Arginase-1 and Treg Profile Appear to Modulate Inflammatory Process in Patients with Chronic Gastritis: IL-33 May Be the Alarm Cytokine in H. pylori-Positive Patients.

Helicobacter pylori (H. pylori) is a highly prevalent bacterium in our environment, directly involved in various upper digestive tract diseases, such as gastritis, peptic ulcer, and gastric cancer. Several molecules activating the immune system have been reported to be involved in containing H. pylori infection. This study is aimed at analyzing the mRNA expression of the cytokines IFN-γ, IL-17, IL-10, TGF-ß, IL-6, IL-22, IL-23, and IL-33; transcription factors T-bet, RORC, and FOXP3; enzymes ARG1, ARG2, and NOS2; and neuropeptides VIP and TAC and their respective receptors VIPR1 and TACR1 in the stomach lining of patients with severe digestive disorders. One hundred and twenty six patients have been evaluated, presenting with symptoms in the upper digestive tract, with the clinical indication for an Upper Digestive Endoscopy exam. Two fragments of the mucosa of the gastric body and antrum have been collected for anatomopathological examination and to analyze the expression of enzymes, cytokines, and transcription factors using qPCR. Expression of the ARG1 gene was seen as significantly higher in the group of patients with chronic inactive gastritis than in the control group. Expression of the TGF-ß gene and its FOXP3 transcription factor was significantly higher in the group of chronic inactive gastritis patients than in the control. Expression of IFN-γ, IL-17, IL-10, and TGF-ß and the transcription factors, T-bet and RORC, in the presence or absence of H. pylori showed no significant difference. However, the expression of FOXP3 was significantly lower in H. pylori-positive patients than that in H. pylori-negative patients. ARG1 and Treg profile appeared to be modulating the inflammatory process, protecting patients from the tissue lesions with chronic inactive gastritis. Furthermore, we suggest that IL-33 may be a crucial mediator of the immune response against an infection, after gastric mucosal damage.

Levels of Pro and Anti-inflammatory Citokynes and C-Reactive Protein in Patients with Chronic Periodontitis Submitted to Nonsurgical Periodontal Treatment

Aim: to compare the levels of IFN-γ, TGF-ß and C-reactive protein (CRP) in healthy patients (HP) and chronic periodontitis patients (CP) before and seven days after the last session of Non-Surgical Periodontal Treatment (NSPT). Materials and Methods: 40 subjects were divided into two groups: healthy (n= 20), and with chronic periodontitis (n = 20). Serum and gingival crevicular fluid (GCF) were collected from each patient and quantified for IFN-γ, TGF-ß and CRP using the enzyme-linked immunosorbent assay (ELISA). Results: IFN-γ was found to be higher in the GCF of the CP group before NSPT in relation to the HP group (p<0.05), and it had significant higher levels after seven days of NSPT (p<0.05). The levels of TGF-ß in the GCF of CP patients before NSPT were significantly higher when compared to HP (p<0.05), but they decreased after seven days of NSPT (p>0.05). Serum CRP levels did not show statistical difference between CP and HP before or after NSPT. Conclusion: Therefore, our results demonstrated for the first time that NSPT causes early exacerbation of the immune response at the local level represented by increased levels of IFN-γ and decreased levels of TGF-ß in the gingival crevicular fluid after seven days of treatment.

COMPARATIVE ANALYSIS OF IMMUNOLOGICAL PROFILES IN WOMEN UNDERGOING CONVENTIONAL AND SINGLE-PORT LAPAROSCOPIC CHOLECYSTECTOMY.

Background: Surgical trauma triggers an important postoperative stress response characterized by significantly elevated levels of cytokines, an event that can favor the emergence of immune disorders which lead to disturbances in the patient's body defense. The magnitude of postoperative stress is related to the degree of surgical trauma. Aim: To evaluate the expression of pro-inflammatory (TNF-α, IFN-γ, IL-1ß, and IL-17) and anti-inflammatory (IL-4) cytokines in patients submitted to conventional and single-port laparoscopic cholecystectomy before and 24 h after surgery. Methods: Forty women with symptomatic cholelithiasis, ranging in age from 18 to 70 years, participated in the study. The patients were divided into two groups: 21 submitted to conventional laparoscopic cholecystectomy and 19 to single-port laparoscopic cholecystectomy. Results: Evaluation of the immune response showed no significant difference in IFN-γ and IL-1ß levels between the groups or time points analyzed. With respect to TNF-α and IL-4, serum levels below the detection limit (10 pg/ml) were observed in the two groups and at the time points analyzed. Significantly higher postoperative expression of IL-17A was detected in patients submitted to single-port laparoscopic cholecystectomy when compared to preoperative levels (p=0.0094). Conclusions: Significant postoperative expression of IL-17 was observed in the group submitted to single-port laparoscopic cholecystectomy when compared to preoperative levels, indicating that surgical stress in this group was higher compared to the conventional laparoscopic cholecystectomy.

Racional: O trauma cirúrgico induz resposta de estresse pós-operatório significativo, evidenciado pelos níveis elevados de citocinas, podendo favorecer o surgimento de distúrbios imunológicos. A magnitude de estresse está relacionada ao grau do trauma cirúrgico. Objetivos: Avaliar a expressão das citocinas pró-inflamatórias (TNF-α, IFN-γ, IL-1ß, IL-17) e da anti-inflamatória (IL-4) no pré e pós-operatório de pacientes submetidas à colecistectomia laparoscópica por dois métodos: convencional e por portal único. Métodos: Quarenta mulheres com colecistolitíase sintomática foram operadas, sendo 21 por procedimento laparoscópico convencional e 19 por portal único. As citocinas TNF-α, IFN-γ, IL-1ß, IL-4, e IL-17 presentes no plasma foram quantificadas pelo método de ELISA em dois momentos: no pré-operatório e após 24 h da operação. Resultados: A avaliação da resposta imune não mostrou diferença estatisticamente significante das citocinas IFN-γ e IL-1ß na comparação entre os grupos e tempos analisados. Em relação às citocinas TNF-α e IL-4 os níveis séricos estavam abaixo dos níveis de detecção (10 pg/ml) em ambos os grupos e tempos analisados. Observou-se que as operadas por portal único apresentaram expressão significativa da IL-17A do período pré para o pós-operatório (p=0,0094). Conclusões: A expressão aumentada da IL-17A no pós-operatório do grupo de portal único pode indicar que o estresse cirúrgico foi maior em comparação ao da colecistectomia laparoscópica convencional.

Comparative analysis of immunological profiles in women undergoing conventional and single-port laparoscopic cholecystectomy / Perfil imunológico de mulheres submetidas à colecistectomia laparoscópica convencional e por portal único

ABSTRACT Background: Surgical trauma triggers an important postoperative stress response characterized by significantly elevated levels of cytokines, an event that can favor the emergence of immune disorders which lead to disturbances in the patient's body defense. The magnitude of postoperative stress is related to the degree of surgical trauma. Aim: To evaluate the expression of pro-inflammatory (TNF-α, IFN-γ, IL-1β, and IL-17) and anti-inflammatory (IL-4) cytokines in patients submitted to conventional and single-port laparoscopic cholecystectomy before and 24 h after surgery. Methods: Forty women with symptomatic cholelithiasis, ranging in age from 18 to 70 years, participated in the study. The patients were divided into two groups: 21 submitted to conventional laparoscopic cholecystectomy and 19 to single-port laparoscopic cholecystectomy. Results: Evaluation of the immune response showed no significant difference in IFN-γ and IL-1β levels between the groups or time points analyzed. With respect to TNF-α and IL-4, serum levels below the detection limit (10 pg/ml) were observed in the two groups and at the time points analyzed. Significantly higher postoperative expression of IL-17A was detected in patients submitted to single-port laparoscopic cholecystectomy when compared to preoperative levels (p=0.0094). Conclusions: Significant postoperative expression of IL-17 was observed in the group submitted to single-port laparoscopic cholecystectomy when compared to preoperative levels, indicating that surgical stress in this group was higher compared to the conventional laparoscopic cholecystectomy.

RESUMO Racional: O trauma cirúrgico induz resposta de estresse pós-operatório significativo, evidenciado pelos níveis elevados de citocinas, podendo favorecer o surgimento de distúrbios imunológicos. A magnitude de estresse está relacionada ao grau do trauma cirúrgico. Objetivos: Avaliar a expressão das citocinas pró-inflamatórias (TNF-α, IFN-γ, IL-1β, IL-17) e da anti-inflamatória (IL-4) no pré e pós-operatório de pacientes submetidas à colecistectomia laparoscópica por dois métodos: convencional e por portal único. Métodos: Quarenta mulheres com colecistolitíase sintomática foram operadas, sendo 21 por procedimento laparoscópico convencional e 19 por portal único. As citocinas TNF-α, IFN-γ, IL-1β, IL-4, e IL-17 presentes no plasma foram quantificadas pelo método de ELISA em dois momentos: no pré-operatório e após 24 h da operação. Resultados: A avaliação da resposta imune não mostrou diferença estatisticamente significante das citocinas IFN-γ e IL-1β na comparação entre os grupos e tempos analisados. Em relação às citocinas TNF-α e IL-4 os níveis séricos estavam abaixo dos níveis de detecção (10 pg/ml) em ambos os grupos e tempos analisados. Observou-se que as operadas por portal único apresentaram expressão significativa da IL-17A do período pré para o pós-operatório (p=0,0094). Conclusões: A expressão aumentada da IL-17A no pós-operatório do grupo de portal único pode indicar que o estresse cirúrgico foi maior em comparação ao da colecistectomia laparoscópica convencional.

IMMUNOLOGICAL EVALUATION OF PATIENTS WITH TYPE 2 DIABETES MELLITUS SUBMITTED TO METABOLIC SURGERY / AVALIAÇÃO IMUNOLÓGICA DE PACIENTES COM DIABETE MELITO TIPO 2 SUBMETIDOS À CIRURGIA METABÓLICA

Background: Immunological and inflammatory mechanisms play a key role in the development and progression of type 2 diabetes mellitus. Aim: To raise the hypothesis that alterations in immunological parameters occur after duodenojejunal bypass surgery combined with ileal interposition without gastrectomy, and influences the insulin metabolism of betacells. Methods: Seventeen patients with type 2 diabetes mellitus under clinical management were submitted to surgery and blood samples were collected before and six months after surgery for evaluation of the serum profile of proinflammatory (IFN-γ, TNF-α, IL-17A) and anti-inflammatory cytokines (IL-4, IL-10). In addition, anthropometric measures, glucose levels and insulin use were evaluated in each patient. Results: No changes in the expression pattern of proinflammatory cytokines were observed before and after surgery. In contrast, there was a significant decrease in IL-10 expression, which coincided with a reduction in the daily insulin dose, glycemic index, and BMI of the patients. Early presentation of food to the ileum may have induced the production of incretins such as GLP-1 and PYY which, together with glycemic control, contributed to weight loss, diabetes remission and the consequent good surgical prognosis of these patients. In addition, the control of metabolic syndrome was responsible for the reduction of IL-10 expression in these patients. Conclusion: These findings suggest the presence of low-grade inflammation in these patients during the postoperative period, certainly as a result of adequate glycemic control and absence of obesity, contributing to a good outcome of surgery.

Racional: Mecanismos imunológicos e inflamatórios desempenham papel-chave no desenvolvimento e progressão do diabete melito tipo 2. Objetivo: Levantar a hipótese de que alterações nos parâmetros imunológicos ocorrem após operação duodenojejunal combinada com interposição ileal sem gastrectomia, e influenciam o metabolismo da insulina das células beta. Métodos: Dezessete pacientes com diabete melito tipo 2 sob manejo clínico foram submetidos à cirurgia e amostras de sangue foram coletadas antes e seis meses após para avaliação do perfil de sorológico de citocinas pró-inflamatórias (IFN-γ, TNF-α, IL-17A) e anti-inflamatórias(IL-4, IL-10). Além disso, parâmetros antropométricos, glicemia e uso de insulina foram avaliados em cada paciente. Resultados: Não ocorreram alterações no padrão de expressão de citocinas pró-inflamatórias observadas antes e depois da operação. Em contraste, houve diminuição significativa na expressão de IL-10, que coincide com redução da dose diária de insulina, com o controle glicêmico e redução do IMC dos pacientes. Apresentação precoce de alimentos para o íleo pode ter induzido a produção das incretinas tais como GLP-1 e PYY, que, juntamente com o controle da glicemia, contribuíram para a perda de peso, remissão do diabete e o bom prognóstico consequente cirúrgico. Além disso, o controle de síndrome metabólica foi responsável pela redução da expressão de IL-10 nestes doentes. Conclusão: Baixo grau de inflamação estava presente nesses pacientes no pós-operatório, certamente pelo adequado controle glicêmico e ausência de obesidade, o que contribuiu para bom resultado cirúrgico.

IMMUNOLOGICAL EVALUATION OF PATIENTS WITH TYPE 2 DIABETES MELLITUS SUBMITTED TO METABOLIC SURGERY.

BACKGROUND: Immunological and inflammatory mechanisms play a key role in the development and progression of type 2 diabetes mellitus. AIM: To raise the hypothesis that alterations in immunological parameters occur after duodenojejunal bypass surgery combined with ileal interposition without gastrectomy, and influences the insulin metabolism of betacells. METHODS: Seventeen patients with type 2 diabetes mellitus under clinical management were submitted to surgery and blood samples were collected before and six months after surgery for evaluation of the serum profile of proinflammatory (IFN-γ, TNF-α, IL-17A) and anti-inflammatory cytokines (IL-4, IL-10). In addition, anthropometric measures, glucose levels and insulin use were evaluated in each patient. RESULTS: No changes in the expression pattern of proinflammatory cytokines were observed before and after surgery. In contrast, there was a significant decrease in IL-10 expression, which coincided with a reduction in the daily insulin dose, glycemic index, and BMI of the patients. Early presentation of food to the ileum may have induced the production of incretins such as GLP-1 and PYY which, together with glycemic control, contributed to weight loss, diabetes remission and the consequent good surgical prognosis of these patients. In addition, the control of metabolic syndrome was responsible for the reduction of IL-10 expression in these patients. CONCLUSION: These findings suggest the presence of low-grade inflammation in these patients during the postoperative period, certainly as a result of adequate glycemic control and absence of obesity, contributing to a good outcome of surgery.

[Association between the plasma levels of TNF-alpha, IFN-gamma, IL-10, nitric oxide and specific IgG isotypes in the clinical forms of chronic Chagas disease]. / Associação entre os níveis plasmáticos de TNF-alpha, IFN-gamma, IL-10, óxido nítrico e os isotipos de IgG específicos nas formas clínicas da doença de Chagas crônica.

Chagas disease is an important chronic parasitic disease that affects around 9-11 million people in Latin America. A combination of parasite and host-related factors are probably responsible for pathogenesis in the chronic phase of the disease. Among the host-related factors, the immunological response is a parameter of special interest. Our aim here was to evaluate the plasma levels of the cytokines interferon gamma, interleukin 10 and tumor necrosis factor alpha and the immunoglobulins total IgG and its subclasses 3 and 4, by means of ELISA, and the levels of nitric oxide by means of the Griess reaction, among individuals who were seropositive for Trypanosoma cruzi, presenting the cardiac, indeterminate and digestive clinical forms of the disease, and among seronegative individuals. The seropositive individuals produced significantly higher levels of total IgG and IgG-3. Individuals with the digestive form presented higher levels of IgG-4 and interleukin 10. However, these individuals presented lower levels of nitric oxide than the controls did. The results suggest that the higher levels of interleukin 10 observed among individuals with the digestive form may contribute towards the higher levels of the specific IgG-4 that were seen.

Associação entre os níveis plasmáticos de TNF-α, IFN-γ, IL-10, óxido nítrico e os isotipos de IgG específicos nas formas clínicas da doença de Chagas crônica / Association between the plasma levels of TNF-α, IFN-γ, IL-10, nitric oxide and specific IgG isotypes in the clinical forms of chronic Chagas disease

A doença de Chagas é uma importante doença parasitária crônica, que acomete cerca de 9-11 milhões de pessoas na América Latina. Provavelmente, uma combinação de fatores relacionados ao parasito e ao hospedeiro podem ser os responsáveis pela patogênese na fase crônica da doença. Dentre os fatores relacionados ao hospedeiro, a resposta imunológica é um parâmetro de especial interesse. Objetivamos avaliar os níveis plasmáticos das citocinas interferon gama, interleucina 10, fator de necrose tumoral alfa e das imunoglobulinas G total, 3 e 4, por ELISA e do óxido nítrico, pela reação de Griess, entre indivíduos soronegativos e soropositivos para Trypanosoma cruzi, com as formas clínicas cardíaca, indeterminada e digestiva. Os indivíduos soropositivos para Trypanosoma cruzi produziram níveis significativamente mais elevados de imunoglobulinas G total e G3. Indivíduos com a forma digestiva apresentam níveis mais elevados de imunoglobulina G4 e interleucina 10. Entretanto, tais indivíduos apresentaram menores níveis de óxido nítrico do que controles. Os resultados sugerem que os maiores níveis de IL-10 observados nos indivíduos com a forma digestiva poderiam contribuir com os maiores níveis de IgG4 específicos observados.

Chagas disease is an important chronic parasitic disease that affects around 9-11 million people in Latin America. A combination of parasite and host-related factors are probably responsible for pathogenesis in the chronic phase of the disease. Among the host-related factors, the immunological response is a parameter of special interest. Our aim here was to evaluate the plasma levels of the cytokines interferon gamma, interleukin 10 and tumor necrosis factor alpha and the immunoglobulins total IgG and its subclasses 3 and 4, by means of ELISA, and the levels of nitric oxide by means of the Griess reaction, among individuals who were seropositive for Trypanosoma cruzi, presenting the cardiac, indeterminate and digestive clinical forms of the disease, and among seronegative individuals. The seropositive individuals produced significantly higher levels of total IgG and IgG-3. Individuals with the digestive form presented higher levels of IgG-4 and interleukin 10. However, these individuals presented lower levels of nitric oxide than the controls did. The results suggest that the higher levels of interleukin 10 observed among individuals with the digestive form may contribute towards the higher levels of the specific IgG-4 that were seen.

Analysis of the cellular immune response in patients with the digestive and indeterminate forms of Chagas' disease.

The commitment of the digestive organs observed in patients during the chronic phase of Chagas' disease is mainly attributed to neuronal damage induced by immune and inflammatory processes elicited by the presence of Trypanosoma cruzi. Here we compare the cellular immune response in patients with the digestive and indeterminate forms of Chagas' disease on the basis of lymphocyte proliferation and cytokine production after antigen or mitogen stimulation. No significant differences between patients groups were observed on proliferative response or on tumor necrosis factor (TNF)-alpha and interleukin (IL)-10 levels, although IL-10 achieves higher levels than TNF-alpha after T. cruzi antigen stimulation. Interferon (IFN)-gamma basal production was significantly higher in the digestive form and IL-4 was significantly higher in patients with megaesophagus when compared with patients with megacolon. These results indicated that patients with the digestive form of Chagas' disease do not suffer immune suppression and that the cytokine balance favors a strong inflammatory reaction in patients with the digestive form, which may contribute to lesions of the mioenteric nervous system.