Evaluation of Maternal Serum sHLA-G Levels for Trisomy 18 Fetuses Screening at Second Trimester.

Human leukocyte antigen-G (HLA-G) has been widely acknowledged to play critical roles in fetal-maternal maintenance. However, the significance of using maternal serum sHLA-G to detect prenatal chromosomal abnormality has not been investigated. In China, prenatal screening using maternal α-fetoprotein (AFP), unconjugated estriol (uE3), and free ß subunit human chorionic gonadotropin (ß-hCG) in the second trimester has been widely applied. In this study, we evaluated the use of sHLA-G as a screening marker, compared with traditional second trimester prenatal screening. Serum samples from 1,019 singleton women in their second trimester were assessed. Among them, 139 infants were confirmed with trisomy 21 (T21) by karyotyping, 83 were confirmed with trisomy 18 (T18), and the remaining 797 infants had no abnormalities. The sHLA-G levels in maternal sera were significantly lower in pregnant women with T18 fetuses (median: 47.8 U/ml, range: 9.8-234.2 U/ml) and significantly higher in those with T21 fetuses (median: 125.7 U/ml, range: 28.7-831.7 U/ml), compared with the normal controls (median: 106.3 U/ml, range: 50.5-1136.4 U/ml) (p < 0.001). The risk values of the screening of T21 or T18 fetuses were assessed using mean and standard deviation log10 analyte multiples of median (MoM) which showed that the predictive values of sHLA-G were the same as free ß-hCG, and superior to AFP and uE3 for T18 screening. Logistic regression analysis revealed that sHLA-G MoM was the highest risk factor associated with pregnant women carrying T18 fetuses [Exp(B): 171.26, 95% CI: 36.30-807.97, p < 0.001]. Receiver operating characteristic (ROC) analysis revealed that the area under ROC curve for sHLA-G MoM was 0.915 (95% CI, 0.871-0.959, p < 0.001), for AFP MoM was 0.796 (95% CI, 0.730-0.861, p < 0.001), for free ß-hCG MoM was 0.881 (95% CI, 0.829-0.934, p < 0.001), and for uE3 MoM was 0.876 (95% CI, 0.828-0.923, p < 0.001) in the T18 group. sHLA-G MoM demonstrated the best sensitivity and negative predictive value. For the first time, our findings reveal that sHLA-G is a better second trimester screening marker for the detection of T18 fetuses and the combined application of sHLA-G with AFP, free ß-hCG, and uE3 could improve clinical screening for T18 fetuses.

Transfection of chondromodulin I into human breast cancer cells and its effect on the inhibition of cancer cell growth.

Breast cancer affects one in every eight women, and has been associated with higher rates of female mortality than any other cancer type, with the exception of lung cancer. It has been reported that chondromodulin I (ChM-I) was able to suppress tumor angiogenesis and growth in vivo. In order to investigate the antitumor action of ChM­I on human breast cancer cells, a plasmid expressing ChM­I was constructed and transfected into human breast cancer cells using an adenoviral vector. Reverse transcription­polymerase chain reaction detected ChM­I expression in human breast cancer cell lines, whereas no expression was detected in the control groups. In order to assess the effect of ChM­I on human breast cancer cells, cell counting kit­8 (CCK­8) and colony formation analyses were used to detect tumor cell proliferation, and the proliferation of ChM­I­transfected cells was significantly reduced, as compared with the control. In addition, the mRNA expression levels of cell cycle­associated genes in ChM­I­transfected cells were significantly decreased, as compared with the control, which suggested that ChM­I transfection was able to inhibit the expression of genes associated with the cell cycle. The results of the present study indicated that ChM­I was able to inhibit the growth of breast cancer cells; thus suggesting that ChM-I may have potential clinical applications in the treatment of breast cancer.

Tumor-specific upregulation of human leukocyte antigen-G expression in bladder transitional cell carcinoma.

Human leukocyte antigen-G (HLA-G) is a potent immunosuppressive molecule that induces functional silencing of both innate and adaptive immune responses. The relevance of the aberrant HLA-G expression in malignant contexts has been intensively investigated. However, its expression status and clinical significance in bladder cancer remain to be elucidated. In the current study, HLA-G expression in 75 primary bladder transitional cell carcinoma (TCC) lesions was analyzed with immunohistochemistry, and relationship between HLA-G expression and clinical parameters, including disease stage was evaluated. Plasma soluble HLA-G levels were analyzed in 15 TCC patients and 109 normal controls. Data revealed that HLA-G was expressed in 68.0% (51/75) primary TCC lesions, whereas it was undetectable in adjacent normal bladder tissues. The proportion of HLA-G expression in TCC samples varied from negative to 100%, and no significant association was observed for the HLA-G expression status with the patient age, gender, and disease stage. Furthermore, no significance for sHLA-G levels was observed between the TCC patients and normal controls (median 10.75 vs 8.69 U/ml, p = 0.578). Given its immunotolerant properties, our finding suggested that lesion HLA-G expression upregulated in bladder TCC lesions might be an additional mechanism for tumor cells evading from host immunosurveillance; however, its clinical relevance needs further investigation.

Possible roles of KIR2DL4 expression on uNK cells in human pregnancy.

PROBLEM: To investigate possible roles of the natural killer (NK) cell receptor killer immunoglobulin-like receptor (KIR)2DL4 expressed on uterine NK (uNK) cells during pregnancy, we investigated KIR2DL4 expression on uNK cells isolated from patients with early recurrent spontaneous abortion (RSA) and normal early pregnancy women, and functions of KIR2DL4 was analyzed in vitro. METHODS OF THE STUDY: Semi-quantitative RT-PCR analysis was introduced to detect KIR2DL4 messenger RNA (mRNA) expression on uNK cells. Cytotoxicity and cytokine production as the result of interaction of KIR2DL4 and its ligand human leukocyte antigen (HLA)-G were analyzed in vitro with lactic dehydrogenase releasing method and enzyme-linked immunosorbent assay, respectively. RESULTS: No significant difference in KIR2DL4 mRNA expression was observed, while the KIR2DL4 protein level in isolated uNK cells is much higher in normal controls than that in RSA patients. Data showed that HLA-G transfection could not reverse the lysis of uNK against HLA-G transfected K562 cells but induced cytokine production. Furthermore, we demonstrated that, via KIR2DL4, membrane-bound HLA-G could induce high cytotoxicity and cytokine production in a high cytotoxic, IL-2 dependent human NK cell line NK-92 cells. CONCLUSION: Our data suggest that KIR2DL4 might play a crucial implication for human pregnancy.