RTA and LANA Competitively Regulate let-7a/RBPJ Signal to Control KSHV Replication

The recombination signal binding protein for immunoglobulin kappa J region (RBPJ) has a dual effect on Kaposi’s sarcoma-associated herpesvirus (KSHV) replication. RBPJ interaction with replication and transcription activator (RTA) is essential for lytic replication, while the interaction with latency-associated nuclear antigen (LANA) facilitates latent infection. Furthermore, our previous study found that LANA decreased RBPJ through upregulating miRNA let-7a. However, it is unclear whether RTA regulates the expression of RBPJ. Here, we show RTA increases RBPJ by decreasing let-7a. During KSHV replication, the RBPJ expression level was positively correlated with the RTA expression level and negatively correlated with the LANA expression level. The let-7a expression level was inverse to RBPJ. Knockdown of RBPJ inhibited the self-activation of RTA promoter and LANA promoter and weakened LANA’s inhibition of RTA promoter. Collectively, these findings indicate that RTA and LANA compete for let-7a/RBPJ signal to control the KSHV replication. Regulating the RBPJ expression level by RTA and LANA plays an important role during KSHV replication.

The immunoglobulin kappa variable cluster is differentially expressed in human epithelial ovarian cancer.

Epithelial ovarian cancer (EOC) is the most lethal gynecologic cancer (1). We performed discovery of genes associated with epithelial ovarian cancer and of the high-grade serous ovarian cancer (HGSC) subtype, using published and public microarray data (2, 3) to compare global gene expression profiles of normal ovary or fallopian tube with that of primary tumors from women diagnosed with epithelial ovarian cancer or HGSC. We identified genes encoded by the immunoglobulin kappa variable cluster, IGKV, as among the genes whose expression was most different in epithelial ovarian cancer as compared to the normal fallopian tube. IGKV1D-8 expression was significantly higher in high-grade serous ovarian tumors relative to normal fallopian tube. These data indicate that expression of IGKV genes is perturbed in epithelial ovarian cancers broadly and in ovarian cancers of the HGSC subtype. IGKV may be relevant to pathways underlying ovarian cancer initiation (transformation) or progression.

Association of a Functional Single Nucleotide Polymorphism (rs874040) in the RBPJ Gene with Susceptibility to Rheumatoid Arthritis in Iranian Population

Background: Rheumatoid Arthritis (RA) is a progressive, heterogeneous, and common multifactorial autoimmune disease. Several Genome-Wide Association Studies (GWASs) have revealed more than 100 risk loci for RA. One of these loci is a functional single nucleotide polymorphism (rs874040; G>C) near the recombination signal-binding protein for the immunoglobulin kappa J region (RBPJ) gene. RBPJ can convert into a transcriptional activator upon activation of the canonical Notch pathway. Notch signaling has recently emerged as an important regulator of immune responses in inflammation and autoimmune diseases. In the present study, the possible association between SNP rs874040 (G>C) upstream of the RBPJ gene with RA risk was assessed in Iranian population. Methods: A case-control study including 60 RA patients and 44 control subjects was conducted to estimate rs874040 genotypes using real‑time polymerase chain reaction High Resolution Melting (HRM) method. Results: Logistic regression analysis indicated that homozygous CC and heterozygous GC genotypes increase the risk of RA compared with GG genotype (CC vs. GG; OR=11.36; 95% CI [3.93-33.33] and CG vs. GG; OR=3.78; 95% CI [1.30-10. 98]). Besides, subjects with C allele were more frequently affected with RA than subjects with G allele (OR=10.42; 95% CI [5.21-20.83]). Furthermore, in the patient group, a significant correlation was found between C-reactive protein concentrations and rs874040 polymorphism (p<0.05). Conclusion: Our findings propose a substantial correlation between rs874040 polymorphism and RA risk in Iranian population.

Assesment Of Protein Profile In Vitreus Samples Of Patients With Age-Related Macular Degeneration By Proteomic Approaches

Background: To investigate the protein content in patients with age-related macular degeneration (AMD) with the aim of revealing the pathogenesis of the disease. Methods: Two groups were formed: 13 patients as the AMD group and 11 patients as the healthy control group. Vitreous samples were taken from both groups under sterile conditions and sent to the proteomics laboratory for proteomics analysis. In this study, we evaluated the proteome of vitreous samples of AMD and control groups using two-dimensional gel electrophoresis (2DE) coupled with MALDI-TOF/TOF.Results: We detected 11 proteins differentially regulated in the vitreous of AMD patients relative to healthy controls. The only protein that was up-regulated was Apolipoprotein E. We observed that Alpha 1-acid glycoprotein (AAG), Leucine-rich alpha-2-glycoprotein (LRG-1), Alpha-2-HS-glycoprotein, Haptoglobin, Alpha-Crystalline A Chain, Alpha- Crystalline B Chain, Immunoglobulin kappa constant, Beta-crystallin B2, Beta-crystallin A3, Beta-crystallin S levels were significantly decreased in patients with AMD. Conclusion: The detected proteins are related to biological regulation, retinal protection, and regulation of inflammation and angiogenesis processes. We believe that investigating these proteins will help to reveal the pathological mechanisms of AMD.

Predictive Markers of Missed Miscarriage

The aim of this study was to find useful the serological markers for missed miscarriage (MM) in order to predict the outcome of pregnancy. The study included 141 pregnant women aged between 18 and 45 years at gestational age under 11 weeks. All women were divided into 3 groups. Group 1 included 68 women with MM; Group 2 included 43 women with spontaneous miscarriage; Group 3 included 30 pregnant women without pathology. Proteomic analysis of the blood serum was performed using liquid chromatography-mass spectrometry. The results of our study show that immunoglobulin kappa variable 3-15 (KV315) can be considered as the most promising serologic marker for MM in early gestation. The potential role of KV315 as the serological marker is very important for predicting the course of pregnancy.

Isotyping and Semi-Quantitation of Monkey Anti-Drug Antibodies by Immunocapture Liquid Chromatography-Mass Spectrometry

There is an urgent demand to develop new technologies to characterize immunogenicity to biotherapeutics. Here, we developed an immunocapture LC-MS assay to isotype and semi-quantify monkey anti-drug antibodies (ADAs) to fully human monoclonal antibody (mAb) drugs. ADAs were isolated from serum samples using an immunocapture step with the Fab of the full-length mAb cross-linked to magnetic beads to minimize matrix interference. A positive monoclonal antibody control against the human immunoglobulin kappa light chain was used as a calibration standard for ADA quantitation. The final LC-MS method contains 17 multiple reaction monitoring (MRM) transitions and an optimized 15-min LC method. The results suggested that IgG1 was the most abundant isotype in ADA-positive samples. IgG2 and IgG4 were identified at lower levels, whereas IgG3 and IgA levels were only observed at very minor levels. In addition, levels of total ADA measured by the LC-MS assay were comparable to results obtained using a traditional ligand binding assay (LBA). The LC-MS ADA assay enabled rapid immunogenicity assessment with additional isotype information that LBAs cannot provide.

ENTIERE IDENTITIES BETWEEN OPHIOCOMINA NIGRA IGKAPPA GENE AND HUMAN IMMUNOGLOBULIN KAPPA LOCUS.NEW ASPECTS OF INVERTEBRATE IGKAPPA GENES (IPA)

Entiere identities between Invertebrate Ophiocomina nigra IGKappa gene and Human IGK gene are confirmed, in the present work, at the level of immunoglobulin domains (constant and variable). The transcriptome of the Ophuirid : Ophiocomina nigra IGKappa gene was discovered recently(1).Since it was synthesized de novo and cloned in a pUC-GW-Kan plasmid (2) which was a gift of Bo Huang laboratories. The original sequence of the IGKappa gene, after cloning, was the following in 5’-3’ :

IGKC, the immunoglobulin kappa constant gene segment, is differentially expressed in brain metastatic breast cancer.

Metastasis to the brain is a clinical problem in patients with breast cancer (1-3). We mined published microarray data (4-6) to compare primary and metastatic tumor transcriptomes to discover genes associated with brain metastasis in patients with metastatic breast cancer. We found that the immunoglobulin light chain kappa constant locus encoded by IGKC was among the genes whose expression was most different in the brain metastases of patients with metastatic breast cancer as compared to primary tumors of the breast. We observed a significant correlation between IGKC expression in primary tumors of the breast and distant-metastasis free survival, as well as overall survival. Down-regulation of IGKC may be an important event for metastasis of primary tumor-derived cancer cells to the brain in humans with metastatic breast cancer.