Protein microarray for profiling antibody responses to Yersinia pestis live vaccine

Laboratory of Analytical Microbiology, National Center for Biomedical Analysis, Army Center for Microbial Detection and Research, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China.
Infection and Immunity (Impact Factor: 4.16). 07/2005; 73(6):3734-9. DOI: 10.1128/IAI.73.6.3734-3739.2005
Source: PubMed

ABSTRACT A protein microarray representing 149 Yersinia pestis proteins was developed to profile antibody responses in EV76-immunized rabbits. Antibodies to 50 proteins were detected. There are 11 proteins besides F1 and V antigens to which the predominant antibody response occurred, and these proteins show promise for further evaluation as candidates for subunit vaccines and/or diagnostic antigens.

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Available from: Zongmin Du, Aug 14, 2015
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    • "These proteins were considered to be immunogens for humans because they can induce humoral responses during Y. pestis infection. Except for 10 proteins that were discovered to be immunogens in previous studies (Anderson et al. 1996; Andrews et al. 1996; Benner et al. 1999; Leary et al. 1999; Li et al. 2005b, 2008; Tanabe et al. 2006), the immunogenicities of the other 11 proteins ( "
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    ABSTRACT: Yersinia pestis is a bacterium that is transmitted between fleas, which have a body temperature of 26 °C, and mammalian hosts, which have a body temperature of 37 °C. To adapt to the temperature shift, phenotype variations, including virulence, occur. In this study, an antigen microarray including 218 proteins of Y. pestis was used to evaluate antibody responses in a pooled plague serum that was unadsorbed, adsorbed by Y. pestis cultivated at 26 °C, or adsorbed by Y. pestis cultivated at 26 and 37 °C to identify protein expression changes during the temperature shift. We identified 12 proteins as being expressed at 37 °C but not at 26 °C, or expressed at significantly higher levels at 37 °C than at 26 °C. The antibodies against 7 proteins in the serum adsorbed by Y. pestis cultivated at 26 and 37 °C remained positive, suggesting that they were not expressed on the surface of Y. pestis in LB broth in vitro or specifically expressed in vivo. This study proved that protein microarray and antibody profiling comprise a promising technique for monitoring gene expression at the protein level and for better understanding pathogenicity, to find new vaccine targets against plague.
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    • "Therefore, discovery of complementary diagnostic targets is urgently needed. Previous studies in our laboratory (Li et al., 2005) showed that among about 200 known or predicted proteins of Y. pestis, more than 10 of them had strong immunogenicity to induce antibody production, which could be detected in the sera of a variety of plague infected animals by the protein chip method. Ten of these proteins were chosen for developing a double-antigen sandwich TC-UPT-LF assay to detect their corresponding antibodies in order to lay a foundation to evaluate more serum samples from different animals or humans with plague infections for screening new plague diagnostic markers. "
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    ABSTRACT: In this study, a 10-channel up-converting phosphor technology-based lateral flow (TC-UPT-LF) assay was developed to profile antibodies against Yersinia pestis. Ten expressed Y. pestis proteins were covalently conjugated with an up-converting phosphor particle to develop double-antigen sandwich immunochromatographic strips to detect corresponding antibodies. After optimization one by one, each strip was integrated into a TC-UPT-LF disc for simultaneously detection of different antibodies. A scanning biosensor was also developed to acquire the results. The performance of the TC-UPT-LF assay was evaluated by using standard samples and plague monkey serum samples. Fifty-one patient serum samples were detected by the TC-UPT-LF assay. The TC-UPT-LF disc could be stable for 10 days at 37°C with an average CV of 10.3%. Its sensitivity and qualitative results are comparable to those of ELISA. Its linearity fitting coefficient of determination (R2) for different antibody detection is between 0.93 and 0.99. Besides F1 antibody, the LcrV and YopD antibodies also showed higher positive ratio than the other seven antibodies, as 100% (13/13) and 92% (12/13) in monkey sera and 86.3% (44/51) and 66.7% (34/51) in patient sera, respectively. It is suggested that the TC-UPT-LF assay has been successfully developed for multi-detection and LcrV and YopD can be the potential diagnostic markers of the plague.
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    • "2,3 The Yunnan Province includes two plague foci, Focus F and Focus E, 4 the Rattus flavipectus plague focus and the Apodemus chevrieri and Eothenomys miletus plague focus, respectively. Y. pestis isolated from these foci belongs to biovar Orientalis and Antiqua, re- spectively. 2 In October 2005, five people in the Yulong county of Yunnan province, where no plague had been reported in the last 100 years, experienced fever, cough, and hard breath. "
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    ABSTRACT: Plague is a deadly infectious disease caused by the gram-negative bacterium, Yersinia pestis. In 2005, five plague patients were confirmed in the Yulong County of the Yunnan Province, China. In this study, the serologic survey of > 2,900 serum samples from domestic dogs and cats in and around the county, where human plague occurred, confirmed that domestic dogs and cats could serve as sentinel animals for plague surveillance. Meanwhile, the antibody responses in the infected dogs and cats were profiled by microarray containing 218 proteins of Y. pestis. In addition to F1, LcrV, YPCD1.28c, and YPO2118 induced humoral responses in all or most of the individuals, providing complementary candidates to F1 antigen for diagnostic markers of plague.
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