Hongmei Zhu

Accumulation of invariant NKT cells with increased IFN-γ production in persistent high-risk HPV-infected high-grade cervical intraepithelial neoplasia.

BackgroundPersistent high-risk human papillomavirus (HR-HPV) infection has been implicated in the development of high-grade cervical intraepithelial neoplasia (CIN) and cervical cancer. Invariant natural killer T (iNKT) cells produce large amounts of cytokines to regulate immune responses. However, the role of iNKT cells in human persistent HPV-infected cervical tissues is unknown.MethodsIn our study, 201 patients with diagnoses ranging from normal ectocervical tissue to CINIII from June 2010 to May 2012 were enrolled. HPV DNA and HPV types were detected using the hybrid capture-2 HPV DNA test. Flow cytometry was used to investigate iNKT and CD3+ T cell infiltration into cervical tissues. Real-time quantitative reverse transcription-polymerase chain reaction was used to study IFN-γ expression and immunohistochemistry was used to determine CD3+ T cell distribution.ResultsA significant increase in iNKT cells was observed in HPV-positive cervical tissues (p < 0.05), especially in CINII-III (p < 0.01). IFN-γ expression was also increased in HPV-positive cervical tissues (p < 0.05). CD3+ T cells were detected among both epithelium and stromal layers in cervical tissues, and the percentage of CD3+ T cells in HPV-positive cervical tissues was similar to that in HPV-negative cervical tissues (p > 0.05).ConclusionsThe iNKT cell aggregation in cervical tissues during the progression from HPV infection to CIN indicates that iNKT cells might play an important role in suppressing immunity. IFN-γ expression could also be related to the HPV infection status. Preventing the accumulation or functioning of iNKT cells in cervical tissues may be a viable method to prevent the development of CIN.Virtual slidesThe virtual slides for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/2521874671514142

Pure non-gestational ovarian choriocarcinoma in a 45,XO/46,XX SRY-negative true hermaphrodite.

Non‐gestational ovarian choriocarcinoma (NGCO) is an extremely rare malignant tumor with a poor prognosis and is difficult to distinguish from gestational choriocarcinoma. True hermaphrodite (TH) is genetically a heterogenous condition causing ovarian and testicular tissue development in the same individual. We report here the first case of pure NGCO in the right ovotestis of a 23‐year‐old 45,XO/46X,X sex‐determining region Y chromosome (SRY)‐negative TH. The diagnosis of non‐gestational origin was confirmed by testing five short tandem repeats (STR). The patient responded well to radical surgery with bleomycin, etoposide, cisplatin (BEP) regimen. We also hypothesize that some mutations of an X‐linked or autosomal gene lead to testicular determination in SRY‐negative TH patients.

Identification of copy number variations among fetuses with ultrasound soft markers using next-generation sequencing

A prospective analysis investigating the associations between pathogenic copy number variations (pCNVs) and ultrasound soft markers (USMs) in fetuses and evaluating the clinical value of copy number variation sequencing (CNV-seq) in such pregnancy studies was carried out. 3,398 unrelated Chinese women with singleton pregnancies and undergone amniocentesis at 18–36 weeks of gestation for fetal CNV-seq were included. According to the prenatal fetal ultrasound screening results, the samples were divided into 3 groups: normal ultrasound (n = 2616), solitary USM (n = 663), and two or more USMs (n = 119). CNV-seq was performed successfully using all samples. The prevalence of pCNVs in fetuses with normal ultrasound and USMs was 3.03% (79/2616) and 2.94% (23/782), respectively. The risk of segmental aneuploidies was significantly higher in the two or more USMs group (5/119, 4.20%) than in the normal ultrasound (27/2616, 1.04%) or solitary USM (9/663, 1.36%) groups (p = 0.002 and p = 0.031, respectively). Assuming that the resolution of karyotyping is ~5 Mb, a cytogenetic analysis would miss 33 of 102 (32.35%) pCNVs in these samples. Our results suggest an association between pCNVs and fetal USMs; multiple USMs indicate an increased risk of fetal segmental aneuploidies. In prenatal diagnostic testing, CNV-Seq identified additional, clinically significant cytogenetic information.

Preliminary study of protein changes in trisomy 21 fetus by proteomics analysis in amniocyte

To discover the candidate biomarker proteins of trisomy 21 (T21) in amniocytes.

Expression of V‑domain immunoglobulin suppressor of T cell activation is associated with the advanced stage and presence of lymph node metastasis in ovarian cancer

V-domain immunoglobulin suppressor of T cell activation (VISTA) is a novel negative immune checkpoint that belongs to the B7 family. VISTA is primarily expressed on hematopoietic cells and inhibits T cell proliferation and cytokine production. The blockade of VISTA has demonstrated promising results in certain murine tumor models. In the present study, an immunohistochemical analysis of VISTA expression on tumor cells, intratumoral immune cells and vascular endothelial cells was performed in a cohort of 65 patients with ovarian cancer (OC). The associations between VISTA expression and different clinicopathological characteristics were evaluated using Fishers exact test, and the analysis of overall survival in different groups was performed by the construction of Kaplan-Meier curves. The results indicated that high expression of VISTA on tumor cells or ICs was significantly associated with advanced tumor stage and the presence of lymph node metastasis (LNM). However, the percentage of cases with high expression of VISTA on tumor cells (24.6%) was decreased compared with those with high expression on ICs (44.6%). There was no association between VISTA expression and the 5-year overall survival rate, and advanced-stage disease was the only independent predictor of poor prognosis based on multivariate Cox regression analysis. In general, VISTA expression increased with advanced disease stage and LNM, indicating that VISTA expression is involved in the progression of OC. More importantly, these data implicate VISTA as a candidate immunotherapeutic target in OC.

Prospective chromosome analysis of 3429 amniocentesis samples in China using copy number variation sequencing.

BACKGROUND: Next‐generation sequencing is emerging as a viable alternative to chromosome microarray analysis for the diagnosis of chromosome disease syndromes. One next‐generation sequencing methodology, copy number variation sequencing, has been shown to deliver high reliability, accuracy, and reproducibility for detection of fetal copy number variations in prenatal samples. However, its clinical utility as a first‐tier diagnostic method has yet to be demonstrated in a large cohort of pregnant women referred for fetal chromosome testing. OBJECTIVE: We sought to evaluate copy number variation sequencing as a first‐tier diagnostic method for detection of fetal chromosome anomalies in a general population of pregnant women with high‐risk prenatal indications. STUDY DESIGN: This was a prospective analysis of 3429 pregnant women referred for amniocentesis and fetal chromosome testing for different risk indications, including advanced maternal age, high‐risk maternal serum screening, and positivity for an ultrasound soft marker. Amniocentesis was performed by standard procedures. Amniocyte DNA was analyzed by copy number variation sequencing with a chromosome resolution of 0.1 Mb. Fetal chromosome anomalies including whole chromosome aneuploidy and segmental imbalances were independently confirmed by gold standard cytogenetic and molecular methods and their pathogenicity determined following guidelines of the American College of Medical Genetics for sequence variants. RESULTS: Clear interpretable copy number variation sequencing results were obtained for all 3429 amniocentesis samples. Copy number variation sequencing identified 3293 samples (96%) with a normal molecular karyotype and 136 samples (4%) with an altered molecular karyotype. A total of 146 fetal chromosome anomalies were detected, comprising 46 whole chromosome aneuploidies (pathogenic), 29 submicroscopic microdeletions/microduplications with known or suspected associations with chromosome disease syndromes (pathogenic), 22 other microdeletions/microduplications (likely pathogenic), and 49 variants of uncertain significance. Overall, the cumulative frequency of pathogenic/likely pathogenic and variants of uncertain significance chromosome anomalies in the patient cohort was 2.83% and 1.43%, respectively. In the 3 high‐risk advanced maternal age, high‐risk maternal serum screening, and ultrasound soft marker groups, the most common whole chromosome aneuploidy detected was trisomy 21, followed by sex chromosome aneuploidies, trisomy 18, and trisomy 13. Across all clinical indications, there was a similar incidence of submicroscopic copy number variations, with approximately equal proportions of pathogenic/likely pathogenic and variants of uncertain significance copy number variations. If karyotyping had been used as an alternate cytogenetics detection method, copy number variation sequencing would have returned a 1% higher yield of pathogenic or likely pathogenic copy number variations. CONCLUSION: In a large prospective clinical study, copy number variation sequencing delivered high reliability and accuracy for identifying clinically significant fetal anomalies in prenatal samples. Based on key performance criteria, copy number variation sequencing appears to be a well‐suited methodology for first‐tier diagnosis of pregnant women in the general population at risk of having a suspected fetal chromosome abnormality.

Comment on “Invariant NKT Cells in Hyperplastic Skin Induced a Local Immune Suppressive Environment by IFN-γ Production”

We read the article by Mattarollo et al. ([1][1]) with great interest. They identified that CD1d-recruited invariant NKT (iNKT) cells had the ability to suppress immune effector mechanisms after specific attraction to persistent HPV16-E7–expressing hyperplastic lesions in a murine model. Several