Recombinant Burkholderia thailandensis Translocator protein BipB (bipB)

Basic Research Questions

• What is Burkholderia thailandensis Translocator protein BipB and what is its role in bacterial pathogenesis?

  BipB is a translocator protein component of the Bsa type III secretion system (T3SS) found in Burkholderia thailandensis. As a full-length protein (1-624 amino acids), BipB functions as part of the molecular machinery that enables bacteria to inject effector proteins directly into host cells .

  BipB shares approximately 46% amino acid identity with the Salmonella translocator protein SipB, and is secreted in a bsaZ-dependent manner . In the context of pathogenesis, BipB is likely involved in forming pores in host cell membranes that allow for the translocation of bacterial effector proteins. This process is critical for bacterial invasion, intracellular survival, and immune evasion.

  While B. thailandensis is less pathogenic than its close relatives B. pseudomallei and B. mallei, studying BipB in this species provides valuable insights into virulence mechanisms across the Burkholderia genus, particularly as B. thailandensis serves as a safer model organism for laboratory research .

• How is Recombinant BipB protein typically produced and purified for research purposes?

  Recombinant BipB is typically produced through heterologous expression in E. coli systems. The general methodology involves:

  1. Cloning: The bipB gene (BTH_II0841) is amplified from B. thailandensis genomic DNA and cloned into an expression vector with an N-terminal His-tag .

  2. Expression: The recombinant plasmid is transformed into an E. coli expression strain, and protein production is induced under optimized conditions .

  3. Purification: The protein is purified using affinity chromatography, typically with Ni-NTA resin that binds the His-tag .

  4. Quality control: The purified protein undergoes SDS-PAGE analysis to confirm purity (typically >90%) .

  5. Storage: The protein is stored in Tris/PBS-based buffer with 6% trehalose at pH 8.0, often with the addition of glycerol (up to 50%) for long-term storage at -20°C or -80°C .

  For reconstitution, the lyophilized protein is typically dissolved in deionized sterile water to a concentration of 0.1-1.0 mg/mL, with glycerol added for stability .

• What sequence conservation exists between BipB proteins across different Burkholderia species?

  BipB proteins demonstrate high sequence conservation among Burkholderia pseudomallei complex (Bpc) species, but less conservation with other bacterial species. Key conservation patterns include:

  Species Comparison	Sequence Identity	Notes
  Among B. pseudomallei isolates	≥98%	High conservation within species
  Between B. pseudomallei and B. mallei	≥98%	Extremely high conservation
  Between B. pseudomallei and B. thailandensis	85-93%	Moderate to high conservation
  BipB and Salmonella SipB	~46%	Functional homology across genera

  This high degree of conservation among B. pseudomallei and B. mallei isolates suggests that BipB plays a critical role in these organisms, and that antibodies or vaccines targeting BipB might be effective across multiple Burkholderia strains . The lower conservation with B. thailandensis reflects the evolutionary divergence between these species, though the proteins likely maintain similar functional properties .

• What are the structural characteristics of the BipB protein?

  The BipB protein from B. thailandensis has several key structural features:

  • Full protein length: 624 amino acids

  • Domains: BipB likely contains transmembrane domains that facilitate its role in forming membrane pores, similar to its homolog SipB .

  • Secondary structure: The protein likely contains alpha-helical regions involved in membrane insertion and pore formation, based on structural similarities with other T3SS translocator proteins .

  • Functional regions: The C-terminal region is likely involved in insertion into host cell membranes, while the N-terminal domain may interact with other components of the secretion apparatus .

  Detailed structural studies using X-ray crystallography or cryo-electron microscopy would provide more precise information about the three-dimensional structure of BipB, which would be valuable for understanding its mechanism of action.

• How does BipB function in the type III secretion system?

  BipB functions as a translocator protein in the Bsa type III secretion system (T3SS) of Burkholderia, with several key roles:

  1. Pore formation: As a translocator, BipB likely participates in forming a pore in the host cell membrane, creating a conduit for the delivery of bacterial effector proteins .

  2. Secretion regulation: In B. pseudomallei, the T3SS appears to be regulated by several factors including BsaP and translocon proteins like BipD. Mutants lacking BipD were observed to hypersecrete the effector protein BopE, suggesting a role in controlling the timing and magnitude of effector protein secretion .

  3. Pathogenesis contribution: BipB likely contributes to virulence by facilitating the injection of bacterial effector proteins into host cells, which can modulate host cell functions such as cytoskeletal organization and immune responses .

  4. Species-specific regulation: In B. thailandensis, the expression of T3SS components including BipB may be regulated differently than in B. pseudomallei. The presence of the arabinose assimilation operon in B. thailandensis causes down-regulation of T3SS-3 genes, which may account for some of the differential virulence between these species .

  The function of BipB must be studied in the context of the entire T3SS, as its activity depends on interactions with other T3SS components and is regulated by complex signaling mechanisms.

Advanced Research Questions

Recombinant Burkholderia thailandensis Translocator Protein BipB (bipB): Research FAQs

This comprehensive FAQ collection provides researchers with evidence-based answers about Recombinant Burkholderia thailandensis Translocator protein BipB (bipB). The content addresses fundamental concepts and advanced research applications, focusing on methodological approaches essential for scientific investigation of this important bacterial protein.

Basic Research Questions

• What is Burkholderia thailandensis Translocator protein BipB and what is its role in bacterial pathogenesis?

  BipB is a translocator protein component of the Bsa type III secretion system (T3SS) in Burkholderia thailandensis. This 624-amino acid protein functions as part of the molecular machinery that enables bacteria to inject effector proteins directly into host cells .

  BipB shares approximately 46% amino acid identity with the Salmonella translocator protein SipB, and is secreted in a bsaZ-dependent manner . In the context of pathogenesis, BipB likely participates in forming pores in host cell membranes that allow for the translocation of bacterial effector proteins, which is critical for bacterial invasion, intracellular survival, and immune evasion.

  While B. thailandensis is less pathogenic than its close relatives B. pseudomallei (the causative agent of melioidosis) and B. mallei (the causative agent of glanders), studying BipB in this species provides valuable insights into virulence mechanisms while offering a safer model organism for laboratory research .

• How is Recombinant BipB protein typically produced and purified for research purposes?

  Recombinant BipB is typically produced through heterologous expression in E. coli systems. The methodology involves:

  1. Cloning: The bipB gene (BTH_II0841) is amplified from B. thailandensis genomic DNA and cloned into an expression vector with an N-terminal His-tag .

  2. Expression: The recombinant plasmid is transformed into an E. coli expression strain, and protein production is induced under optimized conditions .

  3. Purification: The protein is purified using affinity chromatography with Ni-NTA resin that binds the His-tag, followed by additional purification steps if needed .

  4. Quality control: The purified protein undergoes SDS-PAGE analysis to confirm purity (typically >90%) .

  5. Storage: The protein is stored in Tris/PBS-based buffer with 6% trehalose at pH 8.0, often with glycerol (up to 50%) for long-term storage at -20°C or -80°C .

  For reconstitution, the lyophilized protein is typically dissolved in deionized sterile water to a concentration of 0.1-1.0 mg/mL, with glycerol added for stability. Repeated freeze-thaw cycles should be avoided to maintain protein integrity .

• What sequence conservation exists between BipB proteins across different Burkholderia species?

  BipB proteins demonstrate high sequence conservation within the Burkholderia pseudomallei complex (Bpc) species, which has important implications for research and vaccine development:

  Species Comparison	Sequence Identity	Notes
  Among B. pseudomallei isolates	≥98%	High conservation within species
  Between B. pseudomallei and B. mallei	≥98%	Extremely high conservation
  Between B. pseudomallei and B. thailandensis	85-93%	Moderate to high conservation
  BipB and Salmonella SipB	~46%	Functional homology across genera

  This high degree of conservation among B. pseudomallei and B. mallei isolates suggests that BipB plays a critical role in these organisms. The conservation pattern indicates that antibodies or vaccines targeting BipB might be effective across multiple Burkholderia strains . The lower conservation with B. thailandensis reflects the evolutionary divergence between these species, though the proteins likely maintain similar functional properties .

• What experimental approaches can help determine the immunogenicity of BipB protein?

  Several methodological approaches can be used to evaluate BipB's immunogenicity:

  1. Antibody response measurement:

     • ELISA assays can detect anti-BipB antibodies in patient sera or in experimental animals immunized with recombinant BipB .

     • Western blot analysis to confirm antibody specificity and reactivity with native protein.

     • Studies have shown that human melioidosis patients develop antibodies to BipB, though diagnostic ELISAs had low accuracy in endemic regions, possibly due to previous exposure to B. pseudomallei .

  2. T-cell activation assays:

     • Isolation of peripheral blood mononuclear cells (PBMCs) from previously exposed individuals.

     • Stimulation of cells with recombinant BipB and measurement of T-cell activation markers.

     • Quantification of cytokine production, particularly IFN-γ from CD4+ T cells and granzyme B from CD8+ T cells .

     • Analysis of T-cell proliferation in response to BipB stimulation.

  3. Dendritic cell presentation studies:

     • Generation of monocyte-derived dendritic cells (moDCs) from human donors.

     • Pulsing moDCs with recombinant BipB protein.

     • Co-culture with autologous T cells to assess antigen presentation.

     • Studies with related Burkholderia proteins have shown that antigen-pulsed moDCs can trigger T-cell responses from seropositive donors .

  4. In vivo immunization studies:

     • Immunization of mice with recombinant BipB using various adjuvants.

     • Analysis of antibody titers, isotype distribution, and T-cell responses.

     • Challenge studies to assess protection against bacterial infection.

     • Prior studies with related T3SS proteins (BipB, BipC, BipD) did not show protection when used as single-antigen vaccines, suggesting combination approaches may be needed .

  These approaches provide complementary data on how the immune system recognizes and responds to BipB, informing both diagnostic and vaccine development efforts.

• How do researchers confirm the functional activity of recombinant BipB protein?

  Confirming the functional activity of recombinant BipB requires multiple approaches:

  1. Structural integrity assessment:

     • Circular dichroism spectroscopy to verify proper secondary structure.

     • Size exclusion chromatography to confirm monomeric state or appropriate oligomerization.

     • Limited proteolysis to assess domain folding and stability.

  2. Membrane interaction studies:

     • Liposome binding assays to measure association with lipid membranes.

     • Fluorescence-based assays to detect membrane insertion.

     • Liposome leakage assays to assess pore-forming activity.

  3. Heterologous expression systems:

     • Expression in engineered bacterial strains (similar to the approach used for BipD in Shigella) .

     • Complementation of BipB-deficient mutants to restore type III secretion function.

     • Immunofluorescence microscopy to verify incorporation into secretion apparatus structures.

  4. Host cell interaction assays:

     • Red blood cell hemolysis assays to measure pore formation.

     • Cell permeabilization assays using fluorescent dyes.

     • Electrophysiology to characterize pore properties when inserted into membranes.

  5. Effector protein translocation:

     • Measurement of effector protein delivery in the presence or absence of functional BipB.

     • Reporter assays to quantify translocation efficiency.

  Researchers should note that BipB may not be detectable under standard laboratory growth conditions, as studies with B. pseudomallei and B. mallei found that BipB was not expressed at detectable levels under several test conditions, in contrast to its homologs in Shigella and Salmonella .

Advanced Research Questions

• What molecular mechanisms govern BipB's role in type III secretion system assembly and function?

  The molecular mechanisms of BipB function within the T3SS involve several sophisticated interactions:

  1. Secretion signal recognition:

     • BipB contains N-terminal secretion signals recognized by the T3SS apparatus.

     • The secretion mechanism is dependent on the ATPase complex that powers the T3SS.

     • Experimental approach: Site-directed mutagenesis of potential secretion signals followed by secretion assays to identify critical residues.

  2. Assembly pathway:

     • BipB likely follows a hierarchical assembly pathway, requiring proper formation of the needle complex.

     • The timing of BipB secretion is regulated to ensure correct assembly sequence.

     • Experimental approach: Time-course studies of T3SS assembly using immunofluorescence or electron microscopy with antibodies against BipB and other components.

  3. Molecular triggers for insertion:

     • BipB insertion into host membranes may require specific environmental signals or host contact.

     • pH changes or calcium concentration may serve as molecular triggers.

     • Experimental approach: In vitro assays with purified BipB under varying conditions to assess conformational changes and membrane insertion.

  4. Regulatory interactions:

     • In B. pseudomallei, the T3SS is regulated by several factors including BsaP and BipD, with mutants lacking BipD hypersecreeting the effector protein BopE .

     • This suggests a role in controlling the timing and magnitude of effector protein secretion.

     • Experimental approach: Protein-protein interaction studies to identify regulatory partners and signaling cascades affecting BipB function.

  5. Conformational changes:

     • BipB likely undergoes significant conformational changes during the transition from secretion to membrane insertion.

     • These changes expose hydrophobic regions for membrane interaction.

     • Experimental approach: Structural studies using hydrogen-deuterium exchange mass spectrometry to identify dynamic regions of the protein.

  Understanding these mechanisms requires integrated biochemical, structural, and cellular approaches, with careful attention to the native context of the T3SS apparatus.

• How can recombinant antibody technology be applied to study BipB function and develop therapeutics?

  Recombinant antibody technology offers several powerful approaches for studying BipB and developing potential therapeutics:

  1. Development of specific recombinant monoclonal antibodies (R-mAbs):

     • Cloning of heavy and light chain variable regions from hybridomas producing anti-BipB antibodies .

     • Expression using dual promoter plasmids that allow efficient formation of intact IgG molecules .

     • Creating a panel of antibodies targeting different epitopes can provide tools for mapping functional domains.

  2. Engineering antibody fragments for enhanced utility:

     • Production of single-chain variable fragments (ScFvs) by fusing VH and VL regions into a single polypeptide .

     • Generation of Fab fragments that retain antigen binding but have improved tissue penetration.

     • Development of nanobodies (nAbs) based on camelid single-domain antibodies for their small size and stability .

  3. Advanced antibody engineering approaches:

     • Development of bispecific antibodies targeting both BipB and another bacterial component or host receptor .

     • Creation of antibody-drug conjugates for targeted delivery of antimicrobials.

     • Engineering antibodies with enhanced Fc-mediated effector functions to promote bacterial clearance.

  4. Therapeutic applications:

     • Passive immunization strategies using neutralizing antibodies against BipB.

     • Combination antibody cocktails targeting multiple T3SS components.

     • Development of antibody-based diagnostics for detecting Burkholderia infections.

  5. Research applications:

     • Using antibodies to track BipB localization during infection.

     • Blocking specific domains to delineate their functions.

     • Immunoprecipitation to identify interaction partners.

  The advantages of recombinant antibodies include unambiguous identification via DNA sequencing, reliable expression, ease of distribution as DNA sequences or plasmids, and opportunities for engineering to enhance utility . These characteristics make them valuable reagents for both basic research and therapeutic development.

• What strategies can optimize the expression and purification of stable, functional recombinant BipB for structural studies?

  Optimizing expression and purification of stable, functional BipB requires addressing several technical challenges:

  1. Expression system selection:

     Expression System	Advantages	Optimization Strategies
     E. coli	- Fast growth - High yields	- Use specialized strains (BL21, Rosetta, SHuffle) - Lower induction temperature (16-20°C) - Co-express chaperones
     Insect cells	- Better folding - Suitable for membrane proteins	- Optimize MOI and harvest time - Test multiple cell lines (Sf9, Hi5) - Screen various viral promoters
     Cell-free	- Rapid screening - Direct incorporation of labeled amino acids	- Add lipid nanodiscs for membrane proteins - Optimize redox conditions - Supplement with chaperones

  2. Construct design optimization:

     • Test multiple truncation constructs targeting specific domains

     • Incorporate solubility-enhancing fusion tags (MBP, SUMO, thioredoxin)

     • Design constructs avoiding predicted disordered regions

     • Consider surface entropy reduction mutations for crystallization

     • Create chimeric constructs with structurally characterized homologs

  3. Purification strategy refinement:

     • Implement multi-step purification (IMAC, ion exchange, size exclusion)

     • Optimize buffer conditions (pH, salt concentration, additives)

     • Consider on-column refolding for inclusion body purification

     • Use detergent screening for membrane-interacting regions

     • Apply nanodiscs or amphipols to stabilize hydrophobic domains

  4. Stability enhancement:

     • Add stabilizing agents (glycerol, trehalose, arginine)

     • Identify and remove protease-sensitive sites

     • Introduce disulfide bonds to stabilize tertiary structure

     • Test thermostabilizing mutations

     • Screen chemical chaperones and stabilizing ligands

  5. Functional validation methods:

     • Develop liposome binding and pore formation assays

     • Compare recombinant protein to native protein using conformation-specific antibodies

     • Use circular dichroism and thermal shift assays to assess folding

     • Verify activity in complementation assays with bipB-deficient bacteria

  This systematic approach addresses the specific challenges of BipB as a membrane-active protein while providing multiple options for optimization based on experimental outcomes and research goals.

• How can genomic and comparative sequence analysis of BipB inform research on Burkholderia pathogenesis and evolution?

  Genomic and comparative sequence analysis of BipB provides valuable insights into Burkholderia pathogenesis and evolution:

  1. Evolutionary relationship mapping:

     • The high sequence conservation (≥98% identity) between B. pseudomallei and B. mallei BipB proteins reflects their close evolutionary relationship .

     • The moderate conservation (85-93% identity) with B. thailandensis BipB indicates evolutionary divergence that may correlate with differences in virulence .

     • Phylogenetic analysis of BipB sequences can help reconstruct the evolutionary history of the Burkholderia genus.

  2. Functional domain identification:

     • Comparative analysis with homologs like Salmonella SipB (46% identity) helps identify conserved functional domains .

     • Regions under purifying selection (highly conserved) likely represent essential functional domains.

     • Regions under positive selection may indicate host-adaptation signatures.

  3. Host-pathogen co-evolution analysis:

     • Comparing BipB sequences from isolates from different hosts or geographical regions.

     • Identifying selective pressures acting on specific domains that may interact with host factors.

     • Correlating sequence variations with host specificity or tissue tropism.

  4. Virulence prediction models:

     • Developing sequence-based algorithms to predict virulence potential based on BipB sequences.

     • Identifying specific sequence motifs associated with enhanced pathogenicity.

     • Creating databases of BipB sequence variants correlated with clinical outcomes.

  5. Methodological approaches:

     • Multiple sequence alignment using MUSCLE or CLUSTAL algorithms

     • Selection pressure analysis using PAML or HyPhy

     • Structural prediction using homology modeling and ab initio approaches

     • Molecular dynamics simulations to assess the impact of sequence variations

     • Population genetics analysis to study BipB evolution within Burkholderia species

  This comprehensive analysis can guide experimental studies by identifying key regions for mutagenesis, informing vaccine design, and providing insights into the evolutionary trajectories of Burkholderia virulence mechanisms.

• What are the key differences between BipB from B. thailandensis and its homologs in more pathogenic Burkholderia species?

  The differences between B. thailandensis BipB and its homologs in more pathogenic species reveal important aspects of virulence evolution:

  1. Sequence variations:

     • B. thailandensis BipB shares 85-93% amino acid identity with B. pseudomallei BipB .

     • These sequence differences may contribute to the reduced virulence of B. thailandensis compared to B. pseudomallei and B. mallei.

     • Specific amino acid substitutions might affect interactions with host cell components or other T3SS proteins.

  2. Expression regulation differences:

     • The presence of the arabinose assimilation operon in B. thailandensis causes down-regulation of T3SS-3 genes, which may account for some of the differential virulence between B. thailandensis and B. pseudomallei .

     • The expression of BipB in B. thailandensis might be regulated differently in response to environmental cues compared to pathogenic species.

     • Quorum sensing systems, which differ between species, may differentially regulate BipB expression .

  3. Functional differences:

     • While the core function as a translocator is likely conserved, subtle differences may affect efficiency of pore formation or host cell recognition.

     • Differences in post-translational modifications could affect protein stability or interactions.

     • The timing and magnitude of BipB secretion might differ between species, affecting virulence.

  4. Structural implications:

     • Amino acid substitutions may cause conformational differences that affect function.

     • Changes in surface-exposed residues could alter interactions with host immune system components.

     • Differences in membrane-interacting domains might affect tropism for specific host cell types.

  5. Methodological approaches to study these differences:

     • Complementation studies exchanging bipB genes between species

     • Chimeric protein analysis to identify domains responsible for functional differences

     • Site-directed mutagenesis to convert key residues between species

     • Comparative secretion assays under various environmental conditions

     • Differential host cell interaction studies comparing BipB from multiple species

  Understanding these differences provides insights into how B. thailandensis maintains a T3SS with reduced pathogenicity, making it a valuable model organism for studying Burkholderia biology with lower biosafety requirements.

• How can CRISPR-Cas9 technology be applied to study BipB function in Burkholderia infection models?

  CRISPR-Cas9 technology offers powerful approaches for studying BipB function:

  1. Genetic modification of Burkholderia:

     • Generation of precise bipB deletion mutants without polar effects on adjacent genes

     • Introduction of point mutations to study specific functional domains

     • Creation of fluorescently tagged BipB variants for live imaging

     • Engineering of inducible bipB expression systems

     • Methodological consideration: Burkholderia species can be engineered through natural transformation of PCR fragments, which can be combined with CRISPR-Cas9 strategies

  2. Domain function analysis:

     • Systematically target different BipB domains to determine their roles

     • Create truncation libraries to map minimal functional regions

     • Engineer domain swaps between BipB and homologs from other species

     • Introduce epitope tags at different positions to monitor protein processing

     • Methodological approach: Design multiple guide RNAs targeting various regions and screen for functional phenotypes

  3. Host cell factor identification:

     • CRISPR screens in host cells to identify factors required for BipB-mediated effects

     • Knockout of potential host receptors or membrane components

     • Disruption of signaling pathways potentially affected by BipB

     • Modification of host cytoskeletal components to assess their role in BipB function

     • Methodological consideration: Pooled CRISPR screens followed by infection and cell sorting can identify host factors involved in BipB-mediated processes

  4. Regulatory network mapping:

     • Target transcriptional regulators controlling BipB expression

     • Disrupt quorum sensing components that influence T3SS expression

     • Modify two-component systems involved in virulence regulation

     • Methodological approach: CRISPRi (CRISPR interference) can be used for partial repression to study essential genes

  5. In vivo infection models:

     • Generate BipB-modified strains for animal infection studies

     • Create reporter strains to monitor BipB expression during infection

     • Develop tissue-specific knockout models to study host response to BipB

     • Methodological consideration: Different CRISPR delivery systems (plasmid-based, bacteriophage, or direct RNP delivery) may be required depending on the model system

  These approaches require careful consideration of biosafety regulations, particularly when working with more pathogenic Burkholderia species, which may require BSL-3 containment .

• What methodologies are most effective for studying the interactions between BipB and the host immune system?

  Several sophisticated methodologies can effectively investigate BipB-immune system interactions:

  1. Advanced cellular immunology techniques:

     • Single-cell RNA sequencing of immune cells exposed to recombinant BipB

     • Mass cytometry (CyTOF) analysis of multiple immune cell activation markers

     • Multiplexed cytokine profiling using Luminex or similar platforms

     • Real-time imaging of immune cell interactions with BipB-expressing bacteria

     • Methodological consideration: Compare responses between naïve individuals and those with prior Burkholderia exposure to identify memory responses

  2. Epitope mapping approaches:

     • Peptide scanning arrays to identify linear B-cell epitopes

     • HLA-binding prediction algorithms followed by validation with T-cell assays

     • Phage display to identify conformational epitopes

     • X-ray crystallography of antibody-BipB complexes

     • Methodological consideration: Studies have shown that melioidosis patients develop antibodies to BipB, providing a source of convalescent sera for epitope mapping

  3. Host-pathogen interaction analysis:

     • Affinity purification-mass spectrometry to identify BipB binding partners

     • Protein microarrays to screen for interactions with host proteins

     • CRISPR screens to identify host factors required for BipB recognition

     • Biolayer interferometry or surface plasmon resonance to measure binding kinetics

     • Methodological consideration: Controls with related proteins from non-pathogenic bacteria can help identify pathogen-specific interactions

  4. In vivo immune response characterization:

     • Humanized mouse models expressing human immune components

     • In vivo imaging of neutrophil recruitment and activity during infection

     • Adoptive transfer experiments to identify protective immune cell populations

     • Tissue-specific immune profiling during infection with wild-type vs. bipB mutants

     • Methodological consideration: Different infection routes (intravenous, intranasal, subcutaneous) may elicit different immune responses

  5. Immunomodulation studies:

     • Testing if BipB actively suppresses specific immune pathways

     • Examining effects on inflammasome activation and IL-1β/IL-18 processing

     • Investigating potential interference with antigen presentation pathways

     • Assessing impact on phagocyte function and killing mechanisms

     • Methodological consideration: T3SS proteins from related bacteria can induce caspase-1 dependent macrophage death , suggesting BipB might have similar immunomodulatory effects

  These methodologies provide complementary approaches to understand how BipB interacts with the immune system during infection, with implications for both pathogenesis and vaccine development.