Comparative Analysis of the Properties and Ligand Binding Characteristics of CspZ, a Factor H Binding Protein, Derived from Borrelia burgdorferi Isolates of Human Origin

ABSTRACT Borrelia burgdorferi CspZ (BBH06/BbCRASP-2) binds the complement regulatory protein factor H (FH) and additional unidentified serum proteins. The goals of this study were to assess the ligand binding capability of CspZ orthologs derived from an extensive panel of human Lyme disease isolates and to further define the molecular basis of the interaction between FH and CspZ. While most B. burgdorferi CspZ orthologs analyzed bound FH, specific, naturally occurring polymorphisms, most of which clustered in a specific loop domain of CspZ, prevented FH binding in some orthologs. Sequence analyses also revealed the existence of CspZ phyletic groups that correlate with FH binding and with the relationships inferred from ribosomal spacer types (RSTs). CspZ type 1 (RST1) and type 3 (RST3) strains bind FH, while CspZ type 2 (RST2) strains do not. Antibody responses to CspZ were also assessed. Anti-CspZ antibodies were detected in mice by week 2 of infection, indicating that there was expression during early-stage infection. Analyses of sera collected from infected mice suggested that CspZ production continued over the course of long-term infection as the antibody titer increased over time. While antibody to CspZ was detected in several human Lyme disease serum samples, the response was not universal, and the titers were generally low. Vaccination studies with mice demonstrated that while CspZ is immunogenic, it does not elicit an antibody that is protective or that inhibits dissemination. The data presented here provide significant new insight into the interaction between CspZ and FH and suggest that there is a correlation between CspZ production and dissemination. However, in spite of its possible contributory role in pathogenesis, the immunological analyses indicated that CspZ is likely to have limited potential as a diagnostic marker and vaccine candidate for Lyme disease.

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Borrelia burgdorferi Complement Regulator-Acquiring Surface Protein 1 of the Lyme Disease Spirochetes Is Expressed in Humans and Induces Antibody Responses Restricted to Nondenatured Structural Determinants

Infection and Immunity ◽

10.1128/iai.01028-06 ◽

2006 ◽

Vol 74 (12) ◽

pp. 7024-7028 ◽

Cited By ~ 18

Author(s):

Evelyn Rossmann ◽

Veronique Kitiratschky ◽

Heidelore Hofmann ◽

Peter Kraiczy ◽

Markus M. Simon ◽

...

Keyword(s):

Lyme Disease ◽

Borrelia Burgdorferi ◽

Surface Protein ◽

Factor H ◽

Antibody Responses ◽

Dominant Factor ◽

Serum Samples ◽

Structural Determinants ◽

Pathogen Persistence ◽

Complement Regulator

ABSTRACT Borrelia burgdorferi complement regulator-acquiring surface protein 1 (CRASP-1), the dominant factor H and FHL-1-binding protein of the Lyme disease spirochete B. burgdorferi, is implicated in pathogen persistence and was recently reported to be nonimmunogenic in humans. Here we show that serum samples from Lyme disease patients contain antibodies with exclusive specificity for nondenatured structural determinants of CRASP-1.

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Delineation of Species-Specific Binding Properties of the CspZ Protein (BBH06) of Lyme Disease Spirochetes: Evidence for New Contributions to the Pathogenesis of Borrelia spp.

Infection and Immunity ◽

10.1128/iai.00850-07 ◽

2007 ◽

Vol 75 (11) ◽

pp. 5272-5281 ◽

Cited By ~ 34

Author(s):

Elizabeth A. Rogers ◽

Richard T. Marconi

Keyword(s):

Lyme Disease ◽

Ligand Binding ◽

Dna Sequence ◽

Molecular Basis ◽

Serum Proteins ◽

Specific Binding ◽

Coiled Coil ◽

Factor H ◽

Site Directed Mutagenesis ◽

Binding Properties

ABSTRACT Borrelia burgdorferi CspZ (TIGR open reading frame designation, BBH06) is part of a functionally related group of proteins that bind one or more members of the factor H (FH) protein family. In this report we assess the conservation, distribution, properties, and ligand binding abilities of CspZ from the three main Borrelia species associated with Lyme disease infections in humans. CspZ (also referred to as BbCRASP-2 in the literature) was found to be highly conserved at the intraspecies level but divergent at the interspecies level. All CspZ orthologs that originated from B. burgdorferi isolates bound FH from a diverse group of mammals. In contrast, CspZ derived from B. garinii and B. afzelii did not. Regardless of the Borrelia species of origin, all CspZ proteins tested bound to unknown ∼60-kDa serum proteins produced by different mammals. To further define the molecular basis for the differential binding of CspZ orthologs to host proteins, DNA sequence, truncation, and site-directed mutagenesis analyses were performed. DNA sequence analyses revealed that B. garinii and B. afzelii CspZ orthologs possess a 64-amino-acid N-terminal domain that is absent from B. burgdorferi CspZ. However, binding analyses of recombinant proteins revealed that this domain does not in and of itself influence ligand binding properties. Truncation and mutagenesis analyses further revealed that the key determinants required for ligand binding are discontinuous and that the presentation of the ligand binding pocket is dependent on alpha helices with high coiled-coil formation probability. The data presented here provide insight into the molecular basis of CspZ-ligand interactions and suggest that CspZ orthologs from diverse Borrelia species can contribute to the host-pathogen interaction through their interaction with serum proteins.

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Comprehensive Analysis of the Factor H Binding Capabilities of Borrelia Species Associated with Lyme Disease: Delineation of Two Distinct Classes of Factor H Binding Proteins

Infection and Immunity ◽

10.1128/iai.71.6.3597-3602.2003 ◽

2003 ◽

Vol 71 (6) ◽

pp. 3597-3602 ◽

Cited By ~ 96

Author(s):

John V. McDowell ◽

Jill Wolfgang ◽

Emily Tran ◽

Michael S. Metts ◽

Duncan Hamilton ◽

...

Keyword(s):

Lyme Disease ◽

Clinical Course ◽

Regulatory Protein ◽

Comprehensive Analysis ◽

Factor H ◽

Protein Profiles ◽

Complement Regulatory Protein ◽

Binding Ability ◽

Protein Factor ◽

Affinity Ligand

ABSTRACT Some Lyme disease spirochete isolates can bind complement regulatory protein factor H (fH), a process that may allow evasion of complement-mediated killing. Here we demonstrate significant differences in the fH binding capabilities of species of the Borrelia burgdorferi sensu lato complex. The percentages of B. burgdorferi, B. afzelii, and B. garinii bacteria that bound fH in either enzyme-linked immunosorbent assays or affinity ligand binding immunoblot assays were 100, 83, and 29%, respectively. The fH binding protein profiles were examined and found to exhibit variability among isolates and to form two distinct classes. Differences in fH binding ability may contribute to the differences in pathogenesis and clinical course observed upon infection with different species of the B. burgdorferi sensu lato complex.

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Selective Binding of Borrelia burgdorferi OspE Paralogs to Factor H and Serum Proteins from Diverse Animals: Possible Expansion of the Role of OspE in Lyme Disease Pathogenesis

Infection and Immunity ◽

10.1128/iai.74.3.1967-1972.2006 ◽

2006 ◽

Vol 74 (3) ◽

pp. 1967-1972 ◽

Cited By ~ 54

Author(s):

Kelley M. Hovis ◽

Emily Tran ◽

Christina M. Sundy ◽

Eric Buckles ◽

John V. McDowell ◽

...

Keyword(s):

Lyme Disease ◽

Borrelia Burgdorferi ◽

Host Range ◽

Serum Proteins ◽

Factor H ◽

Disease Pathogenesis ◽

Selective Binding

ABSTRACT The binding of Borrelia burgdorferi OspE, OspF, and family 163 (Elp) proteins to factor H/factor H-like protein 1 (FHL-1) and other serum proteins from different animals was assessed. OspE paralogs bound factor H and unidentified serum proteins from a subset of animals, while OspF and Elp proteins did not. These data advance our understanding of factor H binding, the host range of the Lyme spirochetes, and the expanding role of OspE in pathogenesis.

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Faculty Opinions recommendation of West Nile virus nonstructural protein NS1 inhibits complement activation by binding the regulatory protein factor H.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1055710.507623 ◽

2006 ◽

Author(s):

Rino Rappuoli

Keyword(s):

West Nile Virus ◽

Complement Activation ◽

Nonstructural Protein ◽

Regulatory Protein ◽

Factor H ◽

West Nile ◽

Protein Factor

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Complement Regulatory Protein Factor H Is a Soluble Prion Receptor That Potentiates Peripheral Prion Pathogenesis

The Journal of Immunology ◽

10.4049/jimmunol.1701100 ◽

2017 ◽

Vol 199 (11) ◽

pp. 3821-3827 ◽

Cited By ~ 4

Author(s):

Sarah J. Kane ◽

Taylor K. Farley ◽

Elizabeth O. Gordon ◽

Joshua Estep ◽

Heather R. Bender ◽

...

Keyword(s):

Regulatory Protein ◽

Factor H ◽

Complement Regulatory Protein ◽

Protein Factor

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Serum Protein Profiling Reveals a Landscape of Inflammation and Immune Signaling in Early-stage COVID-19 Infection

Molecular & Cellular Proteomics ◽

10.1074/mcp.rp120.002128 ◽

2020 ◽

Vol 19 (11) ◽

pp. 1749-1759 ◽

Cited By ~ 3

Author(s):

Xin Hou ◽

Xiaomei Zhang ◽

Xian Wu ◽

Minya Lu ◽

Dan Wang ◽

...

Keyword(s):

Serum Proteins ◽

Early Stage ◽

Protein Profiling ◽

Cytokine Signaling ◽

Large Set ◽

Serum Samples ◽

Antibody Microarray ◽

Proteomics Analysis ◽

Immune Signaling ◽

Anti Inflammation

Coronavirus disease 2019 (COVID-19) is a highly contagious infection and threating the human lives in the world. The elevation of cytokines in blood is crucial to induce cytokine storm and immunosuppression in the transition of severity in COVID-19 patients. However, the comprehensive changes of serum proteins in COVID-19 patients throughout the SARS-CoV-2 infection is unknown. In this work, we developed a high-density antibody microarray and performed an in-depth proteomics analysis of serum samples collected from early COVID-19 (n = 15) and influenza (n = 13) patients. We identified a large set of differentially expressed proteins (n = 132) that participate in a landscape of inflammation and immune signaling related to the SARS-CoV-2 infection. Furthermore, the significant correlations of neutrophil and lymphocyte with the CCL2 and CXCL10 mediated cytokine signaling pathways was identified. These information are valuable for the understanding of COVID-19 pathogenesis, identification of biomarkers and development of the optimal anti-inflammation therapy.

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The Borrelia burgdorferi 37-Kilodalton Immunoblot Band (P37) Used in Serodiagnosis of Early Lyme Disease Is the flaA Gene Product

Journal of Clinical Microbiology ◽

10.1128/jcm.37.3.548-552.1999 ◽

1999 ◽

Vol 37 (3) ◽

pp. 548-552 ◽

Cited By ~ 15

Author(s):

Robert D. Gilmore ◽

Rendi L. Murphree ◽

Angela M. James ◽

Sarah A. Sullivan ◽

Barbara J. B. Johnson

Keyword(s):

Lyme Disease ◽

Borrelia Burgdorferi ◽

Genomic Library ◽

Disease Patient ◽

Humoral Response ◽

Immunoglobulin M ◽

Leader Peptide ◽

Serum Samples ◽

Present Generation ◽

Periplasmic Flagella

The 37-kDa protein (P37) of Borrelia burgdorferi is an antigen that elicits an early immunoglobulin M (IgM) antibody response in Lyme disease patients. The P37 gene was cloned from aB. burgdorferi genomic library by screening with antibody from a Lyme disease patient who had developed a prominent humoral response to the P37 antigen. DNA sequence analysis of this clone revealed the identity of P37 to be FlaA, an outer sheath protein of the periplasmic flagella. Recombinant P37 expression was accomplished inEscherichia coli by using a gene construct with the leader peptide deleted and fused to a 38-kDa E. coli protein. The recombinant antigen was reactive in IgM immunoblots using serum samples from patients clinically diagnosed with early Lyme disease that had been scored positive for B. burgdorferi anti-P37 reactivity. Lyme disease patient samples serologically negative for theB. burgdorferi P37 protein did not react with the recombinant. Recombinant P37 may be a useful component of a set of defined antigens for the serodiagnosis of early Lyme disease. This protein can be utilized as a marker in diagnostic immunoblots, aiding in the standardization of the present generation of IgM serologic tests.

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