Phenotype changes of oral epithelial stem cells after in vitro culture

Abstract The aim of our study was to isolate populations of keratinocyte stem cells based on the expression of cell surface markers and to investigate whether the culture could affect their phenotype. keratinocytes from human oral mucosa were sorted based on the expression of the epithelial stem cell markers p75NTR and CD71. We also examined the co-expression of other epithelial stem markers such as integrins β1 and α6 and their stem cell-like proprieties in in vitro assays. Three passages after being sorted by MACS, more than 93% of the p75NTR+ve cells lost the expression of p75NTR, while 5.46% of the p75NTR-ve gained it. Within the small population of the p75NTR+ve cells, 88% co-expressed other epithelial stem cell markers such as integrins β1 and α6, while only 28% of p75NTR-ve cells co-expressed these markers. These results were confirmed by sorting cells by FACS. Additionally, when double staining was used for sorting cells, 99% of the p75NTR+veCD71-ve and 33% of the p75NTR-veCD71+ve cells expressed both integrins, but just one week after culture, only 1.74% of the p75NTR+veCD71-ve cells still expressed p75NTR and only 0.32% still expressed CD71. Similar results were obtained when co-culturing p75NTR+ve and p75NTR-ve populations before analysis. Our results suggest that phenotype changes may be part of an intrinsic cellular mechanism to conserve levels of protein expression as they may found in the human body. In addition, in vitro culture may not offer ideal conditions for epithelial stem cell maintenance due to phenotype changes under standard culture conditions.

Serum soluble transferrin receptors in patients with chronic hepatitis C

Recently, it has been found that iron is an important element in the natural history of hepatitis C. Serum markers of iron stores are frequently increased in chronic hepatitis C infected patients and therefore, detection of soluble transferrin receptor[sTfR] may allows for quantitative evaluation of intracellular iron. The aim of the present study was to study serum levels of soluble transferrin receptors in patients with chronic HCV. The study was carried out on 40 patients classified into three groups. Group I enrolled 10 chronic HCV with high serum iron and ferritin, Group II included 10 patients with chronic HCV with normal serum iron and ferritin [Both groups I, and II had no cirrhosis], Group III compromised of2O patients with chronic HCV and liver cirrhosis. Also 10 healthy subjects were taken as control. Soluble transferrin receptors were measured in patient sera using humans TJJ? ELISA Kit. The mean serum sTfR was significantly lower in group III than in groups I, II and IV. Also, it was significantly lower in group I than in groups II and IV On the other hand, no significant difference was found between groups II and IV. Correlation study revealed significant negative correlation between serum sTfR and serum iron and ferritin in groups L II and III. Moreover, in group III significant negative correlation was noticed between serum sTfR and staging [r0.83, 0.00]. sTfR can be regarded as a valuable and specific tool for investigating tissue iron that can be used to predict the degree of hepatocellular damage and progression into advanced fibrosis and subsequent cirrhosis

Expected value of serum soluble transferrin receptor, erythropoietin and ferritin and their correlation among healthy non-anemic Thai children.

Transferrin receptor (TfR) is a glycoprotein which mediates the entry of ferric transferrin from the extracellular compartment into the cells. The measurement of sTfR has become a widely used tool in assessing erythropoiesis but its use has mainly been restricted to research laboratory settings. In Thailand, there are only a few reports concerning the sTfR. The authors studied the expected value of sTFR as well as other basic parameters for monitoring of erythropoiesis as erythropoietin (EPO) and ferritin among a sample of non-anemic healthy Thai children. In addition, correlation was done between each pair of studied parameters. Expected range for sTfR level for the healthy controls in this study was 1.761 to 2.034 mg/L. Expected range for serum EPO level for the healthy controls in this study was 19.445 to 34.176 mU/ml. Expected range for serum ferritin level for the healthy control from this study was 67.895 to 96.692 ng/ml. Of interest, poor correlation among the three studied parameters, sTfR, serum EPO and serum ferritin was observed in this study.

Methodology evaluation of a new immunoturbidimetric method for measuring serum soluble transferrin receptor.

The value of soluble transferrin receptor (sTfR) detected in serum is closely related to erythroid TfR turnover rate. An increased erythropoietic activity causes an increase in the sTfR level. Therefore, it is a useful test for monitoring the erythropoiesis. In this study, a new immunoturbidimetric method for automated measurement of sTfR was evaluated for its performance characteristics. Imprecision studies on patients' sera with 1.01 mg/l and 2.94 mg/l concentrations yielded within-run CVs of 1.16% and 1.27%. Accuracy analysis of the test by using the low and high control kit sera with 1.45 mg/l and 5.41 mg/l concentrations were 89.06% and 95.41%, respectively. The evaluation was also performed in 60 individual pediatric subjects, 30 beta-thalassemia/HbE and 30 control pediatric subjects. There is a statistically significant difference of sTfR between both groups (p < 0.0005, 95% Cl = 9.457-14.124). Ninety-five percent of matched pediatric subjects had sTfR level < or = 2.670 mg/l and 93.33% of patients diagnosed beta-thalassemia/HbE had values > 2.670 mg/l. In conclusion, this immunoturbidimeteric test yields good laboratory performance characteristics in terms of precision and accuracy.

Differences in immunophenotyping of mucosal lymphocytes between ulcerative colitis and Crohn's disease

OBJECTIVES: Immunologic studies have characterized the numbers and types of inflammatory cells in diseased inflammatory bowel disease (IBD) mucosa but have yielded conflicting results regarding intestinal lymphocytes activation in IBD. We investigated the levels of lymphocytes subsets, interleukin-2 receptor, transferrin receptor, and T cell receptors in mainly isolated lamina propria lymphocytes. Including intraepithelial lymphocytes of normal colonic mucosa or IBD (ulcerative colitis and Crohn's disease) mucosa to understand the pathogenesis of IBD. We have results from this study. RESULTS: 1) In comparing ulcerative colitis with control, IL-2R (p < 0.05), TR (p < 0.01), and CD3/HLA-DR (<0.05) showed a significant increase. 2) In comparing Crohn's disease with control, CD3 (P < 0.05), TCR alpha/beta (p < 0.01) and TCR gamma/delta (p < 0.05) showed a significant decrease. 3) In comparing Crohn's disease with ulcerative colitis, CD19 (p < 0.01), TR (p < 0.01), TCR alpha/beta (p < 0.01) and TCR gamma/delta (p < 0.05) showed a significant decrease. CONCLUSION: From these results, there are increased T cell markers, IL-2R, TR, and CD3/HLA-DR in UC, but differently, decreased CD3, TCR alpha/beta and TCR gamma/delta in CD compared with control. In addition, definitive differences in lymphocytes markers, CD19, TR, TCR alpha/beta and TCR gamma/delta, which are higher in UC than in CD, may elucidate the different immunopathogenesis between UC and CD.

Ativaçäo in vivo de linfócitos T em pacientes com esclerose múltipla / In vivo T-lymphocytes activation in patients with multiple sclerosis

Pacientes com esclerose múltipla apresentam flutuaçöes em subtipos de linfócitos no sangue periférico. No presente trabalho, os autores demonstram uma maior incidência de linfócitos ativados em pacientes com atividade clínica ou em surto, detectados através de anticorpos monoclonais contra os receptores de transferrina e inteleucina