Relationships between phenotype and function of blood cd4+ t-cells and ascending thoracic aortic aneurysm: an experimental study

Abstract Introduction: Non-familial ascending thoracic aorta dilation and aneurysms (TAAs) are silent diseases in elderly patients. Histopathology revealed that functionally polarized infiltrating CD4+ T-cells play a key role in aortic wall weakening. Objective: To evaluate the possible associations between phenotype and cytokine production of circulating CD4+ T-lymphocytes and the presence of TAA in patients with aortic valve disease (AVD). Methods: We studied blood samples from 10 patients with TAA and 10 patients with AVD. Flow cytometry was used to quantify: a) CD4+ T-lymphocytes surface expression of CD25, CD28, and chemokine receptors (CCR5, CXCR3, CX3CR1); b) fractions of in vitro stimulated CD4+ T-cells producing cytokines (interferon gamma [IFN-γ], interleukin [IL]-17A, IL-21, IL-10); c) CD4+CD25highFoxP3+ regulatory T-cells (Treg) fraction. Enzyme-linked immunosorbent assays (ELISA) were performed for cytokines (IFN-γ, IL-6, IL-10, IL-17A, IL-23, transforming growth factor beta [TGF-β]) and chemokines (RANTES, CX3CL1). Results: The total CD4+CD28±CD4+/CX3CR1+ T-cells fraction was higher (P=0.0323) in AVD (20.452±4.673) than in TAA patients (8.633±2.030). The frequency ratio of CD4+ T-lymphocytes producing IFN-γ vs. IL-17A+IL-21 cytokine-producing CD4+ T-cells was higher (P=0.0239) in AVD (2.102±0.272) than in TAA (1.365±0.123) patients. The sum of CD4+CD28±CD4+/CX3CR1+ T-cells correlated positively with values of the previous cytokine ratio (P=0.0002, R=0.732). The ratio of CD4+CD28±CD4+/CX3CR1+ T-cells vs. Treg was higher (P=0.0008) in AVD (20.859±3.393) than in TAA (6.367±1.277) patients. Conclusion: Our results show that the presence of TAA in subjects with AVD is associated with imbalance between phenotypic and cytokine-producing subsets of circulating CD4+ T-lymphocytes, prevalently oriented towards a pro-fibrotic and IFN-γ counteracting effect to functional polarization.

Mannose-binding lectin 2 (Mbl2) gene polymorphisms are related to protein plasma levels, but not to heart disease and infection by Chlamydia

The presence of the single nucleotide polymorphisms in exon 1 of the mannose-binding lectin 2 (MBL2) gene was evaluated in a sample of 159 patients undergoing coronary artery bypass surgery (71 patients undergoing valve replacement surgery and 300 control subjects) to investigate a possible association between polymorphisms and heart disease with Chlamydia infection. The identification of the alleles B and D was performed using real time polymerase chain reaction (PCR) and of the allele C was accomplished through PCR assays followed by digestion with the restriction enzyme. The comparative analysis of allelic and genotypic frequencies between the three groups did not reveal any significant difference, even when related to previous Chlamydia infection. Variations in the MBL plasma levels were influenced by the presence of polymorphisms, being significantly higher in the group of cardiac patients, but without representing a risk for the disease. The results showed that despite MBL2 gene polymorphisms being associated with the protein plasma levels, the polymorphisms were not enough to predict the development of heart disease, regardless of infection with both species of Chlamydia.

IL-10 and ET-1 as biomarkers of rheumatic valve disease / IL-10 e ET-1 como biomarcadores de doença valvar reumática

Objective: To evaluate the immunological profile and gene expression of endothelin-1 (ET-1) in mitral valves of patients with rheumatic fever originated from a reference service in cardiovascular surgery. Methods: This was a quantitative, observational and cross-sectional study. Thirty-five subjects (divided into four groups) participated in the study, 25 patients with chronic rheumatic heart disease and ten control subjects. The mean age of the sample studied was 34.5 years. Seventeen of them (48.58%) were male and 18 (51.42%) were female. Inflammatory cytokines (TNF-α, IL-4 and IL-10) were measured and ten mitral valves of patients who underwent first valve replacement were collected for determination of gene expression of endothelin-1 by real time PCR. Results: Among the groups studied (patients vs. controls), there was a statistically significant difference in IL-10 levels (P=0.002), and no differences in other cytokines. Expression of endothelin-1 was observed in 70% of samples. Quantitatively, average of ET-1 expression was 62.85±25.63%. Conclusion: Inflammatory cytokine IL-10 participates in the maintenance of chronicity of rheumatic fever in patients who underwent valve replacement and those who are undergoing medical treatment. The expression of endothelin-1 in heart valve lesions in patients undergoing mitral valve replacement confirms its association with inflammatory activity in rheumatic fever. .

Objetivo: Avaliar o perfil imunológico e a expressão gênica de endotelina-1 em valvas mitrais de pacientes com febre reumática, originados de um serviço de referência em cirurgia cardiovascular. Métodos: Este foi um estudo quantitativo, observacional e transversal. Trinta e cinco indivíduos (divididos em quatro grupos) participaram do estudo, 25 deles com doença cardíaca reumática crônica, além de 10 controles. A média de idade da amostra estudada foi de 34,5 anos. Dezessete (48,58%) dos indivíduos eram homens, e 18 (51,42%) eram mulheres. Foram medidas algumas citocinas inflamatórias (TNF-α, IL-4 e IL-10) e coletadas 10 valvas mitrais de pacientes que se submeteram a primeira troca valvar para determinação da expressão gênica de endotelina-1 pelo PCR real-time. Resultados: Entre os grupos estudados (pacientes e controles), observou-se diferença estatisticamente significante em relação aos níveis de IL-10 (P=0,002), sem diferenças nas outras citocinas. Em relação à endotelina-1, foi observada sua expressão em 70% das amostras. Quantitativamente, a expressão média de endotelina-1 foi de 62,85±25,63%. Conclusão: A citocina inflamatória IL-10 participa da manutenção da cronicidade da febre reumática em pacientes que se submeteram a troca valvar e naqueles que estão em tratamento médico. A expressão de endotelina-1 nas lesões em valvas cardíacas de pacientes que foram submetidos à troca valvar mitral confirma sua relação com a atividade inflamatória na febre reumática. .

Troca valvar aórtica com diferentes próteses: existem diferenças nos resultados da fase hospitalar? / Aortic valve replacement with different types of prosthesis: are there differences in the outcomes during hospital phase?

OBJETIVOS: Analisar dados intra-operatórios e possíveis diferenças na evolução clínica da fase hospitalar de pós-operatório da troca valvar aórtica com diferentes próteses. MÉTODOS: Análise de 60 pacientes, divididos em três grupos: os submetidos a troca valvar por prótese biológica (20); por prótese mecânica (20); e finalmente, por valva homóloga (20). A média da idade foi de 51,1 anos; 60 por cento eram do sexo masculino e 40 por cento do feminino; 86,7 por cento estavam em NYHA II ou III; 63,3 por cento eram hipertensos, 18,3 por cento diabéticos; a etiologia valvar foi degenerativa em 39 por cento, reumática em 36 por cento e endocardite em 15 por cento. RESULTADOS: A mortalidade hospitalar foi de 5 por cento; não houve diferenças entre os grupos na incidência de choque séptico ou cardiogênico, insuficiência renal aguda, arritmias no centro cirúrgico e na unidade de terapia intensiva (UTI), assim como para o tempo de internação na UTI e tempo de ventilação mecânica. Houve diferença estatística nos tempos de circulação extracorpórea (P=0,02) e pinçamento aórtico (P<0,0001) desfavorável aos pacientes com valva homóloga. O tempo de internação na enfermaria foi maior para os pacientes com valva mecânica (P=0,05), assim como o tempo total de internação hospitalar tende a ser maior neste grupo, porém sem significância estatística. Pacientes com hematócrito pré-operatório abaixo de 38,1 por cento utilizaram 2,73 unidades de hemoderivados; e com hematócrito no pós-operatório abaixo de 32 por cento utilizaram 1,79 unidades. Controle ecocardiográfico mostrou mínimas diferenças evolutivas. CONCLUSÃO: A utilização de diferentes próteses, nesta amostra, não gerou diferenças nos resultados da fase hospitalar de pós-operatório; O homoenxerto é uma opção viável e com boa aplicabilidade clínica.

OBJECTIVE: To analyze intraoperative data and possible differences in clinical evolution during postoperative hospital phase for aortic valve replacement surgery using different types of prosthesis. METHODS: Analysis of 60 patients divided into three groups. Valve replacement with bioprosthesis (20), mechanical prosthesis (20) and homologous valve (20). The mean age was 51.1, 60 percent were male and 40 percent female patients; 86.7 percent were in NYHA II or III; 63.3 percent presented arterial hypertension and 18.3 percent had diabetes. Aetiology of valve disease was degenerative for 39 percent, rheumatic for 36 percent and endocardits for 15 percent. RESULTS: The hospital mortality was 5 percent; there were no differences in the incidence of septical or cardiogenic shock, acute renal failure, rhythms disorders during surgery or intensive care, neither for total time in intensive care and mechanical ventilation. However, there was statistical differences as regards the cardiopulmonary bypass total time (P=0.02) and the aortic clamping time (P<0.0001) unfavorable to homograft valve group. The ward admission time was greater for mechanical valve group (P=0.05) as well as for total admission time, but without statistical significance. It was observed that patients with preoperative hematocrit smaller than 38.1 percent used 2.73 units of blood components, and with postoperative hematocrit smaller than 32 percent used 1,79 units of blood components. Echocardiography control showed minimal evolutional differences. CONCLUSION: The use of different types of prosthesis for this study does not cause differences in the results of postoperative hospital phase. The use of homograft valve is a feasible option with good clinical applicability.

Blood conservation in valve surgery.

Blood utilization in 40 patients undergoing elective valve surgery was prospectively studied. The patients had valvular lesions of rheumatic origin with a mean age of 29.1 years and a mean preoperative hematocrit of 35.23 +/- 4.16. Blood was removed from all patients after induction of anesthesia and reinfused after bypass (mean 365.12 +/- 66.96 ml). Membrane oxygenator was used in all the patients. All discard suction was routed through a regionally heparinised collecting and processing system, and the resulting red cell concentrate was transfused. At the conclusion of bypass, all blood remaining in the pump oxygenator was also processed by cell saver and used for subsequent reinfusion. Normovolemic anemia was accepted in hemodynamically stable patients. Thirty two patients (80%) received no bank blood or blood products during their entire hospital course. A total of twelve units of whole blood was transfused into eight patients.

Haptoglobinas em populaçäo normal e em portadores de cardiopatias valvares / Heptoglobins in normal population and in patients with heart valve diseases

Haptoglobina sérica em 100 doadores de sangue foi determinada utilizando-se da capacidade de ligaçäo da haptoglobina à hemoglobina; os valores médios encontrados foram de 211 ñ 82mg/dl; 38 pacientes portadores de valvopatia valvar foram também estudados e apresentaram valores significativamente mais altos (estenose: 310 ñ 128 e insuficiência 409 ñ 137mg/dl) que a populaçäo normal. A atividade da desidrogenase lática também estava aumentada em pacientes com cardiopatia valvar, indicando a presença de doença sistêmica ou eventual hemólise. Estes dados sugerem ter sido a desidrogenase lática mais sensível que a determinaçäo das haptoglobinas em detectar hemólise intravascular leve, nos doentes estudados