As the result of DNA analysis, the average proliferating indices of the various groups examined were measured as follows: in the control group 5.45%, in periapical granuloma 6.64%, in periapical cyst 10.1%. The highest index was observed in periapical cyst.
It has well studied that immune cells are strongly related to tumor progression and tumor suppression. To identify the difference of immune cell between tumor bearing mice and normal mice, we examined systemically the immune cell of CT26 tumor bearing mice on 21 days after tumor cell administration. As previously reported, CD4+ and CD8+ T cells population of tumor bearing mice significantly decreased 38% and 30% on day 21 compared to that of normal mice, respectively. All subpopulation of CD4 and CD8+ T cell significantly decreased, except CD49b+ T cell subpopulation. But, myeloid cell population ($CD11b^{high}$ and all Gr-1+ subpopulation) of tumor bearing mice significantly increased on day 21. Especially, all subpopulation of CD11b+Gr-1+ cell of tumor bearing mice significantly increased on day 21. Also, Foxp3+$CD25^{high}$ CD4 T cell (regulatory T cells) population significantly increased on day 21. These results suggest that tumor can induce the decline of T lymphocyte and the expansion of myeloid cells and regulatory T cells, and provide the basic information for the study of tumor immunology.
Kim, Ki-Woong;Kim, DaeSeong;Won, Yong Lim;Kang, Seong-Kyu
Toxicological Research
/
v.29
no.2
/
pp.115-120
/
2013
To investigate the effects of short-term exposure of beryllium on the human immune system, the proportion of T-lymphocytes such as CD3+, CD4+, CD8+, CD95, and NK cells, and the proportion of B cells and $TNF{\alpha}$ level in peripheral blood and immunoglobulins in the serum of 43 exposed workers and 34 healthy control subjects were studied. External exposure to beryllium was measured by atomic absorption spectrometer as recommended by the NIOSH analytical method 7300. T lymphocyte subpopulation analysis was carried out with flow cytometer. The working duration of exposed workers was less than 3 months and the mean ambient beryllium level was $3.4{\mu}g/m^3$, $112.3{\mu}g/m^3$, and $2.3{\mu}g/m^3$ in molding (furnace), deforming (grinding), and sorting processes, respectively (cited from Kim et al., 2008). However, ambient beryllium level after process change was non-detectable (< $0.1{\mu}g/m^3$). The number of T lymphocytes and the amount of immunoglobulins in the beryllium-exposed workers and control subjects were not significantly different, except for the total number of lymphocytes and CD95 (APO1/FAS). The total number of lymphocytes was higher in the beryllium-exposed individuals than in the healthy control subjects. Multiple logistic regression analysis showed lymphocytes to be affected by beryllium exposure (odd ratio = 7.293; p<0.001). These results show that short-term exposure to beryllium does not induce immune dysfunction but is probably associated with lymphocytes proliferation.
To assess the immunological function of toluene exposed group, the proportions of T lymphocyte, B lymphocyte, CD4 cell, CD8 cell, the ratio of CD4 to CD8(CD4/CD8) in peripheral blood were measured on twenty-one toluene exposed workers and twelve healthy workers who did not have previous history of toluene exposure. In addition, to evaluate the present status of toluene exposure, urinary hippuric acid concenturations were measured in exposed group. The mean concenturation of urinary hippuric acid was 2.84 g/creatinine g in exposed group. The proportions of T lymphocyte, B lymphocyte, CD8 cell and CD4/CD8 of exposed group were slightly lower than non-exposed group except the proportion of CD4 cell which was similar in both groups. But these differences were not statistically different in both groups. The proportions of T lymphocyte and CD4 cell were significantly correlated with the length of duration in exposed group(P<0.05).
Purpose: The purpose of this study is to investigate the effect of Boheo-tang (B) and Boheo-tang plus cervi pantotrichum cornu (B+CP) on immune response in postpartum C57BL/6N mice. Methods: Normal saline(control), B and B+CP (8${\mu}{\ell}$/g) were administerd p.o. twice a day for 20 days. Subpopulation of T and B lymphocyte were accessed by flow cytometric analysis. Results: Splenic T and B lymphocytes were increased by the treatment of B. Subpopulation of cytotoxic T lymphocytes in the spleen, were significantly increased by both treatment of B and B+CP. Subpopulation of cytotoxic T lymphcytes in the thymus, were significantly increased by both treatment of B and B+CP. IL-4 production was significantly increased by the treatment of B+CP. Conclusion: This study shows that treatment of Boheo-tang and Boheo-tang plus cervi pantotrichum cornu can improve postpartum immune response in C57BL/6N mice.
Background : Nowadays asthma is considered to be an inflamatory disease characterized by airborne hyper-responsiveness and pulmonary eosinophilia. Objective : We aimed to identify the effects of Sochongryongtang on blood eosinophil, serum IgE and T lympocyte subpopulation in asthmatic patients. Material and Methods : The subjects consisted of fifteen patients with asthma who had been treated with Sochongryongtang for two weeks from February 2001 through June 2001. Sochongryongtang is herbal decoction which has been used for the traditional therapeutic agent of asthma. Results : The blood eosinophil and serum IgE in a normal controlled group. However, the T lympocyte subpopulation in asthmatic patients was not significantly different from the T lympocyte subpopulation in a normal group. The patients were treated with Sochongryongtang for two weeks. No significant difference in the blood eosinophil, serum IgE and T Iympocyte in the sub population. After treatment with Sochongryongtang for two weeks, FEV 1 increased significantly over 0.5 points out of total scores. Conclusion : This study shows that Sochongryongtang has effects on improvement of pulmonary function and quality of life in asthmatic patients. However, the patients who were treated with Sochongryongtang for two weeks showed no significant difference in the blood eosinophil, serum IgE and T lympocyte subpopulation. Further long-term studies must be made on a large number of asthmatic patients.
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.
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