• Restorative Dentistry and Endodontics
• /
• 제18권2호
• /
• pp.317-340
• /
• 1993

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.

• Restorative Dentistry and Endodontics
• /
• 제17권1호
• /
• pp.55-68
• /
• 1992

Periapical lesions are developed as a result of inflammatory response to irritants from root canal system. Clinicians remove these irritants from root canal system and seal the root canal space to induce healing of the periapical lesions. Immunopathologic responses may play an important role in development and progression of periapical lesions and periapical lesions contain immunocompetent cells. The purposes of the present study were to analys and to compare the distribution of the immunocompetent cells in the human periapical lesions according to the stage of endodontic treatment using indirect immunoperoxdase technique. Obtained 94 human periapical lesions were devided into four groups: Group 1 : no endodontic treatment(28 samples) Group 2 : root canal enlarged and irrigated(28 samples) Group 3 : root canal filled(29 samples) Group 4: unknown(9 samples) Monoclonal antibodies to examine target cells were UCHL-1 for T lymphocytes(1 : 200, Dakopatt, Denmark), L26 for B lymphocytes(1 : 200, Dakopatt, Denmark), OPD4 for helper T lymphocytes(l : 200, Dakopatt, Denmark) and alpha-1-antichymotrypsin for macrophages(l : 2000, Dakopatt, Denmark). The following results were obtained : 1. All the periapical lesions studied were infiltrated by T lymphocytes, plasma cells, B lymphocytes, and macrophages. T lymphocytes were more infiltrated than B lymphocytes, and B lymphocytes and macrophages were less infiltrated than T lymphocytes and plasma cells(P<0.05 : Oneway ANOVA test). 2. In untreated group and canal irrigated and enlarged group of all the periapical lesions, helper T lymphocytes were predominently infiltrated(P>0.05 : Oneway ANOVA test). 3. In canal filled groups of all lesions except periapical cyst, plasma cells were predominently infiltrated. But, in canal filled group of periapical cyst, helper T lymphocytes were the predominent cells(P>0.05 : Oneway ANOVA test). The above results shows that the immunologic responses play important role in pathogenesis of periapical lesions and the immunologic response involved undergoes certain changes after endodontic therapy.

• Restorative Dentistry and Endodontics
• /
• 제22권1호
• /
• pp.374-387
• /
• 1997

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.

• Journal of Periodontal and Implant Science
• /
• 제23권2호
• /
• pp.300-314
• /
• 1993

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.

• Restorative Dentistry and Endodontics
• /
• 제20권1호
• /
• pp.235-249
• /
• 1995

This study was designed to identify the lymphocytes present and to examine the relation between lymphocytes population and clinical symptoms of the pulps clinically diagnosed as normal and irreversible pulpitis. We recorded the history and severity of the pain and performed several clinical tests, before extirpation of vital, irreversibly inflamed pulps in routine endodontic treatment. Then the teeth were divided into two groups. Five teeth, categorized in acute symptom group, had severe spontaneous pain, particularly at night and were extremely sensitive to cold and heat. The other 15 teeth with history of mild to moderate pain and with or without cold or heat responses were categorized as chronic symptom group. Inflamed pulps were also classified into 8 minor groups by presence or absence of signs or symptoms related to the involved teeth, including the presence of pain on percussion, pain on heat and cold stimuli and the periodontal pocket depth. All extirpated pulps were immediately immersed in ultra low-temperature freezer($-74^{\circ}C$), and they were sectioned $6{\mu}m$ in thickness. Specimens were stained using three-stage indirect immunoperoxidase techniques(DAKO, LSAB kit) and monoclonal antibodies for detecting the presence of T lymphocytes(T), B lymphocytes(B) and helper(T4) and suppressor(T8) lymphocytes. Following results were obtained; 1. All the examined normal and inflamed pull) tissues had positive staining for T lymphocytes and T helper and T suppressor cells. But B cells were observed only in inflamed pulp. 2. Statistically more T and B cells were observed in acute symptom group as compared with chronic symptom group(p<0.05). 3. Cell ratio of BIT in acute symptom group were significantly higher than that of chronic symptom group(p<0.05). 4. Only B cells were significantly increased in the percussion positive group than the number of B cells in percussion negative group(p<0.05). 5. No differences were observed in the number of different cell types among other minor groups.

• 대한한방내과학회지
• /
• 제18권2호
• /
• pp.27-39
• /
• 1997

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.

• 한양메디칼리뷰
• /
• 제33권1호
• /
• pp.10-16
• /
• 2013

Follicular helper T cells (Tfh) play a significant role in providing T cell help to B cells during the germinal center reaction, where somatic hypermutation, affinity maturation, isotype class switching, and the differentiation of memory B cells and long-lived plasma cells occur. Antigen-specific T cells with IL-6 and IL-21 upregulate CXCR5, which is required for the migration of T cells into B cell follicles, where these T cells mature into Tfh. The surface markers including PD-1, ICOS, and CD40L play a significant role in providing T cell help to B cells. The upregulation of transcription factor Bcl-6 induces the expression of CXCR5, which is an important factor for Tfh differentiation, by inhibiting the expression of other lineage-specific transcription factors such as T-bet, GATA3, and RORγt. Surprisingly, recent evidence suggests that CD4 T cells already committed to Th1, Th2, and Th17 cells obtain flexibility in their differentiation programs by downregulating T-bet, GATA3, and RORγt, upregulating Bcl-6 and thus convert into Tfh. Limiting the numbers of Tfh within germinal centers is important in the regulation of the autoantibody production that is central to autoimmune diseases. Recently, it was revealed that the germinal center reaction and the size of the Tfh population are also regulated by thymus-derived follicular regulatory T cells (Tfr) expressing CXCR5 and Foxp3. Dysregulation of Tfh appears to be a pathogenic cause of autoimmune disease suggesting that tight regulation of Tfh and germinal center reaction by Tfr is essential for maintaining immune tolerance. Therefore, the balance between Tfh and Tfr appears to be a critical peripheral tolerance mechanism that can inhibit autoimmune disorders.

• 동국한의학연구소논문집
• /
• 제6권2호
• /
• pp.99-107
• /
• 1998

본 실험은 cyclosporin A(CsA)에 의한 시간의 경과에 따른 림프절에서의 세포성 면역억제를 조사하기 위해서 시행된 것으로 BALB/C계 생쥐에 10일동안 CsA(45mg/kg/day) 투여 후 림프절에서의 T 림프구, IL-2 수용기 그리고 자연살해(NK)세포의 분포 변화를 관찰하였다. 대조군의 림프절에서는 L3T4(CD4)에 양성반응을 보이는 도움 T림프구, Ly2(CD8)에 양성반응을 보이는 세포독성 T 림프구 그리고 CD25R에 양성반응을 보이는 IL-2 수용기를 가진 세포는 곁피질(paracortex)과 수질동(medullary sinus)에서 분포하였다. CsA 투여 후 처음 3일까지는 이들 양성반응세포의 분포 변화는 없었으며 양성반응성의 변화도 없었다. 그러나 CsA 투여 7일부터 양성반응 세포수의 감소와 양성반응성의 약화가 관찰되기 시작하였으며 이러한 변화는 시간이 경과하여 14일에 이르렀을 때 가장 큰 감소추세로 나타났다. 한편 NK1.1(CD56)에 양성반응을 보이는 자연살해세포는 피질과 수질에 분포하였으며 CsA 투여 후 시간의 경과에 따라 양성반응 세포수가 감소하였으며, 이러한 감소는 14일에서 가장 큰 것으로 나타났다. 이상의 결과로 미루어보아 CsA 투여는 림프절에서의 IL-2 분비저해를 통해 T 림프구와 NK세포의 활성을 차단하여 선택적이면서 효과적인 세포성 면역억제작용을 하고 있는 것으로 사료된다.

• 대한한방부인과학회지
• /
• 제23권4호
• /
• pp.1-9
• /
• 2010

Purpose: This study was designed to investigate enhancing the immunogenicity effects of Platycodon grandiflorum(PG) on adaptive immune system. Methods: To investigate the effect of PG as an adjuvant, we used the ovalbumin (OVA) as an antigen at first. The proliferation of lymphocytes, the antibody titer, the subisotypes of antibodies and the production of cytokines were measured. Results: The proliferation of lymphocytes and the antibody titer were increased after PG treatment. The increased subisotypes of antibodies were IgG2 and IgG3 induced from T1-helper cells. However IgE induced from T2-helper cells was decreased. The production of cytokines derived from T1-helper cells was increased but that from T2-helper cells was decreased. Conclusion: It is supposed that PG has an immunogenicity effect as an adjuvant on adaptive immune system.

• Parasites, Hosts and Diseases
• /
• 제31권3호
• /
• pp.231-238
• /
• 1993

폐흡충 피낭유충을 BALB/c 마우스에 경구 감염시킨 후 마우스를 시기별(3일, 1주, 2주, 3주, 4주, 5주, 7주, 9주, 13주 및 23주)로 희생시켜 말초혈액내 호산구수와 혈청내 IgE 항체가를 측정하는 한편 비장 림프구를 분리하여 림프구 아군(helper T, cytotoxic / suppressor T, pan T)에 대한 단세포군 항체를 결합시켜 EPICS C fluorocytometer로 분석하였다. 폐흡충에 대한 혈청내 총 IgE 항체가는 감염 3주부터 증가되어 감염 4주에 높게 증가되었으며 13주 이후 23주까지 높은 항체가가 유지되었다. 말초혈액내 호산구수는 감염 2주 부터 증가되어 감염 9주에 $664.1/\textrm{mm}^3$으로 가장 증가 되었으며 그 이후에는 감소하여 23주에는 정상보다 약간 증가되었다. 폐흡충을 감염시키고 감염 3일, 1주, 2주, 3주, 4주, 7주 및 23주에 T 림프구 아군을 분석한 결과 L3T4 helper T 림프구와 Lyt-2 cytotoxic / suppressor T 림프구의 비율은 감염 군에서 감염 초기에 대조군에 비해 약간 감소된 경향을 보였으나 대조군과 유의한 차이를 보이지 않았다. Thy 1.2 pan T 림프구의 비율은 감염 2주와 4주에 대조군에 비해 유의하게 감소되었고(P<0.05) 7주 이후에는 대조군과 비슷하였다. 이러한 결과들로 보아 마우스의 폐흡충감염에서 특이 항원이나 마이토젠의 자극이 없이는 비장킬프구의 아군분포에 큰 변화가 없는 것으로 생각된다. 생각된다.