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Ying Ma,
Jiuping Wang,
Bin Yuan,
Meiliang Wang,
Yun Zhang,
Zhuwei Xu, Chunmei Zhang,
Yusi Zhang,
Bei Liu,
Jing Yi,
Kun Yang,
Angang Yang,
Ran Zhuang,
Boquan Jin
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ABSTRACT: BACKGROUND: Hantaan virus (HTNV) infection in humans is a serious public health concern in Asia. A potent T cell activation peptide vaccine from HTNV structure protein represents a promising immunotherapy for disease control. However, the T cell epitopes of the HTNV restricted by the HLA alleles and the role of epitope-specific T cell response after HTNV infection remain largely unexplored. METHODOLOGYPRINCIPAL FINDINGS: Five well-conserved novel CD8 T-cell epitopes of the HTNV nucleoprotein restricted by the most popular HLA alleles in Chinese Han population were defined with interferon-γ enzyme-linked immunospot assay in 37 patients infected with HTNV during hospitalization. Two epitopes aa129-aa137 and aa131-aa139 restricted by HLA-A2 and B35, respectively, were selected to evaluate the epitope-specific CD8 T-cell response. HLA-peptide pentamer complex staining showed that the frequency of single epitope-specific CD8 T cell could be detected in patients (95% confidence interval for aa129-aa137: 0.080%-0.208%; for aa131-aa139: 0.030%-0.094%). The frequency of epitope-specific pentamer CD8 T-cell response was much higher in mild/moderate patients than in severe/critical ones at the acute stage of the disease. Moreover, the frequency of epitope-specific CD8 T cells at acute stage was inversely associated with the peak level of serum creatinine and was positively associated with the nadir platelet counts during the hospitalization. The intracellular cytokine staining and the proliferation assay showed that the effective epitope-specific CD8 T cells were characterized with the production of interferon-γ, expression of CD69 and the strong capacity of proliferation. CONCLUSIONSIGNIFICANCE: The novel HLA class I restricted HTNV nucleoprotein epitopes-specific CD8 T-cell responses would be closely related with the progression and the severity of the disease, which could provide the first step toward effective peptide vaccine development against HTNV infection in humans.
PLoS Neglected Tropical Diseases 02/2013; 7(2):e2076. · 4.69 Impact Factor
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Xingchun Zhou,
Fan Yang,
Tao Zhang,
Ran Zhuang,
Yuanjie Sun,
Liang Fang, Chunmei Zhang,
Ying Ma,
Gaosheng Huang,
Fucheng Ma,
Chaojun Song,
Boquan Jin
[show abstract]
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ABSTRACT: OBJECTIVE: The goal of this study was to detect the intertumoral heterogeneity of CT10, CT45 and GAGE7 expression and further to analyze their prognostic value. METHODS: The intertumoral heterogeneity of three cancer/testis antigens was examined by immunohistochemistry using 120 samples from patients with infiltrating ductal breast carcinoma. The expression patterns were classified and correlated with the clinicopathologic variables and outcome of the patients. RESULTS: CT10 showed punctate, focal and diffuse expression patterns according to the characteristic of its distribution. CT45 showed cytoplasmic, nuclear or combined cytoplasmic and nuclear expression patterns according to its subcellular location. GAGE7 exhibited nuclear, cytoplasmic and nucleolar expression patterns. Three cancer/testis antigens were also observed coordinately expressed in infiltrating ductal breast carcinoma. Patients with tumors with CT10 expression was significantly correlated with nodal metastases (P < 0.001) and advanced clinical stages (P = 0.001). Patients with tumors with cytoplasmic GAGE7 and with the expression of two or more cancer/testis antigens were significantly correlated with advanced clinical stages (P = 0.001 and P = 0.030). No significant difference was identified between the different expression patterns of CT45 and clinicopathologic variables. In addition, Kaplan-Meier analysis revealed that diffuse CT10 expression and coexpression of three cancer/testis antigens were related to the poor prognosis of patients with infiltrating ductal breast carcinoma. CONCLUSIONS: Diffuse CT10 expression and the coexpression of three cancer/testis antigens can be used as a biomarker to distinguish patients with a poorer outcome of the breast carcinoma. Our finding may provide useful data for evaluating the prognosis of this disease and improving the effectiveness of therapeutic application based on the three cancer/testis antigens.
Japanese Journal of Clinical Oncology 01/2013; · 1.78 Impact Factor
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Guolin Yan,
Yusi Zhang,
Ying Ma,
Jing Yi,
Bei Liu,
Zhuwei Xu,
Yun Zhang, Chunmei Zhang,
Fanglin Zhang,
Zhikai Xu,
Angang Yang,
Ran Zhuang,
Boquan Jin
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ABSTRACT: Hantaan virus (HTNV) of the Bunyaviridae family is a major agent causing hemorrhagic fever with renal syndrome (HFRS), a high-mortality-rate disease threatening approximately 150,000 people around the world yearly. The 3D8 monoclonal antibody displays a neutralizing activity to HTNV infection. In this study, the B-cell epitopes of HTNV glycoproteins (GPs) were finely mapped by peptide scanning. A new B-cell epitope (882)GFLCPEFPGSFRKKC(896) of HTNV, which locates on Gc, has been screened out from a set of 15-mer synthesized peptides covering the full length of HTNV-GPs. It has been shown by the alanine-scanning technique that (885)C, (893)R, (894)K, (895)K, and (896)C are the key amino acids of the binding sites of the GPs. The implications of identifying a novel B-cell epitope for hantavirus immunology and vaccinology are discussed.
Journal of General Virology 08/2012; · 3.36 Impact Factor
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Jing Yi,
Zhuwei Xu,
Ran Zhuang,
Jiuping Wang,
Yusi Zhang,
Ying Ma,
Bei Liu,
Yun Zhang, Chunmei Zhang,
Guolin Yan,
Fanglin Zhang,
Zhikai Xu,
Angang Yang,
Boquan Jin
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[hide abstract]
ABSTRACT: To investigate the role of viral load in the pathogenesis of hemorrhagic fever with renal syndrome, the Hantaan virus RNA load in plasma from 101 patients was quantified, and the relationships between viral load and disease course, severity, and level of specific humoral immunity were analyzed. The viral load, detectable in 79 patients, ranged from 3.43 to 7.33 log(10) copies/mL of plasma. In the early stage of disease, patients in severe/critical group were found to have higher viral loads than those in the mild/moderate group (5.90 vs 5.03 log(10) copies/mL; P = .001), suggesting an association between Hantaan virus load and disease severity.
The Journal of Infectious Diseases 08/2012; · 6.41 Impact Factor
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Chunmei Zhang,
Rende Li,
Yongming Li,
Chaojun Song,
Zhijia Liu,
Yun Zhang,
Zhuwei Xu,
Ran Zhuang,
Jing Yi,
Angang Yang,
Kun Yang,
Boquan Jin
[show abstract]
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ABSTRACT: Traditionally monoclonal antibody (MAb) titer is determined by indirect enzyme-linked immunosorbent assay (ELISA), which is primarily used to evaluate the quality of MAbs. In this study, the titer and affinity of a group of MAbs against ovalbumin (OVA) were tested by indirect ELISA and the ELISA method reported previously. Data showed that there may be great differences between the indirect ELISA antibody titer and affinity value of MAbs. For the first time, a simple and effective reverse direct ELISA (RD-ELISA) was established for the detection of high-affinity MAbs. Among the group of MAbs to OVA, a certain proportion of antibodies with high affinity but low indirect ELISA titer do exist and can be clearly and efficiently detected by RD-ELISA. This study demonstrates that RD-ELISA is an effective method for high-affinity MAb screening.
Hybridoma (2005) 08/2012; 31(4):284-8. · 0.42 Impact Factor
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Zhijia Liu, Chunmei Zhang,
Yongming Li,
Chaojun Song,
Yuanjie Sun,
Yuying Wei,
Zhuwei Xu,
Angang Yang,
Zhikai Xu,
Kun Yang,
Boquan Jin
[show abstract]
[hide abstract]
ABSTRACT: Botulinum neurotoxins (BoNTs) are classified as category A biological threat agents by the Centers for Disease Control and Prevention (CDC) in the United States for its hazardous and potential bioterrorist threat to the public. About 1% naturally occurring botulisms are caused by Botulinum neurotoxin serotype F (BoNT/F). Most of the immunoassays for detecting BoNTs focus on the serotypes A and B, but few methods have been established for the detection of BoNT/F. Recently, the recombinant Hc subunit of botulinum neurotoxin type F (rFHc) was expressed as an effective vaccine against BoNT/F, indicating that this rFHc could be an effective immunogen to raise monoclonal antibodies (MAbs) for the detection and neutralization of BoNT/F. Here we present a novel sandwich enzyme-linked immunosorbent assay (ELISA) based on two MAbs against rFHc, which were FMMU-BTF-8 and FMMU-BTF-29 as capture antibody and detection antibody, respectively. The limit of detection (LOD) of this ELISA reached 12.09 pg/mL, much less than that of the other reported immunoassays. A simple, sensitive ELISA for detecting and quantifying BoNT/F was established, which can be used as a valuable method to detect and quantify BoNT/F.
Hybridoma (2005) 08/2012; 31(4):233-9. · 0.42 Impact Factor
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Zhijia Liu,
Chaojun Song,
Yongming Li,
Fei Liu,
Kui Zhang,
Yuanjie Sun,
Haitao Li,
Yuying Wei,
Zhuwei Xu, Chunmei Zhang,
Angang Yang,
Zhikai Xu,
Kun Yang,
Boquan Jin
[show abstract]
[hide abstract]
ABSTRACT: Botulinum neurotoxins (BoNTs) are the most poisonous substances ever known. The early detection of these toxins could bear more time for appropriate medical intervention. The standard method for detecting BoNTs is the mouse bioassay, which is time consuming (up to 4 days) and requires a large number of laboratory animals. The immunologic detection methods could detect the toxins within a day, but most of these methods are less sensitive compared with the mouse bioassay due to the lack of high-affinity antibodies. Recently, the recombinant H(C) subunit of botulinum neurotoxin type A (rAH(C)) was expressed as an effective vaccine against botulism, indicating that the rAH(C) could be an effective immunogen that raises the monoclonal antibody (mAb) for detecting BoNT/A. After immunized BALB/c mice with rAH(C), 56 mAbs were generated. Two of these mAbs were selected to establish a highly sensitive sandwich chemiluminescence enzyme immunoassay (CLEIA), in which FMMU-BTA-49 and FMMU-BTA-22 were used as capture antibody and detection antibody, respectively. The calculated limit of detection (LOD) based on molecular weight of rAH(C) and BoNT/A reached 0.45 pg mL(-1). This CLEIA can be used in the detection of BoNT/A in matrices such as milk and beef extract. This method has 20-40 fold lower LOD than that of the mouse bioassay and takes only 3 h to complete the detection, indicating that it can be used as a valuable method to detect and quantify BoNT/A.
Analytica chimica acta 07/2012; 735:23-30. · 4.31 Impact Factor
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Ying Ma,
Bin Yuan,
Jing Yi,
Ran Zhuang,
Jiuping Wang,
Yun Zhang,
Zhuwei Xu,
Yusi Zhang,
Bei Liu,
Chao Wei, Chunmei Zhang,
Angang Yang,
Boquan Jin
[show abstract]
[hide abstract]
ABSTRACT: The polymorphism of human leukocyte antigen (HLA), which is a genetic factor that influences the progression of hemorrhagic fever with renal syndrome (HFRS) after Hantaan virus (HTNV) infection, was incompletely understood. In this case-control study, 76 HFRS patients and 370 healthy controls of the Chinese Han population were typed for the HLA-A, -B, and -DRB1 loci. The general variation at the HLA-DRB1 locus was associated with the onset of HFRS (P < 0.05). The increasing frequencies of HLA-DRB1∗09 and HLA-B∗46-DRB1∗09 in HFRS patients were observed as reproducing a previous study. Moreover, the HLA-B∗51-DRB1∗09 was susceptible to HFRS (P = 0.037; OR = 3.62; 95% CI: 1.00-13.18). The increasing frequencies of HLA-B∗46, HLA-B∗46-DRB1∗09, and HLA-B∗51-DRB1∗09 were observed almost in severe/critical HFRS patients. The mean level of maximum serum creatinine was higher in HLA-B∗46-DRB1∗09 (P = 0.011), HLA-B∗51-DRB1∗09 (P = 0.041), or HLA-B∗46 (P = 0.011) positive patients than that in the negative patients. These findings suggest that the allele HLA-B∗46 and haplotypes HLA-B∗46-DRB1∗09 and HLA-B∗51-DRB1∗09 in patients could contribute to a more severe degree of HFRS and more serious kidney injury, which improve our understanding of the HLA polymorphism for a different outcome of HTNV infection.
Clinical and Developmental Immunology 01/2012; 2012:308237. · 1.84 Impact Factor
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Ying Ma,
Bei Liu,
Bin Yuan,
Jiuping Wang,
Haitao Yu,
Yun Zhang,
Zhuwei Xu,
Yusi Zhang,
Jing Yi, Chunmei Zhang,
Xingchun Zhou,
Angang Yang,
Ran Zhuang,
Boquan Jin
[show abstract]
[hide abstract]
ABSTRACT: To investigate the role of vascular endothelial growth factor (VEGF) in the increased permeability of vascular endothelial cells after Hantaan virus (HTNV) infection in humans, the concentration of VEGF in serum from HTNV infected patients was quantified with sandwich ELISA. Generally, the level of serum VEGF in patients was elevated to 607.0 (542.2-671.9) pg/mL, which was dramatically higher compared with healthy controls (P < 0.001). There was a rapid increase of the serum VEGF level in all patients from the fever onset to oliguric stage, at which the serum creatinine reached the peak level of the disease, indicating that VEGF may be involved in the pathogenesis of renal hyper-permeability. Moreover, the serum VEGF level at convalescent stage was positively correlated with the degree of the disease severity. The sustained high level of serum VEGF at convalescence was observed in critical HFRS patients, suggesting that VEGF would probably contribute to the renal recovery after the virus clearance. Taken together, our results suggested that the VEGF would be involved in the pathogenesis of renal dysfunction at the oliguric stage after HTNV infection, but may function as a recovery factor during the convalescence to help the body self-repair of the renal injury.
Clinical and Developmental Immunology 01/2012; 2012:812386. · 1.84 Impact Factor
