[Expression of Tim-3 on Peripheral CD56(+) NK Cells and ItsCorrelation with Liver Fibrosis in Patients withAdvanced Schistosomiasis].

OBJECTIVE: To explore the expression of T cell immunoglobulin and mucin domain protein 3 (Tim-3) on CD56(+) NK cells in peripheral blood and its correlation with liver fibrosis indicators in patients with advanced schistosomiasis. METHODS: Tim-3 expression on CD6(+) NK cells from 28 patients with advanced schistosomiasis and 30 healthy controls was determined by flow cytometry. The serum levels of IFN-y and IL-4 were detected by enzyme-linked immunosorbent assay (ELISA). Fibroscan analyzer was used for liver stiffness measurement (LSM) to determine the extent of liver fibrosis. Four serological indicators of liver fibrosis, collagen type III N-peptide (PIIP N-P), Laminin (LN), collagen IV (CIV) and hyaluronic acid (HA) were measured by the Automated Chemiluminescence Immunoassay Analyzer. RESULTS: Flow cytometry analysis showed that Tim-3 expression on CD56(+) NK cells in advanced schistosomiasis patients was (62.3±11.4)%, significantly higher than that (52.1±6.5)% (P< 0.01). In patients with advanced schistosomiasis and the healthy controls, the levels of PIIIP N-P were (86.5±29.5) ng/mL and (22.0±7.8) ng/mL, LN (49.3±21.5) ng/mL and (20.4±6.3) ng/mL, CIV (67.5±22.3) ng/mL and (22.0±3.9) ng/mL, HA (645.9±483.1) ng/mL and (54.7±27.7) ng/mL, respectively. There were significant differences in all these indicators between the two groups (P<0.05). The levels of IFN-y were (93.9?20.1) ng/L and (107.7?24.6) ng/L, and those of IL-4 were (46.6±11.8) ng/L and (28.9±8.9) ng/L respectively in patients with advanced schistosomiasis and the healthy controls, both with significant differences (P<0.05). Spearman nonparametric correlation analysis showed that Tim-3 expression on CD56(+) NK cells in patients was positively correlated with the levels of LSM, PIIIP N-P, LN, CIV and IL-4 (r=0.528-0.834, P<0.01), but negatively with serum IFN-y (r=-0.501, P<0.01). No correlation was found with the HA level (r=0.352, P>0.05). CONCLUSION: The expression of Tim-3 on peripheral CD56(+) NK cells increases in patients with advanced schistosomiasis compared with the healthy controls, and it positively correlates with the levels of LSM, PIIIP N-P, LN, CIV and IL-4, four indicators of liver fibrosis.

Family-based association study of Tim-1 and Tim-3 gene polymorphisms with childhood asthma in Chinese trios.

BACKGROUND: Asthma is a complex genetic disease, caused by the interaction of multiple genetic and environmental factors. T cell immunoglobulin domain and mucin domain (Tim) genes are located in chromosome 5q31-33, a region repeatedly linked to asthma or asthma-related phenotypes in several populations. Two members of Tim families, Tim-1 and Tim-3, which are expressed on T cell surface and potentially involved in T cell proliferation and differentiation, are good candidate genes for asthma. We investigated whether genetic variants or haplotypes in Tim-1 and Tim-3 genes confer susceptibility to asthma in a Chinese population. METHODS: A total of 9 polymorphisms were selected by using the HapMap Han Chinese population data and a haplotype-tagging single nucleotide polymorphism approach. Polymerase chain reaction fragment length polymorphism was adapted to determine the genotype in 118 complete Chinese trios of asthma. Then, transmission disequilibrium test (TDT), haplotypic relative risk (HRR), linkage disequilibrium and haplotype analysis were performed. RESULTS: The single locus TDT and HRR analysis showed the 9 polymorphisms were not transmitted preferentially to asthma-affected children (all p > 0.05). However, in both the haplotypic TDT and HRR analysis, the haplotype G-A-ins-C-G consisting of 5 Tim-1 polymorphisms was found to be overtransmitted to affected offspring (chi(2) = 4.51, p = 0.03) and the haplotype G-G-G consisting of 3 Tim-3 polymorphisms was found to be undertransmitted to asthma children (chi(2) = 8.24, p = 0.004). CONCLUSIONS: We conclude that it is unlikely that the Tim-1 or Tim-3 polymorphisms influence asthma susceptibility individually, but that the haplotypes of variants may be functional or may be in linkage disequilibrium with other functional single nucleotide polymorphisms.

Development of a SYBR green-based real-time quantitative PCR assay to detect PCV3 in pigs.

Porcine circovirus 3 (PCV3) has been reported in cases of porcine dermatitis and nephropathy syndrome, reproductive failure, cardiac and multi-systemic inflammation. A SYBR green-based real-time quantitative PCR (qPCR) assay was established in this study to detect PCV3 in 203 clinical samples from suckling piglets affected by congenital tremors in China. The limit of detection (LOD) of PCV3 was 1.73×104 copies/µL for gel electrophoresis and 1.73×102 copies/µL for SYBR green-based real-time qPCR. The melt curve analysis showed a single melt peak at 82.5°C.The intra-assay coefficient of variation was up to 1.83% and the inter-assay coefficient of variation was up to 2.27%. The PCV3 positive detection rate of 203 clinical samples for the SYBR green-based real-time qPCR and the conventional PCR was 86.70% (176/203) and 26.60% (54/203), respectively. Each tissue detected in the SYBR green-based real-time qPCR showed a higher positive rate than that detected in the conventional PCR. These results indicated that the SYBR green-based real-time qPCR assay is a powerful method with high sensitivity, specificity and reproducibility for epidemiological investigations of PCV3.

[Expression of Tim-3 on peripheral blood CD4+ T cells and its correlation with liver function damage parameters in advanced schistosomiasis patients].

OBJECTIVE: To explore the expression levels of T cell immunoglobulin and mucin domain protein 3 (Tim-3) on peripheral blood CD4+ T cells and its correlation with liver function damage parameters in advanced schistosomiasis patients. METHOD: Totally 28 advanced schistosomiasis patients were selected as study subjects, and 20 chronic schistosomiasis patients and 30 healthy persons were selected as controls. The expression levels of Tim-3 on CD4+ T cell were detected by flow cytometry, and the serum IFN-γ and IL-4 levels were detected by ELISA. Hitachi 7600 biochemical analyzer was used to analyze the liver function parameters (ALT, γ-GT and TBIL). RESULTS: There was a significant difference in Tim-3 expression among the three groups (F = 4.578,P < 0.05). Tim-3 expression level in advanced schistosomiasis patients was (8.33 ± 2.28)%, which was significantly higher than that of the healthy control group (6.57 ± 1.99)% (t = 3.015, P < 0.01). The Spearman nonparametric correlation analysis showed that the Tim-3 expression level on CD4+ T cells in advanced schistosomiasis patients was positively correlated with the serum ALT(r, = 0.746, P < 0.01), γ-GT(r, = 0.656, P < 0.01) and IL-4(r, = 0.672, P < 0.01) levels, but negatively correlated with the serum IFN-γ levels (r(s) = -0.404, P < 0.05). CONCLUSION: The expression of Tim-3 on peripheral CD4' T cells is increased in advanced schistosomiasis patients, which may regulate the function of CD4' T cells and then be involved in the liver damage process of advanced schistosomiasis.