Heterogenität der kaninen Immunantworten gegen Borrelia burgdorferi in einem Line-Immunoassay mit rekombinantem VlsE und C6-Peptid

Doris Breu; Elisabeth Müller

doi:10.15654/TPK-170058

Tierärztliche Praxis Ausgabe K: Kleintiere / Heimtiere, Table of Contents

Tierarztl Prax Ausg K Kleintiere Heimtiere 2017; 45(05): 295-300
DOI: 10.15654/TPK-170058

Originalartikel

Schattauer GmbH

Heterogenität der kaninen Immunantworten gegen Borrelia burgdorferi in einem Line-Immunoassay mit rekombinantem VlsE und C₆-Peptid

Heterogeneity of canine immune responses to Borrelia burgdorferi in a line immunoassay comprising recombinant VlsE and C₆ peptide

Doris Breu

¹LABOKLIN GmbH & Co KG

,

Elisabeth Müller

¹LABOKLIN GmbH & Co KG

› Author Affiliations

Abstract

Zusammenfassung

Gegenstand: In einem Line-Immunoassay mit sieben Antigenen sowie rekombinantem VlsE (variable major protein like sequence, expressed) und synthetischem C₆-Peptid wurde die Präsenz Borreliaburgdorferi-spezifischer Antikörper (IgG) in kaninen Seren erfasst. Ziel der Studie war, die Antikörperantworten gegen das C₆-Peptid und das VlsE-Protein zu vergleichen und zu evaluieren, ob diese divergieren, da Antikörper gegen die jeweiligen Antigene als spezifisch für eine stattgefundene Infektion mit Borrelien gelten. Material und Methoden: Es wurden 1355 Blutproben von Hunden analysiert, bei denen Verdacht auf eine Infektion mit Borrelien bestand. Die Testung erfolgte mittels eines Line-Immunoassays (IgG) mit neun Antigenen. Ergebnisse: Insgesamt 64,4% der Proben wurden als negativ bewertet, 16,4% als infiziert und 17,4% zeigten ein für eine Impfung spezifisches Bandenmuster. Bei 1,2% der Hunde lag sowohl eine Impfung als auch eine Infektion vor. Die am häufigsten positiv getesteten Banden waren p100 (24,3%), p31/OspA (18,5%), C₆ (16,3%) und VlsE (13,9%). Bei 236 Hunden wurden Antikörper gegen das VlsE-Protein und/oder das C₆-Peptid nachgewiesen. Während 73,3% dieser Hunde übereinstimmende Ergebnisse zwischen VlsE und C₆ aufwiesen, unterschieden sich bei 26,7% die Resultate. Schlussfolgerung und klinische Relevanz: Die Testung mittels Line-Immunoassay erlaubt eine qualitative Analyse der unterschiedlichen IgG-Immunantworten gegen verschiedene Antigene. Die Resultate der Immunantworten gegen die als infektionsspezifisch geltenden Antigene VlsE und C₆ divergierten bei mehr als 26% der positiv getesteten Hunde unserer Population, sodass zum Ausschluss oder der Bestätigung einer Infektion mit Borrelia burgdorferi möglichst unterschiedliche infektionsspezifische Banden berücksichtigt werden sollten.

Summary

Objective: The study aimed to investigate the distribution of specific immune responses (IgG) to Borrelia burgdorferi using a line immunoassay with recombinant VlsE (variable major protein-like sequence, expressed) protein and synthetic C peptide among other antigens. We compared the immune responses to VlsE protein and C₆ peptide, because both antigens have been considered specific for a Borrelia infection. Materials and methods: A total of 1355 blood samples from dogs suspected of Borrelia infection were analysed. The line immunoassay employed nine antigens. Results: A total of 64.4% of all samples tested negative, 16.4% were positive for an infection and 17.4% were positive for vaccination. Band patterns specific for both infection and vaccination were observed in 1.2% of the dogs. The bands that most frequently tested positive were p100 (24.3%), p31/OspA (18.5%), C₆ (16.3%) and VlsE (13.9%). A total of 236 dogs (17.4% of the population) had antibodies to VlsE and/or C₆ peptide. In 73.3% of these dogs, results for VlsE and C₆ peptide were consistent, whereas this was not the case for 26.7% of these animals. Conclusion and clinical relevance: Testing using a line immunoassay allows for qualitative analyses of different immune responses to various antigens used as probes. In our study, > 26% of the dogs displayed discrepant results with regard to VlsE and C₆, the two antigens considered specific for Borrelia burgdorferi infection. To confirm or rule out infection, the results of several band patterns, thought to be specific for infection, need to be taken into consideration.

Schlüsselwörter

Hund - Infektionsmedizin - Labordiagnostik - Borrelien - Western Blot

Keywords

Dog - infectious medicine - laboratory diagnostics - Western blot

Full Text

References

Literatur
1 Barth C, Straubinger RK, Krupka I, Müller E, Sauter-Louis C, Hartmann K. Comparison of different diagnostic assays for the detection of Borrelia burgdorferi specific antibodies in dogs. Vet Clin Pathol 2014; 43 (04) 496-504.
2 Burgdorfer W, Barbour AG, Hayes SF, Benach JL, Grunwaldt E, Davies JP. Lyme disease – a tick-borne spirochetosis?. Science 1982; 216: 1317-1319.
3 Charon NW, Goldstein SF. Genetics of motility and chemotaxis of a fascinating group of bacteria: the spirochetes. Annu Rev Genet 2002; 36: 47-73.
4 Dressler F, Whalen JA, Reinhardt BN, Steere AC. Western blotting in the serodiagnosis of Lyme disease. J Infect Dis 1993; 167 (02) 392-400.
5 Embers ME, Jacobs MB, Johnson JB, Philipp MT. Dominant epitopes of the C6 diagnostic peptide of Borrelia burgdorferi are largely inaccessible to antibody on the parent VlsE molecule. Clin Vaccine Immunol 2007; 14: 931-936.
6 Engstrom SM, Shoop E, Johnson RC. Immunoblot interpretation criteria for serodiagnosis of early Lyme disease. J Clin Microbiol 1995; 33 (02) 419-427.
7 Gerber B, Haug K, Eichenberger S, Reusch CE, Wittenbrink MM. Comparison of a rapid immunoassay for antibodies to the C6 antigen with conventional tests for antibodies to Borrelia burgdorferi in dogs in Europe. Vet Rec 2009; 165 (20) 594-597.
8 Greene CE, Straubinger RK, Levy SA. Borreliosis. In: Infectious Diseases of the Dog and Cat. Greene CE. ed. St. Louis, Missouri: Elsevier; 2012: 447-465.
9 Greene RT, Walker RL, Nicholson WL, Heidner HW, Levine JF, Burgess EC, Wyand M, Breitschwerdt EB, Berkhoff HA. Immunoblot analysis of immunoglobulin G response to the Lyme disease agent (Borrelia burgdorferi) in experimentally and naturally exposed dogs. J Clin Microbiol 1988; 26: 648-653.
10 Hunfeld KP. Borrelien. In: Medizinische Mikrobiologie und Infektiologie. 8. Aufl.. Suerbaum S, Buchard GD, Kaufmann HE, Schulz TF. Hrsg. Berlin: Springer; 2016: 367-374.
11 Indest KH, Howell JK, Jacobs MB, Scholl-Meeker D, Norris SJ, Philipp MT. Analysis of Borrelia burgdorferi vlsE gene expression and recombination in the tick vector. Infect Immun 2001; 69: 7083-7090.
12 Jacek E, Tang KS, Komorowski L, Ajamian M, Probst C, Stevenson B, Wormser GP, Marques AR, Alaedini A. Epitope-specific evolution of human B cell responses to Borrelia burgdorferi VlsE protein from early to late stages of Lyme disease. J Immunol 2016; 196 (03) 1036-1043.
13 Kenedy MR, Lenhardt TR, Akins DR. The role of Borrelia burgdorferi outer surface proteins. FEMS Immunol Med Microbiol 2012; 66 (01) 1-19.
14 Krupka I, Pantchev N, Lorentzen L, Weise M, Straubinger RK. Durch Zecken übertragbare bakterielle Infektionen bei Hunden. Seroprävalenzen von Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato und Ehrlichia canis in Deutschland. Prakt Tierarzt 2007; 88 (10) 776-788.
15 Krupka I, Knauer J, Lorentzen L, O’Connor TP, Saucier J, Straubinger RK. Borrelia burgdorferi sensu lato species in Europe induce diverse immune responses against C6 peptides in infected mice. Clin Vaccine Immunol 2009; 16 (11) 1546-1562.
16 Krupka I, Straubinger RK. Lyme borreliosis in dogs and cats: background, diagnosis, treatment and prevention of infections with Borrelia burgdorferi sensu stricto. Vet Clin Small Anim 2010; 40: 1103-1119.
17 Levy SA, Magnarelli LA. Relationship between development of antibodies to Borrelia burgdorferi in dogs and the subsequent development of limb/joint Borreliosis. J Am Vet Med Assoc 1992; 200 (03) 344-347.
18 Levy SA, O’Connor TP, Hanscom JL, Shields P, Lorentzen L, DiMarco AA. Quantitative measurement of C6 antibody following antibiotic treatment of Borrelia burgdorferi antibody-positive nonclinical dogs. Clin Vaccine Immunol 2008; 15 (01) 115-119.
19 Liang FT, Alvarez AL, Gu Yan, Nowling JM, Ramamoorthy R, Philipp MT. An immunodominant conserved region within the variable domain of VlsE, the variable surface antigen of Borrelia burgdorferi . J Immunol 1999; 163 (10) 5566-5573.
20 Liang FT, Steere AC, Marques AR, Johnson BJ, Miller JN, Philipp MT. 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. J Clin Microbiol 1999; 37 (12) 3990-3996.
21 Liang FT, Aberer E, Cinco M, Gern L, Hu CM, Lobet YN, Ruscio M, Voet PE, Weynant VE, Philipp MT. Antigenic conservation of an immunodominant invariable region of the VlsE lipoprotein among European pathogenic genospecies of Borrelia burgdorferi SL. J Infect Dis 2000; 182 (05) 1455-1462.
22 Liang FT, Jacobsen RH, Straubinger RK, Grooters A, Philipp MT. Characterization of a Borrelia burgdorferi VlsE invariable region useful in canine Lyme disease serodiagnosis by enzyme-linked immunosorbent assay. J Clin Microbiol 2000; 38 (11) 4160-4266.
23 Lin T, Gao L, Edmondson DG, Jacobs MB, Philipp MT, Norris SJ. Central role of the Holliday junction helicase RuvAB in vlsE recombination and infectivity of Borrelia burgdorferi . PLoS Pathog 2009; 5 (12) e1000679.
24 Littmann MP, Goldstein RE, Labato MA, Lappin MR, Moore GE. ACVIM small animal consensus statement on Lyme disease in dogs: diagnosis, treatment, and prevention. J Vet Intern Med 2006; 20: 422-434.
25 Motaleb A, Liu J, Wooten RM. Spirochetal motility and chemotaxis in the natural enzootic cycle and development of Lyme disease. Curr Opin Microbiol 2015; 28: 106-113.
26 O‘Connor TP, Esty KJ, Hanscom JL, Shields P, Philipp MT. Dogs vaccinated with common Lyme disease vaccines do not respond to IR₆, the conserved immunodominant region of the VlsE surface protein of Borrelia burgdorferi . Clin Diagn Lab Immunol 2004; 11 (03) 458-462.
27 Pantchev N, Pluta S, Huisinga E, Nather S, Scheufelen M, Globokar Vrhovec M, Schweinitz A, Hampel H, Straubinger RK. Tick-borne diseases (borreliosis, anaplasmosis, babesiosis) in German and Austrian dogs: status quo and review of distribution, transmission, clinical findings, diagnosis and prophylaxis. Parasitol Res 2015; 114 (Suppl. 01) 19-54.
28 Philipp MT, Bowers LC, Fawcett PT, Jacobs MB, Liang FT, Marques AR, Mitchell PD, Purcell JE, Ratterree MS, Straubinger RK. Antibody response to IR₆, a conserved immunodominant region of the VlsE lipoprotein, wanes rapidly after antibiotic treatment of Borrelia burgdorferi infection in experimental animals and in humans. J Infect Dis 2001; 184 (07) 870-878.
29 Purser JE, Norris SJ. Correlation between plasmid content and infectivity in Borrelia burgdorferi . Proc Natl Acad Sci USA 2000; 97 (25) 13865-13870.
30 Rauter C, Hartung T. Prevalence of Borrelia burgdorferi sensu lato genospecies in Ixodes ricinus ticks in Europe: a metaanalysis. Appl Environ Microbiol 2005; 71 (11) 7203-7216.
31 Seinost G, Golde WT, Berger BW, Dunn JJ, Qiu D, Dunkin DS, Dykhuizen DE, Luft BJ, Dattwyler RJ. Infection with multiple strains of Borrelia burgdorferi sensu stricto in patients with Lyme disease. Arch Dermatol 1999; 135 (11) 1329-33.
32 Sillanpää H, Lahdenne P, Sarvas H, Arnez M, Steere A, Peltomaa M, Seppälä I. Immune responses to borrelial VlsE IR6 peptide variants. Int J Med Microbiol 2007; 297 (01) 45-52.
33 Tijsse-Klasen E, Pandak N, Hengeveld P, Takumi K, Koopmans M, Sprong H. Ability to cause erythema migrans differs between Borrelia burgdorferi sensu lato isolates. Parasit Vectors 2013; 6 (23) 1-8.
34 Tjernberg I, Sillanpää H, Seppälä I, Elisasson I, Forsberg P, Lahdenne P. Antibody responses to Borrelia IR₆ peptide variants and the C6 peptide in Swedish patients with erythema migrans. Int J Med Microbiol 2009; 299 (06) 439-446.
35 Töpfer KH, Straubinger RK. Characterization of the humoral immune response in dogs after vaccination against the Lyme Borreliosis agent. A study with five commercial vaccines using two different vaccination schedules. Vaccine 2007; 25: 314-326.
36 Wagner B, Freer H, Rollins A, Garcia-Tapia D, Erb Hn, Earnhardt C, Marconi R, Meeus P. Antibodies to Borrelia burgdorferi OspA, OspC, OspF, and C6 antigens as markers for early and late infection in dogs. Clin Vaccine Immunol 2012; 19 (04) 527-535.
37 Wang G, van Dam AP, Schwartz I, Dankert J. Molecular typing of Borrelia burgdorferi sensu lato: taxonomic, epidemiological, and clinical implications. Clin Microbiol Rev 1999; 633-653.
38 Wilske B, Fingerle V, Schulte-Spechtel U. Microbiological and serological diagnosis of Lyme borreliosis. FEMS Immunol Med Microbiol 2007; 49: 13-21.
39 Zhang JR, Norris SJ. Kinetics and in vivo induction of genetic variation of VlsE in Borrelia burgdorferi . Infect Immun 1998; 3689-3697.
40 Zhang JR, Hardham JM, Barbour AG, Norris SJ. Antigenetic variation in Lyme disease Borreliae by promiscuous recombination of VMP-like sequence cassettes. Cell 1997; 89: 275-285.