ABSTRACT
To study the effect of small interfering RNA targeting metastasis-associated lung adenocarcinoma transcript1 (si-MALAT1) combining with curcumin on the invasion and migration abilities of human colon cancer SW480 cells, and to explore the involved molecular mechanism. The recombinant lentiviral vector expressing si-MALAT1 was constructed, and its titer was determined by gradient dilution method. The colon cancer SW480 cells with stable expression of si-MALAT1 was established, followed by treatment with curcumin at different concentrations. The effect of curcumin or si-MALAT1 alone and the combination of the two on the cell activity was detected by MTT assay. The cell invasion and migration abilities were detected by transwell and scratch-wound assay. The relative expression level of MALAT1 was detected by RT-qPCR. The protein expression was determined by Western blot analysis. The IC50 of curcumin alone was 77.69 mmol/L, which was 51.17 mol/L when combined with curcumin and random sequence. The IC50 of curcumin was 30.02 mmol/L when combined with si-MALAT1. The increased susceptibility multiples was 2.58. The wound healing rates were 30.9% and 67.5% after treatment with si-MALAT1 combined with curcumin for 24 hrs and 48 hrs, respectively. The numbers of invasion cells were 200±12, 162±13, 66±8, 53±4 and 16±3 after treatment with si-MALAT1 combined with curcumin for 48 hrs. The relative expression level of lncRNA-MALAT1 in the curcumin group was 68%, and the relative expression level of lncRNA-MALAT1 in si-MALAT1group was 56%, and that for the combination treatment group was about 21%. The protein expression levels of β- catenin, c-myc and cyclinD1 were significantly down-regulated upon treatment with certain concentration of si-MALAT1 alone or combined with curcumin.si-MALAT1 could significantly inhibit the invasion and migration of SW480 cells by enhancing the sensitivity of SW480 cells to curcumin. The mechanism involved mignt be related to the down-regulation of β-catenin, c-myc and cyclinD1 proteins.
Subject(s)
Cell Migration Inhibition/drug effects , Colonic Neoplasms , Curcumin/pharmacology , Neoplasms/prevention & control , RNA , RNA, Small Interfering/drug effectsABSTRACT
PURPOSE: The purpose of this study was to investigate the biological role and mechanism of miR-373 targeting of TFIIB-related factor 2 (BRF2) in the regulation of non-small cell lung cancer (NSCLC) cells. MATERIALS AND METHODS: miRNA microarray chip analysis of four paired NSCLC and adjacent non-tumor tissues was performed. Quantitative real-time polymerase chain reaction (qRT-PCR) andwestern blotting were used to detect the expression levels of miR-373 and BRF2 in NSCLC tissues and cell lines. The dual-luciferase reporter method was performed to determine if BRF2 is a target of miR-373. MTT, wound-healing, Transwell, and flow cytometric assays were conducted to examine the proliferation, migration, invasion, and cell cycle progression of NSCLC A549 cells, respectively; western blotting was used to detect the expression of epithelial-mesenchymal transition (EMT)–related proteins. RESULTS: The miRNA microarray chip analysis demonstrated that miR-373 was down-regulated in NSCLC tissues, and this result was confirmed by qRT-PCR. Additionally, miR-373 was confirmed to target BRF2. Moreover, miR-373 expression was inversely correlated with BRF2 expression in NSCLC tissues and cell lines; both miR-373 down-regulation and BRF2 up-regulation were strongly associated with the clinicopathological features and prognosis of NSCLC patients. In vitro, overexpression of miR-373 markedly inhibited cell proliferation, migration, and invasion; up-regulated the expression of E-cadherin; and down-regulated the expression of N-cadherin and Snail in A549 cell. Knockdown BRF2 by siRNA resulted in effects similar to those caused by overexpression of miR-373. CONCLUSION: MiR-373 is decreased in NSCLC, and overexpression of miR-373 can suppress cell EMT, and inhibit the proliferation, migration, and invasion of NSCLC A549 cells by targeting BRF2.
Subject(s)
Humans , Blotting, Western , Cadherins , Carcinoma, Non-Small-Cell Lung , Cell Cycle , Cell Line , Cell Migration Inhibition , Cell Proliferation , Down-Regulation , Epithelial-Mesenchymal Transition , In Vitro Techniques , Methods , MicroRNAs , Prognosis , Real-Time Polymerase Chain Reaction , RNA, Small Interfering , Snails , Up-RegulationABSTRACT
ABSTRACT Objective: To describe the clinical aspects of cases of influenza A(H1N1)pdm09 in Brazil. Methods: A descriptive study of cases reported in Sistema de Informação de Agravos de Notificação (SINAN), 2009-2010. Results: As the final classification, we obtained 53,797 (56.79%) reported cases confirmed as a new influenza virus subtype, and 40,926 (43.21%) cases discarded. Fever was the most common sign, recorded in 99.74% of the confirmed and 98.92% of the discarded cases. Among the confirmed cases, the presence of comorbidities was reported in 32.53%, and in 38.29% of the discarded cases. The case fatality rate was 4.04%; 3,267 pregnant women were confirmed positive for influenza A new viral subtype and 2,730 of them were cured. The case fatality rate of pregnant women was 6.88%. Conclusion: The findings suggested concern of the health system with pregnant women, and patients with comorbidities and quality of care may have favored a lower mortality. We recommend that, when caring for patients with severe respiratory symptoms, with comorbidities, or pregnant women, health professionals should consider the need for hospital care, as these factors make up a worse prognosis of infection by the pandemic influenza virus. .
RESUMO Objetivo: Descrever os aspectos clínicos dos casos de influenza A(H1N1)pdm09 no Brasil. Métodos: Foi desenvolvido um estudo descritivo dos casos notificados no Sistema de Informação de Agravos de Notificação (SINAN) de 2009 a 2010. Resultados: Obtivemos como classificação final 53.797 (56,79%) casos notificados confirmados como influenza por novo subtipo viral e 40.926 (43,21%) descartados. Febre foi o sinal mais frequente, sendo registrada em 99,74% dos casos confirmados e em 98,92% dos descartados. Entre os confirmados, a presença de comorbidades foi notificada em 32,53% dos casos confirmados e entre 38,29% dos casos descartados. A taxa de letalidade foi de 4,04%. Das 3.267 gestantes confirmadas para influenza por novo subtipo viral, 2.730 evoluíram para cura. A taxa de letalidade de gestantes foi de 6,88%. Conclusão: Os achados sugeriram sensibilidade do sistema de saúde para com gestantes e portadores de comorbidades, e que a qualidade do cuidado pode ter favorecido a uma menor mortalidade. Recomendamos aos profissionais de saúde que, diante de casos de influenza pandêmica que apresentem gravidade do quadro clínico, comorbidades ou que estejam gestantes, seja considerada a assistência hospitalar, pois esses fatores compõem um pior prognóstico do quadro da infecção pelo vírus pandêmico da influenza. .
Subject(s)
Humans , Antibodies, Monoclonal, Humanized/adverse effects , Cell Migration Inhibition/drug effects , Crohn Disease/drug therapy , Leukoencephalopathy, Progressive Multifocal/chemically inducedABSTRACT
Gambogic acid (GA) is an anticancer agent in phase ‖b clinical trial in China but its mechanism of action has not been fully clarified. The present study was designed to search the possible target-related proteins of GA in cancer cells using proteomic method and establish possible network using bioinformatic analysis. Cytotoxicity and anti-migration effects of GA in MDA-MB-231 cells were checked using MTT assay, flow cytometry, wound migration assay, and chamber migration assay. Possible target-related proteins of GA at early (3 h) and late stage (24 h) of treatment were searched using a proteomic technology, two-dimensional electrophoresis (2-DE). The possible network of GA was established using bioinformatic analysis. The intracellular expression levels of vimentin, keratin 18, and calumenin were determined using Western blotting. GA inhibited cell proliferation and induced cell cycle arrest at G2/M phase and apoptosis in MDA-MB-231 cells. Additionally, GA exhibited anti-migration effects at non-toxic doses. In 2-DE analysis, totally 23 possible GA targeted proteins were found, including those with functions in cytoskeleton and transport, regulation of redox state, metabolism, ubiquitin-proteasome system, transcription and translation, protein transport and modification, and cytokine. Network analysis of these proteins suggested that cytoskeleton-related proteins might play important roles in the effects of GA. Results of Western blotting confirmed the cleavage of vimentin, increase in keratin 18, and decrease in calumenin levels in GA-treated cells. In summary, GA is a multi-target compound and its anti-cancer effects may be based on several target-related proteins such as cytoskeleton-related proteins.
Subject(s)
Humans , Antineoplastic Agents , Pharmacokinetics , Apoptosis , Breast Neoplasms , Drug Therapy , Metabolism , Calcium-Binding Proteins , Genetics , Cell Line, Tumor , Cell Migration Assays , Cell Migration Inhibition , Cell Proliferation , Computational Biology , Methods , Cytoskeleton , Metabolism , Electrophoresis, Gel, Two-Dimensional , Flow Cytometry , Gene Expression , Keratin-18 , Genetics , Oxidation-Reduction , Protein Biosynthesis , Protein Transport , Proteomics , Methods , Transcription, Genetic , Ubiquitin-Specific Proteases , Pharmacokinetics , Vimentin , Genetics , Xanthones , PharmacokineticsABSTRACT
Immunological response of host and parasite play a key role in developing vaccination and immunization. The present study deals with the immune response and effecter mechanism, which was confirmed by migration inhibition factor [MIF]. The present work was conducted in Parasitological Lab of Postgraduate Department of Zoology, Government Holkar Science College, Indore [M.P.] during 2006-2007. For MIF assay, lymphocytes were separated from heparinized blood of experimental and control mice. Aliquots of cell suspension were placed in four wells cut in a preparation of agarose in a Petri dish. Two wells were filled with soluble test antigen, while rest two wells were filled with medium [control wells]. Petri dish was incubated overnight at 37°C in a humidified environment at 5% CO2 in air. Cells migrated under the agarose in a circle were fixed and stained. Diameters of the migration areas were measured with ocular micrometer. MIF reaction was maximum [44.2%] in the group IVEgESAg5 and minimum [10.8%] in the group IVASoAg1. The maximum MIF reaction was shown by eggs ES antigen and least by adult worm somatic antigen. The interesting observation was that migration inhibition increases as dose increased or we could say the reaction was dose dependent Increased value of MIF response in vaccinated mice suggested the involvement of lymphocytes in cell-mediated immunity. This study also proves that excretory-secretory [ES] antigen of eggs from Trichuris muris was more effective in imparting immunity in mice
Subject(s)
Female , Animals, Laboratory , Lymphocytes/immunology , Cell Migration Inhibition , Trichuriasis , Mice , Vaccination , Antigens, HelminthABSTRACT
Understanding of the pathophysiology of inflammatory bowel disease (IBD) is constantly evolving and, recently, a number of biologic agents have been developed. They selectively target specific molecule or pathways and correct the imbalance of the gut immune system. Among them, antibody to tumor necrosis factor (anti-TNF-alpha) is the first developed drugs, and it dramatically improved the IBD management. However, more than one-third of the patients do not respond to the drugs due to antibody formation. To increase treatment efficacy, enormous effort to develop novel anti-cytokines which can be an alternative to anti-TNF-alpha has been made. They are anti CD4+ T cell cytokine including interleukin (IL)-12/23 and IL-17 blockers, selective anti-adhesion molecule known as natalizumab, vedolizumab and alicaforsen, T-cell proliferation inhibitor, anti-inflammatory cytokine, immune stimulator, growth factor, and mitogen-activated protein kinase inhibitor. The efficacy and safety of each drugs are under investigation. Some drugs reported very promising data, however, others showed disappointing and different results. In addition, most of the trials were done in a very small number of patients, and there is no trial comparing to anti-TNF-alpha. The present paper reviews the action mechanism, short or long term efficacy and safety of variable drugs other than anti-TNF-alpha in IBD.
Subject(s)
Humans , Antibodies, Monoclonal/therapeutic use , Cell Migration Inhibition , Colony-Stimulating Factors/therapeutic use , Cytokines/antagonists & inhibitors , Genetic Therapy , Immunosuppressive Agents/therapeutic use , Inflammatory Bowel Diseases/drug therapy , Intercellular Signaling Peptides and Proteins/therapeutic use , Mitogen-Activated Protein Kinases/antagonists & inhibitors , Stem Cell Transplantation , Tumor Necrosis Factor-alpha/antagonists & inhibitorsABSTRACT
To explore the effect of neamine on cell proliferation, migration, and invasion in H7402 human hepatoma cells. This study was conducted at the Institute of Genetics and Cytology, School of Life Science, Northeast Normal University, Changchun, China between October 2008 and February 2010. First, we employed the MTT [thiazol blue tetraolium bromide] and soft agar assay to detect the effect of neamine on cell proliferation, and investigated the migration and invasion by using a transwell assay in H7402 cells. We, then, investigated nuclear translocation of angiogenin by immunofluorescence staining. Finally, we stable transfected H7402 cells with the plasmids pCI-Ang [+] and pCI-Ang [-], which contain the entire coding region of human angiogenin in the sense and antisense orientations, to obtain angiogenin under-expressing/ over-expressing transfectants, and investigated the effect of neamine on angiogenin induced cell proliferation. The results showed that neamine positively inhibited the proliferation, migration, and invasion of H7402 cells. Nuclear translocation of angiogenin was blocked by neamine, and angiogenin-induced cell proliferation was inhibited by neamine. Neamine positively inhibited H7402 cells. Since the toxicity of neamine is much less than neomycin, and is close to that of streptomycin and kanamycin, it may serve as a lead agent for the development of hepatocellular carcinoma therapeutics
Subject(s)
Humans , Cell Proliferation/drug effects , Cell Migration Inhibition , Cell Movement/drug effects , Carcinoma, Hepatocellular , Neoplasm InvasivenessABSTRACT
The serpin maspin, a tumor suppressor in breast cancer was described as an inhibitor of cell migration and inducer of cell adhesion between the basement membrane and extracellular matrix resulting in inhibition of tumor metastasis. In contrast, overexpression of maspin is correlated with poor prognosis in other types of cancer. Little is known about expression, regulation and function of maspin in canine mammary tumors. It was demonstrated in this study, a loss of maspin expression in malignant canine mammary cells compared with a pool of normal canine mammary tissue, analyzed by quantitative real-time PCR; weak maspin expression in malignant canine mammary tumors were observed by immunohistochemistry. It was also demonstrated that a correlation with nuclear maspin expression and a good prognosis. It is suggested that maspin could be used as a prognostic marker in canine mammary neoplasia.
O serpin maspin, um supressor tumoral no câncer de mama foi descrito como inibidor de migração celular e indutor de adesão celular entre a membrana basal e a matriz extracelular resultando na inibição da metástase tumoral. Por outro lado, a alta expressão do maspin está relacionada com um mau prognóstico em outros tipos de câncer. Pouco se sabe sobre a expressão, regulação e função do maspin nos tumores mamários caninos. Neste estudo, foi demonstrada uma perda da expressão de maspin nas células mamárias malignas de cães quando comparadas com um pool de tecido mamário normal de cães, analisado por PCR quantitativa em tempo real. Houve uma expressão fraca maspin em preparações de tumores mamários malignos observadas por imuno-histoquímica. Também foi verificado que a expressão nuclear do maspin em tumores mamários caninos está relacionada a um bom prognóstico. Assim, o maspin pode ser utilizado como um marcador prognóstico nas neoplasias mamárias em cães.
Subject(s)
Animals , Cell Migration Inhibition , Dogs , Immunohistochemistry , Mammary Neoplasms, Animal , Molecular BiologyABSTRACT
As citocinas são proteínas responsáveis pela interação da quase totalidade do nosso organismo. Seus mecanismos de ação e funções serão revisados dentro de seus aspectos mais importantes. Atuando na imunidade inata e na adaptativa estas pequenas moléculas quando ausentes podem ser responsáveis por doenças graves decorrentes de defeitos importantes na sinalização intra e intercelular. Neste artigo discutiremos a interação do eixo da IL12-23/IFNy e as doenças a elas associadas e relacionadas com mutações em cinco diferentes genes INFGR1, INFGR2, IL12β, IL12Rβ1 e STAT1 que correspondem a pelo menos oito tipos de doenças genéticas e fenotipicamente bem caracterizadas.
Subject(s)
Humans , Cell Communication , Cell Migration Inhibition , Cytokines , Immunoglobulin E , Leukocytes , Mycobacterium , Defense Mechanisms , MethodsABSTRACT
The effect of time of administration of exogenous melatonin (M) at the rate of 100 microg/Kg BW of rat/day for 14 days on immunomodulation to killed Pasteurella multocida (P52 strain) vaccine (KPMV) was investigated in male albino rats during spring season with photoperiod of LL 13: DD 11 h and 25 +/- 2.5 degrees C air temperature and 70 +/- 4% relative humidity. The experiment was conducted at an altitude of 172 mts above mean sea level at latitude 28.20 degrees north, longitude 79.24 degrees east (Bareilly, U.P. India). The experimental animals were divided in-groups of 8 rats each, as KPMV + M at 4.00 h; KPMV + M at 16.00 h; KPMV and their controls M4, M16, PBS respectively. Humoral immune response was monitored at weekly intervals by an indirect ELISA and cellular immunity by leukocyte migration inhibition test (LMIT) and delayed type of hypersensitivity (DTH). As evinced by in-vitro assays and in-vivo protection studies, both humoral and cellular immune responses to KPMV were augmented in rats receiving exogenous melatonin at 4.00 h as compared to slightly reduced responses in rats treated with melatonin at 16.00 h. It was concluded that the circadian timings of melatonin administration modulate immune response in rats.
Subject(s)
Adjuvants, Immunologic/administration & dosage , Animals , Antibody Formation/drug effects , Bacterial Vaccines/immunology , Cell Migration Inhibition , Circadian Rhythm , Enzyme-Linked Immunosorbent Assay , Guinea Pigs , Hypersensitivity, Delayed/immunology , Immunity, Cellular/drug effects , Leukocytes/immunology , Male , Melatonin/administration & dosage , Pasteurella multocida/immunology , Photoperiod , Rats , Seasons , Time FactorsABSTRACT
Biomphalaria glabrata and Schistosoma mansoni relationship was studied by light microscopy (LM) and freeze-fracture replica technique (FFR). We observed very thin cytoplasmic extensions of hemocytes in the LM, which then surround immobilize the miracidia. FFR images showed that the contact site between hemocytes cytoplasmic extensions and the external tegumentary coat involved only superficial layers of miracidia. Numerous vacuoles and filopodia were observed in the hemocyte cytoplasm, the latter binding with those from neighboring cells. In spite of the close interfilopodia contact, no cellular junctions were seen at these sites nor between filopodia-miracidia contact areas. The observed migration of hemocytes and their disposition in layers surrounding the miracidia in vitro correspond to previous studies.
Subject(s)
Animals , Cricetinae , Biomphalaria , Hemocytes , Schistosoma mansoni , Biomphalaria , Cell Migration Inhibition , Freeze Fracturing , Hemocytes , Host-Parasite Interactions , Microscopy, ElectronABSTRACT
The present study investigates the effect of progesterone, a pregnane precursor of neurosteroids, and 4'-chlordiazepam (4'-CD), a specific ligand for mitochondrial diazepam binding inhibitor receptor (MDR) involved in neurosteroidogenesis, on restraint stress (RS)-induced modulation of humoral and cell-mediated immune responses. RS produced a significant reduction in anti-sheep red blood cells (SRBC) antibody titre, a measure of humoral immune response, and % leucocyte migration inhibition (LMI) and foot-pad thickness test, measures of cell-mediated immune responses. These effects of RS on immune responses were effectively blocked by pretreating the animals with progesterone (10 mg/kg, sc) or 4'-CD (0.5 mg/kg, sc) administered just before subjecting the animal to RS. The effect of both progesterone and 4'-CD on RS-induced immune modulation was significantly attenuated by bicuculline (2 mg/kg, ip) but not by flumazenil (10 mg/kg, ip). Unlike its effect on RS-induced immune responsiveness, progesterone (5, 10 mg/kg, sc) when administered to non-stressed animals produced a significant suppression of both humoral and cell-mediated immune responses which was not reversed by bicuculline. However, 4'-CD failed to modulate immune response in naive non-stressed animals. These results suggest that progesterone and 4'-CD affect stress-induced immune responses by modulating GABA-ergic mechanism. However, GABA-A receptor system does not appear to be involved in progesterone-induced immunosuppression in nonstressed animals.
Subject(s)
Animals , Antibody Formation/drug effects , Bicuculline/pharmacology , Cell Migration Inhibition , Diazepam/analogs & derivatives , Diazepam Binding Inhibitor/pharmacology , Dose-Response Relationship, Drug , Edema/chemically induced , GABA Antagonists/pharmacology , Hypnotics and Sedatives/pharmacology , Immunity, Cellular/drug effects , Male , Mice , Progesterone/pharmacology , Rats , Rats, Wistar , Restraint, Physical , Stress, Psychological/immunologyABSTRACT
In dorsolateral hippocampal lesioned, Sham and control animals, the total number of splenocytes was determined and macrophages, B cells and T cells were isolated and their percentage distribution in total splenocytes were determined. The leukocyte migration inhibition was studied in these groups using different fractions of splenocytes namely whole splenocytes, macrophage removed fraction, and nylon wool eluted T cell population. The same groups were also studied after an antigenic challenge. The results indicate alterations in the macrophages, adherent cell population as well as T cell population in lesioned and Sham animals and also in their immunized groups. These altered cell sub-populations may be responsible for the altered migration.
Subject(s)
Animals , B-Lymphocytes/immunology , Cell Adhesion/immunology , Cell Migration Inhibition , Cytokines/physiology , Erythrocytes/immunology , Hippocampus/physiology , Lymphocyte Count , Macrophages/immunology , Male , Neuroimmunomodulation/physiology , Rats , Rats, Wistar , Sheep/immunology , Spleen/cytology , T-Lymphocytes/immunologyABSTRACT
The present study was designed to delineate the role of H1- and H2-histamine receptors in the neuro-immune regulation in rats. The effects of H1- and H2-receptor antagonists on humoral and cell-mediated immune (HI and CMI) responses were investigated after intraperitoneal (i.p.) and intra-cerebroventricular (i.c.v.) administration. HI response was assayed by anti-sheep red blood cell (SRBC) antibody titre in presence and absence of 2-mercaptoethanol (2-ME). The CMI responses were evaluated by delayed type hypersensitivity (DTH) reaction (in vivo), i.e., measurement of footpad thickness, and lymphokine activity such as leucocyte migration inhibition (LMI) test (in vitro). On i.p. administration, both H1- (pheniramine and astemizole) and H2-receptor antagonists (ranitidine and cimetidine) were observed to produce significant enhancement of anti-SRBC antibody response. However, only H2- and not H1-receptor blockers were observed to stimulate CMI response significantly. When administered by icv route, only H2-receptor antagonists caused a statistically significant increase in both HI and CMI responses, while the H1-receptor blockers failed to modify the same. Thus, H2-receptors appear to play a major role in the histaminergic mechanisms involved in immunomodulation both at the level of immunocompetent cells active in the peripheral tissues as well as through the central nervous system structures involved in the central regulation of neuro-immune interaction.
Subject(s)
Animals , Antibody Formation/drug effects , Cell Migration Inhibition , Central Nervous System/physiology , Erythrocytes/immunology , Histamine/pharmacology , Histamine H1 Antagonists/administration & dosage , Histamine H2 Antagonists/administration & dosage , Immunity, Cellular/drug effects , Injections, Intraventricular , Male , Neuroimmunomodulation/physiology , Peripheral Nervous System/drug effects , Rats , Rats, Wistar , Receptors, Histamine H1/drug effects , Receptors, Histamine H2/drug effectsABSTRACT
O objetivo deste trabalho foi estudar a resposta imunológica de ratas diabéticas, submetidas a prenhez. Utilizamos 80 ratas wistar, adultas, virgens, que foram sorteadas para compor os grupos: prenhe e nao prenhe com e sem aloxana. As ratas que receberam aloxana foram separadas de acordo com a glicemia do primeiro dia de prenhez em três subgrupos: sensibilizado (glicemia menor que 120 mg/dl), diábetes moderado (glicemia entre 120 e 199 mg/dl) e diabetes grave (glicemia maior que 200 mg/dl). Avaliou-se nestes grupos: a produçao de anticorpos maternos contra eritrócito de carneiro (SE); a transferência destes anticorpos para os filhotes; a imunidade celular materna e fetal através da técnica de produçao do fator inibidor da migraçao de leucócitos (LIF) e a atividade efetora das células NK materna. Na prenhez de ratas normais nao houve alteraçao de nenhum dos parâmetros imunológicos estudados. As ratas diabéticas apresentaram menor produçao de LIF, indicando supressao na resposta celular, que se refletiu na menor produçao de anticorpos. O diabetes associado à prenhez, além de alterar a resposta imune celular e humoral, aumentou a atividade das células NK nos grupos sensibilizado e diabetes moderado. A hiperglicemia materna suprimiu a resposta celular dos filhotes, com menor produçao do LIF. A deficiência nos níveis de anticorpos dos filhotes decorre da baixa produçao de IgG materna, única classe de anticorpos capaz de ser transferida passivamente através da placenta.
Subject(s)
Animals , Female , Pregnancy , Rats , Immune System , Pregnancy in Diabetics/immunology , Pregnancy/immunology , Alloxan/adverse effects , Antibody Formation , Blood Glucose/analysis , Cell Migration Inhibition , Cytotoxicity, Immunologic , Diabetes Mellitus/chemically induced , Erythrocytes , Fetus/immunology , Immunization , Killer Cells, Natural , Rats, WistarABSTRACT
Pertussis toxin (Ptx) is a hexameric protein with classical AB architecture produced by Bordetella pertussis. The aim of this study was to investigate the effect og Ptx on migration of polymorphonuclear leukocytes to site of inflamation and on cell- dependent edema. Ptx was purified from the supernatant of the culture medium of B. pertussis using hydroxylapatite chromatography and fetuin affinity chromatography. Ptx induced a maximal clusterin of Chinese hamster ovary cells at concentration as low as 0.1 ng/ ml. Intravenous inection of Ptx (400 ng) significantly blocked the neutrophil migration induced by 200 ng of lipopolysaccharide (LPS from E. coli O111:B4; 2.27 ñ 0.13 vs 0.61 ñ 0.16 per 10**6 neutrophils/ml; P < 0.001; N = 5) and by 200 ng of formylmethionyl-leucyl-phenylalanine(fMLP; 2.53 ñ 0.45 vs 0.75 ñ 0.14 per 10**6 neutrophils/ml; P < 0.01; N=6) into the peritoneal cavities of male Wistar rats (eighing 150-180). In addition, Ptx (400ng) pretreatment also blocked the edema induced by intraplantar injection of 100 µg carrageenin ( increase in volume: 0.667 ñ 0.087 vs 0.313 ñ 0.058 ml; P < 0.01; N = 5) but not the edema induced by 100 µg dextran ( increase in volume: 0.537 ñ 0.06 vs 0.385 ñ 0.076 ml; P > 0.05; N = 5). These data demonstrate that Ptx blocked neutrophil migration induced by a direct f MLP stimulus of a site of inflammation. In addition, this toxin blocks the indirect stimulus of LPS on neutrophil migration. Furthermore, Ptx also inhibits the neutrophil-dependent edema induced by carrageenin, but not the edema induced by dextran that is in part dependent on basophil cells. These results warrant further studies on the mechanisms of Ptx inhibition of neutrophil-dependent edema and cell migration
Subject(s)
Animals , Male , Rats , Cell Migration Inhibition , Inflammation/physiopathology , N-Formylmethionine Leucyl-Phenylalanine/pharmacology , Neutrophils/physiology , Pertussis Toxin/pharmacology , Carrageenan/pharmacology , Dextrans/pharmacology , Lipopolysaccharides/pharmacology , Pertussis Toxin/isolation & purificationABSTRACT
The objectives of this study were to evaluate whether a newborn or a neonate is capable of responding immunologically after BCG vaccination and to find out if this immunity persists for one year. Normal infants aged between 0 days-3 months brought to immunization centre were included in the study. In vitro leukocyte migration inhibition test was performed in these children using Phytohemagglutinin and purified protein derivative (PPD). They were grouped based on their age at vaccination, their LMI values and on the time interval after vaccination. The mean values of % LMI (PPD) in all the age groups were positive and there were no significant differences between the newborns, the neonates and other groups. The values were positive and comparable even after 12 months in all the groups. The percentage of infants with positive or negative values to LMI (PHA) and negative values to LMI (PPD) were also comparable at different time intervals in different age groups. The results suggest that newborns or neonates are as capable of eliciting a positive immune response after BCG vaccination, as older infants and the practise of vaccinating a child at birth could be continued.
Subject(s)
Age Factors , BCG Vaccine/administration & dosage , Cell Migration Inhibition , Humans , Immunity, Maternally-Acquired , Immunization Schedule , Infant , Infant, Newborn , Sensitivity and Specificity , Tuberculosis/prevention & controlABSTRACT
In a double-blind placebo-control study the immunomodulating effect of cimetidine treatment for one week and placebo was investigated for cell-mediated immune reactions of 22 patients with herpes zoster (HZ). The mitogen induced leukocyte migration inhibition test (LMIT) and the in vitro proliferation of the patients' lymphocytes to exogenous IL-2 were used. Before any treatment, the mitogen induced leukocyte migration inhibition capacity (LMIC) of HZ patients was found to be significantly reduced (p < 0.02) as compared to healthy blood bank donors (controls). After one week, within the same treatment, the LMIC was significantly improved (p < 0.01). The patients' lymphoproliferative response to IL-2, before any treatment, was not significantly different from that of controls (p < 0.05). However, significantly higher values (p < 0.001) were found in patients tested 7 days after the disease onset as compared to those tested after 12 days. One-week cimetidine treatment significantly improved (p < 0.05) the lymphoproliferative response to IL-2 of initially low responders and had no effect on higher responder patients. In contrast to this, after one week of placebo treatment, a significant decrease in the patients' lymphoproliferative response to IL-2 could be observed as compared to patients' initial responses (p < 0.05) or to those of controls (p < 0.05). Although the number of cases is very small. The data suggest that after cimetidine treatment, as compared to placebo, healing from skin rash and pain was achieved in a significantly shorter time (p < 0.01).
Subject(s)
Adult , Aged , Aged, 80 and over , Case-Control Studies , Cell Migration Inhibition , Cimetidine/therapeutic use , Double-Blind Method , Female , Herpes Zoster/drug therapy , Humans , Immunity, Cellular/drug effects , Interleukin-2/immunology , Male , Middle AgedABSTRACT
The whole amoebic lysate (WAL), Sephadex G-200 eluted fraction-I. (F-I), detergent dissected membrane proteins (DDMP), amoebic membrane glycoprotein (AMG) and amoebal RNA were extracted from E. histolytica. Amoebal RNA recognized 98.21% where as F-I, AMG, DDMP and WAL recognised 92.85, 91.07, 89.28 and 75% of total 56 clinically proven and amoebic serology positive amoebic liver abscess cases respectively. Intensity of recognition of individual fraction i.e., leucocyte migration inhibition index of each fraction was found to be highly significant (P < 0.001) when compared to whole amoebic lysate. This indicate the leucocyte migration inhibition index responses of patients of amoebic liver abscess to various antigenic fractions of E. histolytica with variations.