Published Research

The following articles published in peer-reviewed journals resulted from research projects funded in whole or in part by Global Lyme Alliance.

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Feng J, Zhang S, Shi W, Zhang Y. (2017) Activity of sulfa drugs and their combinations against stationary phase B. burgdorferi in vitro. Antibiotics 6:pii:E10. Doi: 10.3390/antibiotics6010010.

Synopsis: About 10-20% of Lyme disease patients treated with antibiotics continue to suffer debilitating symptoms, known as post-treatment Lyme disease. It is not understood whether this is due to an inappropriate immune response or continued Borrelia bacterial replication. In vitro laboratory experiments have showed that persister forms of bacteria are difficult to eliminate with antibiotics. This is because persisters are very slow-growing, and most antibiotics target actively-replicating bacteria. Limited clinical observations suggested some efficacy of dapsone, a sulfa drug, which belongs to a class of antibiotics not commonly used to treat Lyme disease. To test this drug against persister forms, Dr. Ying Zhang’s group studied sulfamethoxazole, dapsone, and sulfachlorpyridazine singly as well as in combination. By themselves, these drugs had similar efficacy as doxycycline, with 59-92% of stationary bacteria still alive after treatment. Their bacteria-killing activity was weak compared to daptomycin, which when used at the same concentrations, resulted in 26-52% of remaining cell viability. As combinations, they could not completely eradicate persister bacteria, with 74-77% bacteria remaining viable after treatment. But when they were combined with non-sulfa drugs like doxycycline, cefuroxime, ciprofloxacin, rifampin, and azithromycin, the rate of bacterial killing was higher. The most effective combinations included daptomycin. Further research will identify effective combinations that will more completely eradicate persister bacteria in vitro and in patients. Topics: chronic Lyme

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Feng J, Zhang S, Shi W, Zhang Y. (2016) Ceftriaxone pulse dosing fails to eradicate biofilm-like microcolony B. burgdorferi persisters which are sterilized by daptomycin/doxycycline/cefuroxime drug combination without pulse dosingFront Microbiol. Doi: 10.3389/fmicb.2016.01744.

Synopsis:  At least 10% of patients treated with antibiotic therapy for Lyme disease continue to suffer persisting symptoms. The reason for this is unknown, but one suspect is the continued survival of persisting forms of B. burgdorferi, the bacterium responsible for Lyme disease. In the test tube, Zhang and colleagues have grown one form of persisting bacteria known as biofilms, and used it to test various combinations of antibiotics. They also compared single continuous dosing versus pulse-dosing, which consists of repeated cycles of adding and removing antibiotics. They found that pulse-dosing was not effective for all antibiotics, and in fact reduced the activity of daptomycin against persisters. The best strategy they found against biofilm-like persisters was a single dose of a combination of daptomycin, doxycycline, and cefuroxime. They also showed that bacteriocidal drugs, which kill bacteria, are better used in pulse dosing than bacteriostatic drugs, which block bacterial replication but do not directly kill them. These promising in vitro data pave the way for animal model studies. If the findings prevail, they may suggest that future human trials are warranted. Topics: chronic Lyme

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Uhde M, Ajamian M, Li X, Wormser GP, Marques A, Alaedini A. (2016) Expression of C-reactive protein and serum amyloid A in early to late manifestations of Lyme Disease. Clin Infect Dis. first published online September 1, 2016doi:10.1093/cid/ciw599.

Synopsis:  The human immune response to invasive microbes consists of two major arms: the innate and the adaptive response. The innate response occurs rapidly after infection, when the host tries to contain or inactivate the microbe. Two proteins involved in this process are C-reactive protein (CRP) and serum amyloid A (SAA), which are often elevated in the blood during an inflammatory response. In this study, the researchers analyzed CRP and SAA in the blood of people with varying stages of Lyme disease. They found that compared to healthy people, CRP was elevated in early Lyme disease, when patients had erythema migrans (EM) skin rashes. In contrast, CRP levels in patients with early or late neurological symptoms and arthritis were similar to healthy controls. However, patients with post-treatment Lyme disease syndrome (PTLDS) had higher CRP levels that those who recovered, suggesting ongoing inflammation in patients with long-term Lyme disease. SAA was elevated in patients with EM rashes, but was similar to healthy controls in all other patients, including those with PTLDS. These findings suggest that persisting symptoms after antibiotic therapy are associated with elevated CRP, and pave the way for further studies to understand long-term inflammatory processes in these patients. Topics: chronic Lyme

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Aucott JN, Soloski MJ, Rebman AW, Crowder LA, Lahey LJ, Wagner CA, Robinson WH, Bechtold KT. (2016) CCL19 as a Chemokine Risk Factor for Post Treatment Lyme Disease Syndrome: A Prospective Clinical Cohort Study. Clin Vaccine Immunol. June 29. Pii: CVI.00071-16. (Epub ahead of print)

Synopsis:  An estimated 10-20% of Lyme disease patients who are treated with antibiotics continue to suffer persisting symptoms 6 to 12 months later. This is known as post-treatment Lyme disease syndrome, because it may affect various organs. Symptoms include chronic pain, overwhelming fatigue, and neurological or psychiatric symptoms. It is not known why this occurs in a subset of treated patients, but an aberrant immune response has long been suspected. In a new study led by Dr. John Aucott of Johns Hopkins University, elevated CCL19 was identified as associated with persistent symptoms. During acute infection, all patients diagnosed with Lyme disease (defined by physician-diagnosed erythema migrans rash) had elevated CCL19. However, by one month after diagnosis and treatment, those with serum concentrations greater than 112 pg/ml were 13 times more likely to develop post-treatment symptoms at 6 and 12 months later. Other patients who recovered, or who had symptoms without functional impairment had lower CCL19 levels after one month. CCL19 is produced by activated dendritic cells, which are sentinel cells that recruit immune cells to the site of an infection. The authors propose that sustained, elevated CCL19 is linked to continued inflammation and may account for ongoing symptoms, but point out that further studies are needed to understand the mechanistic basis and to identify other relevant immune pathways. Topics: chronic Lyme

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Feng J, Shi W, Zhang S, Sullivan D, Auwaerter PG, Zhang Y. (2016) A drug combination screen identifies drugs active against amoxicillin-induced round bodies of in vitro Borrelia burgdorferi persisters from an FDA drug library. Front Microbiol. May 23;7:743. doi: 10.3389/fmicb.2016.00743.

Synopsis:  In lab culture, Borrelia burgdorferi, the bacterium that causes Lyme disease, can survive amoxicillin treatment by transforming into a round-body form. Such round bodies persist in a slowly-growing state and are tolerant of antibiotics. In this study, the authors tested 1581 drugs that are already approved by the FDA, for antimicrobial activity against amoxicillin-induced round-bodied B. burgdorferi. They found 23 drugs with activity against the round bodies when used singly. In addition, they identified artemisinin, a malaria drug, in combination with cefoperazone and doxycycline, as well as sulfachlorpyridazine together with daptomycin and doxycycline as potent combinations that reduced round bodies. Importantly, these combinations also had some durable inhibitory effect, because re-growth after subculture was low. These findings provide evidence that the continued search for effective drug combinations against persister bacteria is warranted. Topics: treatment, chronic Lyme

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Bouquet J, Soloski MJ, Swei A, Cheadle C, Federman S, Billaud JN, Rebman AW, Kabre B, Halpert R, Boorgula M, Aucott JN, Chiu CY. (2016) Longitudinal Transcriptome Analysis Reveals a Sustained Differential Gene Expression Signature in Patients Treated for Acute Lyme Disease. mBio. Feb 12;7(1). pii: e00100-16. doi: 10.1128/mBio.00100-16.

Synopsis:  The genes that are expressed in sick people differ from those who are healthy. Using advanced sequencing techniques, these researchers found that immune cells in the blood of people with Lyme disease express different genes from those of healthy controls. Genes that are linked to a strong immune response were reduced in Lyme patients compared to those with other bacterial or viral infections. People with persisting symptoms attributed to Lyme disease shared some gene expression patterns with patients suffering chronic illnesses caused by immune system dysfunction. Topics: diagnostics, treatment, chronic Lyme

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Feng J, Weitner M, Shi W, Zhang S, Zhang Y. (2016) Eradication of Biofilm-Like Microcolony Structures of Borrelia burgdorferi by Daunomycin and Daptomycin but not Mitomycin C in Combination with Doxycycline and Cefuroxime. Front Microbiol. Feb 10;7:62. doi: 10.3389/fmicb.2016.00062.

Synopsis:  The persistence of symptoms in Lyme patients treated with antibiotics may be due to the bacteria becoming tolerant to drugs. In lab culture, persistent Lyme bacteria grown into biofilm form were exposed to various antibiotic combinations. The authors found that double antibiotic combinations such as doxycycline and cefoperazone were not enough for biofilm eradication. The addition of either daptomycin or daunomycin was required for bacterial elimination. Topics: treatment, chronic Lyme

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Jacek E, Tang KS, Komorowski L, Ajamian M, Probst C, Stevenson B, Wormser GP, Marques AR, Alaedini A. (2016) Epitope-Specific Evolution of Human B Cell Responses to Borrelia burgdorferi VlsE Protein from Early to Late Stages of Lyme Disease J Immunol 196:1036-43.

Synopsis:  Lyme disease patients make antibodies that recognize Borrelia burgdorferi, the bacteria that cause illness. When the disease progresses from early to late stages, these antibodies may change. A protein on the outside of B. burgdorferi is called VlsE, and antibodies that bind to a portion of it called the membrane-proximal domain illustrate this. These specific antibodies were low in patients with early disease, and were increased in patients with symptoms typical of later illness, such as neurological symptoms. Topics: diagnostics, treatment, chronic Lyme

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Lahey LJ, Panas MW, Mao R, Delanoy M, Flanagan JJ, Binder SR, Rebman AW, Montoya JG, Soloski MJ, Steere AC, Dattwyler RJ, Arnaboldi PM, Aucott JN, Robinson WH. (2015) Development of a Multiantigen Panel for Improved Detection of Borrelia burgdorferi Infection in Early Lyme Disease. J Clin Microbiol 53(12):3834-41. Epub 2015 Oct 7.

Synopsis:  The current blood test for Lyme disease has low accuracy. In this study, the authors designed and tested a new blood test that is more accurate. They used portions of proteins, called peptides, from Borrelia burgdorferi, the bacteria that cause Lyme disease. For a patient to be scored as positive, their blood had to contain antibodies that would recognize at least 2 of the 10 bacterial peptides. This test was more sensitive than the current blood test, and could be developed into a newer generation of diagnostic test for Lyme disease. Topics: diagnostics

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Feng J, Shi W, Zhang S, Zhang Y. (2015) Persister mechanisms in Borrelia burgdorferi: implications for improved intervention. Emerg Microbes Infect Aug 4 (e51). Epub 2015 Aug 19.

Synopsis:  Lyme disease is caused by the bacteria called Borrelia burgdorferi. In lab cultures of B. burgdorferi that are treated with antibiotics, most are killed. However, some become persister bacteria that survive. The authors used RNA sequencing to study how the genes that are expressed may differ in persister compared to drug-free bacteria. They found that expression changes in 675 genes in persister bacteria. Topics: treatment, chronic Lyme

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Ajamian M, Cooperstock M, Wormser GP, Vernon SD, Alaedini A. (2015) Anti-neural antibody response in patients with post-treatment Lyme disease symptoms versus those with myalgic encephalomyelitis/chronic fatigue syndrome. Brain Behav Immun 48;354-355.

Synopsis:  Patients with persisting Lyme symptoms were previously found to have higher and more frequent anti-neural antibodies compared to people who were healthy or who had recovered from Lyme disease. Patients with chronic fatigue syndrome (CFS) and persisting Lyme disease often have similar symptoms. This study found that CFS patients do not have elevated anti-neural antibodies. Thus, the common symptoms of persisting Lyme and CFS are not likely due to anti-neural antibodies. Topics: diagnostics, chronic Lyme

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Timmaraju VA, Theophilus PAS, Balasubramanian K, Shakih S, Luecke DF, Sapi E. (2015) Biofilm formation by Borrelia burgdorferi sensu lato. FEMS Microbiol Lett 362:fnv120.

Synopsis:  The authors studied whether B. afzelii and B. garinii, two bacteria associated with Lyme disease in Europe, are capable of growing as biofilms in lab culture. Using chemical stains and microscopy, they showed that these two bacteria form biofilms, similar to B. burgdorferi, the North American species, when grown under specific lab culture conditions. Topics: treatment, chronic Lyme

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Feng J, Auwaerter PG, Zhang Y. (2015) Drug combinations against Borrelia burgdorferi persisters in vitro: eradication achieved by using daptomycin, cefoperazone and doxycycline. PLoS One. 10(3): e0117207.

Synopsis:  The persistence of Borrelia burgdorferi in the form of drug-tolerant bacteria may be one reason why some Lyme disease patients may continue to suffer symptoms despite antibiotic therapy. The authors determined which drug combinations were effective in eradicating persistent microcolonies of B. burgdorferi, and identified daptomycin, doxycycline and cefoperazone together as an effective way of eliminating bacteria. Even subculturing bacteria treated with this combination to fresh media did not result in bacterial growth, suggesting durable killing. Topics: treatment, chronic Lyme

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Tang KS, Klempner MS, Wormser GP, Marques AR, Alaedini A. (2015) Association of immune response to endothelial cell growth factor with early disseminated and late manifestations of Lyme disease but not posttreatment Lyme disease syndrome. Clin Infec Dis 61:1703-6.

Synopsis:  The persistence of symptoms in patients treated for Lyme disease remains unexplained. One possible reason could be immune system dysfunction, including antibodies that react against the patient. To study this possibility, these researchers studied whether antibodies targeting endothelial cell growth factor, a normal human protein, were associated with post-treatment Lyme disease syndrome. They found that while antibodies were observed in patients with early and later disease symptoms, they were mostly absent in patients with post-treatment Lyme syndrome. Topics: chronic Lyme

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Sharma B, Brown AV, Matluck NE, Hu LT, Lewis K. (2015) Borrelia burgdorferi, the causative agent of Lyme disease, forms drug-tolerant persister cells. Antimicrob Agents Chemother 59(8):4616-24. Epub 2015 May 26.

Synopsis:  Persisting bacteria survive antibiotic treatment in lab culture. Here, the authors showed that antibiotics killed most of lab-grown B. burgdorferi, with a small surviving population that tolerated drug treatment. They found that mitomycin C, a cancer drug, efficiently killed persisters. They also showed how pulse-dosing, wherein four repetitive cycles that included treating bacterial cultures with ceftriaxone, followed by washing out the drug, resulted in eradication of persistent bacteria. Topics: treatment, chronic Lyme

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Divan A, Budd RC, Tobin RP, Newell-Rogers MK. (2015) γδ T Cells and dendritic cells in refractory Lyme arthritis. J Leukoc Biol 97(4):653-663. Epub 2015 Jan 20.

Synopsis:  This is a review about the function of γδT cells, a specialized subset of T cells that are important in coordinating the immune response when it transitions from the innate, or immediate response, to the adaptive, more specific immune reaction. It focuses on these cells during refractory Lyme arthritis. Topics: chronic Lyme

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Feng J, Wang T, Zhang S, Shi W, Zhang Y. (2014) An optimized SYBR Green I/PI assay for rapid viability assessment and antibiotic susceptibility testing for Borrelia burgdorferi. PLoS One. 9(11)e111809.

Synopsis:  The authors analyzed various methods to efficiently and accurately count live and dead Borrelia burgdorferi in lab culture. This is important for calculating the efficacy of antibiotics in the killing of bacteria. They adapted a method that stains live bacteria green, and dead ones red, allowing simple, reproducible, and high-throughput means to count bacteria. Topics: treatment, chronic Lyme

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Fallon BA, Pavlicova M, Coffino SW, Brenner C. (2014) A comparison of Lyme disease serologic test results from 4 laboratories in patients with persistent symptoms after antibiotic treatment. Clin Infect Dis 59:1705-10.

Synopsis:  Diagnosis of Lyme disease in late-stage patients is often erroneous. Fallon and colleagues compared Lyme disease test results from four different testing labs, using blood samples from previously diagnosed Lyme patients and healthy controls. Overall, they found variation in results, but no statistically significant difference between labs. However, there was a higher positivity rate for the C6 test, than for whole bacterial sonicate used in the diagnostic test. Topics: diagnostics, chronic Lyme

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Feng J, Wang T, Shi W, Zhang S, Sullivan D, Auwaerter PG, Zhang Y. (2014) Identification of novel anti-persister activity against Borrelia burgdorferi from an FDA-approved drug library. Emerg Microbes Infect Jul 3 (e49). Epub Jul 2.

Synopsis:  A library of drug compounds that has already been approved by the FDA was tested for efficacy against persister forms of Borrelia burgdorferi that were grown in lab culture. They identified daptomycin, clofazimine, sulfa drugs, and cefoperazone as potential drugs worth pursuing in further studies, which may include animal models of Lyme disease. Topics: treatment, chronic Lyme

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Soloski MJ, Crowder LA, Lahey LJ, Wagner CA, Robinson WH, Aucott JN. (2014) Serum inflammatory mediator as markers of human Lyme disease activity. PLoS One. 9(4):e93243.

Synopsis:  The authors compared the blood of patients suffering more severe symptoms of Lyme disease against those with less. They found that the pattern of chemokines, which are chemical messengers produced by immune cells, varies between the two groups. This work may lead to better diagnostic tools and clinical treatments, and shows a biological difference between these two groups of patients. Topics: diagnostics, treatment, chronic Lyme
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Zhang Y. (2014) Persisters, persistent infections and the Yin–Yang model. Emerg Microbes Infect 3(e3).

Synopsis:  In this review, Dr. Zhang discusses the differences between B. burgdorferi persister and growing cells, and proposes a model for bacterial transition between the two states, with the balance tipped depending on the environment and stresses such as antibiotics. Topics: treatment, chronic Lyme

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Jacek E, Fallon BA, Chandra A, Crow MK, Wormser GP, Alaedini A. (2013) Increased IFN-α activity and differential antibody response in patients with a history of Lyme disease and persistent cognitive deficits. J Neuroimmunol. 255:85–91.

Synopsis:  Some patients treated for Lyme disease continue to suffer neurological symptoms. In these patients compared to healthy controls, there was increased interferon-α (IFNα) activity observed in the blood. IFNα is a cytokine, or chemical messenger, and together with the observation of elevated antibodies against B. burgdorferi and neural proteins, suggests an ongoing inflammatory response in these patients. Topics: treatment, chronic Lyme

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Batheja S, Nields JA, Landa A, Fallon BA.(2013). Post-treatment lyme syndrome and central sensitization. J Neuropsychiatry Clin Neurosci. Summer;25(3):176-86.

Synopsis: A significant subset of patients treated for Lyme disease continue to have symptoms that persist for months or years. These may include pain, fatigue and neurocognitive problems, and collectively has been termed post-treatment Lyme disease syndrome. Whether these patients have ongoing infection is difficult to assess using current technology, and remains an open question. It is also possible that a post-infectious immune-mediated process may be responsible for continued symptoms. Here, the authors review central sensitization syndrome, in which pain and sensory processing is elevated. They discuss whether central sensitization may be related to continued symptoms in post-treatment Lyme disease.Topics: chronic Lyme

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Goessling LS, Allan BF, Mandelbaum RS, Thach RE. (2012) Development of a mitochondrial 12S rDNA analysis for distinguishing Sciuridae species with potential to transmit Ehrlichia and Borrelia species to feeding Amblyomma americanum (Acari: Ixodidae). J Med Entomol 49(3):772-6.

Synopsis:  The authors used a new DNA identification technique to observe that Lone Star ticks in Missouri harbor DNA from both pathogenic bacteria (Ehrlichia and Borrelia) as well as from the species of squirrels on which they feed, the Eastern gray squirrel and the generic squirrel. It was not previously known that these squirrels are hosts for disease-carrying ticks. Topics: prevention

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Schutzer SE, Angel TE, Liu T, Schepmoes AA, Clauss TR, Adkins JN, Camp DG II, Holland BK, Bergquist J, Coyle PK, Smith RD, Fallon BA, Natelson BH. (2011) Distinct cerebrospinal fluid proteomes differentiate post-treatment Lyme disease from chronic fatigue syndrome. PLoS One. 6(2):e17287.

Synopsis:  Chronic fatigue syndrome patients and post-treatment Lyme disease patients share some similar neurological symptoms. This study showed that the protein profiles in cerebrospinal fluid from the two groups were distinctive, suggesting that they are two different diseases. Topics: diagnostics, treatment, chronic Lyme

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Chandra A, Latov N, Wormser GP, Marques AR, Alaedini A. (2011) Epitope mapping of antibodies to VlsE protein of Borrelia burgdorferi in post-Lyme disease syndrome. Clin Immunol 141:103-110.

Synopsis:  The immune response to B. burgdorferi infection includes the production of antibodies that recognize bacterial surface proteins, one of which is called VlsE. Proteins form complex three-dimensional shapes, and only parts of them, called epitopes, are recognized by antibodies. Alaedini’s group mapped these on VlsE. The antibodies against VlsE differ in patients with later compared to earlier stages of Lyme disease. Topics: diagnostics, chronic Lyme

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Newell MK, Tobin RP, Cabrera JH, Sorensen MB, Huckstep A, Villalobos-Menuey EM, Burnett M, McCrea E, Harvey CP, Buddiga A, Bar-Or A, Freedman MS, Nalbantoglu J, Arbour N, Zamvil SS, Antel JP. (2010) TLR-mediated B cell activation results in ectopic CLIP expression that promotes B cell-dependent inflammation. J Leuk Biol 88:779-789.

Synopsis:  In this article, the authors used a mouse model to study B cells, which when activated, produce antibodies and release inflammatory cytokines, or chemical messengers. They found that a protein called CLIP is important in controlling B cells, and speculated that it shows a potential pathway to control chronic immune activation diseases such as Crohn’s disease and post-treatment Lyme disease. Topics: treatment, chronic Lyme

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