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1.
Article in English | WPRIM | ID: wpr-880368

ABSTRACT

BACKGROUND@#Asbestos fibers possess tumorigenicity and are thought to cause mesothelioma. We have previously reported that exposure to asbestos fibers causes a reduction in antitumor immunity. Asbestos exposure in the mixed lymphocyte reaction (MLR) showed suppressed induction of cytotoxic T lymphocytes (CTLs), accompanied by a decrease in proliferation of CD8@*METHODS@#For MLR, human peripheral blood mononuclear cells (PBMCs) were cultured with irradiated allogenic PBMCs upon exposure to chrysotile B asbestos at 5 μg/ml for 7 days. After 2 days of culture, IL-15 was added at 1 ng/ml. After 7 days of MLR, PBMCs were collected and analyzed for phenotypic and functional markers of CD8@*RESULTS@#IL-15 addition partially reversed the decrease in CD3@*CONCLUSION@#These findings indicate that CTLs induced upon exposure to asbestos possess dysfunctional machinery that can be partly compensated by IL-15 supplementation, and that IL-15 is more effective in the recovery of proliferation and granzyme B levels from asbestos-induced suppression of CTL induction compared with IL-2.


Subject(s)
Asbestos/adverse effects , CD8-Positive T-Lymphocytes/metabolism , Humans , Interleukin-15/pharmacology , Lymphocyte Activation/immunology , T-Lymphocytes, Cytotoxic/metabolism
2.
Article in Chinese | WPRIM | ID: wpr-880131

ABSTRACT

OBJECTIVE@#To analyze the changes in the gene expression profile of T cells in CML patients after TCRζ up-regulation expression, and to explore the molecular mechanism of T cell reactivation after transgenic up-regulation of TCRζ.@*METHODS@#The peripheral blood mononuclear cells(PBMCs) from 3 newly untreated chronic-stage CML patients were collected, and the CD3@*RESULTS@#A total of 2248 differentially-expressed genes were obtained, including 553 up-regulated genes and 1695 down-regulated genes in experimental group as compared with those in control group (P<0.05) . The GO and KEGG enrichment analyses showed that differentially expressed genes involved in the biological processes related to T cell immune function, such as TCR signaling pathway, T cell proliferation and activation. Some of core genes involved in promoting the TCR signaling pathway, T cell proliferation, activation and apoptosis pathways were significantly up-regulated, while some core genes involved in inhibiting T cell activation were significantly down-regulated.@*CONCLUSION@#The molecular mechanism of the significantly improved T cell activation and proliferation ability in CML patients after TCRζ up-regulation may be related to the differential transcripts mediated signaling pathways of T cell activation, proliferation and apoptosis.


Subject(s)
Humans , Leukocytes, Mononuclear , Lymphocyte Activation , Receptors, Antigen, T-Cell/genetics , T-Lymphocytes , Up-Regulation
3.
Rev. moçamb. ciênc. saúde ; 6(1): 8-13, Out. 2020. Tab, ilus, graf
Article in Portuguese | RSDM, AIM | ID: biblio-1343979

ABSTRACT

Introdução: Moçambique é um dos países endémicos à malária. Em 2011, a estimativa de prevalência desta doença era de 40­80% em crianças dos 2­9 anos e 90% em crianças menores de 5 anos. Estas altas prevalências podem ser devido à dificuldade das crianças em desenvolverem uma resposta imune eficaz. São necessários mais estudos para entender a resposta imune nestas crianças. Este estudo teve como objectivo descrever as características imuno-hematológicas em crianças menores de 15 anos infectadas por Plasmodium falciparum. Metodologia: Foram recrutadas crianças de 2-15 anos, infectadas por P. falciparum. Em cada criança, cujo tutor legal consentiu que participasse no estudo, colheu-se 5 ml de sangue venoso para um tubo com anticoagulante K3EDTA. O sangue foi usado para a contagem automática de células por citometria de fluxo. Os resultados foram agrupados por idade, dos 2-8 anos e 9-15 anos. Resultados: Das 50 crianças incluídas no estudo, 84% tinham idades entre os 2-8 anos, 70% do sexo masculino e 4% com serologia positiva para HIV. O nível de hemoglobina foi mais elevado no grupo de 9-15 anos (10,3g/dL) em relação ao grupo de 2-8 anos (8,7g/dL). A contagem absoluta de linfócitos T-CD4 foi maior no grupo de 2-8 anos (819 cél./µl). A activação celular não apresentou diferenças entre os grupos. Conclusão: A maioria dos casos de malária e anemia aguda foi observada em crianças dos 2 aos 8 anos, predominantemente do sexo masculino. Os valores absolutos de linfócitos foram mais elevados nas crianças dos 2-8 anos, mas os valores percentuais linfocitários não diferiram entre os grupos.


Introduction: Mozambique is one of the endemic countries to malaria. In 2011, the estimated prevalence of this disease was 40­80% in children aged from 2­9 years and 90% in children under 5 years. These high rates may be due to the difficulty of children in building an effective immune response. Further studies are needed to understand the immune response mounted by children in the presence of Plasmodium. This study aimed to describe the immuno-haematological characteristics of children under 15 years infected with Plasmodium falciparum. Methodology: Children aged from 2-15 years, infected with P. falciparum, were recruited for the study. In each child, whose legal guardian consented to take part of the study, was collected 5 ml of venous blood to a K3EDTA anticoagulant tube. The samples were tested using automatic full blood cell counting and flow cytometry. The results were grouped by age, 2-8 years and 9-15 years. Results: From the 50 children included in the study, 84% were aged 2-8 years, 70% were male and 4% were HIV positive. The haemoglobin level was higher in the 9-15 year old group (10.3g/dL) compared to the 2-8 year old group (8.7g/dL). The absolute T-CD4 lymphocytes levels were higher in the 2-8 year old group (819 cells/µl). The T-CD8 lymphocytes activations levels were similar in both groups. Conclusion: The majority of the children attended in the Paediatric Emergency who diagnosed malaria were 2 to 8 years old. These children were predominantly male and presented acute anaemia. The absolute T-CD4 and T-CD8 lymphocytes levels were higher in children aged 2-8 years, but the percentage levels of lymphocytes did not differ between groups.


Subject(s)
Humans , Infant , Child , Plasmodium falciparum , Lymphocytes , Child , Malaria , Lymphocyte Activation , Prevalence , Flow Cytometry , Vector Borne Diseases , Host-Parasite Interactions , Anemia
4.
Rio de Janeiro; s.n; ilus; 2020. 173 p. ilus.
Thesis in Portuguese | LILACS | ID: biblio-1252813

ABSTRACT

As leishmanioses são causadas por cerca de 20 espécies de Leishmania e se apresentam em diversas formas clínicas, que podem variar de branda a muito grave. Elas afetam milhões de pessoas e até então não existem medidas de controle eficazes. Assim, a imunização da população seria uma alternativa eficiente de controle. Evidências sugerem que a resposta imune que se estabelece após a cura clínica de leishmanioses deva constituir um padrão de resposta imunológica associado à proteção, e que uma preparação vacinal deveria estimulá-la preferencialmente. Nosso grupo demonstrou que antígenos de L. (Viannia) naiffi (espécie considerada benigna) conseguem induzir respostas bem moduladas, e que soros de voluntários curados de leishmaniose tegumentar cutânea reconheceram frações desse antígeno consideradas imunodominantes. Experimentos anteriores demonstraram em hamster que a imunização intranasal com antígenos totais dessa espécie são capazes de induzir uma imunidade protetora contra L. (V.) braziliensis. O mesmo grupo já havia obtido a mesma resposta protetora utilizando o antígeno total de L. (L.) amazonensis. No entanto, sabe-se que nem todos os componentes presentes nesses antígenos estão associados à proteção e que a identificação de proteínas que sejam imunogênicas é uma etapa indispensável na formulação de uma vacina.


O objetivo deste trabalho foi identificar as proteínas imunodominantes presentes nas frações solúveis de L. (L.) amazonensis (LaAg(s)) e L.(V.) naiffi (LnAg(s)), que sejam conservadas dentro do gênero Leishmania, para formular uma vacina pan específica para o controle das leishmanioses. Primeiramente, os antígenos LnAg(s) e LaAg(s) foram subfracionados em gel de poliacrilamida e as bandas com peso molecular entre 35 e 100KDa foram extraídas e submetidas à análise por espectrometria de massa para análise proteômica. Desta análise, foram obtidas as sequências de aminoácidos, o tamanho das sequências, e a abundância das proteínas. As proteínas mais abundantes foram submetidas à análise de similaridade com proteínas de hospedeiros. As proteínas de LnAg(s) e LaAg(s) com baixa similaridade (<30%) às proteínas humanas foram analisadas por preditores de epítopos reconhecidos por linfócitos B.


Em paralelo, a predição de epítopos presentes em LnAg(s) e LaAg(s), capazes de se ligar com alta afinidade a moléculas HLA classes I e II, assim como a promiscuidade de ligação a alelos de HLA para cada proteína, foram avaliados dentre aquelas que apresentaram maior abundância e baixa similaridade. Por fim, após a análise de homologia das proteínas entre as espécies de Leishmania, foram identificadas 11 proteínas com mais homólogos. Elas apresentaram potencial de reconhecimento por linfócitos B, assim como também por componentes da resposta imune celular (como predito para a apresentação por HLA classes I e II). O potencial de ativação de linfócitos T pelos epítopos presentes nas 11 proteínas foi incrementado pela ampla capacidade de apresentação antigênica na população humana, devido à alta promiscuidade quanto à capacidade de ligação destes epítopos a alelos de HLA. Esses resultados contribuem para a identificação de antígenos com potencial de compor um protótipo vacinal capaz de induzir uma resposta imune protetora pan específica em humanos, com características imunodominantes e indutoras de resposta humoral e celular, para serem empregados no controle das leishmanioses. (AU)


Subject(s)
Humans , Leishmania mexicana , Lymphocyte Activation , Leishmaniasis , Immunization , Epitopes, T-Lymphocyte
5.
Article in English | WPRIM | ID: wpr-761933

ABSTRACT

Antiepileptic drugs (AEDs) can induce severe cutaneous adverse reactions (SCARs) such as Stevens-Johnson syndrome (SJS), toxic epidermal necrolysis (TEN), and drug reaction with eosinophilia and systemic symptoms (DRESS) syndrome. We performed HLA genotyping and lymphocyte activation tests (LATs) for five AED-induced SCAR patients (three males and two females; aged 40–66 years old). Three patients were treated with carbamazepine (CBZ) for pain control, one was treated with phenytoin (PHT) for seizure prevention, and one was treated with valproic acid (VPA) for seizure prevention. One patient was diagnosed with CBZ-induced DRESS syndrome and the remaining patients were diagnosed with SJS. All patients recovered from SCARs after stopping suspicious drugs and supportive care. LATs were conducted to confirm the culprit drug responsible for inducing SCARs; and LAT results were positive for the suspected culprit drugs, in all except in one case. HLA-A,


Subject(s)
Alleles , Anticonvulsants , Carbamazepine , Cicatrix , Drug Hypersensitivity Syndrome , Female , HLA-A Antigens , Humans , Long-Acting Thyroid Stimulator , Lymphocyte Activation , Lymphocytes , Male , Methods , Phenytoin , Seizures , Stevens-Johnson Syndrome , Valproic Acid
6.
Article in English | WPRIM | ID: wpr-772938

ABSTRACT

The activation mechanism of chimeric antigen receptor (CAR)-engineered T cells may differ substantially from T cells carrying native T cell receptor, but this difference remains poorly understood. We present the first comprehensive portrait of single-cell level transcriptional and cytokine signatures of anti-CD19/4-1BB/CD28/CD3ζ CAR-T cells upon antigen-specific stimulation. Both CD4 helper T (T) cells and CD8 cytotoxic CAR-T cells are equally effective in directly killing target tumor cells and their cytotoxic activity is associated with the elevation of a range of T1 and T2 signature cytokines, e.g., interferon γ, tumor necrotic factor α, interleukin 5 (IL5), and IL13, as confirmed by the expression of master transcription factor genes TBX21 and GATA3. However, rather than conforming to stringent T1 or T2 subtypes, single-cell analysis reveals that the predominant response is a highly mixed T1/T2 function in the same cell. The regulatory T cell activity, although observed in a small fraction of activated cells, emerges from this hybrid T1/T2 population. Granulocyte-macrophage colony stimulating factor (GM-CSF) is produced from the majority of cells regardless of the polarization states, further contrasting CAR-T to classic T cells. Surprisingly, the cytokine response is minimally associated with differentiation status, although all major differentiation subsets such as naïve, central memory, effector memory, and effector are detected. All these suggest that the activation of CAR-engineered T cells is a canonical process that leads to a highly mixed response combining both type 1 and type 2 cytokines together with GM-CSF, supporting the notion that polyfunctional CAR-T cells correlate with objective response of patients in clinical trials. This work provides new insights into the mechanism of CAR activation and implies the necessity for cellular function assays to characterize the quality of CAR-T infusion products and monitor therapeutic responses in patients.


Subject(s)
Antigens , Metabolism , CTLA-4 Antigen , Metabolism , Cell Differentiation , Cell Line , Cytokines , Metabolism , Cytotoxicity, Immunologic , Granulocyte-Macrophage Colony-Stimulating Factor , Pharmacology , Humans , Lymphocyte Activation , Allergy and Immunology , Lymphocyte Subsets , Metabolism , Phenotype , Proteomics , Receptors, Chimeric Antigen , Metabolism , Single-Cell Analysis , Methods , T-Lymphocytes, Regulatory , Metabolism , Th1 Cells , Cell Biology , Th2 Cells , Cell Biology , Transcription, Genetic , Up-Regulation
7.
Article in Chinese | WPRIM | ID: wpr-772044

ABSTRACT

OBJECTIVE@#To investigate the effect of sputum ubiquitin ligase (Cbl-b) gene known-down on the cytotoxicity of H9 T lymphocytes against human laryngeal squamous cancer Hep-2 cells and explore the underlying mechanism.@*METHODS@#CD4 T lymphocytes isolated from 12 patients with laryngeal squamous carcinoma and 12 healthy individuals were examined for Cbl-b mRNA expressions using RT-PCR. H9 T lymphocytes cultured in 96-well plates were transfected with Cbl-b siRNA via liposomes followed by treatment with an anti-IL-2 monoclonal antibody, with H9 T lymphocytes transfected with a scrambled sequence as the negative control. The expressions of Cbl-b mRNA and protein in the cells were detected using real-time fluorescent quantitative PCR and Western blotting, respectively. The killing effect of the treated T lymphocytes against Hep-2 cells was assessed using the cell counting kit (CCK-8). The positive expression rates of CD69 and CD25 on the surface of H9 T lymphocytes were determined using flow cytometry, and the levels of interleukin-2 (IL-2) and interferon-gamma (INF-γ) in the culture supernatants of H9 T lymphocytes were detected with ELISA.@*RESULTS@#The CD4 T lymphocytes from patients with laryngeal squamous carcinoma showed significantly increased Cbl-b mRNA level compared with those from healthy individuals ( < 0.05). Transfection of H9 T lymphocytes with Cbl-b siRNA significantly reduced the expression levels of Cbl-b mRNA and protein ( < 0.05), which were not significantly affected by subsequent treatment of the cells with the anti-IL-2 antibody (>0.05). At different target-effector ratios, the Cbl-b siRNA-transfected cells showed significantly higher Hep-2 cell killing rates and higher positivity rates of CD69 and CD25 expressions than the blank and negative control cells and the cells with both Cbl-b siRNA transfection and anti-IL-2 treatment ( < 0.05). Cbl-b silencing in H9 T lymphocytes resulted in significantly increased levels of IL-2 and INF-γ in the supernatant as compared with those in the blank and negative control groups ( < 0.05).@*CONCLUSIONS@#Cbl-b gene silencing effectively enhances the killing effect of H9 T lymphocytes against Hep-2 cells probably as the result of enhanced IL-2 secretion and T lymphocyte activation.


Subject(s)
Carcinoma, Squamous Cell , Genetics , Therapeutics , Gene Silencing , Humans , Laryngeal Neoplasms , Genetics , Therapeutics , Lymphocyte Activation , RNA, Small Interfering , T-Lymphocytes
8.
Frontiers of Medicine ; (4): 69-82, 2019.
Article in English | WPRIM | ID: wpr-771259

ABSTRACT

Cytokine-activated T cells (CATs) can be easily expanded and are widely applied to cancer immunotherapy. However, the good efficacy of CATs is rarely reported in clinical applications because CATs have no or very low antigen specificity. The low-efficacy problem can be resolved using T cell antigen receptor-engineered CAT (TCR-CAT). Herein, we demonstrate that NY-ESO-1 HLA-A*02:01-specific high-affinity TCR (HAT)-transduced CATs can specifically kill cancer cells with good efficacy. With low micromolar range dissociation equilibrium constants, HAT-transduced CATs showed good specificity with no off-target killing. Furthermore, the high-affinity TCR-CATs delivered significantly better activation and cytotoxicity than the equivalent TCR-engineered T cells (TCR-Ts) in terms of interferon-γ and granzyme B production and in vitro cancer cell killing ability. TCR-CAT may be a very good alternative to the expensive TCR-T, which is considered an effective personalized cyto-immunotherapy.


Subject(s)
Cell Line, Tumor , Cytokines , Metabolism , Cytotoxicity, Immunologic , Genetic Engineering , HLA-A2 Antigen , Metabolism , Humans , Immunotherapy, Adoptive , Methods , Lymphocyte Activation , Receptors, Antigen, T-Cell , Genetics , Allergy and Immunology , T-Lymphocytes , Allergy and Immunology
9.
Protein & Cell ; (12): 121-129, 2018.
Article in English | WPRIM | ID: wpr-756967

ABSTRACT

Novel biologics that redirect cytotoxic T lymphocytes (CTLs) to kill tumor cells bearing a tumor associated antigen hold great promise in the clinic. However, the ability to safely and potently target CD3 on CTL toward tumor associated antigens (TAA) expressed on tumor cells remains a challenge of both technology and biology. Herein we describe the use of a Half DVD-Ig format that can redirect CTL to kill tumor cells. Notably, Half DVD-Ig molecules that are monovalent for each specificity demonstrated reduced non-specific CTL activation and conditional CTL activation upon binding to TAA compared to intact tetravalent DVD-Ig molecules that are bivalent for each specificity, while maintaining good drug like properties and appropriate PK properties.


Subject(s)
Animals , Antibodies, Bispecific , Allergy and Immunology , Antibodies, Monoclonal , Allergy and Immunology , Pharmacokinetics , CD3 Complex , Metabolism , Cell Line, Tumor , Cytotoxicity, Immunologic , ErbB Receptors , Metabolism , Female , Humans , Lymphocyte Activation , Allergy and Immunology , Mice, SCID , Neoplasms , Allergy and Immunology , Pathology , Rats, Sprague-Dawley , T-Lymphocytes, Cytotoxic , Allergy and Immunology
10.
Asia Pacific Allergy ; (4): e26-2018.
Article in English | WPRIM | ID: wpr-750150

ABSTRACT

Both immediate and nonimmediate type hypersensitivity reactions (HRs) with a single dose of quinolone in the same patient have not been previously reported. A 47-year-old female patient referred to us because of the history of a nonimmediate type HR to radio contrast agent and immediate type HR to clarithromycin. She experienced anaphylaxis in minutes after the second dose of 50 mg when she was provocated with moxifloxacin. She was treated immediately with epinephrine, fluid replacement and methylprednisole and pheniramine. On the following day she came with macular eruptions, and she was treated with methylprednisolone. The positive patch test performed with moxifloxacin as well as the lymphocyte transformation test proved the T-cell mediated HR. In order to prove the immediate type HR, basophil activation test was performed but was found negative. This case report presents for the first time the 2 different types of HRs in a patient with a test dose of quinolone.


Subject(s)
Anaphylaxis , Basophils , Clarithromycin , Epinephrine , Female , Humans , Hypersensitivity , Lymphocyte Activation , Methylprednisolone , Middle Aged , Patch Tests , Pheniramine , T-Lymphocytes
11.
Intestinal Research ; : 273-281, 2018.
Article in English | WPRIM | ID: wpr-714183

ABSTRACT

BACKGROUND/AIMS: Mesalazine is an effective drug for treating ulcerative colitis (UC), but causes allergic symptoms in a few cases. Therefore, the objective of this study was to evaluate the usefulness of the drug-induced lymphocyte stimulation test (DLST) for the diagnosis of mesalazine allergy. METHODS: Patients with UC treated with mesalazine with or without a history of associated adverse events (AEs) were enrolled at Kyorin University Hospital from July 2016 to April 2017. RESULTS: The DLST was performed in 104 patients with UC, of which 24 had a history of AEs due to mesalazine treatment. The control value of DLST was 337.4±296.3 counts per minute (cpm) in the AE+ group and 408.0±371.9 cpm in the AE− group. The measured value of DLST was 578.8±424.7 cpm in the AE+ group and 476.5±471.8 cpm in the AE− group. The stimulation index (SI) was 243.9%±291.1% in the AE+ group and 119.8%±53.0% in the AE− group. The SI value and DLST positivity were significantly higher in the AE+ group than in the AE− group (P=0.030 and P=0.029, respectively). The test sensitivity and specificity were 0.240 and 0.805, respectively, and the false-positive and false-negative rate was 0.195 and 0.760, respectively. CONCLUSIONS: The DLST for mesalazine showed low sensitivity and high specificity, suggesting that it may be useful for the definitive diagnosis of allergy to mesalazine.


Subject(s)
Colitis, Ulcerative , Diagnosis , Humans , Hypersensitivity , Lymphocyte Activation , Lymphocytes , Mesalamine , Sensitivity and Specificity
12.
Article in English | WPRIM | ID: wpr-715546

ABSTRACT

PURPOSE: Little is known about the clinical value of peripheral blood immune profiling. Here, we aimed to identify colorectal cancer (CRC)-related peripheral blood immune cells and develop liquid biopsy-based immune profiling models for CRC diagnosis. METHODS: Peripheral blood from 131 preoperative patients with CRC and 174 healthy controls was analyzed by flow cytometry and automated hematology. CRC-related immune factors were identified by comparing the mean values of immune cell percentages and counts. Subsequently, CRC diagnostic algorithms were constructed using binary logistic regression. RESULTS: Significant differences were observed in percentages and counts of white blood cells, lymphocytes, neutrophils, regulatory T cells, and myeloid-derived suppressor cells (MDSCs) of patients and controls. The neutrophil/lymphocyte and Th1/Th2 ratios were also significantly different. Likewise, the percentages and counts of peripheral blood programed death 1, cytotoxic T lymphocyte antigen 4, B-and T-lymphocyte attenuator, and lymphocyte activation gene-3 were higher in patients with CRC. The binary logistic regression model included 12 variables, age, CD3+%, NK%, CD4+CD279+%, CD4+CD25+%, CD4+CD152+%, CD3+CD366+%, CD3+CD272+%, CD3+CD223+%, CD158b−CD314+CD3−CD56+%, Th2%, and MDSCs cells/µL, for the prediction of cancer. Results of retrospective and prospective evaluation of the area under the curve, sensitivity, and specificity were 0.980 and 0.940, 91.53% and 85.80%, and 93.50% and 86.20%, respectively. CONCLUSION: Peripheral blood immune profiling may be valuable in evaluating the immunity of CRC patients. Our liquid biopsy-based immune diagnostic method and its algorithms may serve as a novel tool for CRC diagnosis. Future largescale studies are needed for better characterization of its diagnostic value and potential for clinical application.


Subject(s)
Blood Cells , Colorectal Neoplasms , CTLA-4 Antigen , Diagnosis , Early Detection of Cancer , Flow Cytometry , Hematology , Humans , Immunologic Factors , Leukocytes , Logistic Models , Lymphocyte Activation , Lymphocytes , Methods , Neutrophils , Prospective Studies , Retrospective Studies , Sensitivity and Specificity , T-Lymphocytes , T-Lymphocytes, Regulatory
13.
Journal of Experimental Hematology ; (6): 1765-1771, 2018.
Article in Chinese | WPRIM | ID: wpr-774388

ABSTRACT

OBJECTIVE@#To investigate the effects of different stimultors (PHA, PMA and IL-2) and culture systems (PBMC and whole blood) on the proliferation of human peripheral blood lymphocyte subsets, so as to provide the experimental basis for selecting the appropriate system according to the experimental purposes.@*METHODS@#A total of 10 ml serum samples were collected from healthy volunteers (n=6). The 300 μl whole blood was directly used to detect lymphocyte subsets by flow cytometry. The 400 μl whole blood were inoculated respectively with 3 different stimulators at 37℃ and 5% CO2 for 60 h; Three different stimulators were also added to the PBMC which were isolated from 2 ml whole blood. Then the proliferation ability of lymphocyte subsets was analyzed by flow cytometry.@*RESULTS@#After the PBMC were stimulated with PHA, CD4CD8CD3 lymphocytes were the most subset; The proportion of CD3CD4 T lymphocytes and CD3CD19 B lymphocytes decreased after being stimulated by PMA (P<0.01, P<0.05); the lymphocyte subset ratio had no significant change after being stimulated by IL-2. After the whole blood system was stimulated with PHA, the CD4/CD8 T lymphoblasts were main subsets, the counts of B lymphocytes and NK cells were reduced; after being stimulated with PMA, the number of CD8CD3 T lymphoblast and CD4CD8T lymphocytes increased, the B/NK cells were not distinguished with the surface markers; after the whole blood system was stimulated with IL-2, the proportion of NK cells significantly increased (P<0.05).@*CONCLUSION@#Lymphocyte proliferation stimulated by PMA is the fastest, while the effect of IL-2 on the lymphocyte subset proportion stimulated by IL-2 is the minimal. After being stimulated by PHA the division cycles of lymphocyte are the most.


Subject(s)
Cell Proliferation , Flow Cytometry , Humans , Killer Cells, Natural , Lymphocyte Activation , Lymphocyte Count , Lymphocyte Subsets
15.
Ann. hepatol ; 16(2): 308-311, Mar.-Apr. 2017. tab
Article in English | LILACS | ID: biblio-887237

ABSTRACT

ABSTRACT A 75-year old male patient had been regularly visiting our hospital for the management of his type 2 diabetes mellitus since he was diagnosed at age 64 years. When he developed hypoglycemic episodes with sulfonylurea, ipragliflozin (50 mg/day) was started to replace the sulfonylurea therapy. However, 49 days after starting ipragliflozin, his AST increased from 13 to 622 U/L, ALT increased from 9 to 266 U/L, ALP increased from 239 to 752 U/L, and γ-GTP increased from 19 to 176 U/L. ZTT was 3.5 U, TTT was 0.4 U, and total bilirubin was 0.7 mg/dL. IgM hepatitis A antibody, hepatitis B antigen, hepatitis C virus antibody, IgM CMV antibody, and IgM EB VCA antibody were negative, whereas a lymphocyte transformation test for ipragliflozin was positive. Abdominal CT scan showed mild fatty liver but no sign of nodular lesions. Following admission to our hospital, he received liver supportive therapy with the discontinuation of ipragliflozin therapy. He was discharged from the hospital 18 days later with AST and ALT levels reduced to 20 U/L and 13 U/L, respectively. Based on the clinical presentation of this patient, it is highly important to monitor liver function along with other possible clinical complications (e.g., dehydration, ketosis, and urinary tract infection) associated with SGLT2 inhibitor therapy.


Subject(s)
Humans , Male , Aged , Lymphocyte Activation/drug effects , Diabetes Mellitus, Type 2/drug therapy , Chemical and Drug Induced Liver Injury/diagnosis , Chemical and Drug Induced Liver Injury/etiology , Chemical and Drug Induced Liver Injury/immunology , Glucosides/adverse effects , Hypoglycemic Agents/adverse effects , Thiophenes/adverse effects , Predictive Value of Tests , Risk Factors , Diabetes Mellitus, Type 2/diagnosis , Diabetes Mellitus, Type 2/blood , Chemical and Drug Induced Liver Injury/therapy , Liver Function Tests
16.
Article in English | WPRIM | ID: wpr-174155

ABSTRACT

Lichenoid drug eruption (LDE) is a rare form of delayed-type drug eruption. Among anti-tuberculosis (Tb) agents, cycloserine (CS) has been reported as a rare cause of LDE. Positive results on the lymphocyte transformation test (LTT) have not been reported in patients with LDE. In the present case, we performed LTT and a patch test, and successfully proved CS as the offending drug in this patient, who had been treated with multiple anti-Tb drugs. These observations suggest that CS should be considered a possible cause of LDE and that LTT can be an option for the diagnosis of LDE.


Subject(s)
Cycloserine , Diagnosis , Drug Eruptions , Drug Hypersensitivity , Humans , Lichenoid Eruptions , Lymphocyte Activation , Lymphocytes , Patch Tests
17.
Article in English | WPRIM | ID: wpr-172330

ABSTRACT

Allopurinol-induced severe cutaneous adverse reactions (SCARs) such as Stevens-Johnson syndrome (SJS), toxic epidermal necrolysis (TEN), and drug reaction with eosinophilia and systemic symptoms (DRESS) syndrome are reportedly associated with the HLA-B*58:01 genotype. Three patients who developed SCARs after allopurinol administration were subjected to HLA-B genotyping and lymphocyte activation test (LAT) to evaluate genetic risk and to detect the causative agent, respectively. All three patients given allopurinol to treat gout were diagnosed with DRESS syndrome. Symptom onset commenced 7-24 days after drug exposure; the patients took allopurinol (100–200 mg/d) for 2-30 days. HLA-B genotyping was performed using a polymerase chain reaction (PCR)-sequence-based typing (SBT) method. All patients had a single HLA-B*58:01 allele: HLA-B*13:02/*58:01 (a 63-year-old male), HLA-B*48:01/*58:01 (a 71-year-old female), and HLA-B*44:03/*58:01 (a 22-year-old male). Only the last patient yielded a positive LAT result, confirming that allopurinol was the causative agent. These findings suggest that patients with HLA-B*58:01 may develop SCARs upon allopurinol administration. Therefore, HLA-B genotyping could be helpful in preventing serious problems attributable to allopurinol treatment, although PCR-SBT HLA-B genotyping is time consuming. A simple genotyping test is required in practice. LAT may help to identify a causative agent.


Subject(s)
Aged , Alleles , Allopurinol , Cicatrix , Drug Hypersensitivity Syndrome , Genotype , Gout , HLA-B Antigens , Humans , Lymphocyte Activation , Lymphocytes , Methods , Middle Aged , Polymerase Chain Reaction , Stevens-Johnson Syndrome , Young Adult
18.
Chinese Medical Journal ; (24): 1211-1217, 2017.
Article in English | WPRIM | ID: wpr-330641

ABSTRACT

<p><b>BACKGROUND</b>Triggering receptor expressed on myeloid cell-1 (TREM-1) may play a vital role in mammalian target of rapamycin (mTOR) modulation of CD8+ T-cell differentiation through the transcription factors T-box expressed in T-cells and eomesodermin during the immune response to invasive pulmonary aspergillosis (IPA). This study aimed to investigate whether the mTOR signaling pathway modulates the proliferation and differentiation of CD8+ T-cells during the immune response to IPA and the role TREM-1 plays in this process.</p><p><b>METHODS</b>Cyclophosphamide (CTX) was injected intraperitoneally, and Aspergillus fumigatus spore suspension was inoculated intranasally to establish the immunosuppressed IPA mouse model. After inoculation, rapamycin (2 mg.kg-1.d-1) or interleukin (IL)-12 (5 μg/kg every other day) was given for 7 days. The number of CD8+ effector memory T-cells (Tem), expression of interferon (IFN)-γ, mTOR, and ribosomal protein S6 kinase (S6K), and the levels of IL-6, IL-10, galactomannan (GM), and soluble TREM-1 (sTREM-1) were measured.</p><p><b>RESULTS</b>Viable A. fumigatus was cultured from the lung tissue of the inoculated mice. Histological examination indicated greater inflammation, hemorrhage, and lung tissue injury in both IPA and CTX + IPA mice groups. The expression of mTOR and S6K was significantly increased in the CTX + IPA + IL-12 group compared with the control, IPA (P = 0.01; P= 0.001), and CTX + IPA (P = 0.034; P= 0.032) groups, but significantly decreased in the CTX + IPA + RAPA group (P < 0.001). Compared with the CTX + IPA group, the proportion of Tem, expression of IFN-γ, and the level of sTREM-1 were significantly higher after IL-12 treatment (P = 0.024, P= 0.032, and P= 0.017, respectively), and the opposite results were observed when the mTOR pathway was blocked by rapamycin (P < 0.001). Compared with the CTX + IPA and CTX + IPA + RAPA groups, IL-12 treatment increased IL-6 and downregulated IL-10 as well as GM, which strengthened the immune response to the IPA infection.</p><p><b>CONCLUSIONS</b>mTOR modulates CD8+ T-cell differentiation during the immune response to IPA. TREM-1 may play a vital role in signal transduction between mTOR and the downstream immune response.</p>


Subject(s)
Animals , CD8-Positive T-Lymphocytes , Cell Biology , Metabolism , Cell Differentiation , Genetics , Physiology , Female , Interferon-gamma , Metabolism , Invasive Pulmonary Aspergillosis , Metabolism , Lymphocyte Activation , Genetics , Physiology , Mice , Mice, Inbred BALB C , Myeloid Cells , Cell Biology , Metabolism , Ribosomal Protein S6 Kinases , Metabolism , TOR Serine-Threonine Kinases , Genetics , Metabolism , Tissue Culture Techniques
19.
Article in English | WPRIM | ID: wpr-226321

ABSTRACT

BACKGROUND: Traditional medicines have been leveraged for the treatment and prevention of obesity, one of the fastest growing diseases in the world. However, the exact mechanisms underlying the effects of traditional medicine on obesity are not yet fully understood. METHODS: We produced the transcriptomes of epididymal white adipose tissue (eWAT), liver, muscle, and hypothalamus harvested from mice fed a normal diet, high-fat-diet alone, high-fat-diet together with green tea, or a high-fat-diet together with Taeumjowitang, a traditional Korean medicine. RESULTS: We found tissue-specific gene expression patterns as follows: (i) the eWAT transcriptome was more significantly altered by Taeumjowitang than by green tea, (ii) the liver transcriptome was similarly altered by Taeumjowitang and green tea, and (iii) both the muscle and hypothalamus transcriptomes were more significantly altered by green tea than Taeumjowitang. We then applied integrated network analyses, which revealed that functional networks associated with lymphocyte activation were more effectively regulated by Taeumjowitang than by green tea in the eWAT. In contrast, green tea was a more effective regulator of functional networks associated with glucose metabolic processes in the eWAT. CONCLUSIONS: Taeumjowitang and green tea have a differential tissue-specific and pathway-specific therapeutic effect on obesity.


Subject(s)
Adipose Tissue, White , Animals , Diet , Gene Expression , Gene Regulatory Networks , Glucose , Hypothalamus , Liver , Lymphocyte Activation , Medicine, Traditional , Metabolism , Mice , Obesity , Sequence Analysis, RNA , Tea , Transcriptome
20.
Article in English | WPRIM | ID: wpr-49038

ABSTRACT

Allergy to antibiotics is an important worldwide problem, with an estimated prevalence of up to 10% of the population. Reaction patterns for different antibiotics have changed in accordance with consumption trends. Most of the allergic reactions to antibiotics have been reported for betalactams, followed by quinolones and macrolides and, to a lesser extent, to others, such as metronidazole clindamycin and sulfonamides. The diagnostic procedure includes a detailed clinical history, which is not always possible and can be unreliable. This is usually followed by in vivo, skin, and drug provocation tests. These are not recommended for severe, potentially lifethreaten reactions or for drugs that are known to produce a high rate of false positive results. Given the limitations of in vivo tests, in vitro test can be helpful for diagnosis, despite having suboptimal sensitivity. The most highly employed techniques for diagnosing immediate reactions to antibiotics are immunoassays and basophil activation tests, while lymphocyte transformation tests are more commonly used to diagnose non-immediate reactions. In this review, we describe different in vitro techniques employed to diagnose antibiotic allergy.


Subject(s)
Anti-Bacterial Agents , Basophils , Clindamycin , Diagnosis , Diagnostic Tests, Routine , Drug Hypersensitivity , Hypersensitivity , Immunoassay , In Vitro Techniques , Lymphocyte Activation , Macrolides , Metronidazole , Prevalence , Quinolones , Skin , Sulfonamides
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