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.
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.
Jeong-Su Park;Ik-Joo Chung;Hye-Ran Kim;Chang-Duk Jun
IMMUNE NETWORK
/
v.23
no.3
/
pp.29.1-29.23
/
2023
Cholesterol (CL) is required for various biomolecular production processes, including those of cell membrane components. Therefore, to meet these needs, CL is converted into various derivatives. Among these derivatives is cholesterol sulfate (CS), a naturally produced CL derivative by the sulfotransferase family 2B1 (SULT2B1), which is widely present in human plasma. CS is involved in cell membrane stabilization, blood clotting, keratinocyte differentiation, and TCR nanocluster deformation. This study shows that treatment of T cells with CS resulted in the decreased surface expression of some surface T-cell proteins and reduced IL-2 release. Furthermore, T cells treated with CS significantly reduced lipid raft contents and membrane CLs. Surprisingly, using the electron microscope, we also observed that CS led to the disruption of T-cell microvilli, releasing small microvilli particles containing TCRs and other microvillar proteins. However, in vivo, T cells with CS showed aberrant migration to high endothelial venules and limited infiltrating splenic T-cell zones compared with the untreated T cells. Additionally, we observed significant alleviation of atopic dermatitis in mice injected with CS in the animal model. Based on these results, we conclude that CS is an immunosuppressive natural lipid that impairs TCR signaling by disrupting microvillar function in T cells, suggesting its usefulness as a therapeutic agent for alleviating T-cell-mediated hypersensitivity and a potential target for treating autoimmune diseases.
Reappraisal measurements of apparent half survival time of red cell by $^{51}Cr$ method was made and effects of blood-letting over red cell survival were observed. The study was performed on 53 normal male subjects under three different experimental conditions. 1. Group 1 Mean $^{51}Cr$ red cell half survival by ACD wash method was 29.7 days. $T\frac{1}{2}$ of Ascorbic acid method was 29.0 days in group with 100 mg dose and 29.1 days in group with 50 mg dose respectively. There was no difference between these two methods in regards to red cell half survival. No difference were noted in amount of ascorbic acid administered. 2. Group 2 As daily amount of blood loss is increased the shortening of red cell half survival was noted. Rapid phase was seen when blood loss ranged 10 to 25 ml per day, while slow phase noted when more loss amounted 25 ml or more daily. Thus, it was clear that there was more than an exponential relation between $T\frac{1}{2}$ and the amount of blood loss. 3. Group 3 $T\frac{1}{2}$ measured by cpm per whole blood was within normal range and $T\frac{1}{2}$ measured by cpm per red cell mass showed shortening tendency when compared with the former in the group measured after blood loss (from 25 ml daily up to 100 ml daily in 10 days). In the group with rather constant blood loss of 100 ml daily for 10 consecutive days revealed the significant difference in two measurements (P<0.01). 4. $T\frac{1}{2}$ in non-steady state When red cell production is increased compared with red cell destruction, $T\frac{1}{2}$ measured by cpm per red cell mass being shorter than that by cpm per whole blood. Shortening of $T\frac{1}{2}$ measured by cpm per whole blood is more prominent. if red cell destrction is enhanced and exceeds production. 5. It is clear that when expressing red cell destruction rate, $T\frac{1}{2}$ measured by cpm per whole blood is more adequate and production more consistent with cpm red cell mass. 6. $T\frac{1}{2}$ measured during blood-letting, when corrected by amount of blood loss, it remains normal. It is erroneous to use conventional equational when measuring $T\frac{1}{2}$ in non-steady. $T\frac{1}{2}$ measured by cpm per whole blood is considred more applicable in clinical evaluation.
Immortalization of primary corneal cells has influence on pharmacy, medical and biological fields. Especially, investigation of immortalization mechanism using viral oncoproteins is useful for medical treatments, and these cell lines will be useful materials for toxic test of medical supplies and cell biological experiments. Rabbit corneal fibroblasts in culture undergo a finite number of divisions before they reach a terminally non-proliferating state known as replicative senescence. Therefore, we attempted to induce immortalization of rabbit corneal fibroblasts with SV 40 large T antigen. As a result of experiment, expression of SV 40 large T antigen was confirmed, and expression of proteins related to cell cycle repressor was decreased in the transfection group compared with non-transfection group. According to the results of cell cycle phase distribution test, SV 40 large T antigen-transfected cells had obtained higher proliferation rate than primary cells. It was confirmed that during induction of immortalization, SV 40 large T antigen was not able to increase telomerase activity. In conclusion, we made a rabbit corneal fibroblast cell line with SV40 large T antigen. This cell line will be useful for further studies of mammalian fibroblast biology, particularly with regard to angiogenesis and malignant transformation. In addition, this cell line offers opportunity for testing potential therapeutics and can be used for toxicity tests of materials or cosmetics. In the future, our cell line can potentially be utilized in a wide range of biology related fields.
For 16 years after the finding of HIV as an agent of AIDS in 1981, HIV therapeutic drugs of reverse transcriptase inhibitors (AZT, ddI, ddC, d4T) and protease inhibitors have been developed. Recent studies also were focused on a combination therapy by using HIV therapeutic drugs or natural compounds. Korean red ginseng (KRG) of natural compounds has been well known as a good reinforcement agent in Asia. The percentage of CD3+CD4+ T cell in nine HIV-infected patients without KRG treatment averaged 17.8% on baseline and decreased 15.8% after 6 months, whereas the percentage of the cell in fifteen HIV-infected patients with KRG treatment averaged 15.3% on baseline and increased up to 18.9% after the same period. The average percentage of CD3+CD8+ T cell of KRG-nontreated and KRG-treated HIV patients increased after 6 months 47.8% to 50.7% and 44.7% to 51.4%, respectively; and the average percentage of B and NK cell in the KRG-nontreated and KRG-treated HIV patients decreased 9.4% to 7.9% and 13.0% to 9.7%, 8.9% to 8.5% and 16.2% to 11.6%, respectively. KRG, therefore, didn't have any effects on the CD3+CD8+ T cell, B cell, and NK cell. However, it seems that KRG has a potential activity for stimulatiing the CD3+CD4+ T cell and some inhibition on destroying of this cell with no significance.
To develop hybridomas secreting monoclonal antibodies to be used as unlimited sources of reagents indispensable for the diagnosis and treatement of leukemic malignancy, a monoclonal antibody was generated to human pre-T leukemia cells (Jurkat). Hybridomas were produced against Jurkat cell line by fusing spleen cells from hyperimmunized mice with murine plasmacytoma cells (P3$\times$63Ag8. V653). One monoclonal antibody derived from this fusion, designated DMJ-2 was reactive with T-cell lines (Jurkat, Molt-4 and RPMI-8402) and normal peripheral E-rosette forming T cells, but unreactive with B-cell lines (Daudi, Nalm-6) and non-T, non-B cell line (K562). Conclusively DMJ-2 reactive with mature and immature T-lineage lymphoid cells.
The results of electron microscopic studies on the cell fine structure of Nocardia sp the location of tellurite-reducing enzyme and the reduction part of T. T. C. (Triphenyl tetrazonium chloride) were summarized as follows. As the fine structure of the cell, the membrane-like structure with unit membrane was distributed in the cytoplasm. The membrane-like structure had complicate forms: some of membrane-like structure appeared spiral form. As the metal tellurium salt appeared in the cytoplasm, it is obvious that tellurite and tellurate-reducing enzymes are present in the cytoplasm. Reduction of T. T. C. took place in the cell membrane and the intracellular membrane-like structure. Therefore, it was thought that reduction of tellurate and T. T. C. took place in different parts. T. T. C. formazane formed in the cell was reoxidized by osmic acid which was used as a fixation reagent for the electron microscopic specimen preparation. As 95% T. T. C. formazane was soluble in ethanol and embedding materials and removed out of the cell, an originally formed formazane appeared as electron light part on the electron microscopic image.
Receptor activator of NF-${\kappa}B$ ligand (RANKL) is an essential cytokine for osteoclast differentiation, activation and survival. T lymphocytes such as $T_{17}$ cells, a subset of T helper cells that produce IL-17, play an important role in rheumatoid arthritic bone resorption by producing inflammatory cytokines and RANKL. It has not yet been clearly elucidated how T cell activation induces RANKL expression. T cell receptor activation induces the activation of nuclear factor of activated T cell (NFAT) and expression of its target genes. In this study, we examined the role of NFAT in T cell activation-induced RANKL expression. EL-4, a murine T lymphocytic cell line, was used. When T cell activation was induced by phorbol 12-myristate 13-acetate (PMA) and ionomycin, RANKL expression increased in a time-dependent manner. In the presence of cyclosporin, an inhibitor of NFAT activation, this PMA/ionomycin-induced RANKL expression was blocked. Overexpression of either NFATc1 or NFATc3 induced RANKL expression. Chromatin immunoprecipitation results demonstrated that PMA/ionomycin treatment induced the binding of NFATc1 and NFATc3 to the mouse RANKL gene promoter. These results suggest that NFATc1 and NFATc3 mediates T cell receptor activation-induced RANKL expression in T lymphocytes.
IL-15 belongs to the common gamma chain cytokine family and has pleiotropic immunological functions. IL-15 is a homeostatic cytokine essential for the development and maintenance of NK cells and memory CD8+ T cells. In addition, IL-15 plays a critical role in the activation, effector functions, tissue residency, and senescence of CD8+ T cells. IL-15 also activates virtual memory T cells, mucosal-associated invariant T cells and γδ T cells. Recently, IL-15 has been highlighted as a major trigger of TCR-independent activation of T cells. This mechanism is involved in T cell-mediated immunopathogenesis in diverse diseases, including viral infections and chronic inflammatory diseases. Deeper understanding of IL-15-mediated T-cell responses and their underlying mechanisms could optimize therapeutic strategies to ameliorate host injury by T cell-mediated immunopathogenesis. This review highlights recent advancements in comprehending the role of IL-15 in relation to T cell responses and immunopathogenesis under various host conditions.
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