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1.
Arch. endocrinol. metab. (Online) ; 61(2): 108-114, Mar.-Apr. 2017. tab, graf
Article in English | LILACS | ID: biblio-838426

ABSTRACT

ABSTRACT Objectives The presence of thyroglobulin (Tg) in needle washouts of fine needle aspiration biopsy (Tg-FNAB) in neck lymph nodes (LNs) suspected of metastasis has become a cornerstone in the follow-up of patients with papillary thyroid carcinoma (PTC). However, there are limited data regarding the measurement of anti-Tg antibodies in these washouts (TgAb-FNAB), and it is not clear whether these antibodies interfere with the assessment of Tg-FNAB or whether there are other factors that would more consistently justify the finding of low Tg-FNAB in metastatic LNs. Materials and methods We investigated 232 FNAB samples obtained from suspicious neck LNs of 144 PTC patients. These samples were divided according to the patient’s serum TgAb status: sTgAb- (n = 203 samples) and sTgAb+ (n = 29). The TgAb-FNAB levels were measured using two different assays. Tg-FNAB was also measured using two assays when low levels (< 10 ng/mL) were identified in the first assay of the metastatic LNs from the sTgAb+ samples. Results The TgAb-FNAB results were negative in both assays in all samples. Low levels of Tg-FNAB were identified in 11/16 of the metastatic LNs of the sTgAb+ patients and 16/63 of the sTgAb- patients (p < 0.05) using assay 1. The measurement of the Tg-FNAB levels using assay 2 indicated additional metastases in 5 LNs of the sTgAb+ patients. Conclusions Factors other than the presence of TgAb-FNAB may contribute to the higher number of metastatic LNs with undetectable Tg-FNAB in the sTgAb+ group. In addition, the measurement of Tg-FNAB using different assays was useful to enhance the diagnosis of metastatic LNs, particularly when cytological and Tg-FNAB results are discordant.


Subject(s)
Humans , Autoantibodies/blood , Thyroglobulin/blood , Thyroid Neoplasms/blood , Carcinoma/blood , Lymph Nodes/immunology , Reference Values , Carcinoma/immunology , Carcinoma/pathology , Carcinoma, Papillary , Fluoroimmunoassay/methods , Predictive Value of Tests , Biopsy, Fine-Needle/instrumentation , Biopsy, Fine-Needle/methods , Lymph Nodes/pathology , Lymphatic Metastasis/immunology , Lymphatic Metastasis/pathology , Neck
2.
Biol. Res ; 50: 8, 2017. graf
Article in English | LILACS | ID: biblio-838960

ABSTRACT

BACKGROUND: CD4+ T cells play an important role in the initiation of an immune response by providing help to other cells. Among the helper T subsets, interferon-γ (IFN-γ)-secreting T helper 1 (Th1) and IL-17-secreting T helper 17 (Th17) cells are indispensable for clearance of intracellular as well as extracellular pathogens. However, Th1 and Th17 cells are also associated with pathogenesis and contribute to the progression of multiple inflammatory conditions and autoimmune diseases. RESULTS: In the current study, we found that BJ-1108, a 6-aminopyridin-3-ol analogue, significantly inhibited Th1 and Th17 differentiation in vitro in a concentration-dependent manner, with no effect on proliferation or apoptosis of activated T cells. Moreover, BJ-1108 inhibited differentiation of Th1 and Th17 cells in ovalbumin (OVA)-specific OT II mice. A complete Freund's adjuvant (CFA)/OVA-induced inflammatory model revealed that BJ-1108 can reduce generation of proinflammatory Th1 and Th17 cells. Furthermore, in vivo studies showed that BJ-1108 delayed onset of disease and suppressed experimental autoimmune encephalomyelitis (EAE) disease progression by inhibiting differentiation of Th1 and Th17 cells. CONCLUSIONS: BJ-1108 treatment ameliorates inflammation and EAE by inhibiting Th1 and Th17 cells differentiation. Our findings suggest that BJ-1108 is a promising novel therapeutic agent for the treatment of inflammation and autoimmune disease.


Subject(s)
Animals , Female , Mice , Cell Differentiation/drug effects , Th1 Cells/drug effects , Encephalomyelitis, Autoimmune, Experimental/drug therapy , Th17 Cells/drug effects , Aminopyridines/pharmacology , Aniline Compounds/pharmacology , Spleen/immunology , CD4-Positive T-Lymphocytes/drug effects , CD4-Positive T-Lymphocytes/immunology , Reproducibility of Results , Apoptosis/drug effects , Apoptosis/immunology , Th1 Cells/immunology , Cell Proliferation/drug effects , Disease Models, Animal , Encephalomyelitis, Autoimmune, Experimental/immunology , Th17 Cells/immunology , Flow Cytometry , Aminopyridines/immunology , Aniline Compounds/immunology , Lymph Nodes/immunology , Mice, Inbred C57BL
3.
Article in English | IMSEAR | ID: sea-159279

ABSTRACT

Lymphoid malignancies (LM) are a heterogeneous group of disorders that are broadly divided into Hodgkin disease (HD) and Non-Hodgkin Lymphoma (NHL). Diagnosing lymphoid malignancies based on morphology in conjunction with immunohistochemistry (IHC) forms the basis of WHO classification and this has prognostic implications.With this background this study was designed thus including all the lymphoid malignancies both NHL and HD. Materials and Methods: This study was conducted at a tertiary centre in Uttarakhand and included a total of 116 cases of lymphoid malignancies. Of these 41 cases were of Hodgkin disease and 75 cases were of NHL. These cases were initially diagnosed on morphology employing Hematoxylin and Eosin (H&E) and special stains like Reticulin. Subsequently, a preliminary panel of monoclonal antibodies using CD3, CD15, CD20, CD30, and CD45 were employed. All the cases were then classified using WHO classification. Results: HD- Of the 41 cases of Hodgkin’s disease the commonest subtype was Nodular Sclerosis seen in 26 cases (48.78%). Reed Sternberg in reactive milieu is diagnostic of Hodgkin disease. In all cases except one Reed Sternberg cells exhibited positivity for both CD15 and CD30. NHL – Of the 75 cases of NHL an initial classification based on morphology was done. All the cases were classified according to International Working Formulation initially. Subsequently, IHC was employed using CD3, CD15, CD20 and CD45. The disease was then classified according to WHO classification and broadly divided into B or T cell types. B cell expression was seen in 60 cases (80%) and T cell expression in 15 cases (20%). The commonest B cell subtype was Diffuse Large B cell Lymphoma (26.4%).


Subject(s)
Adolescent , Adult , Child , Female , Hodgkin Disease/immunology , Humans , Immunohistochemistry/methods , Lymph Nodes/immunology , Lymphocytes/immunology , Lymphoma/immunology , Lymphoma, Non-Hodgkin/immunology , Male , Middle Aged , Young Adult
4.
Indian J Exp Biol ; 2015 Feb; 53(2): 82-92
Article in English | IMSEAR | ID: sea-158381

ABSTRACT

Toll-like receptors (TLR) are a family of pattern recognition receptors identifying pathogen associated molecular patterns (PAMPs). They play a critical role in the innate immune response during the initial interaction between the infecting microorganism and phagocytic cells. Here, we verified the presence of TLR-2 in spleen, lymph node and thymus of Swiss albino mice and their modulation after infection with Staphylococcus aureus and Lipopolysaccharide (LPS) challenge. It was seen that TLR-2 gene transcribed to its respective mRNA on S. aureus infection, in thymus, spleen and lymph node of mice but their levels and mode of expression varied. When challenged with LPS no prominent changes in the expression of TLR-2 receptor was observed but its expression increased gradually with time in the thymus, spleen and lymph node of S. aureus infected mice. TLR-2 expression was also found enhanced in infected splenic macrophages. By studying the serum cytokine profile the functionality of the receptor was measured. The results indicate the presence of TLR-2 in thymus, spleen and lymph node of Swiss albino strain of mice and that they are modulated by S. aureus.


Subject(s)
Animals , Anti-Bacterial Agents/pharmacology , Cytokines/blood , Cytokines/immunology , Gene Expression/drug effects , Gene Expression/immunology , Host-Pathogen Interactions/immunology , Lipopolysaccharides/immunology , Lipopolysaccharides/pharmacology , Lymph Nodes/immunology , Lymph Nodes/metabolism , Lymph Nodes/microbiology , Macrophages/immunology , Macrophages/metabolism , Macrophages/microbiology , Male , Mice , Microbial Sensitivity Tests , Reverse Transcriptase Polymerase Chain Reaction , Spleen/immunology , Spleen/metabolism , Spleen/microbiology , Staphylococcal Infections/blood , Staphylococcal Infections/immunology , Staphylococcal Infections/microbiology , Staphylococcus aureus/drug effects , Staphylococcus aureus/immunology , Staphylococcus aureus/physiology , Thymus Gland/immunology , Thymus Gland/metabolism , Thymus Gland/microbiology , Time Factors , Toll-Like Receptor 2/genetics , Toll-Like Receptor 2/immunology , Toll-Like Receptor 2/metabolism
5.
Rev. bras. enferm ; 67(5): 744-751, Sep-Oct/2014. tab
Article in Portuguese | LILACS, BDENF | ID: lil-731214

ABSTRACT

Este estudo verificou a prevalência e fatores associados ao distress de pacientes oncológicos, na opinião de familiares. Foram entrevistados 140 familiares responsáveis pelo cuidado de pacientes com câncer. O Termômetro de Distress foi adaptado para uso em familiares. Estes consideraram que 72,9% dos pacientes estavam com distress relacionado a preocupações (80,4%), nervosismo (78,4%), tristeza (74,5%), dor (67,6%), fadiga (67,6%) e problemas com alimentação (57,8%). Modelos de regressão logística hierárquica mostraram que, enquanto familiares do sexo masculino (OR=0,025) e idades mais avançadas (OR=0,006 a 0,059) tiveram menor risco de perceber o distress, indivíduos protestantes, comparados a católicos, tiveram chance 12,77 vezes maior de percebê-lo. Quanto aos fatores associados, nervosismo (OR=10,8) contribuiu significativamente mais com a percepção de distress pelos familiares quando comparado a fadiga (OR=3,38) ou ter plano de saúde privado (OR=2,55). Familiares podem ser grandes aliados na avaliação e acompanhamento do distress de pacientes com câncer.


The study aimed to verify the opinion of family members about distress on cancer patients and the factors associated with it. Interviews with 140 family members of cancer patients were conducted. The Distress Thermometer was adapted to be used with family members. Approximately 72.9% of patients were considered in distress, related to concern (80.4%), nervousness (78.4%), sadness (74.5%), pain (67.6%), fatigue (67.6%) and problems with eating (57.8%). The hierarchical logistic regression models showed that while male (OR=0.025) and older ages (OR=0.006 to 0.059) had lower risk of perceiving the distress, individuals Protestants, compared to Catholics, were 12.77 times more likely to perceive it. About the associated factors, nervousness (OR=10.8) contributed significantly more to the perception of distress for family members when compared to fatigue (OR=3.38) or have private health insurance (OR=2.55). Family can be great allies in the evaluation and monitoring of distress in patients with cancer.


Este estudio examinó la prevalencia y los factores asociados con el distress de los pacientes de cáncer, de acuerdo con los familiares. Fueran entrevistados 140 cuidadores familiares de pacientes con cáncer. El Termómetro de Distress fue adaptado para el uso en los familiares. Ellos encontraron que el 72,9% de los pacientes estaban con distress relacionado con preocupaciones (80,4%), nerviosismo (78,4%), tristeza (74,5%), dolor (67,6%), fatiga (67, 6%) y problemas con la alimentación (57,8%). Modelos de regresión logística jerárquica mostraran que, mientras los familiares de sexo masculino (OR=0,025) y de edades más avanzadas (OR=0,006 a 0,059) tuvieron un menor riesgo de percibir el distress, los individuos protestantes, comparados a los católicos, tuvieron oportunidad 12,77 veces mayor para percibirlo. En cuanto a los factores asociados, el nerviosismo (OR=10,8) contribuyó significativamente más a la percepción del distress de los familiares, en comparación con la fatiga (OR=3,38) o tener un seguro de salud privado (OR=2,55). Familiares pueden ser grandes aliados en la evaluación y el seguimiento de sufrimiento en los pacientes con cáncer.


Subject(s)
Animals , Male , Female , Rats , Burns/immunology , Burns/pathology , Lipopolysaccharides/pharmacokinetics , Apoptosis , Lipopolysaccharides/blood , Lymph Nodes/immunology , Lymph Nodes/pathology , Lymphocytes/pathology , Rats, Wistar , Spleen/immunology , Spleen/pathology
6.
Indian J Cancer ; 2014 Jul-Sep; 51(3): 267-271
Article in English | IMSEAR | ID: sea-154377

ABSTRACT

Background: The histological detection of axillary lymph node tumor metastases in cases of breast carcinoma is of major prognostic significance, but may be difficult when metastases are of microscopic size. The micrometastases can be detected either by immunohistochemistry (IHC) or serial sectioning. Aims: We investigated whether immunohistochemical techniques and serial sectioning can increase the accuracy of metastatic detection and compared the efficacy of both. Materials and Methods: Thirty cases of breast carcinoma were studied in all of whom the axillary lymph nodes had been reported as free of metastases. Blocks from these cases were serially sectioned and stained with hematoxylin and eosin and a single section was stained with monoclonal antibody to cytokeratin AE1/AE3 and epithelial membrane antigen. The positivity for micrometastases was correlated with size, number, grade and histological type of primary tumor, lymph node size and number. Results and Conclusion: In 5/30 previously unsuspected cases, micrometastases were revealed by IHC and in 1/30 by serial sectioning. These findings suggested that serial sectioning is a labor intensive, time consuming and impractical procedure. Micrometastases were more frequently detected with age of patient >50 years, Grade 2/3 tumor, tumor size >5 cm and more than one primary tumor. Immunohistochemical analysis can be recommended as a routine procedure or an adjunct to routine histological procedures for the correct staging of breast carcinoma and use of adjuvant chemotherapy, especially in the high risk group.


Subject(s)
Breast Neoplasms/diagnosis , Breast Neoplasms/surgery , Female , Humans , Immunohistochemistry/methods , Lymph Nodes/chemistry , Lymph Nodes/immunology , Lymph Nodes/surgery , Microtomy/methods , Neoplasm Metastasis/diagnosis
7.
Medicina (B.Aires) ; 74(3): 185-188, jun. 2014.
Article in Spanish | LILACS, BINACIS | ID: biblio-1165184

ABSTRACT

In cancer, B cells have been classically associated with antibody secretion, antigen presentation and T cell activation. However, a possible role for B lymphocytes in impairing antitumor response and collaborating with tumor growth has been brought into focus. Recent reports have described the capacity of B cells to negatively affect immune responses in autoimmune diseases. The highly immunogenic mouse tumor MCC loses its immunogenicity and induces systemic immune suppression and tolerance as it grows. We have previously demonstrated that MCC growth induces a distinct and progressive increase in B cell number and proportion in the tumor draining lymph nodes (TDLN), as well as a less prominent increase in T regulatory cells. The aim of this research was to study B cell characteristics and function in the lymph node draining MCC tumor and to analyze whether these cells may be playing a role in suppressing antitumor response and favoring tumor progression. Results indicate that B cells from TDLN expressed increased CD86 and MHCII co-stimulatory molecules indicating activated phenotype, as well as intracellular IL-10, FASL and Granzyme B, molecules with regulatory immunosuppressive properties. Additionally, B cells showed high inhibitory upon T cell proliferation ex vivo, and a mild capacity to secrete antibodies. Our conclusion is that even when evidence of B cell-mediated activity of the immune response is present, B cells from TDLN exhibit regulatory phenotype and inhibitory activity, probably contributing to the state of immunological tolerance characteristic of the advanced tumor condition.


Subject(s)
Animals , Sarcoma/immunology , B-Lymphocytes, Regulatory/immunology , Immune Tolerance/immunology , Lymph Nodes/immunology , Antigens, Neoplasm/immunology , Phenotype , Sarcoma/pathology , Cell Count , T-Lymphocytes, Regulatory/immunology , Cell Line, Tumor , Cell Proliferation/physiology , Flow Cytometry , Lymph Nodes/pathology , Mice, Inbred BALB C
8.
J. bras. pneumol ; 38(3): 321-330, maio-jun. 2012. ilus, tab
Article in Portuguese | LILACS | ID: lil-640755

ABSTRACT

OBJETIVO: Investigar o significado de marcadores de imunidade celular e de componentes elásticos/colágeno da matriz extracelular em estruturas granulomatosas em biópsias de pacientes com sarcoidose pulmonar ou extrapulmonar. MÉTODOS: Determinações qualitativas e quantitativas de células inflamatórias, de fibras de colágeno e de fibras elásticas em estruturas granulomatosas em biópsias cirúrgicas de 40 pacientes com sarcoidose pulmonar e extrapulmonar foram realizadas por histomorfometria, imuno-histoquímica, e técnicas de coloração com picrosirius e resorcina-fucsina de Weigert. RESULTADOS: A densidade de linfócitos, macrófagos e neutrófilos nas biópsias extrapulmonares foi significativamente maior do que nas biópsias pulmonares. Os granulomas pulmonares apresentaram uma quantidade significativamente maior de fibras de colágeno e menor densidade de fibras elásticas que os granulomas extrapulmonares. A quantidade de macrófagos nos granulomas pulmonares correlacionou-se com CVF (p < 0,05), ao passo que as quantidades de linfócitos CD3+, CD4+ e CD8+ correlacionaram-se com a relação VEF1/CVF e com CV. Houve correlações negativas entre CPT e contagem de células CD1a+ (p < 0,05) e entre DLCO e densidade de fibras colágenas/elásticas (r = -0,90; p = 0,04). CONCLUSÕES: A imunofenotipagem e o remodelamento apresentaram características diferentes nas biópsias dos pacientes com sarcoidose pulmonar e extrapulmonar. Essas diferenças correlacionaram-se com os dados clínicos e espirométricos dos pacientes, sugerindo que há duas vias envolvidas no mecanismo de depuração de antígenos, que foi mais eficaz nos pulmões e linfonodos.


OBJECTIVE: To investigate the significance of cellular immune markers, as well as that of collagen and elastic components of the extracellular matrix, within granulomatous structures in biopsies of patients with pulmonary or extrapulmonary sarcoidosis. METHODS: We carried out qualitative and quantitative evaluations of inflammatory cells, collagen fibers, and elastic fibers in granulomatous structures in surgical biopsies of 40 patients with pulmonary and extrapulmonary sarcoidosis using histomorphometry, immunohistochemistry, picrosirius red staining, and Weigert's resorcin-fuchsin staining. RESULTS: The extrapulmonary tissue biopsies presented significantly higher densities of lymphocytes, macrophages, and neutrophils than did the lung tissue biopsies. Pulmonary granulomas showed a significantly higher number of collagen fibers and a lower density of elastic fibers than did extrapulmonary granulomas. The amount of macrophages in the lung samples correlated with FVC (p < 0.05), whereas the amount of CD3+, CD4+, and CD8+ lymphocytes correlated with the FEV1/FVC ratio and VC. There were inverse correlations between TLC and the CD1a+ cell count (p < 0.05), as well as between DLCO and collagen/elastic fiber density (r = -0.90; p = 0.04). CONCLUSIONS: Immunophenotyping and remodeling both showed differences between pulmonary and extrapulmonary sarcoidosis in terms of the characteristics of the biopsy samples. These differences correlated with the clinical and spirometric data obtained for the patients, suggesting that two different pathways are involved in the mechanism of antigen clearance, which was more effective in the lungs and lymph nodes.


Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Extracellular Matrix/immunology , Immunity, Cellular , Immunophenotyping/methods , Sarcoidosis/immunology , Analysis of Variance , Biopsy , Collagen/immunology , Elastic Tissue/immunology , Elastic Tissue/pathology , Extracellular Matrix/pathology , Granuloma, Respiratory Tract/immunology , Granuloma, Respiratory Tract/pathology , Lung/immunology , Lung/pathology , Lymph Nodes/immunology , Lymph Nodes/pathology , Lymphocytes/immunology , Macrophages, Alveolar/immunology , Sarcoidosis, Pulmonary/immunology , Sarcoidosis, Pulmonary/pathology , Sarcoidosis/pathology
9.
Article in English | IMSEAR | ID: sea-140198

ABSTRACT

Background: Oral squamous cell carcinoma is the most common neoplasm and comprises of approximately 80% of the cancers occurring in the oral cavity. The role of the host response to this neoplasm has been recognized, and for many years the regional lymph node in tumor-bearing hosts has been considered as an anatomic barrier to the systematic dissemination of tumor cells. Morphological evaluation of the regional nodes has aided in understanding the immune response. Aim: The current study was carried out to observe the morphological changes occurring in the regional lymph nodes and to evaluate whether these features could be helpful in assessing the immunological status of the patient, and thereby, the prognosis of the patient. Materials and Methods: The study was based on lymph nodes from 63 patients with oral squamous cell carcinoma, who underwent radical neck dissection or modified neck dissection. In the lymph node, four morphological patterns were observed that included lymphocyte predominance, germinal center predominance, mixed pattern (sinus Histiocytosis), and an unstimulated pattern. The cases were then divided into four groups according to the predominant immunoreactivity pattern based on the World Health Organization (WHO) standardized system for reporting human lymph node morphology. Results: Revealed that risk of metastases to cervical lymph nodes in patients with lymphocyte predominance was less (28.6%) when compared to the high risk of metastases with germinal center predominance (68%), and these results were statistically significant (P < 0.05). Patients with a mixed pattern showed less risk of metastases (45.4%), while those with an unstimulated pattern had increased risk of metastases (66.6%), but the results were not statistically significant. It was also found that in the positive nodes, germinal center hyperplasia (50.2%) was the predominant pattern. Conclusion: The present study revealed that patients with lymphocyte predominance had less risk of metastases and patients with germinal center predominance had a high risk of metastases to the lymph node.


Subject(s)
Capillaries/pathology , Carcinoma, Squamous Cell/immunology , Carcinoma, Squamous Cell/pathology , Carcinoma, Squamous Cell/secondary , Endothelial Cells/pathology , Endothelium, Vascular/pathology , Forecasting , Germinal Center/pathology , Histiocytosis, Sinus/pathology , Humans , Hyperplasia , Lymph Nodes/immunology , Lymph Nodes/pathology , Lymphatic Metastasis/immunology , Lymphatic Metastasis/pathology , Lymphocytes/pathology , Macrophages/pathology , Mouth Neoplasms/immunology , Mouth Neoplasms/pathology , Neck Dissection/methods , Prognosis , Risk Factors
10.
Rev. argent. microbiol ; 43(1): 9-17, ene.-mar. 2011. graf, tab
Article in Spanish | LILACS | ID: lil-634672

ABSTRACT

El objetivo de este trabajo fue evaluar un ELISA indirecto desarrollado para medir la respuesta inmune humoral en carneros vacunados contra la linfoadenitis caseosa (LC) y/o desafiados con una cepa de Corynebacterium pseudotuberculosis homóloga. Se distribuyeron corderos de 4 meses clínicamente sanos en 4 grupos: grupo 1, corderos vacunados (G1, n = 5); grupo 2, corderos vacunados e inoculados (G2, n = 8); grupo 3, corderos inoculados (G3, n = 2); y grupo 4, control (G4, n = 2). Los animales del G1 y del G2 recibieron dos dosis de una bacterina experimental; los del G2 y del G3 fueron desafiados con una cepa de C. pseudotuberculosis cuatro semanas posvacunación. Se estudiaron por ELISA los títulos serológicos durante 7 meses y se efectuaron las necropsias en los grupos G2, G3 y G4. Se tomaron muestras de pulmón y linfonódulos para efectuar estudios bacteriológicos e histopatológicos. La cepa inoculada en los animales del G2 y del G3 reprodujo las lesiones macroscópicas y microscópicas típicas de la LC; ésta fue aislada del sitio de inoculación, de linfonódulos o de pulmón en 7/8 animales del G2 y en 2/2 animales del G3. La prueba de ELISA, con una sensibilidad del 98% y una especificidad del 100%, detectó diferencias significativas entre los serorreactores de los diferentes grupos experimentales y permitió establecer una relación con el tipo de tratamiento aplicado. Se concluye que el ELISA desarrollado puede ser una herramienta útil para identificar animales infectados y con clínica positiva a la LC.


The aim of this study was to evaluate an indirect specific ELISA developed for the detection of humoral immune response in vaccinated sheep and/or challenged with a Corynebacterium pseudotuberculosis strain. Healthy 4 month-old lambs were distributed into 4 groups: Group 1 immunized (G1, n = 5), Group 2 vaccinated/inoculated (G2, n = 8), Group 3 inoculated (G3, n = 2) and Group 4 control (G4, n = 2). Groups G1 and G2 received two doses of an experimental bacterin. Four weeks postvaccination, G2 and G3 groups were challenged with a C. pseudotuberculosis strain. Serological titers were studied by ELISA for 7 months and pathological studies were performed in groups G2, G3 and G4 by taking lung and lymph node samples for bacteriology and histopathology. The inoculated strain in G2 and G3 animals reproduced the macroscopic and microscopic lesions typical of caseous lymphadenitis (CL) and was isolated from the inoculation site, lymph nodes and/or lung in 7/8 animals from G2, and 2/2 animals of G3. The developed ELISA test had sensitivity and specificity of 98% and 100% respectively, detected significant differences between serological reactors of different experimental groups and allowed to establish a relationship with the type of treatment. We conclude that the developed ELISA may be a useful tool to identify infected animals with positive clinical CL.


Subject(s)
Animals , Antibodies, Bacterial/analysis , Bacterial Vaccines/immunology , Corynebacterium Infections/veterinary , Corynebacterium pseudotuberculosis/immunology , Enzyme-Linked Immunosorbent Assay/veterinary , Lymphadenitis/veterinary , Sheep Diseases/immunology , Sheep/immunology , Vaccination/veterinary , Corynebacterium Infections/immunology , Corynebacterium Infections/microbiology , Corynebacterium Infections/prevention & control , Enzyme-Linked Immunosorbent Assay/methods , Lung/immunology , Lymph Nodes/immunology , Lymphadenitis/immunology , Lymphadenitis/microbiology , Lymphadenitis/prevention & control , Random Allocation , Sensitivity and Specificity , Sheep Diseases/microbiology , Sheep Diseases/prevention & control
11.
The Korean Journal of Parasitology ; : 115-123, 2011.
Article in English | WPRIM | ID: wpr-47951

ABSTRACT

Toxoplasma gondii Korean isolate (KI-1) tachyzoites were inoculated intraduodenally to BALB/c mice using a silicon tube, and the course of infection and immune responses of mice were studied. Whereas control mice, that were infected intraperitoneally, died within day 7 post-infection (PI), the intraduodenally infected mice survived until day 9 PI (infection with 1x10(5) tachyzoites) or day 11 PI (with 1x10(6) tachyzoites). Based on histopathologic (Giemsa stain) and PCR (B1 gene) studies, it was suggested that tachyzoites, after entering the small intestine, invaded into endothelial cells, divided there, and propagated to other organs. PCR appeared to be more sensitive than histopathology to detect infected organs and tissues. The organisms spread over multiple organs by day 6 PI. However, proliferative responses of splenocytes and mesenteric lymph node (MLN) cells in response to con A or Toxoplasma lysate antigen decreased significantly, suggesting immunosuppression. Splenic CD4+ and CD8+ T-lymphocytes showed decreases in number until day 9 PI, whereas IFN-gamma and IL-10 decreased slightly at day 6 PI and returned to normal levels by day 9 PI. No TNF-alpha was detected throughout the experimental period. The results showed that intraduodenal infection with KI-1 tachyzoites was successful but did not elicit significant mucosal immunity in mice and allowed dissemination of T. gondii organisms to systemic organs. The immunosuppression of mice included reduced lymphoproliferative responses to splenocytes and MLN cells to mitogen and low production of cytokines, such as IFN-gamma, TNF-alpha, and IL-10, in response to T. gondii infection.


Subject(s)
Animals , Mice , Cell Proliferation , Cytokines/metabolism , Disease Models, Animal , Duodenum/immunology , Endothelial Cells/parasitology , Histocytochemistry , Immune Tolerance , Lymph Nodes/immunology , Mice, Inbred BALB C , Polymerase Chain Reaction , Rodent Diseases/immunology , T-Lymphocyte Subsets/immunology , Toxoplasma/immunology , Toxoplasmosis, Animal/immunology
12.
The Korean Journal of Parasitology ; : 245-254, 2011.
Article in English | WPRIM | ID: wpr-182109

ABSTRACT

Many immune down-regulatory molecules have been isolated from parasites, including cystatin (cystain protease inhibitor). In a previous study, we isolated and characterized Type I cystatin (CsStefin-1) of the liver fluke, Clonorchis sinensis. To investigate whether the CsStefin-1 might be a new host immune modulator, we induced intestinal inflammation in mice by dextran sodium sulfate (DSS) and treated them with recombinant CsStefin-1 (rCsStefin-1). The disease activity index (DAI) increased in DSS only-treated mice. In contrast, the DAI value was significantly reduced in rCsStefin-1-treated mice than DSS only-treated mice. In addition, the colon length of DSS only-treated mice was shorter than that of rCsStefin-1 treated mice. The secretion levels of IFN-gamma and TNF-alpha in the spleen and mesenteric lymph nodes (MLNs) were significantly increased by DSS treatment, but the level of TNF-alpha in MLNs was significantly decreased by rCsStefin-1 treatment. IL-10 production in both spleen and MLNs was significantly increased, and IL-10+F4/80+ macrophage cells were significantly increased in the spleen and MLNs of rCsStefin-1 treated mice after DSS treatment. In conclusion, rCsStefin-1 could reduce the intestinal inflammation occurring after DSS treatment, these effects might be related with recruitment of IL-10 secreting macrophages.


Subject(s)
Animals , Female , Mice , Antigens, Differentiation/analysis , Clonorchis sinensis/enzymology , Colon/pathology , Cystatins/metabolism , Cytokines/metabolism , Dextran Sulfate/toxicity , Helminth Proteins/metabolism , Immunologic Factors/metabolism , Inflammation/chemically induced , Interleukin-10/analysis , Intestines/drug effects , Lymph Nodes/immunology , Macrophages/chemistry , Mice, Inbred C57BL , Severity of Illness Index , Spleen/immunology
13.
Mem. Inst. Oswaldo Cruz ; 105(5): 698-705, Aug. 2010. ilus, tab
Article in English | LILACS | ID: lil-557233

ABSTRACT

Protection against Fasciola hepatica in goats immunized with a synthetic recombinant antigen from Schistosoma mansoni fatty acid-binding protein 14 (rSm14) was investigated by assessing worm burdens, serum levels of hepatic enzymes, faecal egg count and hepatic damage, which was evaluated using gross and microscopic morphometric observation. The nature of the local immune response was assessed by examining the distribution of CD2+, CD4+, CD8+ and γ´+ T lymphocytes along with IgG+, IL-4+ and IFN-γ+ cells in the liver and hepatic lymph nodes (HLN). The goats used consisted of group 1 (unimmunized and uninfected), group 2 [infected control - immunized with Quillaia A (Quil A)] and group 3 (immunized with rSm14 in Quil A and infected), each containing seven animals. Immunization with rSm14 in Quil A adjuvant induced a reduction in gross hepatic lesions of 56.6 percent (p < 0.001) and reduced hepatic and HLN infiltration of CD2+, CD4+, CD8+ and γ´+ T lymphocytes as well as IL-4+ and IFN-γ+ cells (p < 0.05). This is the first report of caprine immunization against F. hepatica using a complete rSm14 molecule derived from S. mansoni. Immunization reduced hepatic damage and local inflammatory infiltration into the liver and HLN. However, considering that Quil A is not the preferential/first choice adjuvant for Sm14 immunization, further studies will be undertaken using the monophosphoryl lipid A-based family of adjuvants during clinical trials to facilitate anti-Fasciolavaccine development.


Subject(s)
Animals , Antigens, Helminth/immunology , Fasciola hepatica/immunology , Fascioliasis/immunology , Fatty Acid Transport Proteins/immunology , Goat Diseases , Helminth Proteins/immunology , Fascioliasis , Fatty Acid Transport Proteins , Goats , Goat Diseases/immunology , Helminth Proteins , Liver/immunology , Liver , Lymph Nodes/immunology , Lymph Nodes , Vaccines/immunology
14.
Mem. Inst. Oswaldo Cruz ; 105(3): 263-268, May 2010. ilus, graf, tab
Article in English | LILACS | ID: lil-547296

ABSTRACT

Experimental autoimmune encephalomyelitis (EAE) is mediated by CD4+ Th1 cells that mainly secrete IFN-γ and TNF-α, important cytokines in the pathophysiology of the disease. Spontaneous remission is, in part, attributed to the down regulation of IFN-γ and TNF-α by TGF-β. In the current paper, we compared weight, histopathology and immunological parameters during the acute and recovery phases of EAE to establish the best biomarker for clinical remission. Female Lewis rats were immunised with myelin basic protein (MBP) emulsified with complete Freund's adjuvant. Animals were evaluated daily for clinical score and weight prior to euthanisation. All immunised animals developed the expected characteristics of EAE during the acute phase, including significant weight loss and high clinical scores. Disease remission was associated with a significant reduction in clinical scores, although immunised rats did not regain their initial weight values. Brain inflammatory infiltrates were higher during the acute phase. During the remission phase, anti-myelin antibody levels increased, whereas TNF-α and IFN-γ production by lymph node cells cultured with MBP or concanavalin A, respectively, decreased. The most significant difference observed between the acute and recovery phases was in the induction of TNF-α levels in MBP-stimulated cultures. Therefore, the in vitro production of this cytokine could be used as a biomarker for EAE remission.


Subject(s)
Animals , Female , Rats , Encephalomyelitis, Autoimmune, Experimental/immunology , Interferon-gamma/biosynthesis , Lymph Nodes/metabolism , Spleen/immunology , Tumor Necrosis Factor-alpha/biosynthesis , Acute Disease , Biomarkers/analysis , Encephalomyelitis, Autoimmune, Experimental/pathology , Lymph Nodes/cytology , Lymph Nodes/immunology , Myelin Basic Protein , Rats, Inbred Lew , Spleen/cytology , Time Factors , Weight Loss
15.
Medicina (B.Aires) ; 68(6): 423-427, nov.-dic. 2008. tab
Article in English | LILACS | ID: lil-633581

ABSTRACT

The purpose of this study was to characterize and quantify cells involved in immune response in metastasis-free regional lymph nodes (RLNs) draining different human epithelial tumors and compare them (by immunohistochemistry) with control lymph nodes from patients with non malignant diseases. We showed that T cells number was decreased in RLNs as compared to the controls with reduction in both CD4+ T cells and CD8+ T cells subsets and an inverted ratio (CD4+: CD8+). B lymphocytes and follicular dendritic cells were decreased with respect to the controls. S100+ dendritic cells (DCs) and mature DCs were detected in T dependent areas. Their mean number was significantly lower as compared to control. Immature DCs were significantly diminished compared to RLN and control nodes. CD57+ cells, follicular T helper cells and/or NK cells, were localized in the clear zone of germinal centres and their mean number was significantly increased. There were no CD57+ cells in hypoplastic follicles. In this study we show that RLNs draining human cancer present reduction in almost all immune cells, except CD57+ cells. These findings may be related to the deficient anti-tumor immune response in patients with cancer and subsequent tumor progression.


El objetivo del trabajo fue caracterizar y cuantificar utilizando inmuno-histoquímica, las células involucradas en la respuesta inmune en ganglios linfáticos regionales (GLRs) que drenan distintos tumores epiteliales malignos humanos y compararlas con ganglios controles (GLCs) provenientes de pacientes sin enfermedad neoplásica maligna. Determinamos que los GLRs presentaban una marcada depleción de linfocitos B y T, células dendríticas (CD) foliculares y CD interdigitantes maduras respecto a los controles. En los linfocitos T, además de estar disminuidos, se observó una inversión de la relación T CD4+: T CD8+, a favor de los T CD8+. La depleción de CD inmaduras fue mayor respecto a las maduras. Las células CD57+, células foliculares T helper y/o células NK, localizadas en las zonas claras de los centros germinativos, presentaron un marcado incremento en los GLRs comparados con los GLCs, excepto en los casos de ganglios linfáticos con folículos hipoplásicos. En este estudio, demostramos que los GLRs que drenan carcinomas humanos presentan una significativa reducción en casi todas las células de la respuesta inmune, excepto las células NK. Estos hallazgos podrían estar relacionados con la deficiente respuesta antitumoral de los pacientes con cáncer y la subsiguiente progresión tumoral.


Subject(s)
Adult , Aged , Humans , Middle Aged , Dendritic Cells/cytology , Dendritic Cells/immunology , Lymph Nodes/immunology , Lymphocyte Subsets/immunology , Neoplasms/immunology , T-Lymphocytes/cytology , B-Lymphocytes/immunology , Case-Control Studies , Immunity, Cellular , Lymph Nodes/pathology , Lymphatic Metastasis/immunology , Lymphatic Metastasis/pathology , Neoplasms/pathology , T-Lymphocytes/immunology
16.
The Korean Journal of Parasitology ; : 95-102, 2007.
Article in English | WPRIM | ID: wpr-169039

ABSTRACT

The mRNA expression of several cytokines was evaluated in splenocytes and mesenteric lymph node (MLN) cells of rats infected with Capillaria hepatica by reverse-transcription (RT)-PCR until week 12 after infection. IgG1 and IgG2a, which are associated with Th1 and Th2 response, respectively, were also assessed by ELISA. The results indicated that the majority of cytokines, including the Th1 (IL-2 and IFN-gamma) and Th2 cytokines (IL-4, IL-5 and IL-10) were expressed at maximal levels during the early stage of infection (after week 1-2), and the ELISA data also evidenced a similar pattern of changes in IgG1 and IgG2a. Th1 and Th2 cytokines responded in a similar fashion in this rat model. The expression of cytokines in splenocytes was significantly higher than that in MLN cells, thereby indicating that cytokine production is controlled more by spleen than by MLN. In addition, the observation that IFN-gamma expression increased unexpectedly at the time of maximal egg production (6 weeks after infection) indicated that IFN-gamma is a cytokine reacting against egg production. However, increased IL-5 expression occurring in tandem with worm activity indicated that the activity of C. hepatica might be controlled by IL-5 expression.


Subject(s)
Animals , Rats , Antibodies, Helminth/blood , Capillaria/immunology , Cytokines/biosynthesis , Disease Models, Animal , Enoplida Infections/immunology , Enzyme-Linked Immunosorbent Assay , Gene Expression Regulation , Immunoglobulin G/blood , Lymph Nodes/immunology , Lymphocytes/immunology , RNA, Messenger/analysis , Rats, Sprague-Dawley , Reverse Transcriptase Polymerase Chain Reaction , Spleen/cytology , Th1 Cells/immunology , Th2 Cells/immunology
17.
Journal of Korean Medical Science ; : 696-700, 2000.
Article in English | WPRIM | ID: wpr-171767

ABSTRACT

CD44 is a cell adhesion molecule with numerous isoforms created by mRNA alternative splicing. Expression of CD44 variants has been suggested to play a potential role in tumor progression and metastasis. We designed primers CD44V, CD44V6/7, CD44R1 and CD44V6-10 to analyze and compare the roles of each CD44 variants. Expressions of CD44 variants were investigated in normal colonic mucosa, the lymph nodes which was histopathologically free of cancer cell, and cancer tissues of 44 human colorectal cancer patients by RT-PCR method. The expression of CD44V was observed in 28 out of 39 (71.8%) tumors and 7 out of 11 (63.6%) N1 normal regional lymph nodes, and CD44V6/7 was observed in 28 out of 39 (71.8%) tumors and 9 out of 11 (81.8%) N1 normal regional lymph nodes. The expressions of CD44V and CD44V6/7 were most frequently observed compared with any other CD44 variants. In normal colonic mucosa, the expression of CD44 variants are low but in cancer tissue and its regional lymph node, the expression of CD44V and CD44V6/7 were significantly higher and more frequent than any other CD44 variants (p<0.05). These results suggest that CD44V and CD44V6/7 can be a molecular marker for colorectal cancer and its micrometastasis to the regional normal lymph node.


Subject(s)
Humans , Alternative Splicing , Hyaluronan Receptors/genetics , Colorectal Neoplasms/pathology , Colorectal Neoplasms/immunology , Gene Expression , Lymph Nodes/pathology , Lymph Nodes/immunology , Protein Isoforms/genetics
18.
Journal of Veterinary Science ; : 39-48, 2000.
Article in English | WPRIM | ID: wpr-103272

ABSTRACT

The studies have provided the first comprehensive comparison of the factors regulating activation and proliferation of WC1+ and WC1- gammadelta T cells. The investigation has shown that accessory molecules essential for activation and function of WC1+ and WC1- gammadelta T cells and the sources and roles of cytokines in activation of gammadelta T cells through the T cell receptor (TCR). The study has also shown that the role of cytokines in activation and function of gammadelta T cells activated indirectly through cytokines secreted by ab T cells, accessory cells and antigen presenting cells (APC). Cytokines were differentially produced by subpopulations of gammadelta T cells under different conditions of activation. The investigation obtained in this study has revealed that factors account for activation and proliferation of gammadelta T cells in cultures designed to study MHC-restricted responses to antigens. Evidence obtained here has shown there is biological relevance to activation under these culture conditions that points to potential regulatory and effector functions of gammadelta T cells. The investigations have also provided the information needed to begin identifying and characterizing antigens recognized by the TCR repertoires of WC1+ and WC1- gammadelta T cells. Finally, the investigations have provided the information needed to begin analysis of the mechanisms by which gammadelta T cells modulate MHC restricted immune responses to pathogens and derived vaccines.


Subject(s)
Animals , Cattle , Base Sequence , Concanavalin A , Cytokines/genetics , DNA Primers , Immunophenotyping , Lymph Nodes/immunology , Lymphocyte Activation , Receptors, Antigen, T-Cell, gamma-delta/immunology , Reverse Transcriptase Polymerase Chain Reaction , T-Lymphocytes/classification
19.
Journal of Korean Medical Science ; : 600-606, 1999.
Article in English | WPRIM | ID: wpr-10211

ABSTRACT

We investigated the expression of CD99 in 35 hyperplastic perigastric lymph nodes, which were resected for gastric carcinoma or chronic peptic ulcer. Essentially, all lymphocytes in lymph nodes expressed CD99, but there were two populations with respect to the intensity of CD99 expression--CD99high and CD99low cells. We showed CD99high cells were distributed in paracortical and medullary cords by immunohistochemical study while germinal center cells were CD99low. Using three-color flow cytometric analysis with CD3, CD4, CD8, CD19, CD23, CD45RA, CD45RO, CD69, CD138, IgM, IgD, and IgG, most of CD99high cells were shown to be activated/memory T cells. CD4+CD45RO+ T cells were the subset revealing the highest intensity of CD99 expression while CD4+CD45RA+ T cells were CD99low. Among B cells, IgG+ B cells revealed a higher level of CD99 molecules than IgM+ B cells. These results suggest that CD99 is one of activation-related molecules which are upregulated in recently activated lymphocytes.


Subject(s)
Adult , Aged , Humans , Antigens, CD/analysis , B-Lymphocytes/immunology , Cell Adhesion Molecules/analysis , Flow Cytometry , Germinal Center/immunology , Immunohistochemistry , Immunologic Memory/immunology , Lymph Nodes/immunology , Middle Aged , Peptic Ulcer/immunology , Stomach Neoplasms/immunology , T-Lymphocytes/immunology
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