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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:

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                  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



=====*=====


I.    LYMENET: Co-infection, Medscape Coverage Highlight LDF Conference
-----------------------------------------------------------------------
Sender: Marc Gabriel <a229@Lehigh.edu>
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


=====*======


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.



=====*=====


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. rks4@cornell.edu
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.


=====*=====


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. rbg9@cdc.gov
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.



=====*=====


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.



=====*=====


VI.   ABOUT THE LYMENET NEWSLETTER
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