Academic literature on the topic 'Th1 and Th17 cells'

The activation of naïve T cells results in their proliferation and differentiation into a particular T-helper (Th) phenotype, namely Th1, Th2 or Th17 cells. This thesis focuses on the role of pro-inflammatory Th1 and Th17 cells in the induction of autoimmune disease of the central nervous system (CNS), using murine experimental autoimmune encephalomyelitis (EAE) as the model. Classically, EAE has been considered to be a Th1-mediated disease. However, since the emergence of the Th17 cells, there has been a paradigm shift towards Th17 cells being the key pathogenic subset in autoimmune disease. This thesis established robust protocols for the differentiation of naïve T cells into myelin-reactive Th1 or Th17 cells, producing ‘clean’ populations devoid of any contaminating cells. Passive T cell-transfer experiments revealed that myelin-reactive Th1 cells could induce EAE, whereas Th17 cells could not. This lack of disease correlated with the inability of the Th17 cells to accumulate in the non-inflamed CNS. Myelin-reactive Th1 cells did have this capability and only once inflammation was established, could Th17 cells be identified in the CNS, potentially exacerbating the disease. After these differences were observed, the project investigated two main aims: 1) to identify differences in homing molecule expression on Th1 and Th17 cells which could explain the difference in their ability to home to the CNS, and to investigate the functional significance of such differences, by molecular blockade; 2) to investigate the requirements for three key cytokines in EAE pathogenesis in passive T cell transfer models, investigating IFN-γ,IL-17 and TNF-α. P-selectin glycoprotein ligand-1 appeared to be important for the initial entry of inflammatory T cells into the CNS. Th1 cells deficient in IFN-γ were capable of IFNinducing EAE. A proportion of the mice developed “atypical” clinical signs, which correlated with T cell infiltration predominantly of the brain, rather than the spinal cord. This atypical EAE may be IL-17 dependent. In conclusion, this thesis indicates the importance of not focusing all resources and therapeutic approaches on Th17- induced inflammation as Th17 cells may not play such a major role as previously thought.

Introdução: O Papilomavírus Humano está muito bem associado com diversos tipos de cânceres humanos, como câncer anogenital e oral. Alguns estudos demonstram que o aparecimento de lesões e a progressão para o câncer estão relacionados ao tipo de resposta imune do hospedeiro. Deste modo, evidências indicam que a resposta imune do hospedeiro tem um papel muito importante para o curso da infecção pelo HPV. Objetivo: Avaliar a resposta imune específica in vitro ao Papilomavírus Humano (HPV) em homens com lesões causadas por HPV e sem lesão por HPV. Material e Métodos: Foram recrutados 31 pacientes e 11 voluntários, que formaram 4 grupos de estudo; sendo 12 pacientes no Grupo A (HIV +/ HPV +); 09 pacientes no Grupo B (HIV-/HPV+); 10 pacientes no Grupo C (HIV+/ HPV-); e 11 indivíduos saudáveis no Grupo D (HIV-/HPV-). Foram realizados ensaios de cultura celular para mensurar a resposta celular específica \"in vitro\" do tipo Th1/Th2/Th17 (INF-y, IL-2, TNFalfa, IL-4, IL-10 e IL-17) sob o estímulo da vacina quadrivalente do HPV (HPV 6, 11, 16 e 18) e à proteína E7 de HPV-16. Resultados: O grupo coinfectado (HIV +/ HPV+) apresentou níveis mais elevados de citocinas, principalmente do perfil Th2, comparando-se com os dados dos demais grupos de estudo. O grupo coinfectado apresentou níveis elevados de IL-6 e IL-10 (Perfil Th2) em relação ao grupo controle (HIV-/HPV-), com significância estatística (p < 0.0001 e p < 0.0001, respectivamente). Conclusão: Foi demonstrada uma elevada produção de citocinas no grupo HPV+/HIV+, sugerindo uma forte imunomodulação pela coinfecção HIV/HPV. Entretanto, novos estudos devem ser realizados para comprovar estes dados. Além de apresentar um perfil essencialmente Th2 do grupo coinfectado, principalmente pelos níveis elevados de IL-6 e IL-10 apresentados, sugerindo que estas duas citocinas possam servir como biomarcadores para persistência viral, uma vez que, os pacientes soropositivos para HIV apresentam maior persistência de HPV, e monitorar a progressão para lesões mais graves<br>Introduction: Human Papillomavirus is associated with different types of human cancers, such as anogenital and oral cancer. Some studies show that the appearance of lesions and progression to cancer are related to the type of host immune response. Thus, evidence indicates that the host immune response has a role key in the course of HPV infection. Objective: To evaluate the specific immune response in vitro to HPV in men with lesions caused by HPV and without injury caused by HPV. Methods: We recruited 31 patients and 11 volunteers, who formed four groups, with 12 patients in Group A (HIV+/HPV+); 09 patients in Group B (HIV-/HPV+); 10 patients in Group C (HIV+/HPV-) and 11 healthy subjects in Group D (HIV-/HPV-). Cells culture assay was performed to measure the specific immune response \"in vitro\" Th1/Th2/Th17 (IFN-y, IL-2, TNF-alfa, IL-4, IL-10 and IL-17) under the stimulation of quadrivalent HPV vaccine (HPV 6, 11, 16 and 18) and the E7 protein of HPV-16. Results: The coinfected group (HIV+/HPV+) had higher levels of cytokines, especially Th2 profile, compared with data from the other study groups. The coinfected group showed high levels of IL-6 and IL-10 (Th2 profile) compared to the control Group (HIV- /HPV-), with statistical significance (p < 0.0001 and p < 0.0001, respectively). Conclusion: This study demonstrated a high production of cytokines in the coinfected group, suggesting a strong immunomodulation by coinfection HIV/HPV. However, further studies should be conducted to confirm these data. In addition to presenting essentially a Th2 profile, especially by high levels of IL-6 and IL-10 presented, suggesting that these two cytokines may serve as biomarkers for viral persistence, since HIV seropositive patients have a higher persistence of HPV, and monitor the progression to more serious injuries

BACKGROUND: Recently, it is found that T-helper (Th) 22 cells are involved in different types of autoimmune and tumor diseases. But, till now, no study has been carried out to understand the involvement of these cells in cervical cancer (CC). METHODS: Flow cytometry was used to determine the expression of interferon gamma (IFN-gamma), Interleukin-22 (IL-22), IL-17 in the peripheral blood of healthy controls (HC), CIN and cervical cancer patients. From peripheral blood mononuclear cells (PBMCs), mRNA expression levels of Aryl hydrocarbon receptor (AHR), RAR-related orphan receptor C (RORC), TNF-alpha and IL-6 were respectively determined. Using the method of ELISA, plasma concentrations of IL-22, IL-17 and TNF-alpha were examined. RESULTS: Th22 and Th17 cells were elevated in CC and CIN patients. Th1 cells and the plasma concentrations of IL-22 in CC patients were significantly increased compared with HC. In CC patients, an increased prevalence of Th22 cells was associated with lymph node metastases. There was a positive correlation between Th22 and Th17 cells, but an approximately negative correlation between Th22 and Th1 cells in CC patients. The mRNA expression of RORC, TNF-alpha and IL-6 was significantly high in CC patients. CONCLUSIONS: Our results indicate that there is a higher circulatory frequency of Th22, Th17 and Th1 cells in CC which may conjointly participate in the pathogenesis and growth of CC.

Avaliou-se a capacidade dos cimentos endodônticos: Sealapex®, Activ-GP® e AH-Plus® de alterarem o perfil das citocinas nas respostas Th1, Th2 e Th17, após a implantação destes em subcutâneo de camundongos. A quantificação das citocinas IL-2, IL-6, TNF-&alpha;, IFN-&gamma;, IL-4, IL-10 e IL-17 foi realizada in vivo, no tecido reacional circundante aos implantes, os quais foram confeccionados a partir de sondas nasogástricas estéreis e apirogênicas de cloreto de polivinila preenchidas com os cimentos, tendo um grupo controle com sondas vazias. Utilizou-se camundongos isogênicos da linhagem C57BL/6 machos de 6/8 semanas de vida sendo que cada um recebeu 2 implantes na região dorsal (lado direito e esquerdo). Após os períodos experimentais de 7, 21 e 63 dias, com os camundongos anestesiados, os implantes foram removidos juntamente com o tecido circundante, e os animais sacrificados em seguida por deslocamento cervical. As amostras de cada grupo foram divididas sendo: duas, contendo implante/tecido, processadas histotecnicamente e as demais com apenas tecido (sem implante) foram homogeneizadas e centrifugadas com solução formada por tampão RIPA e inibidor de protease. O sobrenadante, fruto deste processo, foi coletado e a dosagem das citocinas realizada através do kit CBA mouse-Th1/Th2/Th17 Cytokine Kit (BD Cytometric Bead Array, San Jose, CA, USA) em análise por citometria de fluxo. Os parâmetros avaliados foram a concentração da citocina em função do cimento testado em cada período experimental. Submeteu-se os resultados à análise estatística por meio do teste t com a correção de Welch\'s. Para todos os testes o nível de significância foi de 5%. Com relação à IL-2 houve diferenças estatísticas significantes entre os grupos Activ-GP® e AH-Plus® (p=0,0391). No período de 21 dias foram detectadas diferenças entre o grupo controle e AH-Plus® (p=0,0402) e entre o grupo Sealapex® e AH-Plus® (p=0,0244). O AH-Plus® induziou um maior aumento na IL-6, aos 7 dias em relação ao Activ-GP® (p=0,0286) e aos 21 dias entre o grupo controle (p=0.0402) e Activ-GP® (p=0.0244). Os níveis de TNF-&alpha; foram estatisticamente superiores após 7 dias quando comparou-se o grupo AH-Plus® com os demais. Observou-se que no grupo controle aos 7 e 21 dias ocorreram diferenças estatísticas em relação ao Sealapex® e AH-Plus® respectivamente quando avaliadas as concentrações de IFN-&gamma;. Houve também diferenças estatísticas entre o grupo controle e Sealapex® (p=0,0158) no período de 7 dias para a citocina IL-4. Os valores de IL-10 foram estatisticamente superiores para o grupo controle em relação ao Activ-GP® no período de 21 dias (p=0,0471). Com relação a IL-17 no período de 21 dias, observou-se os maiores valores para o grupo controle, seguido pelo Sealapex®, Activ-GP® e AH-Plus®. Foram detectadas diferenças entre os grupos controle e AH-Plus® (p=0,0121), controle e Activ-GP® (p=0,0262) e entre Sealapex® e Activ GP® (p=0,0314). Baseado nesses resultados podê-se concluir que: os cimentos endodônticos são capazes de modular as respostas Th1, Th2 e Th17 através da inibição ou estimulação da liberação das citocinas testadas. O cimento AH-Plus® promoveu as maiores diferenças no perfil de resposta Th1.<br>It was evaluated the capacity of the following endodontic sealers: Sealapex, Activ-GP and AH-Plus to modify the cytokine profile in Th1, Th2 and Th17 responses, after their implantation in the subcutaneous tissue of mice. Quantification of IL-2, IL-6, TNF-&alpha;, IFN-&gamma;, IL-4, IL-10 and IL-17 was performed in vivo, in the reactional tissue surrounding the implants, which were made from sterile nasogastric probes and apyrogenic of polyvinyl chloride filled with sealer, and a control group of empty probes. It was used isogenic mice of C57BL/6 lineage, 6/8 weeks old males, each of which received two implants in the dorsal region (left and right). After the experimental time of 7, 21 and 63 days, the mice were anesthetized and the implants were removed along with the surrounding tissue, the animals were then sacrificed by cervical dislocation. Samples from each group were divided as follows: two containing implant / tissue processed histologically and with only the remaining tissue (without implant) were mixed and centrifuged with a solution formed by RIPA buffer and protease inhibitors. The supernatant result of this process was collected and cytokine dosage accomplished by mouse-Th1/Th2/Th17 Cytokine CBA Kit Kit (BD cytometric Bead Array, San Jose, CA, USA) for flow cytometry analysis. The evaluated parameters were the cytokine concentration in function of sealer tested in each trial. The results were submitted to statistical analysis using the t test with Welch\'s correction. For all tests the significance level was 5%. With respect to IL-2 there were significant statistical differences between groups-Activ GP and AH-Plus (p=0.0391). In the period of 21 days differences were found between the control group and AH-Plus (p=0.0402) and between the group Sealapex and AH-Plus (p=0.0244). The AH-Plus induced a greater increase in IL-6, at 7 days compared to Activ-GP (p=0.0286) and at 21 days between the control group (p=0.0402) and Activ-GP (p=0.0244). The levels of TNF-&alpha; were significantly higher after 7 days when the AH-Plus group was compared with others. It was observed that in the control group at 7 and 21 days there were statistical differences in relation to Sealapex and AH-Plus respectively when evaluated concentrations of IFN-&gamma;. There were also significant differences between the control group and Sealapex (p=0.0158) within 7 days for the cytokine IL-4. The amounts of IL-10 were statistically higher in the control group compared to the Activ GP in a period of 21 days (p=0.0471). With respect to IL-17 in a period of 21 days, it was observed the highest values for the control group, followed by Sealapex, Activ-GP and AH-Plus. Differences were found between the control groups and AH-Plus (p=0.0121), control and Activ-GP (p=0.0262) and between Sealapex and Activ-GP (p=0.0314). Based on the presented results theendodontic sealers are able to promote changes in the response cytokine profile Th1, Th2 and Th17; Sealer AH-Plus produced the greatest changes, in the Th1 response profile.

Produzida por macrófagos e células dendríticas, a IL-27 é uma citocina heterodimérica capaz de induzir células Tr1 produtoras de IL-10 e consequentemente regular linfócitos Th1, Th2 e Th17, dependendo da doença envolvida. Partindo-se do pressuposto de que a infecção causada por Trypanosoma cruzi normalmente induz miocardite refletida pela migração intensa de linfócitos Th1 para o tecido cardíaco, nós analisamos o papel regulador da IL-27 nesta condição inflamatória. Nós inicialmente verificamos que a IL-27 foi prontamente induzida in vitro em células infectadas com T. cruzi. Para gerar miocardite intensa coordenada por linfócitos Th1, nós polarizamos linfócitos T naïves para o padrão Th1 na ausência de moléculas relacionadas ao perfil Th17 (camundongos IL-17R-/-, IL-23-/- e IL-6-/-). Como esperado, a inflamação cardíaca intensa e o dano tecidual foram observados na ausência das moléculas do padrão Th17, o que contribuiu para a morte prematura dos animais IL-17R-/-, IL-23-/- e IL-6-/-, precisa e notoriamente pela indução da migração excessiva de linfócitos Th1 para o tecido cardíaco via CXCL-9 e CXCL-10. Para explorar os mecanismos pelos quais a IL-27 controla a miocardite induzida pelo T. cruzi, nós encontramos um recrutamento substancial de macrófagos produtores de IL-27 para o tecido cardíaco, o qual foi mediado pelas quimiocinas CCL3 e CCL4 na ausência de moléculas do padrão Th17. Para determinar quais os receptores necessários para a produção de IL-27, nós observamos que macrófagos derivados da medula óssea de camundongos deficientes de TLR4-/-, TLR9-/- e NLRP3-/- aboliram completamente a produção desta citocina após a infecção in vitro com T. cruzi, enquanto o receptor TLR2 foi dispensável. Nós também verificamos que macrófagos produtores de IL-27 suprimiram linfócitos Th1 através da indução de células Tr1 produtoras de IL-10 após a infecção com T. cruzi. Em seguida, nós avaliamos se a IL-27 foi correlacionada com a proteção cardíaca durante a doença de Chagas. Nós observamos níveis séricos elevados de IL-27 tanto em pacientes com a forma clínica indeterminada ou cardíaca leve, enquanto pacientes com cardiomiopatia moderada ou grave produziram níveis reduzidos de IL-27. Neste estudo, nós descrevemos um novo mecanismo regulador desempenhado por macrófagos produtores de IL-27 no controle da miocardite induzida por T. cruzi. Macrófagos produtores de IL-27 podem suprimir processos inflamatórios desencadeados por linfócitos Th1, os principais vilões na doença de Chagas.<br>IL-27 is a heterodimeric cytokine produced by macrophages and dendritic cells known to induce IL-10-producing Tr1 cells and to regulate Th1, Th2, and Th17 lymphocytes, depending on the underlying disease. Because the infection caused by Trypanosoma cruzi normally induces myocarditis mirrored by an outstanding migration of Th1 cells to the heart tissue, we analyzed the regulatory role of IL-27 in this inflammatory condition. We firstly verified that IL-27 was promptly induced by in vitro T. cruzi-infected spleen cells. To generate a robust myocarditis coordinated by Th1 lymphocytes, we polarized lymphocytes to a Th1 pattern by infecting mice in the absence of Th17-related molecules (IL-17R-/-, IL-23-/-, and IL-6-/- mice). As expected, an impressive cardiac inflammation and damage was observed in the absence of Th17-related molecules, leading IL-17R-/-, IL-23-/-, and IL-6-/- mice to the premature death, precisely and notably by inducing an exuberant Th1 migration to the heart tissue via CXCL9 and CXCL10 chemokines. To explore the mechanisms by which IL-27 controls T. cruzi-induced myocarditis, we found a striking recruitment of IL-27-producing macrophages to the heart tissue mediated by increased levels of CCL3 and CCL4 chemokines in the absence of Th17-associated molecules. To gain further insights into the receptors required to IL-27 production, we observed that bone marrow-derived macrophages from TLR4-/-, TLR9-/-, and NLRP3-/- mice completely abolished IL-27 production after in vitro T. cruzi infection, while TLR2 was dispensable. We also verified that IL-27-producing macrophages supressed Th1 lymphocytes by inducing IL-10-producing Tr1 cells after T. cruzi infection. We next assessed whether IL-27 was correlated to cardiac protection during Chagas Disease. We observed augmented serum levels of IL-27 in either patients with indeterminate (asymptomatic) form or mild cardiac form, whereas patients with moderate or severe cardiomyopathy were poor producers of IL-27. Here, we described a novel regulatory mechanism developed by IL-27-producing macrophages in the control of T. cruzi-induced myocarditis. IL-27-producing macrophages can suppress inflammatory processes caused by Th1 lymphocytes, the bona fide culprits of Chagas Disease.

Anti-TNF agents have revolutionised the treatment of rheumatoid arthritis (RA), ankylosing spondylitis (AS) and psoriatic arthritis (PsA), however a significant proportion of patients respond inadequately. Studies in murine and human arthritis have paradoxically shown that anti-TNF treatment can increase circulating Th17 and Th1 cells but the relationship of these changes to treatment response remains unclear. The aim of the work in this thesis was to conduct a prospective, longitudinal investigation of patients with inflammatory arthritis during anti-TNF treatment and using clinical, ultrasound and immunological assessments to gain an understanding of the immune correlates of treatment response. Patients with RA (n=25), AS (n=15) and PsA (n=8) were recruited and followed over the first 12 weeks of treatment. Improvement in validated disease activity scores defined treatment responders and non-responders. Power Doppler ultrasound (PDUS) provided a quantitative assessment of changes in synovial thickening and vascularity during treatment, with synovial vascularity showing faster and greater reduction with treatment than synovial thickening. PBMCs testing using IL17 and IFNy ELISpot assays and flow cytometry consistently showed increased frequencies of circulating Th1 and Th17 cells in all three disease groups during anti-TNF therapy. Multiplex cytokine testing demonstrated a decrease in serum levels of proinflammatory cytokines and chemokines. Analyses of relationships between clinical, ultrasonographic and T-cell immunological changes revealed significant negative correlations between the increased frequency of Th1 and Th17 cells and reduction in synovial thickening and vascularity from baseline to 12 weeks on treatment in the RA group. Higher numbers of circulating Th17 cells at baseline in the RA group were associated with poorer anti-TNFa treatment response as defined by DAS28 score and ultrasonographic measures. This is the first study to link changes in T-cell immunopathology evaluated by cellular assays with morphological changes in arthritis assessed by PDUS during anti-TNFa treatment.

Experiências vivenciadas durante o período pré-natal são determinantes para a saúde do feto. A ocorrência de infecções maternas e a consequente ativação do sistema imune da mãe ocasionam uma série de alterações estruturais e funcionais no cérebro da prole, podendo predispor o indivíduo a transtornos psiquiátricos na vida pós-natal, como esquizofrenia e autismo. No entanto, estudos que investigam as alterações imunes na prole ainda são escassos na literatura. Dessa forma, o objetivo do presente estudo foi avaliar, na prole, o impacto da ativação imune materna sobre a atividade imune periférica, a resposta imune-inflamatória no sistema nervoso central (SNC), e sobre o desenvolvimento da encefalomielite autoimune experimental (EAE), o modelo murino de Esclerose Múltipla. Camundongos fêmeas prenhes receberam uma administração de salina ou lipopolissacarídeo (LPS) ao final da gestação (dia gestacional 17) e, quando adulta, a prole foi submetida a 3 experimentos principais, analisando: (1) produção de citocinas, atividade de células da periferia e desenvolvimento da hipersensibilidade do tipo tardia; (2) produção de mediadores inflamatórios por células residentes do SNC e; (3) desenvolvimento dos sintomas clínicos e da resposta imune no decorrer da EAE. Nossos resultados mostraram que a ativação imune materna provocou na prole alterações imunes periféricas, como aumento da produção de Interleucina(IL)- 12 e exacerbação da resposta de hipersensibilidade do tipo tardia; potencialização da produção das citocinas IL-1&#946; e IL-6 em cultura primária de células residentes do SNC e; piora na severidade dos sintomas clínicos causados pela EAE, que coincide com aumento do infiltrado de linfócitos e macrófagos no SNC e ativação imuneinflamatória das células da glia. Tomados em seu conjunto, os dados do presente trabalho sugerem que condições inflamatórias durante a gestação, particularmente durante o final da gestação, podem predispor o feto a distúrbios autoimunes e neurodegenerativos na vida adulta.<br>Prenatal period experiences are crucial for the fetal health. The occurrence of maternal infections and subsequent maternal immune system activation cause a number of structural and functional changes in the brain of the offspring that may predispose individuals to psychiatric disorders in post-natal life, such as schizophrenia and autism. However, studies investigating offspring´s immune alterations are still scarce in the literature. The aim of this study was to evaluate, in mice offspring taken from LPS-treated dams, the impact of maternal immune activation on peripheral immune cell activity, central nervous system (CNS) inflammatory response, and development of experimental autoimmune encephalomyelitis (EAE), the murine model of multiple sclerosis. Pregnant female mice received a dose of either saline or lipopolysaccharide (LPS) during late gestation (gestational day 17), and offspring were used in three experiments to analyze: (1) cytokine production and activity by peripheral immune cells and development of delayed type hypersensitivity, (2) production of inflammatory mediators by resident CNS cells and, (3) development of clinical symptoms and immune response during the course of EAE. Our results showed that maternal immune activation resulted in immune alterations in the offspring, such as increased peripheral production of interleukin (IL) -12 and exacerbated response of delayedtype hypersensitivity; enhancement of IL-1&#946; and IL-6 productions in primary CNS resident cells culture and; increased severity of EAE clinical symptoms, which is positively correlated with the increased lymphocytes and macrophages infiltration within the CNS and also with the immune-inflammatory activation of glial cells. Taken together, the data from this study suggest that inflammatory conditions during pregnancy, especially during the late pregnancy, may predispose the fetus to autoimmune and neurodegenerative disorders in adulthood.