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Bacterial ghosts (BGs): A promising approach as candidate vaccine

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Abstract

Bacterial ghosts (BGs) are an emerging vaccine platform produced by completely removing the cytoplasmic contents of bacterial cells while preserving their native surface architecture. This unique structural integrity enables BGs to function simultaneously as safe, non-living vaccines and efficient delivery vehicles. Recent advances in both genetic (phage lysis gene E) and chemical “sponge-like” protocols have expanded the range of microorganisms from which BGs can be produced, improving safety, scalability, and antigenic stability. This review summarizes current progress in BG technology with a focus on their innovative applications as antibacterial, antifungal, and anticancer vaccines; adjuvants; and carriers for DNA, proteins, and bioactive molecules. Particular emphasis is placed on emerging directions such as yeast and fungal ghosts, novel characterization methods, and the development of BG-based nano-vaccines. Future prospects highlight the need for standardized production, improved clinical translation, and comparative evaluation with related platforms such as membrane vesicles. Together, these advancements position BGs as a promising next-generation vaccine and drug-delivery strategy with significant potential for translational impact.

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No datasets were generated or analysed during the current study.

Abbreviations

Abbreviation:

Full Term

APC:

Antigen-presenting cells

BG:

Bacterial Ghosts

GFP:

Green fluorescent protein

GMO:

Genetically manipulated organisms

MAP:

Model amphiphilic peptide

MGC:

Minimum growth concentration

MIC:

Minimum inhibitory concentration

PAMP:

Pathogen-associated molecular patterns

PRR:

Pattern recognition receptors

SLP:

Sponge-Like-protocol

TLR:

Toll-like receptors

References

  • Abd El-Baky N, Amara AA (2014) Newcastle disease virus (LaSota strain) as a model for virus ghosts preparation using H2O2 bio-critical concentration. Int Sci Invest J 3:38–50

    Google Scholar 

  • Abdelnaby MA, Shoueir KR, Ghazy AA, Abdelhamid SM, El Kemary MA, Mahmoud HE, Baraka K, Abozahra RR (2022) Synthesis and evaluation of metallic nanoparticles-based vaccines against Candida albicans infections. J Drug Deliv Sci Technol 68:102862

    Article  CAS  Google Scholar 

  • Abdullah NA, Mahmoud HE, El-Nikhely NA, Hussein AA, El-Khordagui LK (2023) Carbon Dots labeled Lactiplantibacillus plantarum: a fluorescent multifunctional biocarrier for anticancer drug delivery, frontiers in bioengineering and biotechnology, Volume 11–2023

  • Abtin A, Kudela P, Mayr UB, Koller VJ, Mildner M, Tschachler E, Lubitz W (2010) Escherichia coli ghosts promote innate immune responses in human keratinocytes. Biochem Biophys Res Commun 400:78–82

    Article  CAS  PubMed  Google Scholar 

  • Ahmad TA, Haroun M, Hussein AA, Ashry SHE el, and, El-Sayed LH (2012) Development of a new trend conjugate vaccine for the prevention of Klebsiella pneumoniae, Infect Dis Rep, 4: e33

  • Akira S, Hemmi H (2003) Recognition of pathogen-associated molecular patterns by TLR family. Immunol Lett 85:85–95

    Article  CAS  PubMed  Google Scholar 

  • Al-Fattah Amara A (2017) Cell evacology: could it be a new science. Cell Mol Med 3:9

    Google Scholar 

  • Ali RH, Ali ME, Samir R (2023) Production and characterization of bacterial ghost vaccine against neisseria meningitidis. Vaccines 11:37

    Article  CAS  Google Scholar 

  • Amara AAAF (2013) Cracking the microbial Cell Wall. In Biol Sci, 363 – 69

  • Amara A (2015) Saccharomyces cerevisiae ghosts using the Sponge-Like Re-Reduced protocol. SOJ Biochem 2:1–4

    Article  Google Scholar 

  • Amara A (2017) Smart green technology for microbial ghosts preparation. Iioab J 8:53–54

    Google Scholar 

  • Amara A (2020) E. coli BL21 bacterial and Newcastle Pestikal lasota viral ghosts preparation, DNA, RNA elimination/purification/isolation/quantification using agarose gel entrapment bubble method. J Protein Res Bioinf 2:010

    Google Scholar 

  • Amara A, Salem-Bekhet M, Alanazi F (2013a) Preparation of bacterial ghosts for E. coli JM109 using Sponge-like reduced protocol. Asian J Biol Sci 6:363–369

    Article  CAS  Google Scholar 

  • Amara AA, Salem-Bekhit MM, Alanazi FK (2013b) Sponge-like: a new protocol for preparing bacterial ghosts, scientific world journal, 2013: 545741

  • Amro AA, Neama AJ, Hussein A, Hashish EA, Sheweita SA (2014a) Evaluation the Surface Antigen of the Salmonella typhimurium ATCC 14028 Ghosts Prepared by SLRP, The Scientific World Journal, 2014: 840863

  • Amro AA, Salem-Bekhit MM, Alanazi FK (2014b) Plackett-Burman randomization method for bacterial ghosts Preparation form E. coli JM109. Saudi Pharm J 22:273–279

    Article  PubMed  Google Scholar 

  • Anwer M, Bhaliya K, Munn A, Wei MQ (2025) Bacterial ghosts: A breakthrough approach to cancer vaccination. Biomed Pharmacother 182:117766

    Article  CAS  PubMed  Google Scholar 

  • Batah AM, Ahmad TA (2020) The development of ghost vaccines trials. Expert Rev Vaccines 19:549–562

    Article  CAS  PubMed  Google Scholar 

  • Bezbaruah R, Chavda VP, Nongrang L, Alom S, Deka K, Kalita T, Ali F, Bhattacharjee B, Vora L (2022) Nanoparticle-Based Delivery Systems for Vaccines, Vaccines (Basel), 10

  • Bläsi U, Linke RP, Lubitz W (1989) Evidence for membrane-bound oligomerization of bacteriophage phi X174 lysis protein-E. J Biol Chem 264:4552–4558

    Article  PubMed  Google Scholar 

  • Cai K, Gao X, Li T, Hou X, Wang Q, Liu H, Xiao L, Tu W, Liu Y, Shi J, Wang H (2010) Intragastric immunization of mice with enterohemorrhagic Escherichia coli O157:H7 bacterial ghosts reduces mortality and shedding and induces a Th2-type dominated mixed immune response. Can J Microbiol 56:389–398

    Article  CAS  PubMed  Google Scholar 

  • Chen J, Li N, She F (2014) Helicobacter pylori outer inflammatory protein DNA vaccine-loaded bacterial ghost enhances immune protective efficacy in C57BL/6 mice. Vaccine 32:6054–6060

    Article  CAS  PubMed  Google Scholar 

  • Chen H, Ji H, Kong X, Lei P, Yang Q, Wu W, Jin L, Sun D (2021) Bacterial Ghosts-Based Vaccine and Drug Delivery Systems, Pharmaceutics, 13: 1892

  • Chu W, Zhuang X, Lu C (2008) Generation of Aeromonas hydrophila ghosts and their evaluation as oral vaccine candidates in Carassius auratus gibelio. Wei Sheng Wu Xue Bao 48:202–6

    CAS  PubMed  Google Scholar 

  • D’Amico C, Fontana F, Cheng R, Santos HA (2021) Development of vaccine formulations: past, present, and future. Drug Deliv Transl Res 11:353–72

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Dalabehera M, Ghosh A, Mohanty S, Chellappan DK, Chaudhari S, Ale Y, Poonia N, Subudhi RN, Khanna MK, Lim HG (2025) mRNA delivery systems 2.0: engineering extrahepatic delivery for non-vaccine therapeutics. Mater Today Bio 35:102584

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Dentovskaya SV, Vagaiskaya AS, Trunyakova AS, Kartseva AS, Ivashchenko TA, Gerasimov VN, Platonov ME, Firstova VV, Anisimov AP (2025) Genetically Engineered Bacterial Ghosts as Vaccine Candidates Against Klebsiella pneumoniae Infection, Vaccines (Basel), 13

  • Dillard JA, Taft-Benz SA, Knight AC, Anderson EJ, Pressey KD, Parotti B, Martinez SA, Diaz JL, Sarkar S, Madden EA, De la Cruz G, Adams LE, Dinnon KH, Leist SR, Martinez DR, Schäfer A, Powers JM, Yount BL, Castillo IN, Morales NL, Burdick J, Evangelista MKD, Ralph LM, Pankow NC, Linnertz CL, Lakshmanane P, Montgomery SA, Ferris MT, Baric RS, Baxter VK, Heise MT (2024) Adjuvant-dependent impact of inactivated SARS-CoV-2 vaccines during heterologous infection by a SARS-related coronavirus. Nat Commun 15:3738

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ding L, Liang X, Ma J, Liu X, Zhang Y, Long Q, Wen Z, Teng Z, Jiang L, Liu G (2024) Sono-triggered biomimetically nanoantibiotics mediate precise sequential therapy of MRSA-induced lung infection. Adv Mater 36:2403612

    Article  CAS  Google Scholar 

  • Dobrovolskienė N, Pašukonienė V, Darinskas A, Kraśko JA, Žilionytė K, Mlynska A, Gudlevičienė Ž, Mišeikytė-Kaubrienė E, Schijns V, Lubitz W, Kudela P, Strioga M (2018) Tumor lysate-loaded bacterial ghosts as a tool for optimized production of therapeutic dendritic cell-based cancer vaccines. Vaccine 36:4171–4180

    Article  PubMed  Google Scholar 

  • Ebensen T, Paukner S, Link C, Kudela P, de Domenico C, Lubitz W, Guzmán CA (2004) Bacterial ghosts are an efficient delivery system for DNA vaccines1. J Immunol 172:6858–65

    Article  CAS  PubMed  Google Scholar 

  • Edgar RH, Viator JA (2024) “Ghost-Particle-Enhanced Photoacoustic Flow Cytometry for Identification of Bacterial Cells in Body Fluids.” In: Xia Wenfeng (ed) Biomedical Photoacoustics: Technology and Applications. Springer Nature Switzerland, Cham

  • Ekong EE, Okenu DN, Mania-Pramanik J, He Q, Igietseme JU, Ananaba GA, Lyn D, Black C, Eko FO (2009) A Vibrio cholerae ghost-based subunit vaccine induces cross-protective chlamydial immunity that is enhanced by CTA2B, the nontoxic derivative of cholera toxin. FEMS Immunol Med Microbiol 55:280–91

    Article  CAS  PubMed  Google Scholar 

  • Felnerova D, Kudela P, Bizik J, Haslberger A, Hensel A, Saalmuller A, Lubitz W (2004) T cell-specific immune response induced by bacterial ghosts. Med Sci Monit 10:Br362–Br370

  • Frosch M, Meyer TF (1992) Transformation-mediated exchange of virulence determinants by co-cultivation of pathogenic Neisseriae. FEMS Microbiol Lett 100:345–9

    Article  CAS  PubMed  Google Scholar 

  • Gong S, Nan N, Sun Y, He Z, Li J, Chen F, Li T, Ning N, Wang J, Li Z, Luo D, Wang H (2020) Protective immunity elicited by VP1 chimeric antigens of bacterial ghosts against hand-foot-and-mouth disease virus. Vaccines. https://doi.org/10.3390/vaccines8010061

    Article  PubMed  PubMed Central  Google Scholar 

  • Groza D, Gehrig S, Kudela P, Holcmann M, Pirker C, Dinhof C, Schueffl HH, Sramko M, Hoebart J, Alioglu F, Grusch M, Ogris M, Lubitz W, Keppler BK, Pashkunova-Martic I, Kowol CR, Sibilia M, Berger W, Heffeter P (2018) Bacterial ghosts as adjuvant to oxaliplatin chemotherapy in colorectal carcinomatosis. Oncoimmunology 7:e1424676

    Article  PubMed  PubMed Central  Google Scholar 

  • Haidinger W, Mayr UB, Szostak MP, Resch S, Lubitz W (2003) Escherichia coli ghost production by expression of lysis gene E and Staphylococcal nuclease. Appl Environ Microbiol 69:6106–13

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hajam IA, Dar PA, Won G, Lee JH (2017) Bacterial ghosts as adjuvants: mechanisms and potential. Vet Res 48:37

    Article  PubMed  PubMed Central  Google Scholar 

  • Haslberger AG, Kohl G, Felnerova D, Mayr UB, Fürst-Ladani S, Lubitz W (2000) Activation, stimulation and uptake of bacterial ghosts in antigen presenting cells. J Biotechnol 83:57–66

    Article  CAS  PubMed  Google Scholar 

  • Hensel A, van Leengoed LA, Szostak M, Windt H, Weissenböck H, Stockhofe-Zurwieden N, Katinger A, Stadler M, Ganter M, Bunka S, Pabst R, Lubitz W (1996) Induction of protective immunity by aerosol or oral application of candidate vaccines in a dose-controlled pig aerosol infection model. J Biotechnol 44:171–81

    Article  CAS  PubMed  Google Scholar 

  • Hensel A, Huter V, Katinger A, Raza P, Strnistschie C, Roesler U, Brand E, Lubitz W (2000) Intramuscular immunization with genetically inactivated (ghosts) Actinobacillus pleuropneumoniae serotype 9 protects pigs against homologous aerosol challenge and prevents carrier state. Vaccine 18:2945–2955

    Article  CAS  PubMed  Google Scholar 

  • Hjelm A, Söderström B, Vikström D, Jong WSP, Luirink J, Gier J-Wd (2015) Autotransporter-based antigen display in bacterial ghosts. Appl Environ Microbiol 81:726–35

    Article  PubMed  Google Scholar 

  • Hu J, Dong H, Fu L, Zuo J, Wu X, Mi R, Huang Y, Lu K, Chen Z, Han X, Hu S (2017) Comparison of three methods for preparation of bacterial ghosts from avian pathogenic Escherichia coli. Sheng Wu Gong Cheng Xue Bao 33:2009–16

    CAS  PubMed  Google Scholar 

  • Huter V, Szostak MP, Gampfer J, Prethaler S, Wanner G, Gabor F, Lubitz W (1999) Bacterial ghosts as drug carrier and targeting vehicles. J Control Release 61:51–63

    Article  CAS  PubMed  Google Scholar 

  • Huter V, Hensel A, Brand E, Lubitz W (2000) Improved protection against lung colonization by Actinobacillus pleuropneumoniae ghosts: characterization of a genetically inactivated vaccine. J Biotechnol 83:161–72

    Article  CAS  PubMed  Google Scholar 

  • Igietseme JU, Eko FO, He Q, Black CM (2004) Antibody regulation of Tcell immunity: implications for vaccine strategies against intracellular pathogens. Expert Rev Vaccines 3:23–34

    Article  CAS  PubMed  Google Scholar 

  • Jalava K, Hensel A, Szostak M, Resch S, Lubitz W (2002) Bacterial ghosts as vaccine candidates for veterinary applications. J Controlled Release 85:17–25

    Article  CAS  Google Scholar 

  • Jawale CV, Lee JH (2014a) Comparative evaluation of Salmonella enteritidis ghost vaccines with a commercial vaccine for protection against internal egg contamination with Salmonella. Vaccine 32:5925–5930

    Article  PubMed  Google Scholar 

  • Jawale CV, Lee JH (2014b) A novel approach for the generation of Salmonella gallinarum ghosts and evaluation of their vaccine potential using a prime-booster immunization strategy. Vaccine 32:6776–6782

    Article  CAS  PubMed  Google Scholar 

  • Jeyanathan M, Afkhami S, Smaill F, Miller MS, Lichty BD, Xing Z (2020) Immunological considerations for COVID-19 vaccine strategies. Nat Rev Immunol 20:615–632

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kang S-R, Nguyen D-H, Yoo SW, Min J-J (2022) Bacteria and bacterial derivatives as delivery carriers for immunotherapy. Adv Drug Deliv Rev 181:114085

    Article  CAS  PubMed  Google Scholar 

  • Kawai T, Akira S (2011) Toll-like receptors and their crosstalk with other innate receptors in infection and immunity. Immunity 34:637–650

    Article  CAS  PubMed  Google Scholar 

  • Khalil MS, Makled SO, El-Nikhely N, Hussein AA, El-Khordagui L, Mahmoud HE (2025) Microencapsulation of monascus red pigment using Saccharomyces cerevisiae ghosts. Process Optimization and Characterization, Probiotics and Antimicrobial Proteins

  • Kraśko JA, Žilionytė K, Darinskas A, Strioga M, Rjabceva S, Zalutsky I, Derevyanko M, Kulchitsky V, Lubitz W, Kudela P, Miseikyte-Kaubriene E, Karaman O, Didenko H, Potebnya H, Chekhun V, and V Pašukonienė (2017) Bacterial ghosts as adjuvants in syngeneic tumour cell lysate-based anticancer vaccination in a murine lung carcinoma model. Oncol Rep 37:171–178

    Article  PubMed  Google Scholar 

  • Kudela P, Koller VJ, Lubitz W (2010) Bacterial ghosts (BGs)—advanced antigen and drug delivery system. Vaccine 28:5760–5767

    Article  CAS  PubMed  Google Scholar 

  • Langemann T, Koller VJ, Muhammad A, Kudela P, Mayr UB, Lubitz W (2010) The bacterial ghost platform system: production and applications. Bioeng Bugs 1:326–336

    Article  PubMed  PubMed Central  Google Scholar 

  • Lavelle EC, McEntee CP (2024) Vaccine adjuvants: tailoring innate recognition to send the right message. Immunity 57:772–89

    Article  CAS  PubMed  Google Scholar 

  • Laver WG, Air GM, Webster RG, Smith-Gill SJ (1990) Epitopes on protein antigens: misconceptions and realities. Cell 61:553–556

    Article  CAS  PubMed  Google Scholar 

  • Lee C, Jeong E, Sung KH, Park SJ, Choi Y (2025) The influence of bioactive glass (BGS-7) on enamel remineralization: an in vitro study. Restor Dent Endod 50:e33

    Article  PubMed  PubMed Central  Google Scholar 

  • Li Z, Wang Y, Liu J, Rawding P, Bu J, Hong S, Hu Q (2021) Chemically and biologically engineered Bacteria-Based delivery systems for emerging diagnosis and advanced therapy. Adv Mater 33:2102580

    Article  CAS  Google Scholar 

  • Ling D, Jia X, Wang K, Yan Q, Yuan B, Du L, Li M, Jin Y (2024) Cancer cell membrane-coated bacterial ghosts for highly efficient paclitaxel delivery against metastatic lung cancer. Acta Pharm Sin B 14:365–377

    Article  CAS  PubMed  Google Scholar 

  • Lubitz P, Mayr UB, Lubitz W (2009) “Applications of Bacterial Ghosts in Biomedicine.” In: Guzmán Carlos A., Feuerstein Giora Z. (eds) Pharmaceutical Biotechnology. Springer New York, New York, NY

  • Ma Y, Cui L, Wang M, Sun Q, Liu K, Wang J (2021) A novel and efficient high-yield method for preparing bacterial ghosts. Toxins 13:420

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ma J, Ding L, Peng X, Jiang L, Liu G (2024) Recent advances of engineered cell membrane-based nanotherapeutics to combat inflammatory diseases. Small 20:e2308646

  • Ma Y, Wang S, Hong B, Feng L, Wang J (2024) Construction and mechanism exploration of highly efficient system for bacterial ghosts preparation based on engineered phage ID52 lysis protein E. Vaccines 12:472

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ma J, Teng Z, Ding L, Peng X, Xie Y, Long Q, Jiang L, Huo S, Liang X, Liu G (2025) Ultrasound-triggered nano-bomb: bacteria-targeted oxygen-supplying liposomes as seed bubbles for enhanced mechano-sonodynamic therapy of Pseudomonas aeruginosa biofilms. Adv Funct Mater 35:2501347

    Article  CAS  Google Scholar 

  • Mader HJ, Szostak MP, Hensel A, Lubitz W, Haslberger AG (1997) Endotoxicity does not limit the use of bacterial ghosts as candidate vaccines. Vaccine 15:195–202

    Article  CAS  PubMed  Google Scholar 

  • Makino K, Ishii K, Yasunaga T, Hattori M, Yokoyama K, Yutsudo CH, Kubota Y, Yamaichi Y, Iida T, Yamamoto K, Honda T, Han CG, Ohtsubo E, Kasamatsu M, Hayashi T, Kuhara S, Shinagawa H (1998) Complete nucleotide sequences of 93-kb and 3.3-kb plasmids of an enterohemorrhagic Escherichia coli O157:H7 derived from Sakai outbreak. DNA Res 5:1–9

    Article  CAS  PubMed  Google Scholar 

  • Mayr UB, Haller C, Haidinger W, Atrasheuskaya A, Bukin E, Lubitz W, Ignatyev G (2005) Bacterial ghosts as an oral vaccine: a single dose of Escherichia coli O157:H7 bacterial ghosts protects mice against lethal challenge. Infect Immun 73:4810–7

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mayr UB, Walcher P, Azimpour C, Riedmann E, Haller C, Lubitz W (2005) Bacterial ghosts as antigen delivery vehicles. Adv Drug Deliv Rev 57:1381–91

    Article  CAS  PubMed  Google Scholar 

  • Medzhitov R, Janeway CJ (2000) Innate immune recognition: mechanisms and pathwaysabstract. Immunol Rev 173:89–97

    Article  CAS  PubMed  Google Scholar 

  • Mendel S, Holbourn JM, Schouten JA, Bugg TDH (2006) Interaction of the transmembrane domain of Lysis protein E from bacteriophage phiX174 with bacterial translocase MraY and peptidyl-prolyl isomerase SlyD. Microbiol Reading 152:2959–2967

    Article  Google Scholar 

  • Michalek J, Hezova R, Turanek-Knötigova P, Gabkova J, Strioga M, Lubitz W, Kudela P (2017) Oncolysate-loaded Escherichia coli bacterial ghosts enhance the stimulatory capacity of human dendritic cells. Cancer Immunol Immunother 66:149–159

    Article  CAS  PubMed  Google Scholar 

  • Minor PD (2015) Live attenuated vaccines: historical successes and current challenges. Virology 479–480:379–92

    Article  PubMed  Google Scholar 

  • Mm M, Hussein A, Ghazy A, Sheweita S, Amara A (2017) Klebsiella pneumoniae ghosts as vaccine using sponge like reduced protocol. Open access, Cellular & Molecular Medicine, p 03

    Google Scholar 

  • Moghanian A, Zohourfazeli M, Mahdi Tajer MH, Miri Z, Hosseini S, Rashvand A (2021) Preparation, characterization and in vitro biological response of simultaneous co-substitution of Zr + 4/Sr + 2 58S bioactive glass powder. Ceram Int 47:23762–23769

    Article  CAS  Google Scholar 

  • Mohamed AAAFA, Khalif A (2015) Kostenlos Viral Ghosts, Bacterial Ghosts, Microbial Ghosts and More. Schüling Verlag

    Google Scholar 

  • Muhammad A, Kassmannhuber J, Rauscher M, Falcon AA, Wheeler DW, Zhang AA, Lubitz P, Lubitz W (2019) Subcutaneous immunization of dogs with Bordetella bronchiseptica bacterial ghost vaccine. Front Immunol 10:1377

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Narayanan S, Baburajan AP, Muhammad M, Joseph A, Vemula PK, Bhat SG (2024) Demonstrating the immunostimulatory and cytokine-augmentation effects of bacterial ghosts on natural killer cells and caenorhabditis elegans. Biotechnol Bioeng 121:959–970

    Article  CAS  PubMed  Google Scholar 

  • Nunberg JH, Westover JB, York J, Jung KH, Bailey KW, Boardman KM, Li M, Furnell RS, Wasson SR, Murray JS, Kaundal R, Thomas AJ, Gowen BB (2024) Restoration of virulence in the attenuated Candid#1 vaccine virus requires reversion at both positions 168 and 427 in the envelope glycoprotein GPC. J Virol 98:e0011224

    Article  PubMed  PubMed Central  Google Scholar 

  • Palm-Apergi C, Hällbrink M (2008) A new rapid cell-penetrating peptide based strategy to produce bacterial ghosts for plasmid delivery. J Control Release 132:49–54

    Article  CAS  PubMed  Google Scholar 

  • Panoff J-M, Chuiton C (2004) Horizontal gene transfer: a universal phenomenon. Hum Ecol Risk Assess 10:939–943

    Article  CAS  Google Scholar 

  • Park S-Y (2023) Chemically induced bacterial ghosts: a novel approach for advancing biomedical applications. Molecular & Cellular Toxicology 19:657 − 65

    Article  Google Scholar 

  • Paukner S, Kohl G, Jalava K, Lubitz W (2003) Sealed bacterial ghosts–novel targeting vehicles for advanced drug delivery of water-soluble substances. J Drug Target 11:151–161

    CAS  PubMed  Google Scholar 

  • Paukner S, Kudela P, Kohl G, Schlapp T, Friedrichs S, Lubitz W (2005) DNA-loaded bacterial ghosts efficiently mediate reporter gene transfer and expression in macrophages. Mol Ther 11:215–223

    Article  CAS  PubMed  Google Scholar 

  • Plotkin SA (2005) Vaccines: past, present and future. Nat Med 11:S5–11

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Quevedo-Diaz MA, Song C, Xiong Y, Chen H, Wahl LM, Radulovic S, Medvedev AE (2010) Involvement of TLR2 and TLR4 in cell responses to Rickettsia akari. J Leukoc Biol 88:675–85

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rabea S, Salem-Bekhit MM, Alanazi FK, Yassin AS, Moneib NA, Hashem AEM (2018) A novel protocol for bacterial ghosts’ preparation using tween 80. Saudi Pharm J 26:232–237

    Article  PubMed  Google Scholar 

  • Rabea S, Alanazi FK, Ashour AE, Salem-Bekhit MM, Yassin AS, Moneib NA, Hashem AEM, Haq N (2020) Salmonella-innovative targeting carrier: loading with doxorubicin for cancer treatment. Saudi Pharm J 28:1253–62

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ran X, Meng XZ, Geng HL, Chang C, Chen X, Wen X, Ni H (2019) Generation of porcine Pasteurella multocida ghost vaccine and examination of its immunogenicity against virulent challenge in mice. Microb Pathog 132:208–214

    Article  CAS  PubMed  Google Scholar 

  • Riedmann EM, Kyd JM, Cripps AW, Lubitz W (2007) Bacterial ghosts as adjuvant particles. Expert Rev Vaccines 6:241–253

    Article  CAS  PubMed  Google Scholar 

  • Riedmann EM, Lubitz W, McGrath J, Kyd JM, Cripps AW (2011) Effectiveness of engineering the nontypeable <Emphasis Type="Italic">Haemophilus</Emphasis> influenzae antigen Omp26 as an S-layer fusion in bacterial ghosts as a mucosal vaccine delivery. Hum Vaccin 7(Suppl):99–107

    Article  CAS  PubMed  Google Scholar 

  • Sabbaghi A, Miri SM, Keshavarz M, Zargar M, Ghaemi A (2019) Inactivation methods for whole influenza vaccine production. Rev Med Virol 29:e2074

    Article  PubMed  Google Scholar 

  • Saleh N, Mahmoud HE, Eltaher H, Helmy M, El-Khordagui L, Hussein AA (2023) Prodigiosin-functionalized probiotic ghosts as a bioinspired combination against colorectal cancer cells. Probiotics Antimicrob Proteins 15:1271–86

    Article  CAS  PubMed  Google Scholar 

  • Salem-Bekhit MM, Youssof AME, Alanazi FK, Aleanizy FS, Abdulaziz A, Taha EI, Amara A (2021) Bacteria from infectious particles to cell based anticancer targeted drug delivery systems. Pharmaceutics. https://doi.org/10.3390/pharmaceutics13121984

    Article  PubMed  PubMed Central  Google Scholar 

  • Senevirathne A, Hewawaduge C, Park JY, Park S, Lee JH (2020) Parenteral immunization of Salmonella Typhimurium ghosts with surface-displayed Escherichia coli flagellin enhancesTLR-5 mediated activation of immune responses that protect the chicken against Salmonella infection. Microb Pathog 147:104252

    Article  CAS  PubMed  Google Scholar 

  • Senevirathne A, Hewawaduge C, Lee JH (2021) Immunization of chicken with flagellin adjuvanted Salmonella enteritidis bacterial ghosts confers complete protection against chicken salmonellosis. Poult Sci 100:101205

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sheweita SA, Batah AM, Ghazy AA, Hussein A, Amara AA (2019) A new strain of Acinetobacter baumannii and characterization of its ghost as a candidate vaccine. J Infect Public Health 12:831–42

    Article  CAS  PubMed  Google Scholar 

  • Sheweita SA, Amara AA, Gamal H, Ghazy AA, Hussein A, Bahey-El-Din M (2022) Bacterial ghosts of <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> as a promising candidate vaccine and its application in diabetic rats. Vaccines (Basel). https://doi.org/10.3390/vaccines10060910

    Article  PubMed  PubMed Central  Google Scholar 

  • Sridharan G, Shankar AA (2012) Toluidine blue: a review of its chemistry and clinical utility. J Oral Maxillofac Pathol 16:251–255

    Article  PubMed  PubMed Central  Google Scholar 

  • Stein E, Inic-Kanada A, Belij S, Montanaro J, Bintner N, Schlacher S, Mayr UB, Lubitz W, Stojanovic M, Najdenski H (2013) In vitro and in vivo uptake study of Escherichia coli Nissle 1917 bacterial ghosts: cell-based delivery system to target ocular surface diseases. Investigative Opthalmology & Visual Science 54:6326–33

    Article  CAS  Google Scholar 

  • Steinhagen F, Kinjo T, Bode C, Klinman DM (2011) TLR-based immune adjuvants. Vaccine 29:3341–55

    Article  CAS  PubMed  Google Scholar 

  • Sührer I, Langemann T, Lubitz W, Weuster-Botz D, Castiglione K (2015) A novel one-step expression and immobilization method for the production of biocatalytic preparations. Microb Cell Fact 14:180

    Article  PubMed  PubMed Central  Google Scholar 

  • Szostak MP, Hensel A, Eko FO, Klein R, Auer T, Mader H, Haslberger A, Bunka S, Wanner G, Lubitz W (1996) Bacterial ghosts: non-living candidate vaccines. J Biotechnol 44:161–170

    Article  CAS  PubMed  Google Scholar 

  • Tabrizi CA, Walcher P, Mayr UB, Stiedl T, Binder M, McGrath J, Lubitz W (2004) Bacterial ghosts–biological particles as delivery systems for antigens, nucleic acids and drugs. Curr Opin Biotechnol 15:530–7

    Article  CAS  PubMed  Google Scholar 

  • Tian Q-f, Wei Z, Wei S, Fei Y, Xin H, Min Y, Chen L-p, Liu S-g, Yu S-y (2018) Construction of Salmonella Pullorum ghost by co-expression of lysis gene E and the antimicrobial peptide SMAP29 and evaluation of its immune efficacy in specific-pathogen-free chicks. J Integr Agric 17:197–209

    Article  CAS  Google Scholar 

  • Tuntufye HN, Ons E, Pham AD, Luyten T, Van Gerven N, Bleyen N, Goddeeris BM (2012) Escherichia coli ghosts or live E. coli expressing the ferri-siderophore receptors FepA, FhuE, IroN and IutA do not protect broiler chickens against avian pathogenic E. coli (APEC). Vet Microbiol 159:470–8

    CAS  PubMed  Google Scholar 

  • Vinod N, Oh S, Park HJ, Koo JM, Choi CW, Kim SC (2015) Generation of a novel Staphylococcus aureus ghost vaccine and examination of its immunogenicity against virulent challenge in rats. Infect Immun 83:2957–65

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wang S, Li Z, Zhang J, Xi L, Cui Y, Zhang W, Zhang J, Zhang H (2020) A safe non-toxic Brucella abortus ghosts induce immune responses and confer protection in BALB/c mice. Mol Immunol 124:117–24

    Article  CAS  PubMed  Google Scholar 

  • Witte A, Wanner G, Bläsi U, Halfmann G, Szostak M, Lubitz W (1990) Endogenous transmembrane tunnel formation mediated by phi X174 lysis protein E. J Bacteriol 172:4109–4114

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Witte A, Brand E, Mayrhofer P, Narendja F, Lubitz W (1998) Mutations in cell division proteins FtsZ and FtsA inhibit φX174 protein-E-mediated lysis of Escherichia coli. Arch Microbiol 170:259–68

    Article  CAS  PubMed  Google Scholar 

  • Won G, Hajam IA, Lee JH (2017) Improved lysis efficiency and immunogenicity of Salmonella ghosts mediated by co-expression of λ phage holin-endolysin and ɸX174 gene E. Sci Rep 7:45139

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wu X, Ju X, Du L, Yuan J, Wang L, He R, Chen Z (2017) Production of bacterial ghosts from Gram-positive pathogen Listeria monocytogenes. Foodborne Pathog Dis 14:1–7

    Article  PubMed  Google Scholar 

  • Xie S, Zhang P, Zhang Z, Liu Y, Chen M, Li S, Li X (2021) Bacterial navigation for tumor targeting and photothermally-triggered bacterial ghost transformation for spatiotemporal drug release. Acta Biomater 131:172–184

    Article  CAS  PubMed  Google Scholar 

  • Youssof AME, Alanazi FK, Salem-Bekhit MM, Shakeel F, Haq N (2019) Bacterial ghosts carrying 5-fluorouracil: a novel biological carrier for targeting colorectal cancer. AAPS PharmSciTech 20:48

    Article  CAS  PubMed  Google Scholar 

  • Zahid A, Ismail H, Wilson JC, Grice ID (2025) Bioengineering outer-membrane vesicles for vaccine development: strategies, advances, and perspectives. Vaccines 13:767

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhu W, Zhang Y, Liu X (2015) Efficient production of safety-enhanced Escherichia coli ghosts by tandem expression of PhiX 174 mutant gene E and staphylococcal nuclease A gene. Microbiol Res 176:7–13

    Article  CAS  PubMed  Google Scholar 

  • Zhu W, Hao L, Liu X, Borrás-Hidalgo O, Zhang Y (2018) Enhanced anti-proliferative efficacy of epothilone B loaded with Escherichia coli Nissle 1917 bacterial ghosts on the HeLa cells by mitochondrial pathway of apoptosis. Drug Dev Ind Pharm 44:1328–35

    Article  CAS  PubMed  Google Scholar 

  • Zong R, Ruan H, Liu C, Fan S, Li J (2023) Bacteria and bacterial components as natural bio-nanocarriers for drug and gene delivery systems in cancer therapy. Pharmaceutics. https://doi.org/10.3390/pharmaceutics15102490

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This research received no external funding.

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Authors and Affiliations

  1. Division of Microbiology, Immunology and Biotechnology, Department of Natural Products and Alternative Medicine, Faculty of Pharmacy, University of Tabuk, Tabuk, 71491, Saudi Arabia

    Helal F. Hetta

  2. Department of Oral Medicine and Periodontology Diagnosis and Oral Radiology, Assiut Branch, Al- Azhar University, Assiut, 71524, Egypt

    Ibraheem M. Mwafey

  3. Department of Pharmaceutics, Faculty of Pharmacy, Assiut University, 71515, Assiut, Egypt

    Noura H. Abd Ellah

  4. Department of Pharmacology and Toxicology, Faculty of Pharmacy, University of Tabuk, Tabuk, 71491, Saudi Arabia

    Fawaz E. Alanazi

  5. Department of Microbiology and Immunology, Faculty of Pharmacy, Assiut University, Assiut, 71515, Egypt

    Yasmin N. Ramadan

Authors
  1. Helal F. Hetta
  2. Ibraheem M. Mwafey
  3. Noura H. Abd Ellah
  4. Fawaz E. Alanazi
  5. Yasmin N. Ramadan

Contributions

Conceptualization, H.F.H. and Y.N.R.; literature search, data analysis, curation, and visualization, H.F.H. and Y.N.R.; writing—original draft preparation, H.F.H. and Y.N.R.; writing—review and editing, H.F.H., I.M.M., N.H.A.E., F.E.A., and Y.N.R. All authors have read and agreed to the published version of the manuscript.

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Correspondence to Yasmin N. Ramadan.

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Hetta, H.F., Mwafey, I.M., Ellah, N.H.A. et al. Bacterial ghosts (BGs): A promising approach as candidate vaccine. World J Microbiol Biotechnol 42, 69 (2026). https://doi.org/10.1007/s11274-026-04783-7

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  • DOI: https://doi.org/10.1007/s11274-026-04783-7

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