Antibody Profiling of Borrelia burgdorferi Infection in Horses

ABSTRACT Infection with Borrelia burgdorferi is common in horses and ponies from the New England and mid-Atlantic regions of the United States. Here, we evaluated luciferase immunoprecipitation systems (LIPS) for profiling antibody responses against three different antigenic targets for the diagnosis of equine B. burgdorferi infection. LIPS testing of horse serum samples suspected of Lyme infection revealed that approximately 75% of the horse samples (114/159) were seropositive against the synthetic VOVO antigen, comprising repeated immunodominant C6 epitopes as well as OspC immunodominant epitopes. A comparison of VOVO and immunofluorescence assays (IFA) showed that 51% of the samples were positive in both assays (VOVO+/IFA+), 13% were VOVO−/IFA+, 21% were VOVO+/IFA−, and 15% were negative in both. To further understand humoral responses to B. burgdorferi and reconcile the diagnostic differences between IFA and VOVO, two additional B. burgdorferi LIPS tests were performed with DbpA and DbpB. Robust seropositive antibody responses against DbpA and/or DbpB were detected in 98% (79/81) of the VOVO+/IFA+ and 93% (50/54) of the discrepant samples. Additionally, some of the samples negative by both VOVO and IFA showed immunoreactivity against DbpA and/or DbpB. Overall, 94% of the suspected horse samples were seropositive by LIPS, and heat map analysis revealed that seropositive samples often were immunoreactive with at least two of the three antigens. These results suggest that LIPS tests employing multiple recombinant antigens offer a promising approach for the evaluation of antibody responses in Lyme disease.

[1]  M. Spetea,et al.  Introduction of a 6-cyano group in 14-oxygenated N-methylmorphinans influences in vitro and in vivo pharmacological activities , 2011, BMC Pharmacology.

[2]  M. Iadarola,et al.  Serological Studies Confirm the Novel Astrovirus HMOAstV-C as a Highly Prevalent Human Infectious Agent , 2011, PloS one.

[3]  M. Iadarola,et al.  Searching for biomarkers: humoral response profiling with luciferase immunoprecipitation systems , 2011, Expert review of proteomics.

[4]  S. Barthold,et al.  Lyme Neuroborreliosis in 2 Horses , 2011, Veterinary pathology.

[5]  R. Iyer,et al.  BBK07 Immunodominant Peptides as Serodiagnostic Markers of Lyme Disease , 2010, Clinical and Vaccine Immunology.

[6]  M. Iadarola,et al.  Distinct profiles of antibodies to Kaposi sarcoma-associated herpesvirus antigens in patients with Kaposi sarcoma, multicentric Castleman disease, and primary effusion lymphoma. , 2010, The Journal of infectious diseases.

[7]  B. L. Han,et al.  Antibody-profiling technologies for studying humoral responses to infectious agents , 2010, Expert review of vaccines.

[8]  M. Iadarola,et al.  Rapid, Simple, Quantitative, and Highly Sensitive Antibody Detection for Lyme Disease , 2010, Clinical and Vaccine Immunology.

[9]  A. Bojesen,et al.  Seroprevalence of Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum in Danish horses , 2010, Acta veterinaria Scandinavica.

[10]  A. Marques.  Lyme Disease: A Review , 2010, Current allergy and asthma reports.

[11]  S. Telford,et al.  Burden of Tick-borne Infections on American Companion Animals , 2009, Topics in Companion Animal Medicine.

[12]  M. Iadarola,et al.  Antibody profiling by Luciferase Immunoprecipitation Systems (LIPS). , 2009, Journal of visualized experiments : JoVE.

[13]  A. Coleman,et al.  BBK07, a Dominant In Vivo Antigen of Borrelia burgdorferi, Is a Potential Marker for Serodiagnosis of Lyme Disease , 2009, Clinical and Vaccine Immunology.

[14]  M. Iadarola,et al.  A Four-Antigen Mixture for Rapid Assessment of Onchocerca volvulus Infection , 2009, PLoS neglected tropical diseases.

[15]  M. Iadarola,et al.  Four-Antigen Mixture Containing V-Cyclin for Serological Screening of Human Herpesvirus 8 Infection , 2009, Clinical and Vaccine Immunology.

[16]  Pierre Baldi,et al.  A Genome-Wide Proteome Array Reveals a Limited Set of Immunogens in Natural Infections of Humans and White-Footed Mice with Borrelia burgdorferi , 2008, Infection and Immunity.

[17]  Yung-Fu Chang,et al.  Validation of an In-Clinic Enzyme-Linked Immunosorbent Assay Kit for Diagnosis of Borrelia Burgdorferi Infection in Horses , 2008, Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc.

[18]  Barbara J. B. Johnson,et al.  Dominant Epitopes of the C6 Diagnostic Peptide of Borrelia burgdorferi Are Largely Inaccessible to Antibody on the Parent VlsE Molecule , 2007, Clinical and Vaccine Immunology.

[19]  J. Kovacs,et al.  Rapid antibody quantification and generation of whole proteome antibody response profiles using LIPS (luciferase immunoprecipitation systems). , 2007, Biochemical and biophysical research communications.

[20]  H. Sarvas,et al.  Antibodies to recombinant decorin-binding proteins A and B in the cerebrospinal fluid of patients with Lyme neuroborreliosis , 2007, Scandinavian journal of infectious diseases.

[21]  S. Mak,et al.  Distribution and Characterization of Borrelia burgdorferi Isolates from Ixodes scapularis and Presence in Mammalian Hosts in Ontario, Canada , 2006, Journal of medical entomology.

[22]  F. Jongejan,et al.  Borrelia burgdorferi infections with special reference to horses. A review , 2005, The Veterinary quarterly.

[23]  J. Nowakowski,et al.  A Decline in C6 Antibody Titer Occurs in Successfully Treated Patients with Culture-Confirmed Early Localized or Early Disseminated Lyme Borreliosis , 2005, Clinical Diagnostic Laboratory Immunology.

[24]  A. Lauhio,et al.  Diagnosis of Lyme neuroborreliosis with antibodies to recombinant proteins DbpA, BBK32, and OspC, and VlsE IR6 peptide , 2003, Journal of Neurology.

[25]  A. Steere,et al.  Serodiagnosis of Lyme Disease by Kinetic Enzyme-Linked Immunosorbent Assay Using Recombinant VlsE1 or Peptide Antigens of Borrelia burgdorferi Compared with 2-Tiered Testing Using Whole-Cell Lysates , 2003, The Journal of infectious diseases.

[26]  D. Dykhuizen,et al.  Recombinant Chimeric Borrelia Proteins for Diagnosis of Lyme Disease , 2000, Journal of Clinical Microbiology.

[27]  E. Fikrig,et al.  Serologic Diagnosis of Lyme Borreliosis by Using Enzyme-Linked Immunosorbent Assays with Recombinant Antigens , 2000, Journal of Clinical Microbiology.

[28]  M. Ruscio,et al.  Evidence of Dbps (decorin binding proteins) among European strains of Borrelia burgdorferi sensu lato and in the immune response of LB patient sera. , 2000, FEMS microbiology letters.

[29]  A. Steere,et al.  Sensitive and Specific Serodiagnosis of Lyme Disease by Enzyme-Linked Immunosorbent Assay with a Peptide Based on an Immunodominant Conserved Region of Borrelia burgdorferi VlsE , 1999, Journal of Clinical Microbiology.

[30]  E. Fikrig,et al.  Serologic diagnosis of Lyme disease using recombinant outer surface proteins A and B and flagellin. , 1992, The Journal of infectious diseases.

[31]  A. Steere,et al.  2-tiered antibody testing for early and late Lyme disease using only an immunoglobulin G blot with the addition of a VlsE band as the second-tier test. , 2010, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[32]  M. Iadarola,et al.  Virology Journal BioMed Central Methodology , 2009 .

[33]  H. Saxén,et al.  Species-Specific Serodiagnosis of Lyme Arthritis and Neuroborreliosis Due to Borrelia burgdorferi Sensu Stricto, B. afzelii , and B. garinii by Using Decorin Binding Protein A , 2002 .