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Journal articles on the topic 'Bacterial ghosts'

Author: Grafiati

Published: 4 June 2021

Last updated: 19 February 2022

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1

Panthel, Klaus, Wolfgang Jechlinger, Alexander Matis, Manfred Rohde, Michael Szostak, Werner Lubitz, and Rainer Haas. "Generation of Helicobacter pylori Ghosts by PhiX Protein E-Mediated Inactivation and Their Evaluation as Vaccine Candidates." Infection and Immunity 71, no. 1 (January 2003): 109–16. http://dx.doi.org/10.1128/iai.71.1.109-116.2003.

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ABSTRACT Bacterial ghosts are empty cell envelopes, which may be generated by the controlled expression of the PhiX174 lysis gene E in gram-negative bacteria to obtain vaccine candidates. We describe here the application of this technology to Helicobacter pylori. The lysis gene cassette was cloned into an Escherichia coli-Helicobacter pylori shuttle vector and introduced into an H. pylori recipient strain by bacterial conjugation. Temperature induction of the lysis gene cassette revealed a quantitative killing of the H. pylori culture without induction of lysis-resistant bacteria. Biochemical and transmission electron microscopic studies identified structurally intact H. pylori. Prophylactic oral vaccination experiments using these H. pylori ghosts in the BALB/c mouse model showed a significant reduction of the bacterial load in the ghost group, as measured by a quantitative bacterial reisolation procedure. Ten of 10 and 5 of 10 mice were protected, respectively, without the use of a mucosal adjuvant. Coadministration of ghosts with cholera toxin as mucosal adjuvant resulted in a complete protection of 10 of 10 and 8 of 8 mice against H. pylori challenge, with three animals showing a sterile immunity.

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2

Hjelm, Anna, Bill Söderström, David Vikström, Wouter S. P. Jong, Joen Luirink, and Jan-Willem de Gier. "Autotransporter-Based Antigen Display in Bacterial Ghosts." Applied and Environmental Microbiology 81, no. 2 (November 14, 2014): 726–35. http://dx.doi.org/10.1128/aem.02733-14.

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ABSTRACTBacterial ghosts are empty cell envelopes of Gram-negative bacteria that can be used as vehicles for antigen delivery. Ghosts are generated by releasing the bacterial cytoplasmic contents through a channel in the cell envelope that is created by the controlled production of the bacteriophage ϕX174 lysis protein E. While ghosts possess all the immunostimulatory surface properties of the original host strain, they do not pose any of the infectious threats associated with live vaccines. Recently, we have engineered theEscherichia coliautotransporter hemoglobin protease (Hbp) into a platform for the efficient surface display of heterologous proteins in Gram-negative bacteria, HbpD. Using theMycobacterium tuberculosisvaccine target ESAT6 (early secreted antigenic target of 6 kDa), we have explored the application of HbpD to decorateE. coliandSalmonellaghosts with antigens. The use of different promoter systems enabled the concerted production of HbpD-ESAT6 and lysis protein E. Ghost formation was monitored by determining lysis efficiency based on CFU, the localization of a set of cellular markers, fluorescence microscopy, flow cytometry, and electron microscopy. Hbp-mediated surface display of ESAT6 was monitored using a combination of a protease accessibility assay, fluorescence microscopy, flow cytometry and (immuno-)electron microscopy. Here, we show that the concerted production of HbpD and lysis protein E inE. coliandSalmonellacan be used to produce ghosts that efficiently display antigens on their surface. This system holds promise for the development of safe and cost-effective vaccines with optimal intrinsic adjuvant activity and exposure of heterologous antigens to the immune system.

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3

Mayr, Ulrike Beate, Christoph Haller, Wolfgang Haidinger, Alena Atrasheuskaya, Eugenij Bukin, Werner Lubitz, and Georgy Ignatyev. "Bacterial Ghosts as an Oral Vaccine: a Single Dose of Escherichia coli O157:H7 Bacterial Ghosts Protects Mice against Lethal Challenge." Infection and Immunity 73, no. 8 (August 2005): 4810–17. http://dx.doi.org/10.1128/iai.73.8.4810-4817.2005.

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ABSTRACT Enterohemorrhagic Escherichia coli (EHEC) is a bacterial pathogen that is associated with several life-threatening diseases for humans. The combination of protein E-mediated cell lysis to produce EHEC ghosts and staphylococcal nuclease A to degrade DNA was used for the development of an oral EHEC vaccine. The lack of genetic material in the oral EHEC bacterial-ghost vaccine abolished any hazard of horizontal gene transfer of resistance genes or pathogenic islands to resident gut flora. Intragastric immunization of mice with EHEC ghosts without the addition of any adjuvant induced cellular and humoral immunity. Immunized mice challenged at day 55 showed 86% protection against lethal challenge with a heterologous EHEC strain after single-dose oral immunization and 93.3% protection after one booster at day 28, whereas the controls showed 26.7% and 30% survival, respectively. These results indicate that it is possible to develop an efficacious single-dose oral EHEC bacterial-ghost vaccine.

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4

Kim, Sam Woong, Yeon Jo Ha, Kyu Ho Bang, Seungki Lee, Joo-Hong Yeo, Hee-Sun Yang, Tae-Won Kim, Kyu Pil Lee, and Woo Young Bang. "Potential of Bacteriocins from Lactobacillus taiwanensis for Producing Bacterial Ghosts as a Next Generation Vaccine." Toxins 12, no. 7 (July 1, 2020): 432. http://dx.doi.org/10.3390/toxins12070432.

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Bacteriocins are functionally diverse toxins produced by most microbes and are potent antimicrobial peptides (AMPs) for bacterial ghosts as next generation vaccines. Here, we first report that the AMPs secreted from Lactobacillus taiwanensis effectively form ghosts of pathogenic bacteria and are identified as diverse bacteriocins, including novel ones. In detail, a cell-free supernatant from L. taiwanensis exhibited antimicrobial activities against pathogenic bacteria and was observed to effectively cause cellular lysis through pore formation in the bacterial membrane using scanning electron microscopy (SEM). The treatment of the cell-free supernatant with proteinase K or EDTA proved that the antimicrobial activity is mediated by AMPs, and the purification of AMPs using Sep-Pak columns indicated that the cell-free supernatant includes various amphipathic peptides responsible for the antimicrobial activity. Furthermore, the whole-genome sequencing of L. taiwanensis revealed that the strain has diverse bacteriocins, confirmed experimentally to function as AMPs, and among them are three novel bacteriocins, designated as Tan 1, Tan 2, and Tan 3. We also confirmed, using SEM, that Tan 2 effectively produces bacterial ghosts. Therefore, our data suggest that the bacteriocins from L. taiwanensis are potentially useful as a critical component for the preparation of bacterial ghosts.

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5

Riedmann, Eva M., Jennelle M. Kyd, Allan W. Cripps, and Werner Lubitz. "Bacterial ghosts as adjuvant particles." Expert Review of Vaccines 6, no. 2 (April 2007): 241–53. http://dx.doi.org/10.1586/14760584.6.2.241.

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6

Szostak, M., G. Wanner, and W. Lubitz. "Recombinant bacterial ghosts as vaccines." Research in Microbiology 141, no. 7-8 (September 1990): 1005–7. http://dx.doi.org/10.1016/0923-2508(90)90141-c.

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7

Gong, Saisai, Nan Nan, Yakun Sun, Zhili He, Jiajia Li, Fanghong Chen, Tao Li, et al. "Protective Immunity Elicited by VP1 Chimeric Antigens of Bacterial Ghosts against Hand-Foot-and-Mouth Disease Virus." Vaccines 8, no. 1 (February 1, 2020): 61. http://dx.doi.org/10.3390/vaccines8010061.

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This study was designed to evaluate the immunogenicity and protective efficacy of two VP1 chimeric antigens of bacterial ghosts. Inoculation of the two VP1 chimeric antigens of bacterial ghosts into BALB/c mice markedly elicited humoral and mucosal immune responses. The specific antibodies induced by the chimeric ghosts protected mice not only against the virus that causes hand-foot-and-mouth disease but also against E. coli O157:H7 bacterial infection. In comparison with the negative control, immunization with the chimeric ghosts protected mice against two LD50 hand-foot-and-mouth disease viral infection. In addition, this specific immunity also protected the pups of pregnant mice immunized with the VP1 chimeric antigens of bacterial ghosts against 20 MLD E. coli O157:H7 infection. Taken together, the results of this study verify for the first time that the VP1 chimeric antigens of bacterial ghosts are target candidates for a new type of vaccine against hand-foot-and-mouth disease. Additionally, this vaccine strategy also elicited a stronger immune response against E. coli O157:H7.

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8

Doig, P., A. L. Franklin, and R. T. Irvin. "The binding of Pseudomonas aeruginosa outer membrane ghosts to human buccal epithelial cells." Canadian Journal of Microbiology 32, no. 2 (February 1, 1986): 160–66. http://dx.doi.org/10.1139/m86-032.

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The binding of outer membrane (OM) ghosts derived from Pseudomonas aeruginosa strain 492c to human buccal epithelial cells (BECs) was examined. Electron microscopic examination of the binding of OM ghosts to BECs revealed direct OM ghost–BEC interaction. Equilibrium analysis of the binding of OM ghosts to trypsinized BECs employing the Langmuir adsorption isotherm indicated the number of binding sites (iV) to be 1.3 × 10−1 μg protein per BEC with an apparent association constant (Ka) of 3.4 × 10−2 mL/μg protein. The Langmuir analysis of binding of OM ghosts to untrypsinized BECs was complex, suggesting two possible classes of receptors, a high affinity–low copy number class (Ka, 1.8 × 10−2 mL/μg protein; N, 8.6 × 10−5 μg protein per BEC) and a low affinity – high copy number class(Afa, 3.7 × 10−3 mL/μg protein; N, 9.2 × 10−4 μg protein per BEC). Sugar inhibition studies incorporating D-galactose enhanced binding to each BEC type. N-Acetylneuraminic acid and N-acetyl-glucosamine both enhanced binding of OM ghosts to untrypsinized BECs, while inhibiting binding to trypsinized BECs. D-Arabinose inhibited binding to both BEC types. Binding of OM ghosts to both BEC types was greatly inhibited by D-fucose, while L-fucose only greatly inhibited binding to untrypsinized BECs. These sugar inhibition data demonstrated a difference in the binding of OM ghosts to trypsinized and untrypsinized BECs and possibly reveal the nature of the receptor(s), free of possible bacterial metabolic effects. These data indicated that OM ghosts from 492c appear to bind to BECs in a similar manner to the intact bacteria and represent a simple model system to study the adhesion of P. aeruginosa to BECs.

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9

Koller, Verena J., Verena M. Dirsch, Hortenzia Beres, Oliver Donath, Gottfried Reznicek, Werner Lubitz, and Pavol Kudela. "Modulation of bacterial ghosts - induced nitric oxide production in macrophages by bacterial ghost-delivered resveratrol." FEBS Journal 280, no. 5 (January 31, 2013): 1214–25. http://dx.doi.org/10.1111/febs.12112.

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10

Mayr, Ulrike Beate, Petra Walcher, Chakameh Azimpour, Eva Riedmann, Christoph Haller, and Werner Lubitz. "Bacterial ghosts as antigen delivery vehicles." Advanced Drug Delivery Reviews 57, no. 9 (June 2005): 1381–91. http://dx.doi.org/10.1016/j.addr.2005.01.027.

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11

Szostak, Michael P., Andreas Hensel, Francis O. Eko, Reinhard Klein, Tatjana Auer, Horst Mader, Alexander Haslberger, Sebastian Bunka, Gerhard Wanner, and Werner Lubitz. "Bacterial ghosts: non-living candidate vaccines." Journal of Biotechnology 44, no. 1-3 (January 1996): 161–70. http://dx.doi.org/10.1016/0168-1656(95)00123-9.

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12

Haidinger, W., M. P. Szostak, W. Jechlinger, and W. Lubitz. "Online Monitoring of Escherichia coli Ghost Production." Applied and Environmental Microbiology 69, no. 1 (January 2003): 468–74. http://dx.doi.org/10.1128/aem.69.1.468-474.2003.

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ABSTRACT Controlled expression of cloned φX174 gene E in gram-negative bacteria results in lysis of the bacteria by the formation of a transmembrane tunnel structure built through the cell envelope complex. Production of bacterial ghosts is routinely monitored by classical microbiological procedures. These include determination of the turbidity of the culture and the total number of cells and the number of reproductive cells present during the time course of growth and lysis. Although conceptually simple, these methods are labor intensive and time consuming, providing a complete set of results after the determination of viable cell counts. To avoid culturing methods for bacterial growth, an alternative flow cytometric procedure is presented for the quantification of ghosts and polarized, as well as depolarized, nonlysed cells within a culture. For this method, which is based on the discriminatory power of the membrane potential-sensitive dye bis-(1,3-dibutylbarbituric acid) trimethine oxonol, a staining protocol was developed and optimized for the maximum discrepancy in fluorescence between bacterial ghosts and viable cells. The total quantitative analysis procedure takes less than 2 min. The results derived from classical or cytometric analyses correlate with respect to the total cell numbers and the viability of the culture.

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13

Vanlint, Dietrich, Mehari Tesfazgi Mebhratu, Chris W. Michiels, and Abram Aertsen. "Using Mild High-pressure Shock to Generate Bacterial Ghosts of Escherichia coli." Zeitschrift für Naturforschung B 63, no. 6 (June 1, 2008): 765–68. http://dx.doi.org/10.1515/znb-2008-0626.

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In the context of vaccine development, bacterial ghosts are inert cells that retain the capacity to activate the immune system, and that can be used as vaccine or carrier for subunit or DNA vaccines. In this study we provide evidence that increasing the copynumber of the E. coli K12 mrr locus can render naturally occurring and virulent avian pathogenic E. coli (APEC) strains hypersensitive to high pressure. We further demonstrate that mild HP shock generates inactive bacterial ghosts from these cells that have not incurred any microscopically visible structural damage. Possible benefits of high-pressure generated bacterial ghosts as a vaccine are discussed.

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14

Jawale, Chetan V., and John Hwa Lee. "Salmonella enterica Serovar Enteritidis Ghosts Carrying the Escherichia coli Heat-Labile Enterotoxin B Subunit Are Capable of Inducing Enhanced Protective Immune Responses." Clinical and Vaccine Immunology 21, no. 6 (March 26, 2014): 799–807. http://dx.doi.org/10.1128/cvi.00016-14.

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ABSTRACTTheEscherichia coliheat-labile enterotoxin B subunit (LTB) is a potent vaccine adjuvant.Salmonella entericaserovar Enteritidis ghosts carrying LTB (S. Enteritidis-LTB ghosts) were genetically constructed using a novel plasmid, pJHL187-LTB, designed for the coexpression of the LTB and E lysis proteins.S. Enteritidis-LTB ghosts were characterized using scanning electron microscopy to visualize their transmembrane tunnel structures. The expression of LTB inS. Enteritidis-LTB ghost preparations was confirmed by immunoblot and enzyme-linked immunosorbent assays. The parenteral adjuvant activity of LTB was demonstrated by immunizing chickens with eitherS. Enteritidis-LTB ghosts orS. Enteritidis ghosts. Chickens were intramuscularly primed at 5 weeks of age and subsequently boosted at 8 weeks of age. In total, 60 chickens were equally divided into three groups (n= 20 for each): group A, nonvaccinated control; group B, immunized withS. Enteritidis-LTB ghosts; and group C, immunized withS. Enteritidis ghosts. Compared with the nonimmunized chickens (group A), the immunized chickens (groups B and C) exhibited increased titers of plasma IgG and intestinal secretory IgA antibodies. The CD3+CD4+subpopulation of T cells was also significantly increased in both immunized groups. Among the immunized chickens, those in group B exhibited significantly increased titers of specific plasma IgG and intestinal secretory IgA (sIgA) antibodies compared with those in group C, indicating the immunomodulatory effects of the LTB adjuvant. Furthermore, both immunized groups exhibited decreased bacterial loads in their feces and internal organs. These results indicate that parenteral immunization withS. Enteritidis-LTB ghosts can stimulate superior induction of systemic and mucosal immune responses compared to immunization withS. Enteritidis ghosts alone, thus conferring efficient protection against salmonellosis.

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15

Lubitz, W. "Bacterial ghosts as carrier and targeting systems." Expert Opinion on Biological Therapy 1, no. 5 (September 2001): 765–71. http://dx.doi.org/10.1517/14712598.1.5.765.

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16

Mayrhofer, Peter, Chakameh Azimpour Tabrizi, Petra Walcher, Wolfgang Haidinger, Wolfgang Jechlinger, and Werner Lubitz. "Immobilization of plasmid DNA in bacterial ghosts." Journal of Controlled Release 102, no. 3 (February 2005): 725–35. http://dx.doi.org/10.1016/j.jconrel.2004.10.026.

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17

Thanh, Doan Duy, and Tran Xuan Hanh. "Study of chemical-based induced bacterial ghost applied in vaccine production." ENGINEERING AND TECHNOLOGY 10, no. 1 (June 4, 2020): 17–28. http://dx.doi.org/10.46223/hcmcoujs.tech.en.10.1.356.2020.

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Introduction: Bacterial ghosts (BGs), known as the empty cell envelope of gram-negative bacteria lacking cytoplasmic content yet retaining all unaltered morphological and structural features of their living counterparts, are widely studied and used as the platform for the production of the vaccines as well as the transporting drug and gene delivery. However, the study related to the creation of BGs based on gene expression is still limited because of the difference in cell wall structure between microorganisms. Therefore, in the current study, for the aims to determine chemicals combination and minimum inhibition concentration (MIC) to optimize BGs production. Material and method: Salmonella choleraesuis strain was collected from NAVETCO company. The study used critical concentrations from chemical combination to convert salmonella cells to BGs. Chemicals combination and MIC, temperature, shaking speed were optimized using Plakett- Burman matrix and response surface methodology. Cell structure was determined by using a scanning electron microscope, experimental mice were vaccinated and challenged with virulence to determine immune responses of bacterial ghost. Results: The appropriate chemicals for the production of BGs biomass were NaOH 3.125 mg/ml; SDS 1.15 mg/ml, H2O2 8.79 µl/ml, ethanol. The observation of morphology, BGs have remained the structure and shape, which were like the living microbial cells. Conclusions: The conditions of BGs production have been identified to produce large amounts of bacterial ghost biomass to further application in vaccine production and pharma.

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18

Jalava, K. "Bacterial ghosts as vaccine candidates for veterinary applications." Journal of Controlled Release 85, no. 1-3 (December 13, 2002): 17–25. http://dx.doi.org/10.1016/s0168-3659(02)00267-5.

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19

Huter, Veronika, Michael P. Szostak, Jörg Gampfer, Saskia Prethaler, Gerhard Wanner, Franz Gabor, and Werner Lubitz. "Bacterial ghosts as drug carrier and targeting vehicles." Journal of Controlled Release 61, no. 1-2 (August 1999): 51–63. http://dx.doi.org/10.1016/s0168-3659(99)00099-1.

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20

Park, Hyun, Sung Oh, Nagarajan Vinod, Seongmi Ji, Han Noh, Jung Koo, Su Lee, Sei Kim, Ki-Sung Lee, and Chang Choi. "Characterization of Chemically-Induced Bacterial Ghosts (BGs) Using Sodium Hydroxide-Induced Vibrio parahaemolyticus Ghosts (VPGs)." International Journal of Molecular Sciences 17, no. 11 (November 15, 2016): 1904. http://dx.doi.org/10.3390/ijms17111904.

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21

Kudela, Pavol, Verena Juliana Koller, and Werner Lubitz. "Bacterial ghosts (BGs)—Advanced antigen and drug delivery system." Vaccine 28, no. 36 (August 2010): 5760–67. http://dx.doi.org/10.1016/j.vaccine.2010.06.087.

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22

Wu, Xueyou, Xingrong Ju, Lihui Du, Jian Yuan, Lifeng Wang, Rong He, and Zhengxing Chen. "Production of Bacterial Ghosts from Gram-Positive PathogenListeria monocytogenes." Foodborne Pathogens and Disease 14, no. 1 (January 2017): 1–7. http://dx.doi.org/10.1089/fpd.2016.2184.

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23

Amara, Amro A., Mounir M. Salem-Bekhit, and Fars K. Alanazi. "Sponge-Like: A New Protocol for Preparing Bacterial Ghosts." Scientific World Journal 2013 (2013): 1–7. http://dx.doi.org/10.1155/2013/545741.

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Bacterial Ghosts (BGs) received an increasing interest in the recent years for their promising medicinal and pharmaceutical applications. In this study, for the first time we introduce a new protocol for BGs production.E. coliBL21 (DE3) pLysS (Promega) was used as a model to establish a general protocol for BGs preparation. The protocol is based on using active chemical compounds in concentrations less than the Minimum Inhibition Concentration (MIC). Those chemical compounds are SDS, NaOH, and H2O2. Plackett-Burman experimental design was used to map the best conditions for BGs production. Normal and electronic microscopes were used to evaluate the BGs quality (BGQ). Spectrophotometer was used to evaluate the amount of the released protein and DNA. Agarose gel electrophoresis was used to determine the existence of any residue of DNA after each BGs preparation. Viable cells, which existed after running this protocol, were subjected to lysis by inducing the lysozyme gene carried on pLysS plasmid. This protocol is able to produce BGs that can be used in different biotechnological applications.

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Kudela, Pavol, Susanne Paukner, Ulrike Beate Mayr, Dana Cholujova, Gudrun Kohl, Zuzana Schwarczova, Jozef Bizik, Jan Sedlak, and Werner Lubitz. "Effective gene transfer to melanoma cells using bacterial ghosts." Cancer Letters 262, no. 1 (April 2008): 54–63. http://dx.doi.org/10.1016/j.canlet.2007.11.031.

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25

Kudela, Pavol, Verena Juliana Koller, Ulrike Beate Mayr, Johannes Nepp, Werner Lubitz, and Talin Barisani-Asenbauer. "Bacterial Ghosts as antigen and drug delivery system for ocular surface diseases: Effective internalization of Bacterial Ghosts by human conjunctival epithelial cells." Journal of Biotechnology 153, no. 3-4 (May 2011): 167–75. http://dx.doi.org/10.1016/j.jbiotec.2011.03.022.

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26

Meyer-Bahlburg, Almut, Jörg Brinkhoff, Veit Krenn, Karlheinz Trebesius, Jürgen Heesemann, and Hans-Iko Huppertz. "Infection of Synovial Fibroblasts in Culture by Yersinia enterocolitica and Salmonella enterica Serovar Enteritidis: Ultrastructural Investigation with Respect to the Pathogenesis of Reactive Arthritis." Infection and Immunity 69, no. 12 (December 1, 2001): 7915–21. http://dx.doi.org/10.1128/iai.69.12.7915-7921.2001.

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ABSTRACT Synovial fibroblasts were infected with Yersinia enterocolitica or Salmonella enterica serovar Enteritidis and analyzed by electron microscopy and fluorescence in situ hybridization. Intracellular bacterial replication was followed by degradation leading to “ghosts” possessing lipopolysaccharides but not DNA. However, single bacteria survived for more than 2 weeks. Therefore, transient intra-articular infection might be the missing link between initial intestinal infection and late synovial inflammation in the pathogenesis of reactive arthritis.

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27

Ma, Yi, Liu Cui, Meng Wang, Qiuli Sun, Kaisheng Liu, and Jufang Wang. "A Novel and Efficient High-Yield Method for Preparing Bacterial Ghosts." Toxins 13, no. 6 (June 13, 2021): 420. http://dx.doi.org/10.3390/toxins13060420.

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Bacterial ghosts (BGs) are empty cell envelopes possessing native extracellular structures without a cytoplasm and genetic materials. BGs are proposed to have significant prospects in biomedical research as vaccines or delivery carriers. The applications of BGs are often limited by inefficient bacterial lysis and a low yield. To solve these problems, we compared the lysis efficiency of the wild-type protein E (EW) from phage ΦX174 and the screened mutant protein E (EM) in the Escherichia coli BL21(DE3) strain. The results show that the lysis efficiency mediated by protein EM was improved. The implementation of the pLysS plasmid allowed nearly 100% lysis efficiency, with a high initial cell density as high as OD600 = 2.0, which was higher compared to the commonly used BG preparation method. The results of Western blot analysis and immunofluorescence indicate that the expression level of protein EM was significantly higher than that of the non-pLysS plasmid. High-quality BGs were observed by SEM and TEM. To verify the applicability of this method in other bacteria, the T7 RNA polymerase expression system was successfully constructed in Salmonella enterica (S. Enterica, SE). A pET vector containing EM and pLysS were introduced to obtain high-quality SE ghosts which could provide efficient protection for humans and animals. This paper describes a novel and commonly used method to produce high-quality BGs on a large scale for the first time.

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Haidinger, W., U. B. Mayr, M. P. Szostak, S. Resch, and W. Lubitz. "Escherichia coli Ghost Production by Expression of Lysis Gene E and Staphylococcal Nuclease." Applied and Environmental Microbiology 69, no. 10 (October 2003): 6106–13. http://dx.doi.org/10.1128/aem.69.10.6106-6113.2003.

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ABSTRACT The production of bacterial ghosts from Escherichia coli is accomplished by the controlled expression of phage φX174 lysis gene E and, in contrast to other gram-negative bacterial species, is accompanied by the rare detection of nonlysed, reproductive cells within the ghost preparation. To overcome this problem, the expression of a secondary killing gene was suggested to give rise to the complete genetic inactivation of the bacterial samples. The expression of staphylococcal nuclease A in E. coli resulted in intracellular accumulation of the protein and degradation of the host DNA into fragments shorter than 100 bp. Two expression systems for the nuclease are presented and were combined with the protein E-mediated lysis system. Under optimized conditions for the coexpression of gene E and the staphylococcal nuclease, the concentration of viable cells fell below the lower limit of detection, whereas the rates of ghost formation were not affected. With regard to the absence of reproductive cells from the ghost fractions, the reduction of viability could be determined as being at least 7 to 8 orders of magnitude. The lysis process was characterized by electrophoretic analysis and absolute quantification of the genetic material within the cells and the culture supernatant via real-time PCR. The ongoing degradation of the bacterial nucleic acids resulted in a continuous quantitative clearance of the genetic material associated with the lysing cells until the concentrations fell below the detection limits of either assay. No functional, released genetic units (genes) were detected within the supernatant during the lysis process, including nuclease expression.

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29

Ebensen, Thomas, Susanne Paukner, Claudia Link, Pavol Kudela, Carola de Domenico, Werner Lubitz, and Carlos A. Guzmán. "Bacterial Ghosts Are an Efficient Delivery System for DNA Vaccines." Journal of Immunology 172, no. 11 (May 20, 2004): 6858–65. http://dx.doi.org/10.4049/jimmunol.172.11.6858.

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30

Rabea, Sameh, Mounir M. Salem-Bekhit, Fars K. Alanazi, Aymen S. Yassin, Nayera A. Moneib, and Abd Elgawad M. Hashem. "A novel protocol for bacterial ghosts’ preparation using tween 80." Saudi Pharmaceutical Journal 26, no. 2 (February 2018): 232–37. http://dx.doi.org/10.1016/j.jsps.2017.12.006.

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31

Groza, Diana, Sebastian Gehrig, Pavol Kudela, Martin Holcmann, Christine Pirker, Carina Dinhof, Hemma H. Schueffl, et al. "Bacterial ghosts as adjuvant to oxaliplatin chemotherapy in colorectal carcinomatosis." OncoImmunology 7, no. 5 (February 16, 2018): e1424676. http://dx.doi.org/10.1080/2162402x.2018.1424676.

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32

Hoffelner, Herbert, and Rainer Haas. "Recombinant bacterial ghosts: versatile targeting vehicles and promising vaccine candidates." International Journal of Medical Microbiology 294, no. 5 (October 2004): 303–11. http://dx.doi.org/10.1016/j.ijmm.2004.04.003.

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33

Groza, D., S. Gehrig, C. Dinhof, M. Holcman, C. Pirker, H. Schueffl, M. Ogris, et al. "Bacterial ghosts as adjuvant to oxaliplatin chemotherapy in colorectal carcinomatosis." European Journal of Cancer 110 (March 2019): S23—S24. http://dx.doi.org/10.1016/j.ejca.2019.01.076.

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Xie, Songzhi, Shang Li, Zhanlin Zhang, Maohua Chen, Pan Ran, and Xiaohong Li. "Bacterial ghosts for targeting delivery and subsequent responsive release of ciprofloxacin to destruct intracellular bacteria." Chemical Engineering Journal 399 (November 2020): 125700. http://dx.doi.org/10.1016/j.cej.2020.125700.

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35

Jalava, Katri, Francis O. Eko, Eva Riedmann, and Werner Lubitz. "Bacterial ghosts as carrier and targeting systems for mucosal antigen delivery." Expert Review of Vaccines 2, no. 1 (February 2003): 45–51. http://dx.doi.org/10.1586/14760584.2.1.45.

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Haslberger, A. G., G. Kohl, D. Felnerova, U. B. Mayr, S. Fürst-Ladani, and W. Lubitz. "Activation, stimulation and uptake of bacterial ghosts in antigen presenting cells." Journal of Biotechnology 83, no. 1-2 (September 2000): 57–66. http://dx.doi.org/10.1016/s0168-1656(00)00298-4.

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Mader, Horst J., Michael P. Szostak, Andreas Hensel, Werner Lubitz, and Alexander G. Haslberger. "Endotoxicity does not limit the use of bacterial ghosts as candidate vaccines." Vaccine 15, no. 2 (February 1997): 195–202. http://dx.doi.org/10.1016/s0264-410x(96)00141-7.

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Amro, Amara A., Mounir M. Salem-Bekhit, and Fars K. Alanazi. "Plackett–Burman randomization method for Bacterial Ghosts preparation form E. coli JM109." Saudi Pharmaceutical Journal 22, no. 3 (July 2014): 273–79. http://dx.doi.org/10.1016/j.jsps.2013.06.002.

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Zhu, Wenxing, Guiwen Yang, Yuyu Zhang, Jinduo Yuan, and Liguo An. "Generation of Biotechnology-DerivedFlavobacterium columnareGhosts by PhiX174 GeneE-Mediated Inactivation and the Potential as Vaccine Candidates against Infection in Grass Carp." Journal of Biomedicine and Biotechnology 2012 (2012): 1–8. http://dx.doi.org/10.1155/2012/760730.

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Flavobacterium columnareis a bacterial pathogen causing high mortality rates for many freshwater fish species. Fish vaccination with a safe and effective vaccine is a potential approach for prevention and control of fish disease. Here, in order to produce bacterial ghost vaccine, a specificFlavobacteriumlysis plasmid pBV-E-cat was constructed by cloning PhiX174 lysis geneEand thecatgene with the promoter ofF. columnareinto the prokaryotic expression vector pBV220. The plasmid was successfully electroporated into the strainF. columnareG4cpN22 after curing of its endogenous plasmid.F. columnareG4cpN22 ghosts (FCGs) were generated for the first time by geneE-mediated lysis, and the vaccine potential of FCG was investigated in grass carp (Ctenopharyngodon idellus) by intraperitoneal route. Fish immunized with FCG showed significantly higher serum agglutination titers and bactericidal activity than fish immunized with FKC or PBS. Most importantly, after challenge with the parent strain G4, the relative percent survival (RPS) of fish in FCG group (70.9%) was significantly higher than FKC group (41.9%). These results showed that FCG could confer immune protection againstF. columnareinfection. As a nonliving whole cell envelope preparation, FCG may provide an ideal alternative to pathogen-based vaccines against columnaris in aquaculture.

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Huppertz, H. I., and J. Heesemann. "Experimental Yersinia infection of human synovial cells: persistence of live bacteria and generation of bacterial antigen deposits including "ghosts," nucleic acid-free bacterial rods." Infection and immunity 64, no. 4 (1996): 1484–87. http://dx.doi.org/10.1128/iai.64.4.1484-1487.1996.

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Jawale, Chetan V., Atul A. Chaudhari, Byung Woo Jeon, Rahul M. Nandre, and John Hwa Lee. "Characterization of a Novel Inactivated Salmonella enterica Serovar Enteritidis Vaccine Candidate Generated Using a Modified cI857/λ PR/GeneEExpression System." Infection and Immunity 80, no. 4 (January 30, 2012): 1502–9. http://dx.doi.org/10.1128/iai.06264-11.

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ABSTRACTA new strategy to develop an effective vaccine is essential to control food-borneSalmonella entericaserovar Enteritidis infections. Bacterial ghosts (BGs), which are nonliving, Gram-negative bacterial cell envelopes, are generated by expulsion of the cytoplasmic contents from bacterial cells through controlled expression using the modified cI857/λ PR/geneEexpression system. In the present study, the pJHL99 lysis plasmid carrying the mutated lambda pR37-cI857 repressor and PhiX174 lysis geneEwas constructed and transformed inS. Enteritidis to produce a BG. Temperature induction of the lysis gene cassette at 42°C revealed quantitative killing ofS. Enteritidis. TheS. Enteritidis ghost was characterized using scanning and transmission electron microscopy to visualize the transmembrane tunnel structure and loss of cytoplasmic materials, respectively. The efficacy of the BG as a vaccine candidate was evaluated in a chicken model using 60 10-day-old chickens, which were divided into four groups (n= 15), A, B, C, and D. Group A was designated as the nonimmunized control group, whereas the birds in groups B, C, and D were immunized via the intramuscular, subcutaneous, and oral routes, respectively. The chickens from all immunized groups showed significant increases in plasma IgG and intestinal secretory IgA levels. The lymphocyte proliferation response and CD3+CD4+and CD3+CD8+T cell subpopulations were also significantly increased in all immunized groups. The data indicate that both humoral and cell-mediated immune responses are robustly stimulated. Based on an examination of the protection efficacy measured by observations of gross lesions in the organs and bacterial recovery, the candidate vaccine can provide efficient protection against virulent challenge.

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Meitz, Andrea, Patrick Sagmeister, Werner Lubitz, Christoph Herwig, and Timo Langemann. "Fed-Batch Production of Bacterial Ghosts Using Dielectric Spectroscopy for Dynamic Process Control." Microorganisms 4, no. 2 (March 24, 2016): 18. http://dx.doi.org/10.3390/microorganisms4020018.

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Afkhami-Poustchi, Amin, and Maryam M. Matin. "Use of Bacterial Ghosts as Novel Drug Delivery Systems to Improve Cancer Treatment." Cancer Press 2, no. 1 (February 29, 2016): 8. http://dx.doi.org/10.15562/tcp.11.

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Afkhami-Poustchi, Amin, and Maryam M. Matin. "Use of bacterial ghosts as novel drug delivery systems to improve cancer treatment." Journal of Cellular Immunotherapy 1, no. 1-2 (December 2015): 6–7. http://dx.doi.org/10.1016/j.jocit.2015.10.009.

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Liu, Jun, Yi Li, Yang Sun, Xue Ji, Lingwei Zhu, Xuejun Guo, Wei Zhou, et al. "Immune responses and protection induced by Brucella suis S2 bacterial ghosts in mice." Veterinary Immunology and Immunopathology 166, no. 3-4 (August 2015): 138–44. http://dx.doi.org/10.1016/j.vetimm.2015.04.008.

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Paukner, Susanne, Pavol Kudela, Gudrun Kohl, Tobias Schlapp, Sonja Friedrichs, and Werner Lubitz. "DNA-Loaded Bacterial Ghosts Efficiently Mediate Reporter Gene Transfer and Expression in Macrophages." Molecular Therapy 11, no. 2 (February 2005): 215–23. http://dx.doi.org/10.1016/j.ymthe.2004.09.024.

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Amara, Amro A., Mounir M. Salem-Bekh, and Fars K. Alanazi. "Preparation of Bacterial Ghosts for E. coli JM109 Using “Sponge-like Reduced Protocol”." Asian Journal of Biological Sciences 6, no. 8 (November 1, 2013): 363–69. http://dx.doi.org/10.3923/ajbs.2013.363.369.

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Paukner, Susanne, Thomas Stiedl, Pavol Kudela, Jozef Bizik, Firas Al Laham, and Werner Lubitz. "Bacterial ghosts as a novel advanced targeting system for drug and DNA delivery." Expert Opinion on Drug Delivery 3, no. 1 (December 22, 2005): 11–22. http://dx.doi.org/10.1517/17425247.3.1.11.

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Kassmannhuber, Johannes, Mascha Rauscher, Lea Schöner, Angela Witte, and Werner Lubitz. "Functional display of ice nucleation protein InaZ on the surface of bacterial ghosts." Bioengineered 8, no. 5 (January 25, 2017): 488–500. http://dx.doi.org/10.1080/21655979.2017.1284712.

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Tabrizi, Chakameh Azimpour, Petra Walcher, Ulrike Beate Mayr, Thomas Stiedl, Matthias Binder, John McGrath, and Werner Lubitz. "Bacterial ghosts – biological particles as delivery systems for antigens, nucleic acids and drugs." Current Opinion in Biotechnology 15, no. 6 (December 2004): 530–37. http://dx.doi.org/10.1016/j.copbio.2004.10.004.

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