An Enzyme-Linked Immunosorbent Spot Assay Measuring Borrelia burgdorferi B31-Specific Interferon Gamma-Secreting T Cells Cannot Discriminate Active Lyme Neuroborreliosis from Past Lyme Borreliosis: a Prospective Study in the Netherlands.

T Van Gorkom,S U C Sankatsing,W Voet,D W Notermans,M Salomons,A W J Bossink,J J M Bouwman,B J M Vlaminckx,K Kremer,S F T Thijsen,R H Muilwijk,D M Ismail,Nathan A Ledeboer

doi:10.1128/jcm.01695-17

Abstract

ABSTRACTTwo-tier serology testing is most frequently used for the diagnosis of Lyme borreliosis (LB); however, a positive result is no proof of active disease. To establish a diagnosis of active LB, better diagnostics are needed. Tests investigating the cellular immune system are available, but studies evaluating the utility of these tests on well-defined patient populations are lacking. Therefore, we investigated the utility of an enzyme-linked immunosorbent spot (ELISpot) assay to diagnose active Lyme neuroborreliosis. Peripheral blood mononuclear cells (PBMCs) of various study groups were stimulated by using Borrelia burgdorferi strain B31 and various recombinant antigens, and subsequently, the number of Borrelia-specific interferon gamma (IFN-γ)-secreting T cells was measured. We included 33 active and 37 treated Lyme neuroborreliosis patients, 28 healthy individuals treated for an early manifestation of LB in the past, and 145 untreated healthy individuals. The median numbers of B. burgdorferi B31-specific IFN-γ-secreting T cells/2.5 × 105 PBMCs did not differ between active Lyme neuroborreliosis patients (6.0; interquartile range [IQR], 0.5 to 14.0), treated Lyme neuroborreliosis patients (4.5; IQR, 2.0 to 18.6), and treated healthy individuals (7.4; IQR, 2.3 to 14.9) (P = 1.000); however, the median number of B. burgdorferi B31-specific IFN-γ-secreting T cells/2.5 × 105 PBMCs among untreated healthy individuals was lower (2.0; IQR, 0.5 to 3.9) (P ≤ 0.016). We conclude that the Borrelia ELISpot assay, measuring the number of B. burgdorferi B31-specific IFN-γ-secreting T cells/2.5 × 105 PBMCs, correlates with exposure to the Borrelia bacterium but cannot be used for the diagnosis of active Lyme neuroborreliosis.

Highlights

Two-tier serology testing is most frequently used for the diagnosis of Lyme borreliosis (LB); a positive result is no proof of active disease
In this study, we used well-defined patient populations and healthy controls to evaluate the utility of the Borrelia enzyme-linked immunosorbent spot (ELISpot) assay
We found that the number of B. burgdorferi B31-specific IFN-␥-secreting T cells/2.5 ϫ 105 Peripheral blood mononuclear cells (PBMCs) in peripheral blood was significantly elevated in active Lyme neuroborreliosis patients, treated Lyme neuroborreliosis patients, and healthy individuals treated for early manifestations of LB in the past compared to untreated healthy individuals (P Յ 0.016)

Summary

Introduction

Two-tier serology testing is most frequently used for the diagnosis of Lyme borreliosis (LB); a positive result is no proof of active disease. Peripheral blood mononuclear cells (PBMCs) of various study groups were stimulated by using Borrelia burgdorferi strain B31 and various recombinant antigens, and subsequently, the number of Borrelia-specific interferon gamma (IFN-␥)-secreting T cells was measured. We conclude that the Borrelia ELISpot assay, measuring the number of B. burgdorferi B31-specific IFN-␥-secreting T cells/2.5 ϫ 105 PBMCs, correlates with exposure to the Borrelia bacterium but cannot be used for the diagnosis of active Lyme neuroborreliosis. A study among general practitioners (GPs) found a threefold increase of patients reporting tick bites and diagnoses of erythema migrans (EM), an early, localized skin rash, in the period between 1994 and 2009 [1]. The diagnosis of Lyme neuroborreliosis is based on clinical symptoms and needs to be supported by laboratory tests. Active Lyme neuroborreliosis patients were used as a proxy for active disease

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Conclusion