LymeNet Newsletter Volume 7 Issue 04

Volume: 7
Issue: 04
Date: 19-Apr-99

Table of Contents:

I. LYMENET: Co-infection, Medscape Coverage Highlight LDF Conference II. WIEN KLIN WOCHENSCHR: Genospecies and their influence on immunoblot results. III. WIEN KLIN WOCHENSCHR: Clinical manifestations, pathogenesis, and effect of antibiotic treatment on Lyme borreliosis in dogs. IV. J CLIN MICROBIOL: The Borrelia burgdorferi 37-kilodalton immunoblot band (P37) used in serodiagnosis of early lyme disease is the flaA gene product. V. INFECTION: A proposal for the reliable culture of Borrelia burgdorferi from patients with chronic Lyme disease, even from those previously aggressively treated. VI. ABOUT THE LYMENET NEWSLETTER

Newsletter:

*********************************************************************** * The National Lyme Disease Network * * http://www.LymeNet.org/ * * LymeNet Newsletter * ***********************************************************************

Volume 7 / Number 04 / 19-APR-1999 INDEX

I. LYMENET: Co-infection, Medscape Coverage Highlight LDF Conference II. WIEN KLIN WOCHENSCHR: Genospecies and their influence on immunoblot results. III. WIEN KLIN WOCHENSCHR: Clinical manifestations, pathogenesis, and effect of antibiotic treatment on Lyme borreliosis in dogs. IV. J CLIN MICROBIOL: The Borrelia burgdorferi 37-kilodalton immunoblot band (P37) used in serodiagnosis of early lyme disease is the flaA gene product. V. INFECTION: A proposal for the reliable culture of Borrelia burgdorferi from patients with chronic Lyme disease, even from those previously aggressively treated. VI. ABOUT THE LYMENET NEWSLETTER

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I. LYMENET: Co-infection, Medscape Coverage Highlight LDF Conference ----------------------------------------------------------------------- Sender: Marc Gabriel <[email protected]> Date: April 12, 1999

Co-infection with two or more of the three major tick-borne pathogens was one of the top themes at the 12th Annual Lyme Disease Foundation Conference on Lyme Disease and Other Spirochetal and Tick-Borne Disorders Conference held in New York City April 9th and 10th. The Conference, which attracted its 2nd largest turnout ever, focused on clinical management, diagnosis, and the basic sciences.

Clinical presentations emphasized on the need for physician awareness concerning the other two tick-borne illnesses, babesia and ehrlichia. Differentiating between the three diseases and the myriad of combinations can be difficult. During the poster presentations diagnostic labs described their testing methodologies for these emerging illnesses.

The vaccine controversy continued to play itself out. Representatives from SmithKline Beecham presented their data supporting an accelerated vaccination schedule of 0, 1 and 2 months. The only schedule currently approved by the US Food and Drug Administration is 0, 1, 12. A subsequent presentation by Dr. Ronald Schell of the University of Wisconsin reported that OspA, the main protein in the human vaccine, induces severe destructive arthritis in hamsters. He concluded that OspA should not be used to vaccinate humans until further testing is conducted. The vaccine session concluded with a lively debate, which had to be cut short due to time constraints.

For the first time, the conference was covered by Medscape, the Internet web site providing physicians of most specialties with updates in their fields. Full coverage is available at:

http://www.medscape.com/conferences/Lyme99

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II. WIEN KLIN WOCHENSCHR: Genospecies and their influence on immunoblot results. -------------------------------------------------------------- AUTHORS: Wilske B, Hauser U, Lehnert G, Jauris-Heipke S ORGANIZATION: Max von Pettenkofer-Institut fur Hygiene und Medizinische Mikrobiologie, Lehrstuhl Bakteriologie, LMU Munchen, Federal Republic of Germany. REFERENCE: Wien Klin Wochenschr 1998 Dec 23;110(24):882-5 ABSTRACT:

In Europe at least three human pathogenic species of Borrelia burgdorferi sensu lato are the causative agents of Lyme borreliosis. All three species have been isolated or detected by PCR from skin, CSF and synovial fluid of patients with skin lesions, neuroborreliosis and Lyme arthritis respectively. Studies using strains representing the three species as antigen for the immunoblot revealed that interpretation criteria depend strictly on the strain used as antigen. More than using certain species as antigen it is important to use strains (f.e. B. afzelii strain PKo) expressing certain immunodominant antigens like OspC and p17 which may not be expressed by other strains in vitro. Using strain PKo as antigen the two band criterium can be used without loss of too much sensitivity compared to using B. burgdorferi sensu stricto strain PKa2 and B. garinii strain PBi. The use of recombinant antigens allows selection of highly specific and combination of homologous antigens from different strains; however not all desirable antigens have been recombinantly
expressed. Addition of p17 and p58 as antigens may improve the sensitivity of the hitherto described recombinant antigen immunoblots containing the antigens p83/100, p39, OspC and the p41 internal fragment.

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III. WIEN KLIN WOCHENSCHR: Clinical manifestations, pathogenesis, and effect of antibiotic treatment on Lyme borreliosis in dogs. ---------------------------------------------------------------------- AUTHORS: Straubinger RK, Straubinger AF, Summers BA, Jacobson RH, Erb HN ORGANIZATION: James A. Baker Institute for Animal Health, Ithaca, New York, USA. [email protected] REFERENCE: Wien Klin Wochenschr 1998 Dec 23;110(24):874-81 ABSTRACT:

BACKGROUND: Borrelia burgdorferi, the causative agent of Lyme disease, infects humans and animals. In humans, the disease primarily affects the skin, large joints, and the nervous system days to months after infection. Data generated with appropriate animal model help to understand the fundamental mechanisms of the disease. OBJECTIVE: 1) More clearly define the clinical manifestation and pathogenetic mechanisms of Lyme disease in dogs; 2) evaluate the effect of antibiotics in dogs infected with B. burgdorferi; 3) describe the effects of corticosteroids on dogs persistently infected with B. burgdorferi. DESIGN: Specific-pathogen-free beagles were infected with B. burgdorferi using ticks collected in an endemic Lyme disease area. Clinical signs were recorded daily. Antibody titers were measured by ELISA at two-week intervals. B. burgdorferi organisms were detected in tissues by culture and PCR. Synovial fluids were evaluated microscopically and with a chemotaxis cell migration assay. Histological sections were examined for pathological lesions. Specific
cytokine up-regulation in tissues was detected by RT-PCR. INTERVENTIONS: In three separate experiments, B. burgdorferi-infected dogs received antibiotic treatment (amoxicillin; azithromycin; ceftriaxone; doxycycline) for 30 consecutive days. Two subclinical persistently infected dogs received oral prednisone for 14 consecutive days starting at day 420 post-infection. RESULTS: Dogs developed acute arthritis in the joints closest to the tick bites after a median incubation period of 68 days. Synovial membranes of lame and non-lame dogs produced the chemokine IL-8 in response to B. burgdorferi. Antibiotic treatment prevented or resolved episodes of acute arthritis, but failed to eliminate the bacterium from infected dogs. Corticosteroid treatment reactivated Lyme disease in persistently infected dogs, which had not received antibiotics previously. CONCLUSIONS: B. burgdorferi disseminates through tissue by migration following tick inoculation, produces episodes of acute arthritis, and establishes persistent infection. The spirochete survives antibiotic
treatment and disease can be reactivated in immunosuppressed animals.

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IV. J CLIN MICROBIOL: The Borrelia burgdorferi 37-kilodalton immunoblot band (P37) used in serodiagnosis of early lyme disease is the flaA gene product. --------------------------------------------------------------- AUTHORS: Gilmore RD Jr, Murphree RL, James AM, Sullivan SA, Johnson BJ ORGANIZATION: Division of Vector-Borne Infectious Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Public Health Service, U.S. Department of Health and Human Services, Fort Collins, Colorado, USA. [email protected] REFERENCE: J Clin Microbiol 1999 Mar;37(3):548-52 ABSTRACT:

The 37-kDa protein (P37) of Borrelia burgdorferi is an antigen that elicits an early immunoglobulin M (IgM) antibody response in Lyme disease patients. The P37 gene was cloned from a B. burgdorferi genomic library by screening with antibody from a Lyme disease patient who had developed a prominent humoral response to the P37 antigen. DNA sequence analysis of this clone revealed the identity of P37 to be FlaA, an outer sheath protein of the periplasmic flagella. Recombinant P37 expression was accomplished in Escherichia coli by using a gene construct with the leader peptide deleted and fused to a 38-kDa E. coli protein. The recombinant antigen was reactive in IgM immunoblots using serum samples from patients clinically diagnosed with early Lyme disease that had been scored positive for B. burgdorferi anti-P37 reactivity. Lyme disease patient samples serologically negative for the B. burgdorferi P37 protein did not react with the recombinant. Recombinant P37 may be a useful component of a set of defined antigens for the serodiagnosis of early Lyme
disease. This protein can be utilized as a marker in diagnostic immunoblots, aiding in the standardization of the present generation of IgM serologic tests.

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V. INFECTION: A proposal for the reliable culture of Borrelia burgdorferi from patients with chronic Lyme disease, even from those previously aggressively treated. -------------------------------------------------------------------- AUTHORS: Phillips SE, Mattman LH, Hulinska D, Moayad H ORGANIZATION: Greenwich Hospital, CT 06830, USA. REFERENCE: Infection 1998 Nov-Dec;26(6):364-7 ABSTRACT:

Since culture of Borrelia burgdorferi from patients with chronic Lyme disease has been an extraordinarily rare event, clarification of the nature of the illness and proving its etiology as infectious have been difficult. A method for reliably and reproducibly culturing B. burgdorferi from the blood of patients with chronic Lyme disease was therefore sought by making a controlled blood culture trial studying 47 patients with chronic Lyme disease. All had relapsed after long-term oral and intravenous antibiotics. 23 patients with other chronic illness formed the control group. Positive cultures were confirmed by fluorescent antibody immuno-electron microscopy using monoclonal antibody directed against Osp A, and Osp A PCR. 43/47 patients (91%) cultured positive. 23/23 controls (100%) cultured negative. Although persistent infection has been, to date, strongly suggested in chronic Lyme disease by positive PCR and antigen capture, there are major problems with these tests. This new method for culturing B. burgdorferi from patients with chronic Lyme disease
certainly defines the nature of the illness and establishes that it is of chronic infectious etiology. This discovery should help to reestablish the gold standard in laboratory diagnosis of Lyme disease.

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VI. ABOUT THE LYMENET NEWSLETTER ----------------------------------------------------------------------- For the most current information on LymeNet subscriptions, contributions, and other sources of information on Lyme disease, please refer to: http://newsletter.lymenet.org ----------------------------------------------------------------------- To unsubscribe from the LymeNet newsletter, send a message to: [email protected] On the first line of the message, write: unsub lymenet-l ----------------------------------------------------------------------- LymeNet - The Internet Lyme Disease Information Source ----------------------------------------------------------------------- Editor-in-Chief: Marc C. Gabriel <[email protected]> FAX (for contributions ONLY): 908-789-0028 Contributing Editors: Carl Brenner <[email protected]> John Setel O'Donnell <[email protected]> Frank Demarest <[email protected]>
Advisors: Carol-Jane Stolow, Director <[email protected]> William S. Stolow, President <[email protected]> The Lyme Disease Network of New Jersey ----------------------------------------------------------------------- WHEN COMMENTS ARE PRESENTED WITH AN ATTRIBUTION, THEY DO NOT NECESSARILY REPRESENT THE OPINIONS/ANALYSES OF THE EDITORS. ----------------------------------------------------------------------- THIS NEWSLETTER MAY BE REPRODUCED AND/OR POSTED ON BULLETIN BOARDS FREELY AS LONG AS IT IS NOT MODIFIED OR ABRIDGED IN ANY WAY. ----------------------------------------------------------------------- SEND ALL BUG REPORTS TO [email protected] -----------------------------------------------------------------------