ABSTRACT
The immunomodulatory effects of mesenchymal stem cells (MSCs) are an important mediator of their therapeutic effects in stem cell therapy and regenerative medicine. The regulation mechanism of MSCs is orchestrated by several factors in both intrinsic and extrinsic events. Recent studies have shown that the dynamic expression of cytokines secreted from MSCs control T cell function and maturation by regulating the expression of FoxP3, which figures prominently in T cell differentiation. However, there is no evidence that placenta-derived mesenchymal stem cells (PD-MSCs) have strong immunomodulatory effects on T cell function and maturation via FoxP3 expression. Therefore, we compared the expression of FoxP3 in activated T cells isolated from peripheral blood and co-cultured with PD-MSCs or bone marrow-derived mesenchymal stem cells (BM-MSCs) and analyzed their effect on T cell proliferation and cytokine profiles. Additionally, we verified the immunomodulatory function of PD-MSCs by siRNA-mediated silencing of FoxP3. MSCs, including PD-MSCs and BM-MSCs, promoted differentiation of naive peripheral blood T cells into CD4+CD25+FoxP3+ regulatory T (Treg) cells. Intriguingly, the population of CD4+CD25+FoxP3+ Treg cells co-cultured with PD-MSCs was significantly expanded in comparison to those co-cultured with BM-MSCs or WI38 cells (p < 0.05, p < 0.001). Dynamic expression patterns of several cytokines, including anti- and pro-inflammatory cytokines and members of the transforming growth factor-beta (TGF-β) family secreted from PD-MSCs according to FoxP3 expression were observed. The results suggest that PD-MSCs have an immunomodulatory effect on T cells by regulating FoxP3 expression.
Subject(s)
Humans , Cell Differentiation , Cell Proliferation , Cytokines , Mesenchymal Stem Cells , Regenerative Medicine , Stem Cells , T-Lymphocytes , T-Lymphocytes, Regulatory , Therapeutic UsesABSTRACT
Neuropathic pain is a chronic pain disorder caused by nervous system lesions as a direct consequence of a lesion or by disease of the portions of the nervous system that normally signal pain. The spinal nerve ligation (SNL) model in rats that reflect some components of clinical pain have played a crucial role in the understanding of neuropathic pain. To investigate the direct effects of gabapentin on differential gene expression in cultured dorsal root ganglion (DRG) cells of SNL model rats, we performed a differential display reverse transcription-polymerase chain reaction analysis with random priming approach using annealing control primer. Genes encoding metallothionein 1a, transforming growth factor-beta1 and palmitoyl-protein thioesterase-2 were up-regulated in gabapentin-treated DRG cells of SNL model rats. The functional roles of these differentially expressed genes were previously suggested as neuroprotective genes. Further study of these genes is expected to reveal potential targets of gabapentin.
Subject(s)
Animals , Rats , Chronic Pain , Diagnosis-Related Groups , Ganglia, Spinal , Gene Expression , Ligation , Metallothionein , Nervous System , Neuralgia , Spinal Nerve Roots , Spinal NervesABSTRACT
BACKGROUND: ABO genotyping is commonly used in cases of an ABO discrepancy between cell typing and serum typing, as well as in forensic practice for personal identification and paternity testing. We evaluated ABO genotyping via multiplex allele-specific PCR (ASPCR) amplification using whole blood samples without DNA purification. METHODS: A four-reaction multiplex ASPCR genotyping assay was designed to detect specific nucleotide sequence differences between the six ABO alleles A101, A102, B101, O01, O02, and cis-AB01. The ABO genotypes of 127 randomly chosen samples were determined using the new multiplex ASPCR method. RESULTS: The genotypes of the 127 samples were found to be A101/A102 (n=1), A102/A102 (n=9), A101/O01 (n=3), A102/O01 (n=12), A102/O02 (n=14), B101/B101 (n=5), B101/O01 (n=18), B101/O02 (n=15), O01/O01 (n=14), O02/O02 (n=8), O01/O02 (n=14) and A102/B101 (n=14), from which phenotypes were calculated to be A (n=39), B (n=38), O (n=36) and AB (n=14). The multiplex ASPCR assay results were compared with the serologically determined blood group phenotypes and genotypes determined by DNA sequencing, and there were no discrepancies. CONCLUSIONS: This convenient multiplex ASPCR assay, performed using whole blood samples, provides a supplement to routine serological ABO typing and might also be useful in other genotyping applications.
Subject(s)
Humans , ABO Blood-Group System/genetics , Alleles , DNA/blood , Genotype , Polymerase Chain Reaction/methodsABSTRACT
BACKGROUND: There is room for doubt that the reference intervals currently used in many hospitals or health institutions in Korea are appropriate, because some scientists do not agree that the selections of reference individuals were valid universally. If we adopt the inappropriate reference intervals in the decision making of examinees' health status, we are liable to lead to false-negatives or false-positives. METHODS: We selected 555 healthy and 2,134 unhealthy adult samples who took medical check-up at an institution between 2000 and 2004 through semi-stratified random sampling method. Disease groups were divided into 7 subgroups: hepatic, gastrointestinal, obesity, circulatory, endocrine, urogenital and others. RESULTS: Through parametric and non-parametric methods, we produced new reference intervals and compared the newly developed intervals with current ones. Some reference values should be adjusted newly; ALT-male < or =33 IU/L, ALT-female < or =22 IU/L, AST < or =28 IU/L, cholesterol < or =198 mg/dL, triglyceride-male < or =172 mg/dL, triglyceride-female < or =133 mg/dL, fasting blood sugar 65-101 mg/dL. CONCLUSIONS: In order to reduce the rate of false-positives or false-negatives, we suggest that reference ranges of some items might be reestablished or adjusted according to gender through the further studies on current reference ranges.