Searching for Persistence of Infection in Lyme Disease

Lyme disease is the most common vector borne disease in the United States. Although antibiotic therapy is clinically effective in treating the symptoms of Lyme disease for most patients early in the course of disease, a significant number of patients who receive therapy report persistent symptoms. The cause of persistent symptoms after antibiotic therapy for Lyme disease is an area of great controversy. Recent studies have shown that the organism (Borrelia burgdorferi) may persist in animals after antibiotic therapy and can be detected by using the natural tick vector (Ixodes scapularis) to acquire the organism through feeding (xenodiagnosis). Whether this occurs in humans is unknown. Currently available tests for human Lyme disease do not allow determination of persistent infection after antibiotic therapy.

In this proposal, the utility of xenodiagnosis for identifying persistence of B. burgdorferi in treated human Lyme disease will be tested in up to 120 subjects with various stages of Lyme disease with 30 healthy adults to serve as controls. Subjects will be followed for approximately 3 months. In Objective #1, subjects who have the characteristic erythema migrans (EM) rash and have been treated with antibiotics early (within 3 weeks of infection) in the course of Lyme disease will be tested. After completion of antibiotic therapy, 25-30 larval Ixodes ticks will be allowed to feed on the subject and biopsies of the EM site will be performed. Repleted ticks will be collected and tested for the presence of B. burgdorferi. In Objective #2, similar studies will be performed, but enrollment will target subjects with elevated C6 (region 6 of the VlsE surface protein of B. burgdorferi) antibody levels. In Objective #3, patients with persistent symptoms after antibiotic therapy will be evaluated. As an attempt to increase the chances of a positive xenodiagnosis in humans, patients with EM who are in the first 2 days of antibiotic therapy and patients with untreated Lyme arthritis will also be evaluated. Evidence that B. burgdorferi can be recovered by xenodiagnosis after antibiotic therapy in subjects with continued symptoms would change the current paradigm for potential mechanisms of disease and provide researchers and clinicians a tool for identifying patients with persistent infection. In Objective #4 we will assess the safety of the planned xenodiagnostic procedure in humans. All individuals who underwent xenodiagnosis under the study will be assessed for adverse events.

• INCLUSION CRITERIA:

Criteria for the diagnosis of Lyme disease can be found at The clinical assessment, treatment, and prevention of Lyme disease, human granulocytic anaplasmosis, and babesiosis: clinical practice guidelines by the Infectious Diseases Society of America (4).

• Patients With EM (N=35), Post Treatment

• Age 18 or older
• EM diagnosed by the study physician or, with a diagnosis of EM that is thought to be highly likely by the study physician.
• Treatment with at least 1 course of antibiotics that fulfills the Infectious Diseases Society of America guidelines for the recommended therapy for Lyme disease. There must be at least 1 month and up to 4 months between the end of the therapy and the study procedures.
• High C6 Antibody Titer (N=35)

• Age 18 or older
• Diagnosed with confirmed or probable early or late Lyme disease as per Centers for Disease Control and Prevention (CDC) case definition (http://www.cdc.gov/ncphi/disss.nndss/caseded/lyme disease 2008.htm), have received recommended antibiotic therapy and have a high C6 ELISA titer (index above 3) at least 6 months after therapy.
• Post Lyme disease syndrome (N=20)

• Age 18 or older
• Diagnosed with confirmed or probable early or late Lyme disease fulfilling the case definition of Lyme disease by the CDC (http://www.cdc.gov/ncphi/disss.nndss/caseded/lyme disease 2008.htm)
• Received recommended antibiotic therapy, with resolution or stabilization of the objective manifestation(s) of Lyme disease.
• New complaints of fatigue, paresthesias or dysesthesias, widespread musculoskeletal pain or cognitive difficulties within 6 months of the diagnosis of Lyme disease
• Symptoms are persistent or relapsing for at least a 6-month period after completion of antibiotic therapy, causing reduction in previous levels of occupational, educational, social, or personal activities.
• No antibiotic therapy active against Lyme disease in the previous 3 months.
• Patients With EM (N=20), On Treatment

• Age 18 or older
• EM diagnosed by the study physician and receiving antibiotic therapy for less than 48 hours.
• Lyme Arthritis (N=10)

• Age 18 or older
• Lyme arthritis and have not received therapy.
• Healthy Volunteers (N=30)

• Age 18 or older
• No history of Lyme disease
• Negative whole-cell Borrelia ELISA or C6 ELISA
• Living in an endemic area for Lyme disease.

Exclusion criteria

• History of allergy to surgical tape or Nitex nylon mesh
• History of severe reactions to tick bites (granuloma or systemic reactions)
• Inability to maintain the dressing for any reason
• Currently receiving any antibiotic or having received antibiotics in the last month (3 months for patients with post Lyme disease syndrome and high C6 titer) (except patients with EM on treatment)
• Pregnancy or lactation
• Unwillingness to use an effective method of birth control for 3 months after tick placement (women of child-bearing potential only)
• Not able to understand all of the requirements of the study or unable to give informed consent and/or comply with all aspects of the evaluation.
• Investigational therapy during the time of the study and/or in the month prior to signing the informed consent.
• Active severe skin disease, uncontrolled diabetes, cancer other than non-melanoma skin cancers, autoimmune disease requiring immunosuppressive therapy, or history of HIV, chronic viral hepatitis, or syphilis.
• Oral steroids in the previous 2 weeks (nasal steroids and replacement doses of steroids are not exclusions)
• Any other condition that, in the opinion of the investigator, would make the patient unsuitable for enrollment or could interfere with the patient participating in and completing the study.

EXCLUSION FROM SKIN BIOPSY PART OF THE PROTOCOL:

• History of forming large thick scars after skin injuries or surgery
• History of excessive bleeding after cuts or procedures
• Currently taking anticoagulants
• History of allergy to lidocaine
• Biopsies will not be done on face, neck, scalp, or over the tibia