[Receptor-ligand analysis of killer cell Ig-like receptor in Jiangsu Han population].

AIM: To investigate the distribution of killer cell immunoglobulin-like receptor (KIR) and its specific ligand human leukocyte antigen (HLA) in Jiangsu Han population. METHODS: 173 samples from unrelated healthy individuals of Jiangsu Han population were genotyped and observed for KIR, HLA-Cw and HLA-Bw4 using a SYBR Green I real-time PCR and PCR-SSP method, respectively. The number and type of KIR/HLA pairs inherited in each individual were analyzed. RESULTS: In Jiangsu Han population, all four inhibitory KIR (2DL1, 2DL2/3, 3DL1 and 3DL2) that recognize the classical HLA class I molecules HLA-A, -B and -C were present in >92% of the study group. Frequencies of 2DL2/HLA-C1, 2DL3/HLA-C1, 2DL1/HLA-C2 and 3DL1/Bw4 were 0.243, 0.971, 0.457 and 0.590, respectively; frequencies of 2DS1/HLA-C2 and 2DS2/HLA-C1 were 0.162 and 0.231, respectively. 54.3% of the cases expressed KIR2DL1 without HLA-C1, 32.9% inherited 3DL1 without HLA-Bw4 and 5.8% expressed HLA-Bw4 without 3DL1. 27.7% of the individuals had three iKIR/HLA pairs, 26% carried two iKIR/HLA pairs, and 25.4% inherited a single iKIR/HLA pair and no one was deficient in all three iKIR/HLA pairs. CONCLUSION: There was disparity between KIR receptor and HLA ligand in Jiangsu Han population. Inhibitory KIR/HLA pair frequency was higher than stimulatory one. About 1/4 of the study group expressed a single iKIR/HLA pair alone.

[Detection for KIR gene with use of SYBR green I real-time PCR].

AIM: To develope a novel real-time PCR method for KIR genotyping. METHODS: KIR genotyping is performed using 16 real-time PCR reactions, each containing two-three KIR-specific primers, a pair of internal positive control primers and a fluorescence dye, SYBR Green I. By the analysis of the Tm and the characteristic of the melting curve of the amplified products, we identified the presence or absence of 16 KIR genes. A variety of dilution folds were made to detect the sensitivity of this method. RESULTS: KIR genes were effectively genotyped by the analysis of the melting curve. This method can be used to detect KIR genes even from 0.1 ng of DNA. The feasibility of this method was tested by genotyping 10 DNA samples from Peripheral blood and 10 DNA samples from cervical cell. CONCLUSION: We developed a novel real-time PCR assay with SYBR Green I, which is a simple, rapid, sensitive, real-time and environmental method. It provides the possibility of the automated KIR genotyping.

Effect of recombinant human erythropoietin on serum S100B protein and interleukin-6 levels after traumatic brain injury in the rat.

Erythropoietin (EPO) has a neuroprotective effect in the animal model of ischemia/hypoxia, but the mechanisms underlying the EPO effect in traumatic brain injury (TBI) are not well understood. This study examined the potential neuroprotective mechanisms of recombinant human EPO (rhEPO) in rats after TBI. Sixty healthy adult male Sprague-Dawley rats were randomly divided into 5 groups: 1000 U/kg rhEPO-treated, 3000 U/kg rhEPO-treated, 5000 U/kg rhEPO-treated, citicoline, and normal saline (control) groups. The TBI model was based on the modified Feeney's free falling model. Serum samples were collected at 6 hours, 24 hours, 3 days, 5 days, and 7 days after trauma. The serum S100B protein and interleukin-6 (IL-6) levels were measured after treatment in each group with double antibody sandwich enzyme-linked immunosorbent assay. Both serum S100B protein and IL-6 levels were significantly lower in 3000 U/kg rhEPO-treated and 5000 U/kg rhEPO-treated groups (p < 0.001). The decrease in serum S100B protein level was correlated with the dosage of rhEPO. Medium doses of rhEPO achieved the optimum decreases in the serum IL-6 level. Therefore, inhibition of the composition and secretion of S100B protein and IL-6 levels by EPO might be one of the mechanisms involved in decreasing inflammatory reaction in the brain, and may be responsible for the neuroprotective effect after TBI.