Laboratory of Microbial Immunochemistry and Vaccines

Biological chemistry of macromolecules involved in immunological processes – immunochemical studies of bacterial endotoxins. Structural analysis of lipopolysaccharides, polysaccharides and glycoconjugates using NMR spectroscopy and mass spectrometry in combination with chemical and biochemical methods. Investigation of the relationship between the structure of bacterial endotoxins and their biological activities/physical properties.

Studies of glycoconjugate structures and ligand-receptor interactions using high-resolution NMR spectroscopy (HR-MAS NMR, STD-NMR), mass spectrometry (LC-MS, ESI-MS/MSn, MALDI-TOF MS) combined with SPR and FACS. Developments of neoglycoconjugates as antibacterial and antiendotoxin vaccines – synthesis, immunochemical properties, immune response and protective activity.

Laboratory Head

Head: prof. dr hab. Jolanta Łukasiewicz (professor, 0000-0001-8081-7261)

Jolanta Lukasiewicz studied biotechnology and earned a Master of Sciences degree in engineering and biotechnology (1997) at the Wroclaw University of Science and Technology. In 2001 she received a doctoral degree in biological science the thesis entitled: „Structure and biological activity of Plesiomonas shigelloides CNCTC 113/92 lipopolysaccharide (LPS)”. Her expertise covers the structural analysis of bacterial carbohydrate antigens and LPS in particular, comprising studies on LPS structure and biological activity relationships (immunogenicity, toxicity, immune system activation, vaccine development). Among her  major scientific achievements  are: (i) identification of a covalent linkage between enterobacterial common antigen (ECA) and LPS in Shigella sonnei; (ii) characteristics of sepsis-relevant interactions between human complement system (MBL and ficolin-3) with LPS of Gram-negative bacteria; (iii) Identification of new structural modifications that determine diversity of Klebsiella pneumoniae O-antigens relevant for development of prophylactic and therapeutic strategies against infections.

Team

Professor:

Associate Professors:

Assistant Professors:

Technical Staff:

  • Emilia Strąk, MSc, lab technicians

PhD Students:

Most important scientific achievements
  • Identification of new LPS structures and modifications with the emphasise on conservative motifs and modifications for diagnostics and therapeutic strategies development against nosocomial infections caused by Gram-negative bacteria, pertussis, and travellers’ diarrhoea.
  • Development of neoglycoconjugates as new vaccine components or diagnostic tools and novel  therapeutic strategies against infections caused by Klebsiella pneumoniae, Bordetella spp., Shigella sonnei, Escherichia coli and other enteric bacteria.
  • The first evidence for a covalent linkage between enterobacterial common antigen (ECA) and LPS in Shigella sonnei phase II (ECALPS) complemented with development of ECA-LPS-based vaccine against enteric bacteria. Chemotyping of bacterial surface antigens – applicability of HR-MAS NMR for the structural analysis of O-antigens in situ, directly on bacteria.
  • Demonstration of the broad cross-reactivity between human mannan-binding lectin (MBL) of complement system with LPS core regions and  possible implications for the onset and   development of sepsis.
Research methods
  • structural analysis of glycans, glycolipids, polysaccharides and oligosaccharides
  • NMR spectroscopy
  • MALDI-TOF and ESI mass spectrometry
  • Immunochemistry and glycoconjugates synthesis and characteristics
  • microbiology
  • chromatography (LC, HPLC, LC-MS, affinity chromatography)
  • surface plasmon resonance (SPR, Biacore T200)
  • fluorescent labelling of bacterial antigens in situ (FACS)
Key equipment

Most important projects (during the last 10 years)
  • „Nontypeable Klebsiella pneumoniae O antigens – structure and seroepidemiology”, NCN, OPUS 16, 2018/31/B/NZ7/04002, 2019-2022. PI: prof. Jolanta Lukasiewicz
  • „Development of monoclonal antibody-based passive immune therapy and companion diagnostics for severe Klebsiella infections – KLEBSICURE”, European Commission project EUROSTARS,  No. E!7563, 2012-2015, the consortium of: Arsanis Biosciences GmbH, Vienna, Austria (Leader), GATC Biotech AG, Konstanz, Germany (participant), Max Planck Institute for Infection Biology, Berlin, Germany (participant), and Hirszfeld Institute, Wroclaw, Poland (participant). PI for polish tasks: prof. Jolanta Lukasiewicz
  • „Influence of glycosylation of the SARS-CoV-2 Spike protein on complement activation”, NCN, OPUS20, 2020/39/B/NZ6/01660, 2021-2024, The consortium: Institute for Medical Biology (Leader, prof. Maciej Cedzynski), HIIET PAS, Medical University of Warsaw. PI: Prof. dr hab. Maciej Cedzyński. Co-Pis of HIIET PAS: prof. Marcin Czerwiński, prof. Jolanta Lukasiewicz, Dariusz Martynowski, PhD.
  • „Host-pathogen interplay: bacterial outer membrane vesicles as target for complement system.”, NCN, OPUS 16,  2018/31/B/NZ6/03514, 2019-2022. The consortium: Institute for Medical Biology (Leader, prof. Maciej Cedzynski), HIIET PAS, University of Silesia. PI: prof. Anna Świerzko. Co-PI in HIIET PAS: prof. Jolanta Lukasiewicz
  • „Research on innovative vaccine against SARS-CoV-2 virus causing COVID-19 disease”, NCBiR, Wsparcie Szpitali Jednoimiennych 28/2020, 2020-2021. PI: Prof. Andrzej Gamian. Co-PI: prof. Marcin Czerwiński, prof. Jolanta Lukasiewicz et al.
  • „Structural modifications of bacterial endotoxins (lipopolysaccharides) of significant importance to future antibacterial therapeutic strategies. NCN, OPUS8, 2014/15/B/NZ6/03203, 201502019, PI: prof. Jolanta Lukasiewicz
  • „Development of recombinant Hsp60, OMP 40 and LPS-derived subunit vaccines for cattle and pigs “, NCBiR, PBS, PBS3/A8/33/2015, 2015–2018, The consortium: the Wrocław University of Environmental and Life Sciences (Lider, prof. Tadeusz Stefaniak), HIIET PAS, Pure Biologics, IŻ PIB Żerniki Wielkie Sp. z o.o., Genetic Dairy Breeding”Głogówek” Sp. z o.o. Co-PI of HIIET PAS: Wojciech Jachymek, PhD.
  • „New generation of anti-bacterial vaccines: preparation, immunochemical properties and protective activities” commissioned and financed by WCB EIT+ as a part of the project “BioMed – Biotechnologies and advanced medical technologies”. Innovative Economy Operational Programme, No. POIG.01.01.02-02-003/08-00, 2009-2014, PI: Lugowski C.
  • “Molecular basis for interaction between endotoxins and ficolin H and its biological effects investigation”. MSHE, No. N401267339, 2010-2013, PI: prof. Jolanta Lukasiewicz
  • „Bacterial lyses products as potential anticancer preparations. Biochemical characteristics and biological activities analyses. Devising a production engineering of active component”. NCBR, No. NR13008906, 2010-2014, PI: prof. J. Boratynski.
Selected publications
  • Koj S, Ucieklak K, Lugowski C, Niedziela T. Structure and Immunogenicity of the Bordetella pertussis LOS-Derived Oligosaccharides in the Endosomal-Like Pre-Processing Mice Model. Vaccines (Basel). 2021, 9(6):645. https://pubmed.ncbi.nlm.nih.gov/34199173/
  • Gozdziewicz TK, Maciejewska A, Tsybulska A, Lugowski C, Lukasiewicz J. A New Look at the Enterobacterial Common Antigen Forms Obtained during Rough Lipopolysaccharides Purification. Int J Mol Sci. 2021, 22(2):701. https://pubmed.ncbi.nlm.nih.gov/33445776/
  • Ucieklak K, Koj S, Niedziela T. Bordetella holmesii Lipopolysaccharide Hide and Seek Game with Pertussis: Structural Analysis of the O-Specific Polysaccharide and the Core Oligosaccharide of the Type Strain ATCC 51541. International Journal of Molecular Sciences. 2020; 21(17):6433. https://pubmed.ncbi.nlm.nih.gov/32899371/
  • Maciejewska A, Kaszowska M, Jachymek W, Lugowski C, Lukasiewicz J. Lipopolysaccharide-Linked Enterobacterial Common Antigen (ECALPS) Occurs in Rough Strains of Escherichia coli R1, R2, and R4. Int J Mol Sci. 2020, 21;21:6038; https://pubmed.ncbi.nlm.nih.gov/32839412/
  • Artyszuk D, Izdebski R, Maciejewska A, Kaszowska M, Herud A, Szijártó V, Gniadkowski M, Lukasiewicz J. The Impact of Insertion Sequences on O-Serotype Phenotype and Its O-Locus-Based Prediction in Klebsiella pneumoniae O2 and O1. Int J Mol Sci. 2020, 21:E6572; https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7556023/
  • Rollenske T, Szijarto V, Lukasiewicz J, Guachalla LM, Stojkovic K, Hartl K, Stulik L, Kocher S, Lasitschka F, Al-Saeedi M, Schröder-Braunstein J, von Frankenberg M, Gaebelein G, Hoffmann P, Klein S, Heeg K, Nagy E, Nagy G, Wardemann H. Cross-specificity of protective human antibodies against Klebsiella pneumoniae LPS O-antigen. Nature Immunology, 2018;19:617-624; https://pubmed.ncbi.nlm.nih.gov/29760533/
  • Guachalla LM, Stojkovic K, Hartl K, Kaszowska M, Kumar Y, Wahl B, Paprotka T, Nagy E, Lukasiewicz J, Nagy G, Szijártó V. Discovery of monoclonal antibodies cross-reactive to novel subserotypes of K. pneumoniae O3. Scientific Reports, 2017, 7(1):6635; https://pubmed.ncbi.nlm.nih.gov/28747785/
  • Szijártó V, Guachalla LM, Hartl K, Varga C, Banerjee P, Stojkovic K, Kaszowska M, Nagy E, Lukasiewicz J, Nagy G. Both clades of the epidemic KPC-producing Klebsiella pneumoniae clone ST258 share a modified galactan O-antigen type. International Journal of Medical Microbiology, 2016, 306(2):89-98. https://pubmed.ncbi.nlm.nih.gov/26723873/
  • Man-Kupisinska A, Michalski M, Maciejewska A, Swierzko AS, Cedzynski M, Lugowski C, Lukasiewicz J. A new ligand-based method for purifying active human plasma-derived ficolin-3 complexes supports the phenomenon of crosstalk between pattern-recognition molecules and immunoglobulins. PLoS One, 2016, 11(5):e0156691. https://pubmed.ncbi.nlm.nih.gov/27232184/
  • Gozdziewicz TK, Lugowski C, Lukasiewicz J. First evidence for a covalent linkage between enterobacterial common antigen and lipopolysaccharide in Shigella sonnei phase II ECA-LPS. Journal of Biological Chemistry, 2014, 289:2745-2754; https://pubmed.ncbi.nlm.nih.gov/24324266/
  • Kaszowska M, Jachymek W, Lukasiewicz J, Niedziela T, Kenne L, Lugowski C. The unique structure of complete lipopolysaccharide isolated from semi-rough Plesiomonas shigelloides O37 (strain CNCTC 39/89) containing (2S)-O-(4-oxopentanoic acid)-α-D-Glcp (α-D-lenose). Carbohydrate Research, 2013, 378:98-107. https://pubmed.ncbi.nlm.nih.gov/23711248/
  • Swierzko A, Lukasiewicz J, Cedzynski M, Maciejewska A, Jachymek W, Niedziela T, Matsushita M, Lugowski C. New functional ligands for ficolin-3 among lipopolysaccharides of Hafnia alvei. Glycobiology, 2012, 22:267-80; https://pubmed.ncbi.nlm.nih.gov/21890891/
  • Niedziela T, Letowska I, Lukasiewicz J, Kaszowska M, Czarnecka A, Kenne L, Lugowski C. Epitope of the vaccine-type Bordetella pertussis strain 186 lipooligosaccharide and antiendotoxin activity of antibodies directed against the terminal pentasaccharide-tetanus toxoid conjugate. Infect Immun. 2005, 73(11):7381-9. https://pubmed.ncbi.nlm.nih.gov/16239537/
  • Jachymek W, Niedziela T, Petersson C, Lugowski C, Czaja J, Kenne L. Structures of the O-specific polysaccharides from Yokenella regensburgei (Koserella trabulsii) strains PCM 2476, 2477, 2478, and 2494: high-resolution magic-angle spinning NMR investigation of the O-specific polysaccharides in native lipopolysaccharides and directly on the surface of living bacteria. Biochemistry, 1999, 38:11788-95; https://pubmed.ncbi.nlm.nih.gov/10512635/