• Asian Pacific Journal of Cancer Prevention
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• v.13 no.8
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• pp.3815-3819
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• 2012

The CD4+CD25+ regulatory T cell (Treg) is a special kind of T cell subset. Studies have showed that Treg cells are involved in a number of physiological processes and pathologic conditions such as autoimmune diseases, transplantation tolerance and cancer. Tregs with unique capacity for immune inhibition can impair anti-tumour immunity and help tumor cells to escape from immune surveillance. The aim of our study was to investigate whether Tregs are involved in hepatocellular carcinoma (HCC). A BABL/C mouse with HCC in situ model was established to evaluate the Treg existence in carcinoma tissues and the changes of Tregs in spleen using flow cytometry and immunohistochemistry methods. Granzyme B expression in carcinoma tissues was analyzed by immunohistochemistry to investigate the tumor local immune status.The proportion of CD4+CD25+/CD4+ spleen lymphocytes of tumor bearing mice ($18.8%{\pm}1.26%$) was found to be significantly higher than that in normal mice ($9.99%{\pm}1.90%$) (P<0.01 ). Immunohistochemistry of spleen tissue also confirmed that there was an increase in Treg in tumor-bearing mice, while in carcinomas it showed Treg cells to be present in tumor infiltrating lymphocyte areas while Granzyme B was rarely observed. Anti-tumour immunity was suppressed, and this might be associated with the increase of Tregs. Our observations suggest that the CD4+CD25+Treg/CD4+ proportion in spleen lymphocytes can be a sensitive index to evaluate the change of Tregs in hepatocellular carcinoma mice and the Treg may be a promising therapeutic target for cancer.

• Journal of Yeungnam Medical Science
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• v.14 no.2
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• pp.359-369
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• 1997

We studied the expression of the cell surface antigen associated with myeloid and lymphoid leukemias on bone marrow or peripheral blood blast cells from 153 leukemic patients including 61 cases of acute myelogenous leukemias(AML), 46 of acute lymphocytic leukemias(ALL) and 12 of acute leukemias. They were analyzed by direct or indirect immunofluorescence method for reactivity with the monoclonal antibodies to B cells(CD10, CD19, SmIg), T cells(CD2, CD5, CD7, CD3, CD4, CD8), myeloid antigen(CD13, CD14, CD33, CD61) and a nonspecific antigen, HLA-DR. Lymphoid associated markers detected on AML is CD7 32.8%, CD10 14.8%, CD5 13.1%, CD2 6.6% and CD19 1.6%. TdT was positive in 4.9% of AMLs. Hybrid leukemias were 8 cases out 61 AML cases and were mainly composed of monocytic lineage, M4 and M5a. Myeloid markers detected in ALL were CD13 2.2% and CD33 2.2%. In this study, immunologically classified ALLs were composed of 65.2% of CALLA (+) B precursor type, 10.9% of CALLA (-) B precursor pattern, 8.7% of T cell type, 2.2% of B cell type, 4.5% of mixed lymphoid lineage(B&T), 2.2% of undifferentiated leukemia, and 6.5% of hybrid leukemia. Twelve cases of acute leukemias ware finally diagnosed to be 5 cases of hybrid leukemia, 3 cases of B lineage, 3 case of T lineage and 1 case of mixed lymphoid(B&T) leukemia. In summary, we think the best method for typing acute leukemias is by using a combination of FAB classification and immunophenotying.

• The Journal of Korean Medicine
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• v.24 no.1
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• pp.169-180
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• 2003

Background : Allergic asthma is thought to be mediated by $CD4^{+}$ T lymphocytes producing the Th2-associated cytokines. According to investigations of lung biopsies and respiratory secretions from patients, $CD4^{+}$ T cells and eosinophils are the main features of the inflammatory process. Object : This study aimed to find an inhibition effect on allergens induced by JCT (Jungchun-tang) and JCTG (Jungchuntanggagambang) through the change of $CD4^{+}$ T cells and $CD8^{+}$ T cells in BALF of rat, and to see the change of IgE m serum. Materials and Methods : Laboratory rats were primary sensitized with OA (ovalbumin); on day 1, rats of a control group and a sample group (SBP group) were systemically immunized by subcutaneous injection of 1mg OA and 300mg of A1(OH)3 in a total volume of 2 ml saline. The rats of the sample group were orally administered with an SBP water extract for 14 days after primary immunization. On day 14 after the systemic immunization, rats received local immunization by inhaling 0.9% saline aerosol containing 2%(wt/vol) OA. A day after local immunization, BAL fluid and serum were collected from the rats. Total cells, lymphocytes, $CD4^{+}$ T cells, $CD8^{+}$ T cells, and $CD4^{+}$/$CD8^{+}$ ratio in the BALF, and IgE level in serum were measured and evaluated. Results : L Total cell in BALF of rat : JCT was observed to be significantly reduced but JCTG had no significant difference in comparison with the control group. 2. Lymphocytes in BALF of rat : JCT and JCTG were observed to be significantly reduced in comparison with the control group. 3. $CD4^{+}$T cells in BALF of rat : JCT was observed to be more significantly reducing than JCTG in comparison with the control group. 4. $CD8^{+}$T cells in BALF of rat : JCT and JCTG were observed not to be significantly different than in the control group. 5. $CD4^{+}/CD8^{+}$ ratio in BALF of rat : JCT and JCTG were observed not to be significantly different than in the control group. 6. The IgE level in serum : JCT and JCTG were observed to be significantly reduced in comparison with the control group. Conclusion : This study shows that JCT inhibits allergen-induced specially select $CD4^{+}$T cell channel in BALF of rat.

• IMMUNE NETWORK
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• v.13 no.6
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• pp.264-274
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• 2013

The unrestricted population of $CD4^+Foxp3^+$ regulatory T (Treg) cells, which have been known to control the expression of autoimmune diseases and protective immunity to inflammatory reactions, has led to greater appreciation of functional plasticity. Detecting and/or isolating Ag-specific $CD4^+Foxp3^+$ Tregs at the single cell level are required to study their function and plasticity. In this study, we established and compared both MHC class II tetramer and intracellular CD154 staining, in order to detect $CD4^+Foxp3^+$ Treg specific for foreign Ag in acute and chronic infections with lymphocytic choriomeningitis virus (LCMV). Our results revealed that MHC class II tetramer staining showed a lower detection rate of LCMV $GP_{66-77}$-specific $CD4^+$ T cells because most of MHC class II tetramers were unbound and unstable when combined staining was performed with intracellular cytokines. In contrast, intracellular CD154 staining was revealed to be easier and simple for detecting LCMV $GP_{66-77}$-specific $CD4^+$ T cells, compared to MHC class II tetramer staining. Subsequently, we employed intracellular CD154 staining to detect LCMV $GP_{66-77}$-specific $CD4^+Foxp3^+$ Tregs using $Foxp3^{GFP}$ knock-in mouse, and found that LCMV $GP_{66-77}$-specific $CD4^+Foxp3^+$ Tregs and polyclonal $CD4^+Foxp3^+$ Tregs showed differential expansion in mice infected with LCMV Arms or Cl13 at acute (8 and 13 days pi) and chronic phases (35 days pi). Therefore, our results provide insight into the valuable use of intracellular CD154 staining to detect and characterize foreign Ag-specific $CD4^+Foxp3^+$ Treg in various models.

• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.23 no.3
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• pp.266-272
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• 2013

Objectives: This study aimed to evaluate the effects of low-level exposure to toluene on T lymphocytes subpopulations.s. Methods: The study lasted from April to October 2010. The subjects were 390 male workers, among whom 137 were chronically exposed to toluene in video-tape manufacturing factories and 253 were controls had never been occupationally exposed to hazardous chemicals. The subpoupulations of CD4+, CD8+, CD16+ (natural killer cells) and total (CD3+) T lymphocytes were examined by two-color staining using monoclonal antibodies. The general and job characteristics of subjects were assessed through a self-administered questionnaire. Results: There was no significant difference in general and job characteristics between both groups. No significant difference in lipid peroxide level was observed between the control and exposed workers, but the concentration of hydrogen peroxide was significantly higher in the exposed workers. The numbers of CD16+ T lymphocytes in controls were significantly higher than those in exposed workers, but no significant differences were found in CD4+, CD8+ and CD3+ T lymphocytes. Hydrogen peroxide levels showed a significantly negative correlation with CD8+ (r = -0.29, p < 0.01), CD16+ (r = -0.56, p < 0.01) and CD3+(r = -0.22. p < 0.01), and toluene levels was significantly negative correlated with CD3+ (r = -0.29, p < 0.05). Conclusions: Our results suggest that chronic low-level exposure to toluene affects cell-mediated immunity and the effects might mediate through ROSs (Reactive Oxygen Species) such as hydrogen peroxide.

• Korean Journal of Veterinary Research
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• v.61 no.3
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• pp.21.1-21.6
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• 2021

Canine T-zone lymphoma (TZL) is a mature T-cell lymphoma in dogs. The diagnosis and sub-classification are impossible without biopsy or immunophenotyping by flow cytometry. An 11-year-old, spayed, female Golden Retriever presented with lymph node enlargement. Clinical examination was consistent with canine multicentric lymphoma. However, immunophenotyping revealed positive for CD3, CD4, CD5, CD8, CD21, TCRαβ, and MHCII but negative for CD34, CD45, CD79a, and TCRγδ. Histopathology revealed lymphocytes expanding to the cortex-preserving architecture and thinning of the nodal capsule, and CD3 positive but PAX-5 negative. Owing to the indolent nature of TZL, careful monitoring approach without clinical intervention was utilized.

• The Korean Journal of Physiology and Pharmacology
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• v.24 no.4
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• pp.363-372
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• 2020

Gardenia jasminoides (GJ) is a widely used herbal medicine with anti-inflammatory properties, but its effects on the ORAI1 channel, which is important in generating intracellular calcium signaling for T cell activation, remain unknown. In this study, we investigated whether 70% ethanolic GJ extract (GJ_EtOH) and its subsequent fractions inhibit ORAI1 and determined which constituents contributed to this effect. Whole-cell patch clamp analysis revealed that GJ_EtOH (64.7% ± 3.83% inhibition at 0.1 mg/ml) and all its fractions showed inhibitory effects on the ORAI1 channel. Among the GJ fractions, the hexane fraction (GJ_HEX, 66.8% ± 9.95% at 0.1 mg/ml) had the most potent inhibitory effects in hORAI1-hSTIM1 co-transfected HEK293T cells. Chemical constituent analysis revealed that the strong ORAI1 inhibitory effect of GJ_HEX was due to linoleic acid, and in other fractions, we found that genipin inhibited ORAI1. Genipin significantly inhibited I_ORAI1 and interleukin-2 production in CD3/CD28-stimulated Jurkat T lymphocytes by 35.9% ± 3.02% and 54.7% ± 1.32% at 30 μM, respectively. Furthermore, the same genipin concentration inhibited the proliferation of human primary CD4⁺ T lymphocytes stimulated with CD3/CD28 antibodies by 54.9% ± 8.22%, as evaluated by carboxyfluorescein succinimidyl ester assay. Our findings suggest that genipin may be one of the active components of GJ responsible for T cell suppression, which is partially mediated by activation of the ORAI1 channel. This study helps us understand the mechanisms of GJ in the treatment of inflammatory diseases.

• Tuberculosis and Respiratory Diseases
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• v.86 no.4
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• pp.304-318
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• 2023

Background: Cancer-associated fibroblasts (CAFs) are key components of the tumor microenvironment and significantly contribute to immune evasion. We investigated the effects of CAFs on the immune function of CD4+ and CD8+ T cells in non-small cell lung cancer (NSCLC). Methods: We isolated CAFs and normal fibroblasts (NFs) from tumors and normal lung tissues of NSCLC patients, respectively. CAFs were co-cultured with activated T cells to evaluate their immune regulatory function. We investigated the effect of CAF conditioned medium (CAF-CM) on the cytotoxicity of T cells. CAFs were also co-cultured with activated peripheral blood mononuclear cells and further incubated with cyclooxygenase-2 (COX2) inhibitors to investigate the potential role of COX2 in immune evasion. Results: CAFs and NFs were isolated from the lung tissues (n=8) and lymph nodes (n=3) of NSCLC patients. Immune suppressive markers, such as COX2 and programmed death-ligand 1 (PD-L1), were increased in CAFs after co-culture with activated T cells. Interestingly, CAFs promoted the expression of programmed death-1 in CD4+ and CD8+ T cells, and strongly inhibited T cell proliferation in allogenic and autologous pairs of CAFs and T cells. CAF-CM decreased the cytotoxicity of T cells. COX2 inhibitors partially restored the proliferation of CD4+ and CD8+ T cells, and downregulated the expression of COX2, prostaglandin E synthase, prostaglandin E2, and PD-L1 in CAFs. Conclusion: CAFs promote immune evasion by suppressing the function of CD4+ and CD8+ T cells via their effects on COX2 and PD-L1 in NSCLC. The immunosuppressive function of CAFs could be alleviated by COX2 inhibitors.

• Journal of Physiology & Pathology in Korean Medicine
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• v.16 no.4
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• pp.801-809
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• 2002

CSBHT is known to improve immunological response in mice and humans. In this study, CSBHT effect was examined in the context of CD4+ T cells' survival and TCR/CD3 induced activation responses. Spleen cells from 8 week BALB/c mice were cultured in CSBHT containing medium without activation for 24, 48 hr. The MTS assay and revealed that CSBHT did not stimulate spleen lymphocytes as mitogen. Spleen lymphocytes were treated with anti-CD3e/anti-CD28 antibodies for 48hr. Flow cytometry revealed that activity of T cell decreased with CSBHT concentration. CD4+ T cells were isolated and cultured ,in CSBHT containing medium for 48 hr. CSBHT did not affect survival of sorted CD4+ T cells without any involvement of APC. In order to evaluate the direct effect of CSBHT on helper T cells's proliferative capacity prior to activation, CD4+ T cells are isolated after 24hr of culture in CSBHT containing medium and activated with and without anti-CD3e/anti-CD28 activation for 48hr. A higher level of CD69 was observed in 1 ㎍/㎖ of CSBHT treatment than control using flow cytometry. But low CD69 expression was observed in 5㎍/㎖ of CSBHT treatment. Expression of mRNA for cytokines in CD4+ T cell revealed that IL-2 expression was increased in 1 ㎍/㎖. The expression of IL-2R α, INF- γ were increased with concentration. On the other hand mRNA of IL-4 was decreased in dose dependent manner. Results suggest that CSBHT may be desirable for CD4+ T cell's activity in immune responses. Further more, CSBHT may relatively activate Th1 and inactivate Th2.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.14 no.1
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• pp.129-153
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• 2001

Background: SMG (升麻葛根湯加味方) is an herbal medicine which has been used in oriental medicine as a traditional therapeutic agent of pruritus and skin disease. Objective: This study was performed to investigate the effect of SMG on the anti-hypersensitivity and immune response in the murine of type I hypersensitivity induced by the experiment. Materials and Methods: Laboratory rats were primary sensitized with OA (ovalbumin); on day 1, rats of a Control group and Sample group (SMG group) were systemically immunized by subcutaneous injection of 1mg OA and 300mg of Al(OH)3 in a total volume of 2ml saline. The rats of the sample group were orally administered with an SMG water extract for 14 days after primary immunization. On day 14 after the systemic immunization, rats received local immunization by inhaling $0.9\%$ saline aerosol containing $2\%$(wt/vol) OA. A day after local immunization, BAL fluid and peripheral blood were collected from the rats. Total cell, lymphocyte, $CD4^+\;T\;cell,\;CD8^+\;T\;cell,\;CD4^+/CD8^+$ ratio in the BALF, and IgE, $CD4^+\;T\;cell,\;CD8^+$ T cell in the peripheral blood were measured and evaluated. Results: SMG showed a suppressive effect on the immune response in the rats. 1. Total Cells in the BALF decreased in the SMG treated group in comparison group, but statistic differences were not observed. 2. Total lymphocytes in the BALF were statistically decreased in SMG treated group in comparison to the control group. 3. CD4+ T cells in the BALF were statistically decreased in SMG treated group in comparison to the control group. 4. CD8+ T cells in the BALF were decreased in SMG treated group in comparison to the control group, but statistic differences were not observed. 5. The ratio of CD4+/CD8+ in the BALF was statistically decreased in SMG treated group in comparison to the control group. 6. The IgE level in serum was statistically decreased in SMG treated group in comparison to the control group. 7. The ratio of CD4+ and CD8+ in peripheral blood showed undetectable differences between each group of rats. From the experiment cited above, this study shows that SMG has both anti-hypersensitivity effects and immunoregulatory effects when administered to rats. Based on this experiment, it is suggested that SMG could be a useful immunomodulator and anti-allergy agent.