Serum antibodies against Bovrelia afzelii, Borrelia burgdorferi sensu stricto and the 41‐kiloDalton flagellin in patients from a Lyme borreliosis endemic area: analysis by EIA and immunoblot

Abstract

Nilsson, I., Anderson von Rosen, I . Serum antibodies against Borreliu ufzelii, Borreliu hurgdoyfiri sensu stricto and the 41-kiloDalton flagellin in patients from a Lyme borreliosis endemic area: Analysis by EIA and immunoblot. APMIS 104: 907-914, 1996. To evaluate the possible importance of antigenic heterogeneity in the serological diagnosis of Lyme borreliosis a study was performed using antigens from various Lyme Borreliu strains. Serum samples from 102 patients with clinical signs of the infection, all living in an endemic area in southern Sweden, were evaluated by four enzyme immuno assays (EIA). The sera were initially tested for the immunoglobulin G response to antigens from a local Borreliu ufxlii strain (ACAI). Serum samples from healthy blood donors residing in the same region were used to define seropositivity in the ACAI-EIA. Immunoblotting was performed with the ACAI antigen and the reactive bands were analysed. A serum was defined as positive when at least four of the Borreliu specific polypeptides (OspC, OspA, OspB, p39, p41 [flagellin], p83, p94, I 10kDa) were stained. The same sera were then analysed in three other IgG enzyme immunoassays, one based on antigens from Borreliu burgclorjeri sensu stricto B3 1, and another on pooled protein fractions from strains B31 and ACAI. In the third EIA, sera were analysed for antiflagellin reactivity (B. ufzelii strain DK-1). An inconstant immune response was demonstrated in the EIAs and the seropositivity varied between 30-47'%1 when low positive values were excluded, and between 38-73'%1 if all values were included. Fifty sera (50/102) met the criteria for a positive immunoblot, but positive immunoblots were detected with both low positive and negative sera independent of antigen used in the EIAs. Antigens of the local B. ufzelii strain were found to detect a higher number of seropositive individuals, which suggests that the antibody reactivity to Lyme Borreliu increases when antigens from a strain endemic in a particular geographical region are used. Data from this study suggest that EIA alone seems insufficient for the serodiagnosis, and antigenic heterogeneity of Lyme Borreliu spp. influences the performance of serum antibody tests. The reliability of serological assays could be increased when the serum antibodies against antigens of Borrdiu spp. predominant in the local geographical region are measured. hurgdorferi sensu lato (9). The clinical diagnosis is often difficult, owing to the diffuse and non-

Characterization of Lyme Borreliosis Isolates from Patients with Erythema Migrans and Neuroborreliosis in Southern Sweden

“…Use of local LB strains, together with information on the seroreactivity in the population to be tested, should be taken into account when developing diagnostic serological assays for clinical diagnosis (13,36).…”

Positive serology for Lyme disease Borrelias in primary cutaneous B-cell lymphoma: a study in 22 patients; is it a fortuitous finding?

“…22,23 The relation of two cases of acrodermatitis chronica atrophicans to Borrelia afzelii could not be proved on the differential immunoblot, but as much as 20 per cent of cases could be caused by either Borrelia garinii or Borrelia burgdorferi sensu stricto. 25 On the other hand, when antigens of the local strains of Borrelia afzelii are used, a higher number of seropositive individuals is detected; 26 this again con®rms that antibody reactivity to Lyme disease Borrelia increases when antigens from endemic strains are used.…”

Propensity to excessive proinflammatory response in chronic Lyme borreliosis

“…Whole sonificated B. afzelii antigens were prepared as follows. B. afzelii (ACA1 strain isolated from a Swedish acrodermatitis chronicum atrophicans patient (27)) was cultivated in Barbour‐Stoenner‐Kelly medium II with 10% rabbit sera (Sigma, St. Louis, MO) at 33 °C. A whole‐cell lysate of the spirochetes was prepared by washing the organisms in pyrogen‐free phosphate‐buffered saline (PBS) twice, sonicating the pellet on ice until intact organisms were no longer visible, and determining protein content using the Bradford method.…”