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.
The purpose of this study was to examine the distribution of lymphocyte populations in normal, reversibly inflamed and irreversibly inflamed human dental pulp tissues using flow cytometry. Flow cytometry, with specific antibody and fluorochrome reagent allows us to know cellular properties of hematolymphoid cells by measuring fluorescence of stained cells. Before extirpation of pulps in routine endodontic treatment, the clinical diagnosis were performed by symptom. The extirpated pulp tissues were divided into normal pulp group (N=5), reversible pulpit is group(N=10) and irreversible pulpitis group(N=7). The specimen was placed into RPMI 1640 medium, minced into small pieces, and then digested in medium with collagenase. The cell suspension was resuspended in PBS for monoclonal antibody staining of T lymhocytes(CD3+), B lymphocytes (CD19+), T helper cell (CD4+) and T supressor cell (CD8+). The percentages of cells were counted by FACStar(BD) flow cytometer. Following results were obtained; 1. In the most normal and inflamed pulps, the percentages of T lymphocyte, B lymphocytes, T helper cell and T suppressor/cytotoxic cell were less than 1 % in total counted pulpal cells. 2. The higher percentages of T, B, T helper and T suppressor cells were observed in irreversible pulpitis group as compared with the normal pulp and reversible pulpitis group but the differences between groups were not statistically significant (p>0.05). 3. The percentages of T helper cells (CD4 + cells) were greater than that of T suppressor/cytotoxic cells (CD8 + cells) in the inflamed pulps.
To understand the cytotoxic activity of Machilus thunbergii, which has been used as a traditional oriental medicine, the mechanism underlying the cytotoxic effect of its extract on human acute Jurkat T cells was investigated. The methanol extract of roots (3 kg) of M. thunbergii was evaporated, dissolved in, and then extracted by water. The water-extracted active substance was designated MTWE. When Jurkat T cells were treated with MTWE at concentrations of 0, 25, 50, and $100{\mu}g/ml$, the apoptotic phenomenon of cells accompanying several subsequent biochemical reactions, such as mitochondrial cytochrome c release, activation of caspase-3, and ICAD degradation, was detected in the Jurkat T cells. Moverover. the expression of Bcl-xL, which is a suppressor for mitochondrial cytochrome c release pathway, was reduced in the Jurkat T cells. As DUSP6, a growth suppressor of cancer cells, ranged from 0, 25, 50, $100{\mu}g/ml$ of MTWE, the expression level was elevated in the Jurkat T cells. The apoptotic morphological change of the nuclei was observed by DAPI staining. Although the potential involvement of the other factors and DUSP6 is currently being investigated in more detail, these findings support the notion that MTWE is able to achieve the apoptosis of Jurkat T cells, and it seems that MTWE is useful as a method of evaluating a chemotherapeutic agent or tonic materials for human acute leukemia.
Periodontal disease research has been focused on understanding the immunopathologic mechanisms which may operate in the development and maintenance of peiodontal inflammatory changes. Immunologic and inflammatory responses may relate to the etiology and pathogenesis of periodontal disease. In order to research immunopathology of periodontal disease, previous investigators have spent much time on the distribution of lymphocyte subpopulations and NK cells but they have spent less time on the changes of those cells to the periodontal disease severity. The purpose of study was performed to investigate the changes of the distribution of T lymphocytes, B lymphocytes, T lymphocyte subsets, and Natural Killer cells in the gingival epithelium and connective tissue of the periodontal disease with the various clinical parameters including Gingival Index, Sulcular Bleeding Index, and pocket depth. Gingival tissues were obtained from 25 patients with different severity of periodontal disease. Serial cryostat sections displaying a cross section of gingiva were labelled with monoclonal antibody for pan T cells, T cytotoxic/suppressor cells, T helper/inducer cells, pan B cells, and NK cells were develped using an avidin-biotin-peroxidase system. Lymphocyte populations were enumerated in repeatable fields from gingival section. 1. T cells were more increased at grade 1 and 3 than at grade 0 of gingival index (p<0.05). Helper T cells and NK cells were significantly increased at grade 1, 2, 3 than at grade 0(p<0.05). 2. T cells were more decreased at grade 3 and 4 than at grade 1 of sulcular bleeding index (p<0.01, p<0.05). Especially, Natural Killer cells were significantly increased at grade 1, 2, 3, 4 than at grade 0 (p<0.05, p<0.001). 3. The ratios of helper T/suppressor T cells were more decreased at grade 4 than at grade 0 and at grade 4 than at grade 2 of sulcular bleeding index (p<0.05, p<0.05). 4. Helper T cells were significantly decreased at grade II and III than at grade I, however the Natural Killer cells showed a increasing tendency with the increase of the pocket depth, there were no significant differences between each grade of pocket depth. 5. The ratios of helper T/suppressor T cells were tended to be decreased with the increase of the pocket depth, there were no significant differences between each grades of pocket depth. There was a very weak change in the distribution of T lymphocytes, B lymphocytes, T lymphocyte subsets, and Natural Killer cells in the gingival epithelium and connective tissue of the periodontal lesion with the various clinical parameters including gingial index, sulcular bleeding index, and pocket depth. But, the number of T lymphocytes and Natural Killer cells were significantly changed in gingival index and sulcular bleeding index.
Kim, Jay Sik;Lee, Won Kil;Suh, Jang Soo;Song, Kyung Eun;Lee, Joong Won;Lee, Nan Young;Weksler, Marc E.
IMMUNE NETWORK
/
v.1
no.3
/
pp.236-243
/
2001
Background: An immunological approach for aging mechanism appears to be important. Lymphocyte subsets analysis in peripheral blood is widely performed to assess the immune status and to diagnose and monitor various diseases. Some lymphocyte subsets are known to change with age, but only few data about age-related reference ragnes for these subsets in healthy individuals have been reported. So we attempted to report reference ranges for these subsets in each age group and review changes of the results with age for the secondary studies about immune cell function as lymphocyte blast transformation and immunoglobulin gene rearrangement (VDJ) including recombination activating genes (RAG-1 and RAG-2). Methods: Lymphocyte subset analysis was performed on 302 subjects, 189 males and 113 females with age group of all decades of life. Two color direct immunofluorescene flow cytometry (FCM) was done using $Simultest^{TM}$ IMK-Lymphocyte kit (Becton Dickinson, USA), $FACScan^{TM}$ (Becton Dickinson, USA) and $FACSCalibur^{TM}$ (Becton Dickinson, USA). Lymphocyte subsets analysed were T ($CD3^+$) and B cells ($CD19^+$), helper/inducer T ($CD4^+$) and suppressor/cytotoxic T cells ($CD8^+$), helper/suppressor ($CD4^+/CD8^+$) ratio and natural killer (NK) cells ($CD3^-CD16^+/CD56^+$). The absolute numbers of each subset were calculated from total lymphocyte counts. Data collected was analysed using SAS 6.12. A P-value of < 0.05 was considered significant. Results: We reported the counts and percentages of lymphocyte and these subsets in each age group. There were no statistically significant differences between male and female subjects. The percentage of $CD4^+$ T cells, and the count of NK cells did not show the significant difference among the various age groups. The age-related changes observed in our study were as following: 1) a decrease in the percentages of T cells, B cells and $CD8^+$ T cells ; 2) a decrease in the counts of B cells and $CD8^+$ T cells ; 3) an increase in the percentage and count of NK cells ; and 4) an increase in the $CD4^+/CD8^+$ ratio. Conclusion: The characteristics of aging process appeared to be showing a marked decrease of lympocyte subsets T and B cells as well as T8 ($CD8^+$). The age-related increase of the percentage of cells bearing NK marker can be interpreted as a compensatory consequence to cope with the decrease of T cells related to the thymic involution. These changes with age appeared to be for the secondary study about immune cell function as lymphocyte blast transformation and immunoglobulin gene rearrangement.
Park, Hae-Young;Wakefield, Lalage M;Mamura, Mizuko
IMMUNE NETWORK
/
v.9
no.4
/
pp.122-126
/
2009
Transforming growth factor-$\beta$ (TGF-$\beta$) is a highly pleiotropic cytokine playing pivotal roles in immune regulation. TGF-$\beta$ facilitates tumor cell survival and metastasis by targeting multiple cellular components. Focusing on its immunosuppressive functions, TGF-$\beta$ antagonists have been employed for cancer treatment to enhance tumor immunity. TGF-$\beta$ antagonists exert anti-tumor effects through #1 activating effector cells such as NK cells and cytotoxic $CD8^+$ Tcells (CTLs), #2 inhibiting regulatory/suppressor cell populations, #3 making tumor cells visible to immune cells, #4 inhibiting the production of tumor growth factors. This review focuses on the effect of TGF-$\beta$ on T cells, which are differentiated into effector T cells or newly identified tumor-supporting T cells.
Kim, Ji-Hye;Lee, Jong-Hwan;Kim, Young-Ho;Kim, Kwang-Hyeon
Journal of Life Science
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v.19
no.2
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pp.163-168
/
2009
To understand cytotoxic activity of Rubia cordifolia L. (Rubiaceae), which has been used as a traditional oriental medicine, the mechanism underlying cytotoxic effect of its extract on human acute Jurkat T cells was investigated. The methanol extract of roots (3 kg) of R. codifolia was evaporated, dissolved in water, and then extracted by dichloromethane. The substances in the chloroform extract showing the most cytotoxic activity were further purified by a series of preparative HPLC. The extracted active substance (65 mg) was designated as CCH1. When Jurkat T cells were treated with CCH1 at concentration ranging from 0.5 to 2.0 ${\mu}g$/ml, apoptotic phenomena of cells companying several subsequent biochemical reactions such as mitochondria cytochrome c release, activation of casapase-8, -9, and caspase- 3, degradation of PARP and DNA fragmentation occurred via mitochondria-dependent pathway. However, abrogation of apoptosis was observed in an ectopic expression of Bcl-xL, which is a suppressor for mitochondrial cytochrome c release. These results demonstrate that the cytotoxicity of CCH1 against Jurkat T cells is attributable to apoptosis mediated by mitochodria-dependent death-signaling regulated by Bcl-xL. In addition, the CCH1 is more potent to leukemia Jurkat T cell than to human peripheral blood monocyte cells (PBMC).
The effect of immune activation by Echinacea purpurea was investigated by measuring total immunoglobulin (Ig) G, IgM. and the radioprotective effect of immune activation by Echinacea purpurea was investigated by measuring T lymphocyte subsets in the peripheral blood of mice following whole body irradiation. Echinacea purpurea activated macrophages to stimulate $IFN-{\gamma}$ production in association with the secondary activation of T lymphocytes, resulting in a decrease in IgG and IgM production. Cytokines released from macrophages in mouse peripheral blood after Echinacea purpurea administration activated helper T cells to proliferate. In addition, activated macrophages in association with the secondary T lymphocyte activation increased $IFN-{\gamma}$ production and stimulated proliferation of cytotoxic T cells and suppressor T cells, indicating the activation of cell-mediated immune responses.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.29
no.2
/
pp.108-115
/
2003
Suppression of cellular immunity is the host responses to surgical stress. When the body is exposed to surgical stress, decreased immunocyte function is one of the surgical stress-induced biologic responses. In all patients exposed to the surgical stress, peripheral blood lymphocyte numbers and function were suppressed until at least 2 weeks postoperatively. This immunosuppression was mainly due to a decrease of helper-inducer T cells, cytotoxic T cells, natural killer cells, and an increase of suppressor T cells. The blood levels of interleukin-6(IL-6) cytokine increase in response to surgical stress and cause an increase of so-called acute phase reactants, including C-reactive protein(CRP). In the previously damaged patients group, expected to early stress expose, immunosuppression was more developed than other normal groups. Cellular immunosuppression by surgical stress was mainly due to an increase of lymphocyte subsets that depress cellular immunity coupled with a decrease of the subsets that promote it. Overproduction of CRP in response to surgical stress may play an important role in the development of immunosuppression.
The more society has complicated, the more we have met stressful circumstance. And it is found that many physical and mental symptoms induced by stress. Soyosan(SYS) is one of the well-known oriental medicine for the treatment of general syndrome induced by emotional stress. This study was taken to know effects of SYS water extract on immune-depressed mice induced by stress. The results obtained in this study were as follows : 1. SYS inhibited murine weight-loss induced by stress 2. In vivo& in vitro, SYS increased phagocytic activity. 3. SYS enhanced the production of such reactive oxygen intermediates as superoxide and hydrogen peroxide from macrophages. 4. In vitro, SYS little influenced the production of reactive nitrogen intermediates. 5. SYS increased the number of the rosette forming cells of spleen. 6. SYS changed the ratio of helper and suppressor T cells by increasing $CD4^+$ T cells and decreasing $CD8^+$ T cells. 7. SYS increased cytotoxic activity on human lymphoma cell line(K562). 8. SYS increased the plasma level of GH and DHEA. whereas it decreased that of ACTH and cortisol. According to the above results, it might be considered that SYS would be used for immune-depressive disease induced by stress.
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