Pengxiang Zhao

Quantification of Epstein-Barr Virus DNA in Patients with Idiopathic Orbital Inflammatory Pseudotumor

Inflammatory pseudotumors (IPT) are soft tissue tumors that include a diverse group of lesions characterized by inflammatory cell infiltration and variable fibrotic responses. Idiopathic orbital inflammatory pseudotumors (IOIP) are IPTs of unknown etiology that develop in the orbit. Due to the lack of well-defined pathogenic mechanisms, diagnosis and treatment of this disease remain a significant challenge. Epstein-Barr virus (EBV) infection, which causes significant lymphocyte infiltration, has been proposed to be involved in IOIP. This study tries to validate the relationship between EBV infection and the development of IOIP. Sixteen IOIP tissue samples were obtained from patients during surgical resection of the lesion. One Graves ophthalmopathy tissue sample and 20 normal donors plasma serves as controls. The plasma level of five EBV antibodies, including VCA-IgG, VCA-IgA, VCA-IgM, EA-IgG and EBNA1-IgG were examined. All plasma samples were EB-VCA-IgG positive and EB-VCA-IgM negative, suggesting that all people tested had been infected with EBV but not in the acute infection stage. EBV-DNA was detected in 15/16 (94%) of IOIP tissue samples despite different levels of lymphocyte infiltration and 5/16 plasma samples (31%) were detected EBV DNA positive which is higher than the normal controls (10%). Percent of positive plus suspected positive samples with one or more of the three important risk markers (VCA-IgA, EA-IgG, EBV-DNA) is 50% of the patients (8/16) which is much higher compare with the normal controls (20%). The results further reveal the relationship between IOIP and EBV infection.

Detection of Epstein-Barr Virus in Idiopathic Orbital Inflammatory Pseudotumor

Purpose: The etiology of idiopathic orbital inflammatory pseudotumors (IOIP) is unknown. Epstein-Barr virus (EBV) infection, which causes significant lymphocyte infiltration, has been proposed to be involved in IOIP. This study tries to validate the relationship between EBV infection and the development of IOIP. Methods: Eleven surgical samples from IOIP patients and four non-IOIP control samples from surgery removal of eye due to trauma were analyzed by microarray. The data were confirmed by real time RT-PCR. The existence of EBV-DNA segments in IOIP samples was determined by PCR. Results: Hierarchical clustering of microarray data successfully distinguished the IOIP group and the non-IOIP control group. The gene EBI2, usually induced by EBV infection, was significantly up-regulated in the IOIP group (max = 11.7-fold over control). EBV-DNA was detected in 9/10 (90%) of the IOIP samples and in 0/4 of the control samples. Up-regulation of EB12, the immunoglobulin kappa light chain (IGKC) and the immunoglobulin heavy chain gene 1(IGHA1) were detected in the IOIP group compared to the control group. Conclusion: Collectively, these findings suggest that IOIP may be related to EBV infection and IGKC and IGHA1 activation. Furthermore, we hypothesize that the expression of the EBI2 gene may serve as a molecular marker for IOIP. More IOIP patient samples and detection of EBV related genes ( EBER1, EBNA2 and LMP1) expression need to be investigated.

Genetic and Phenotypic Analysis of CRF01_AE HIV-1 env Clones from Patients Residing in Beijing, China.

CRF01_AE is one of the four dominant HIV-1 strains circulating in China. In this study, we performed genetic and phenotypic analyses using a total of 60 full-length envelope gene clones from 14 HIV-1-infected individuals in the Beijing area. Among the 60 sequences analyzed, 32 have a complete open reading frame (ORF), whereas the others contain premature stop codons. The phylogenetic tree analysis suggested that all of the sequences maintained a close relationship with the CRF01_AE strain. Most of the potential N-linked glycosylation sites (PNGS) were located within the V1/V2, V4, C2, or C3 regions. In relation to gp41, the majority of the glycosylation sites were located in the ectodomain. The 32 env genes that contained intact ORFs were used to construct Env-pseudotyped viruses, and eight strains that resulted in high titers were further studied. All the eight strains used CCR5 as the co-receptor for infection, and they were sensitive to neutralization by the broadly neutralizing monoclonal antibodies, including VRC_01, PG9, PG16, and NIH45-46, but they were insensitive to 2G12. Notably, seven of these eight strains lacked a glycan at residues 295 or 332 (or both), suggesting that these two PNGSs play an important role in 2G12 binding and neutralization. In addition, the pseudoviruses were more sensitive to neutralization by plasma isolated from individuals infected with subtypes CRF01_AE and CRF07/08_BC, suggesting the occurrence of a cross-neutralizing antibody profile between these two strains. These findings are likely to have important implications for the design of an effective HIV vaccine and relevant therapeutic drugs.

Genes and pathways associated with the occurrence of malignancy in benign lymphoepithelial lesions

There is increasing evidence concerning the occurrence of malignant lymphoma among people suffering from Mikulicz disease, also termed benign lymphoepithelial lesion (BLEL) and immunoglobulin G4-associated disease. However, the underlying molecular mechanism of the malignant transformation remains unclear. The present study aimed to investigate the gene expression profile between BLEL and malignant lymphoepithelial lesion (MLEL) conditions using tissue microarray analysis, to identify genes and pathways which may be associated with the risk of malignant transformation. Comparing gene expression profiles between BLEL tissues (n=13) and MLEL (n=14), a total of 1,002 differentially expressed genes (DEGs) were identified including 364 downregulated and 638 upregulated DEGs in BLEL. The downregulated DEGs in BLEL were frequently associated with immune-based functions, immune cell differentiation, proliferation and survival, and metabolic functions, whereas the upregulated DEGs were primarily associated with organ, gland and tissue developmental processes. The B cell receptor signaling pathway, the transcription factor p65 signaling pathway, low affinity immunoglobulin γ Fc region receptor II-mediated phagocytosis, the high affinity immunoglobulin ε receptor subunit γ signaling pathway and Epstein-Barr virus infection, and pathways in cancer, were the pathways associated with the downregulated DEGs. The upregulated DEGs were associated with three pathways, including glutathione metabolism, salivary secretion and mineral absorption pathways. These results suggested that the identified signaling pathways and their associated genes may be crucial for understanding the molecular mechanisms underlying malignant transformation from BLEL, and they may be considered to be markers for predicting malignancy among the BLEL group.

TLR7 and TLR8 agonist resiquimod (R848) differently regulates MIF expression in cells and organs

&NA; Since its first description in 1966, macrophage migration inhibitory factor (MIF) was found to play a critical role in inflammatory and immune responses as well as in disease pathogenesis especially in tumor pathogenesis and cancer progression. MIF is expressed in different cell types and is associated with many disease severity and tumor pathogenesis. Here, we investigated the influence of TLR7 and TLR8 agonist resiquimod (R848), an immune response inducer used as a prophylactic agent for several infectious diseases as well as anticancer agents and vaccine adjuvant on MIF expression in cells and organs. Humans, mice and rats cell lines from different tissues (blood, retinal, nasopharynx, brain and liver) and C57BL/6J mice organs (brain, liver and spleen) were used for this investigation. In vitro, R848 induced MIF gene overexpression except in brain and liver cells. Furthermore, it enhanced cells ability to release soluble MIF and differently regulated mRNA expression of MIF‐related receptors (CD74, CXCR4, CXCR2 and CD44). Its influence on MIF gene expression and MIF proteins release was more consistent in cancer cells. In vivo, a strong positive expression of MIF was observed in different regions in brain and spleen in response to R848 treatment; however in liver, increased MIF expression was observed in hepatocytes only. On the other hand, R848 treatment had induced a slight enhancement of MIF concentration in the plasma of C57BL/6J mice. Taken together, these data suggest that R848 differently regulates MIF mRNA expression depending on organ types and could influence MIF concentration in cellular microenvironment. HighlightsR848 differently regulates MIF mRNA and proteins expression in cells and organs.R848 significantly increases MIF release.Activation of TLR7 and TLR8 could induce MIF downstream cascades activation.

Macrophage migration inhibitory factor contributes to the pathogenesis of benign lymphoepithelial lesion of the lacrimal gland

BackgroundBenign Lymphoepithelial Lesion (BLEL) is a rare disease observed in the adult population. Despite the growing numbers of people suffering from BLEL, the etiology and mechanisms underlying its pathogenesis remain unknown.MethodsIn the present study, we used gene and cytokines expression profiling, western blot and immunohistochemistry to get further insight into the cellular and molecular mechanisms involved in the pathogenesis of BLEL of the lacrimal gland.ResultsThe results showed that Macrophage Migration Inhibitory Factor (MIF) was the most highly expressed cytokine in BLEL, and its expression positively correlated with the expression of Th2 and Th17 cells cytokines. MIF was found to regulate biological functions and pathways involved in BLEL pathogenesis, such as proliferation, resistance to apoptosis, MAPK and PI3K/Akt pathways. We also found that MIF promotes fibrosis in BLEL by inducing BLEL fibroblast differentiation into myofibroblasts as well as the synthesis and the deposit of extracellular matrix in BLEL tissues.ConclusionsOur findings demonstrate the contribution of MIF to the pathogenesis of BLEL of the lacrimal gland and suggested MIF as a promising therapeutic target for its treatment.

Gene Expression Profiling of Idiopathic Orbital Inflammatory Pseudotumors

Propose: Idiopathic orbital inflammatory pseudotumors (IOIP) are inflammatory pseudotumors (IPT) of unknown etiology that develop in the orbit. Due to the lack of well-defined pathogenic mechanisms and diagnostic markers, diagnosis and treatment of this disease remain a significant challenge. Therefore, the aims of this study are to define the etiological factors of IOIP and to identify potential gene targets for the treatment of IOIP. Methods: Eleven IOIP tissue samples were obtained from patients during surgical resection; as control tissue, four normal tissue samples were collected from surgery removal of eye due to trauma. Total RNA was extracted and the gene expression profile of both diseased and normal tissues was analyzed using human oligonucleotide microarrays, which contained 32,050 well-characterized human genes. The expression patterns of selected genes were further confirmed by Real-time RT-PCR. Results: Microarray analyses identified 744 genes differentially expressed in IOIP and non-IOIP samples, with at least 21.5-fold changes and P-value < .005, including 552 downregulated and 192 upregulated genes in IOIPs. Principal Component Analysis (PCA) and Hierarchical Clustering of microarray data successfully distinguished the IOIP group from the non-IOIP control group. Functional annotation using the Molecule Annotation System categorized the 744 genes into immune functions, inflammatory responses, signaling molecules in the PI3K and NF-&#954;B pathways, extracellular matrix degradation, and unknown functions. Conclusions: Immune and inflammatory responses and activation of the PI3K and NF-&#954;B pathways are likely associated with IOIPs. Drugs that would suppress these two responses and two signal pathways may offer new therapeutics of IOIP treatment.