The Long-Term Persistence of Borrelia burgdorferi Antigens and DNA in the Tissues of a Patient with Lyme Disease

Abstract

Whether Borrelia burgdorferi, the causative agent of Lyme disease, can persist for long periods in the human body has been a controversial question. The objective of this study was to see if we could find B. burgdorferi in a Lyme disease patient after a long clinical course and after long-term antibiotic treatment. Therefore, we investigated the potential presence of B. burgdorferi antigens and DNA in human autopsy tissues from a well-documented serum-, PCR-, and culture-positive Lyme disease patient, a 53-year-old female from northern Westchester County in the lower Hudson Valley Region of New York State, who had received extensive antibiotic treatments during extensive antibiotic treatments over the course of her 16-year-long illness. We also asked what form the organism might take, with special interest in the recently found antibiotic-resistant aggregate form, biofilm. We also examined the host tissues for the presence of inflammatory markers such as CD3+ T lymphocytes. Autopsy tissue sections of the brain, heart, kidney, and liver were analyzed by histological and immunohistochemical methods (IHC), confocal microscopy, fluorescent in situ hybridization (FISH), polymerase chain reaction (PCR), and whole-genome sequencing (WGS)/metagenomics. We found significant pathological changes, including borrelial spirochetal clusters, in all of the organs using IHC combined with confocal microscopy. The aggregates contained a well-established biofilm marker, alginate, on their surfaces, suggesting they are true biofilm. We found B. burgdorferi DNA by FISH, polymerase chain reaction (PCR), and an independent verification by WGS/metagenomics, which resulted in the detection of B. burgdorferi sensu stricto specific DNA sequences. IHC analyses showed significant numbers of infiltrating CD3+ T lymphocytes present next to B. burgdorferi biofilms. In summary, we provide several lines of evidence that suggest that B. burgdorferi can persist in the human body, not only in the spirochetal but also in the antibiotic-resistant biofilm form, even after long-term antibiotic treatment. The presence of infiltrating lymphocytes in the vicinity of B. burgdorferi biofilms suggests that the organism in biofilm form might trigger chronic inflammation.

Lyme Disease Frontiers: Reconciling Borrelia Biology and Clinical Conundrums

“…In the case of Borrelia, the existence of antibiotic persister cells has been demonstrated by numerous groups, both in vitro and in vivo with animal models and human patients [262][263][264][265][266]. Notably, this persister fraction has been found to tolerate high concentrations of antimicrobials that exceed levels considered clinically achievable [262].…”

“…Although these may not fall under a strict definition of pleomorphism, which tends to focus on the individual cell, they have likewise been implicated in disease, and we include them in briefly in this discussion for illustrative and comparative purposes. Borrelial clusters are routinely observed in cultures of type strains, and community structures have also been reported histologically in human biopsy and autopsy specimens [266,323]. Microbiologically, biofilms are defined as a structure or matrix composed of extracellular polymeric substances (EPS), housing a host of microbial cells, that adheres to a static surface (biotic or abiotic) [324].…”

Detection and Transstadial Passage of Babesia Species and Borrelia burgdorferi Sensu Lato in Ticks Collected from Avian and Mammalian Hosts in Canada

“…Lyme disease is a zoonosis with multisystemic clinical manifestations in humans. Bbsl is pleomorphic with diverse forms (i.e., spirochetes, spherocytes, blebs, granules) and, collectively, as dormant biofilms [9,12,[112][113][114]. Lyme disease spirochetes have an affinity for immune privileged sites, and side-step the immune response, and lodge in niche reservoirs including bone [115], brain [116][117][118], eye [119], muscle [120], collagenous tissues (ligaments, tendons) [121,122], glial and neuronal cells [123][124][125], and fibroblasts/scar tissue [126].…”

“…Lyme disease spirochetes have an affinity for immune privileged sites, and side-step the immune response, and lodge in niche reservoirs including bone [115], brain [116][117][118], eye [119], muscle [120], collagenous tissues (ligaments, tendons) [121,122], glial and neuronal cells [123][124][125], and fibroblasts/scar tissue [126]. Left untreated or inadequately treated, this insidious spirochetosis can be persistent [9][10][11]113,116,[127][128][129], and develop into chronic Lyme disease [12,13]. Often, patients advance to chronic Lyme disease before they get diagnosed and treated.…”

“…Bbsl is normally carried by ixodid ticks; however, this spirochete has other means of transmission. Bbsl is pleomorphic and has diverse forms, and can evade the immune response, and become persistent [9][10][11][12]. If this complex, multisystem zoonosis is not recognized and treated early, it can develop into chronic Lyme disease [12,13].…”