NK and NKT cell dynamics after rituximab therapy for systemic lupus erythematosus and rheumatoid arthritis

Biomarkers of clinical response to rituximab (RTX) therapy and early predictors of outcome are still under investigation. We report a flow cytometric immunophenotyping analysis from peripheral blood leukocyte subpopulations of two patients with systemic lupus erythematosus (SLE) associated thrombocytopenia and one patient with rheumatoid arthritis (RA), before and after 6 weeks of treatment with RTX. Our results show a reduced population of CD19(+) expressing cells (B cells) after RTX treatment in all three patients. Increased frequency of peripheral regulatory CD4(+)CD25(high) T cell subset and the CD3(-)CD16(-)CD56(bright) NK cell subset after RTX therapy were also observed in all patients, the latter being more pronounced in the SLE patient with sustained clinical response. In addition, an increased population of NKT cell subsets was observed in the patients with clinical response. This is the first evaluation of NK and NKT cells as biomarkers of clinical response after rituximab therapy in rheumatic diseases.     

Hypertension control in brazilian publications

Hypertension is a major public health problem due to its high prevalence andcardiovascular complications. Its treatment is aimed at reducing cardiovascularmorbidity and mortality, its goal being to maintain blood pressure levels below140/90 mm Hg. Hypertension control in Brazil is low, and nationwide rates areunknown. The objective of this review was to provide an overview on hypertensioncontrol in Brazil from publications in a database. We identified 45 publications. Inpopulation-based studies, the highest control rate (57.6%) was reported in amulticenter study in 100 municipalities and the city of São José do RioPreto, São Paulo state (52.4%), while the lowest rates (around 10%) wereidentified in microregions of the Rio Grande do Sul state and in the city ofTubarão, Santa Catarina state. In conclusion, the studies assessed showed awide variation in hypertension control rates. It is worth noting that the comparisonbetween studies was a major limiting factor, because of the different methodsused.     

Paraoxonase Gln-Arg(192) and Leu-Met(55) gene polymorphisms and enzyme activity in a population with a low rate of coronary heart disease

OBJECTIVES: To assess whether paraoxonase (PON1) polymorphisms at positions 55 and 192 and/or their phenotypic expressions influence the risk of myocardial infarction (MI) in Spanish population.DESIGN AND METHODS: Two hundred and fifteen male survivors of a MI and their age-matched controls were included in the study. Lipids, apolipoproteins (apo) A-I and B, PON1 activity on paraoxon and phenylacetate and PON1 polymorphisms were determined.RESULTS: Genotype distribution was similar in patients and controls. Enzyme activities were lower in patients, but multiple logistic regression analysis did not show any independent association with a higher risk of MI.CONCLUSION: None of the PON1 polymorphisms or their corresponding measured activities are independent risk factors for MI in our population.     

Cytokeratin-based CTC counting unrelated to clinical follow up

Background

Circulating tumor cells (CTCs) have been reported to be a relevant prognostic biomarker in metastatic patients. However, their clinical use and impact is still under debate. We have thus comparatively and kinetically assessed two CTC detection methods according to the patient’s clinical follow up.

Methods

CTC counting and characterization were repeatedly performed during follow up in a patient with metastatic undifferentiated non-small cell lung cancer by using cytokeratin (CK)-dependent immunomagnetic separation (Miltenyi) and CK-independent, size-based isolation [isolation by size of tumor cells (ISET)] (Rarecells).

Results

Comparison between the two methods showed a parallel increase of CTC detected by ISET and worsening of the clinical status, while CK-dependent CTC numbers were decreasing, misleadingly suggesting a response to treatment. ISET results were in agreement with the clinical follow up showing Circulating tumor microemboli (CTM) and CTC expressing a mesenchymal marker with absence of epithelial markers.

Conclusions

This case report study shows the interest of a comparative and kinetic analysis of different methods for CTCs detection combined with their evaluation according to the clinical follow up. Our results should open up an area for future research and validation in larger clinical cohorts.KEYWORDS : Circulating tumor cells (CTCs), circulating tumor microemboli (CTM), lung cancer, immunomagnetic separation, isolation by size of tumor cells (ISET)     

qPCR detection of Mycobacterium leprae in biopsies and slit skin smear of different leprosy clinical forms

Leprosy, whose etiological agent is Mycobacterium leprae, is a chronic infectious disease that mainly affects the skin and peripheral nervous system. The diagnosis of leprosy is based on clinical evaluation, whereas histopathological analysis and bacilloscopy are complementary diagnostic tools. Quantitative PCR (qPCR), a current useful tool for diagnosis of infectious diseases, has been used to detect several pathogens including Mycobacterium leprae. The validation of this technique in a robust set of samples comprising the different clinical forms of leprosy is still necessary. Thus, in this study samples from 126 skin biopsies (collected from patients on all clinical forms and reactional states of leprosy) and 25 slit skin smear of leprosy patients were comparatively analyzed by qPCR (performed with primers for the RLEP region of M. leprae DNA) and routine bacilloscopy performed in histological sections or in slit skin smear. Considering clinical diagnostic as the gold standard, 84.9% of the leprosy patients were qPCR positive in skin biopsies, resulting in 84.92% sensitivity, with 84.92 and 61.22% positive (PPV) and negative (NPV) predictive values, respectively. Concerning bacilloscopy of histological sections (BI/H), the sensitivity was 80.15% and the PPV and NPV were 80.15 and 44.44%, respectively. The concordance between qPCR and BI/H was 87.30%. Regarding the slit skin smear, 84% of the samples tested positive in the qPCR. Additionally, qPCR showed 100% specificity, since all samples from different mycobacteria, from healthy individuals, and from other granulomatous diseases presented negative results. In conclusion, the qPCR technique for detection of M. leprae using RLEP primers proved to be specific and sensitive, and qPCR can be used as a complementary test to diagnose leprosy irrespective of the clinical form of disease.