The Lyme Enterprise

NO CONTROVERSY


It has been said, that there is "controversy" in the Medical Community
regarding the management; the treatment of Lyme borreliosis (Lyme
disease and associated vector-borne diseases).  That there is a
"Controversy", is a great misunderstanding- there is no controversy at
all.  Rather, there has been a deliberate attempt to curtail the
discovery of specific therapeutics for difficult, and as yet,
impossible to eradicate, spirochetal and rickettsial diseases, and
retain United States Government funding in the development of 
vaccines and diagnostic test kits within commercial biotechnology
enterprise groups, which intend to market possibly  unsafe vaccines
and inaccurate diagnostic testing products.
It is truly next to impossible to create either a vaccine or a
commercial diagnostic test kit for Lyme borreliosis (the Borrelia
burgdorferi group) or other borrelial spirochetal infections due to
the nature of these organisms.  Simlar to the trypansosomes (African
Sleeping Sickness and Chagas Disease), the Borrelia are resistent to
natural immunity and antimicrobial therapy, due to mechanisms of
intracellular and extracellular latency, and antigenic variation.
Antigenic variation is a mechanism of switching the outer surface
profile, or "skin", if you will, of the organism.  In this way, as
antibodies are created against the infection organism, by the host
(human victim), the infecting organism then switches the profile of
its outer membrane components, rendering the antibodies useless for
protection from illness.  In order for a vaccine or testing procedure
to accurately address a specific infectious disease, it would require
the infecting organism to remain stable enough over time, such that
this specific aspect of the infection, the antigenic profile, can be
targeted in the creation or detection of specific antibodies.
 Although the creation of a vaccine or commercial test kit may be
impossible due to antigenic variation, detection of infection is more
easily accomplished than is generally known, either deliberately or by
error.
The essential conceptual basis in the case of the commonest human
borreliosis is that the tick borne borrelial genome consists of a
single linear chromosome, and acquired circular and linear plasmids,
or short versions of DNA.  On these plasmids are encoded some of the
virulence and infectivity factors.  In the case of Borrelia
burgdorferi, the primary surface markers, or outer surface proteins
(Osps) are encoded on these plasmids.  Borrelia can lose plasmids
through generations of cell replication, and still retain some
infectivity.  What was intended to identify the class of Borrelial
infections as "Lyme", was outer surface protein "A", and a slight
difference in the number of flagellar filaments, as compared to the
Relapsing Fever Borrelia (although that a species distinction can be
made on the basis of flagellar filaments is disputable).
What is encoded on these plasmids as lipoproteins changes, as do the
main membrane components, known as Variable Major Proteins-, or
Variable Surface Proteins (Vmps or Vsps).  Ticks parasitize different
mammalian hosts, and each mammalian host will have a different immune
response to the borrelial spirochetal organims. Ticks bear more than
one species or strain of borrelia and numerous other known and unknown
co-infections.  Humans, by nature of their inherited immune
competence, vary in clinical response.  Non-human primates have a much
more variable range of competence and tissue types, even within a
non-human primate species.  The mouse, although a primary host for
ticks and these infections, is a poor model for human clinical
response.  Some coinfections, such as the Erhlichia suppress the
immune reponse.  Burgdorferi borrelial surface proteins have an
immune-suppressing effect, via induction of anti-inflammatory
cytokines (e.g., Interleuken 10).  Infection with Borrelia burgdorferi
appears to activate latent infections of the non-vectored kind (e.g.,
Epstein-Barr).
In summary, variable antigens, variable species and strains of
borrelia, various known and unknown co-infections conferred with tick
attachment, variable mammalian hosts, variable geographic ranges of
the vectors, and variable human immune reponse and competence, make
vaccination assessment and detection of just burgdorferi borreliosis
alone, not likely to be accurate.
United States National Institutes of Health Taxonomy Browser:
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=Taxonomy
Pasteur Institute Borrelia burgdorferi sensu lato Molecular Genetics
Server: http://www.pasteur.fr/recherche/borrelia/Welcome.html
For just Borrellia burgdorferi alone in the United States, the most
accurate description of vector biology and human clinical status was
reported in the Infectious Diseases Society of America's journal,
"Reviews of Infectious Diseases" Volume II, Supplement 6,
September-October 1989.  Some abstracts and excerpts from this series
of articles are mentioned below (beyond the "Controversy" section).
In 1988 and 1989, the first Lyme borreliosis vaccine antigen patents
were applied for in the European and the United States patent
database.  The first, to the best of my knowledge was for a whole
sonicate of Borrelia burgdorferi, issued to Russell C. Johnson, a
microbiologist in the United States.  A second and third patent were
issued to Yale University (New Haven, Connecticut, USA), and Alan G.
Barbour (UCAL, Irvine) and his associates in Sweden and the United
States, and were for the recombinant antigen outer surface protein A
(OspA), of Borrelia burgdorferi.
Once these latter two "vaccines" were advanced beyond Phase I and
Phase II animal trials, the understanding of Human Borreliosis changed
from that described in "Reviews of Infectious Diseases" Volume II,
Supplement 6, September-October 1989.  In the early 1990s, a
"Controversy" appeared in the Medical Community, initiated by former
Yale University researcher Allen C. Steere, now at Harvard University,
Massachusetts, USA.
No longer was Borreliosis the chronic infection it was understood to
be.  It became "curable", without any formal controlled antimicrobial
therapeutics outcomes studies.
 Blood tests for detection of infection had always been difficult and
inaccurate, due to antigenic variation, and the use of efficient, but
not accurate, diagnostic methods, such as the Western Blot for
Borrelia burgdorferi specific antibody determination.  Until the
developmental phases of OspA vaccines were well along, the standard
for diagnosis was detection of new specific antigens, over time:
J Clin Invest 1986 Oct;78(4):934-9	 

Antigens of Borrelia burgdorferi recognized during Lyme disease.
Appearance of a new immunoglobulin M response and expansion of the
immunoglobulin G response late in the illness.

Craft JE, Fischer DK, Shimamoto GT, Steere AC.

Using immunoblots, we identified proteins of Borrelia burgdorferi
bound by IgM and IgG antibodies during Lyme disease. In 12 patients
with early disease alone, both the IgM and IgG responses were
restricted primarily to a 41-kD antigen. This limited response
disappeared within several months. In contrast, among six patients
with prolonged illness, the IgM response to the 41-kD protein
sometimes persisted for months to years, and late in the illness
during arthritis, a new IgM response sometimes developed to a 34-kD
component of the organism. The IgG response in these patients appeared
in a characteristic sequential pattern over months to years to as many
as 11 spirochetal antigens. The appearance of a new IgM response and
the expansion of the IgG response late in the illness, and the lack of
such responses in patients with early disease alone, suggest that B.
burgdorferi remains alive throughout the illness.  PMID: 3531237
[PubMed - indexed for MEDLINE]
 
34kD is OspB
At almost the same time, it was discovered by Steere, that the
population of infected persons who were ill from the infection, was
about equal to the population of persons who were infected, but were
not ill:
J Infect Dis 1986 Aug;154(2):295-300	 

Longitudinal assessment of the clinical and epidemiological features
of Lyme disease in a defined population.

Steere AC, Taylor E, Wilson ML, Levine JF, Spielman A.

From 1979 to 1983, Lyme disease was studied longitudinally in the 162
long-term residents of Great Island, Massachusetts. In retrospect, the
index case occurred in 1962, and the peak years of disease
transmission (about three new cases per 100 residents per year) were
the late 1970s. Thereafter, during the period of active surveillance,
attack rates declined by half. Altogether, 26 (16%) of the 162
residents developed symptoms of the disease. Most of those affected
had erythema chronicum migrans, and when untreated, they subsequently
developed arthritis or, in one instance, myocarditis. A minority of
individuals, mostly children, had arthritis alone. Of 121 asymptomatic
residents who gave blood samples, 10 adults (8%) had high titers of
IgG antibodies to the Lyme disease spirochete; these titers sometimes
persisted for years. From 1981 to 1983, the estimated ratio of
apparent-to-inapparent infection was 1:1. The high frequency of Lyme
disease on Great Island underscores the need for surveillance and
control programs.

PMID: 3722867 [PubMed - indexed for MEDLINE]
 
Half were infected but not ill, or immune competent, and half
succumbed to the infection.  Symptomatically and Asymptomatically
infected, as they were later called.
Scientists Raymond Dattwyler, Benjamin Luft, and Edward Bosler of the
State University of New York, USA, Long Island, Stony Brook, Divisions
of Infectious Diseases and Allergy and Rheumatology, made the same
observations and the same argument in a summary of Conference
discussions held at Cold Spring Harbor Laboratories, also Long Island,
New York, USA, published in 1992, by the Cold Spring Harbor Laboratory
Press, in 1992.  The publication of the conference summary was through
a book edited by one of the conference presenters, Steven E. Schutzer
entitled "Lyme Disease, Molecular and Immunologic Approaches".
In the 10 years after the discovery of the Lyme (so-called, because of
the name of the town in Connecticut USA where a parent, Polly Murray,
noticed a cluster of arthritis, Lyme, Connecticut) spirochete by Swiss
scientist and US immigrant Willy Burgdorfer (Borrelia burgdorferi), an
employee of the National Institutes of Health (Vector-Borne Diseases
Division, Rocky Mountain Laboratories, in Montana, USA), from 1982 to
about 1992, Lyme borreliosis appeared to be similar to Multiple
Sclerosis, with some reactive arthritis in a subset of patients, who
had the genetic predisposition for this condition in response to
bacterial and viral infection localizing in connective tissue.  (In
Europe, a species/strain of borrelial infection manifesting as another
connective tissue, skin, presentattion, was much earlier recognized as
Acrodermatitis Chronicum Atrophicans ?ACA).  In the next ten years, it
became something different, to everyone's astonishment.
Simultaneously, in the first ten years since the discovery of
Burgdorfer's spirochete, Federal Republic of Germany researcher Roland
Martin, was concluding the same:  The neurological presentation of
Lyme borreliosis is clinically indistinguishable from Multiple
Sclerosis.  Currently, Roland Martin is employed at the United States
National Institutes of Health, National Institute of Neurological
Disorders and Stroke (NINDS).
http://accessible.ninds.nih.gov/about_ninds/labs/98.htm
Due to antigenic variation and other variables, specifically, the
limitations of the less expensive methods, there was no way to
adequately detect or vaccinate against the growingly-more-virulent
borrelioses.  There exist better methods of separation and detection
of specific markers of burgdorferi borreliosis infection, across the
spectrum of molecular marker classes, however, the employing these in
rapid commercial test kits with any accuracy would require several
steps, and therefore not be especially "rapid".  These methods would
be cost-prohibitive in large vaccine trials.
The continued evolution of test kit methods expands to address the
difficulties associated with reaching the range of antigens necessary
to detect borreliosis.  A greater range of specific recombinant
antigens, are now being included in testing methods.  Still these
efficient testing methods are limited to the "species" Borrelia
burgdorferi, while more than burgdorferi cause human disease, and due
to global warming, mammalian immediate and intermediate hosts, and
arthropod vector habitat ranges also expand.  These conditions make it
not likely that these infections will ever be stable enough to have an
adequate vaccine or diagnostic test kit.  One approach to addressing
this problem is the development of a vaccine against tick saliva, and
the creation of an immune reponse that counters the local
infection.tick "bite", in which molecules in tick saliva which inhibit
a host response to the bite are inhibited, facilitating the
infections' tranfer.
Given these known problems asociated with assessing outcomes of
Osp-based borreliosis vaccines, vaccines were developed anyway.
Yale had a patent, and Alan Barbour had a patent for OspA.  Allen
Steere was an employee of Yale University, and who was, also, a United
States Centers for Disease Control (CDC) Epidemiological Intelligence
Officer (EIS).   Alan Barbour was also a CDC EIS.
The CDC held a Conference in Dearborn, Michigan, USA, in which several
laboratories were invited to participate in a consensus over what
would be the blood work standard for diagnosis of "Lyme disease". 
Prior to this Conference, however, in 1993, Allen Steere and Frank
Dressler adopted a standard for assessing borreliosis in which the
late infection signs of borreliosis arthritis presentation, in IgG
antibody type, with its many-fold higher antibody concentration
developing in people with this genetic history than occurs in most
other patients was the criteria.  This was the report:
J Infect Dis 1993 Feb;167(2):392-400	 
Western blotting in the serodiagnosis of Lyme disease.

Dressler F, Whalen JA, Reinhardt BN, Steere AC.

Division of Rheumatology/Immunology, Tufts University School of
Medicine, New England Medical Center, Boston, Massachusetts 02111.

There are currently no accepted criteria for positive Western blots in
Lyme disease. In a retrospective analysis of 225 case and control
subjects, the best discriminatory ability of test criteria was
obtained by requiring at least 2 of the 8 most common IgM bands in
early disease (18, 21, 28, 37, 41, 45, 58, and 93 kDa) and by
requiring at least 5 of the 10 most frequent IgG bands after the first
weeks of infection (18, 21, 28, 30, 39, 41, 45, 58, 66, and 93 kDa).
When these definitions were tested in a prospective study of all 237
patients seen in a diagnostic Lyme disease clinic during a 1-year
period and in 74 patients with erythema migrans or summer flu-like
illnesses, the IgM blot in early disease had a sensitivity of 32% and
a specificity of 100%; the IgG blot after the first weeks of infection
had a sensitivity of 83% and a specificity of 95%. Among patients with
indeterminate IgG responses by ELISA, 6 of 9 patients with active Lyme
disease had positive blots compared with 2 of 34 patients with other
illnesses (P < .001). Thus, Western blotting can be used to increase
the specificity of serologic testing in Lyme disease.   PMID: 8380611
[PubMed - indexed for MEDLINE]
The Dearborn Conference was held in October, 1994.  Meanwhile, the
major vaccine trial of Alan Barbour's vaccine, ImuLyme, developed by
Pasteur-Merieux Connaught,
has already been launched, with the first vaccination of the three
injection series, having begun in March 1994.
At the Dearborn Conference, nearly a dozen laboratories submitted
their analyses.  None agreed with Dressler and Steere for IgG antibody
accuracy, except a company called MarDx, which had been given CDC
banked Lyme arthritis patients' blood to qualify their Western Blot
test strips.  MarDx received the contract from both the SmithKline
Beecham vaccine LYMErix, the patent for which was owned by Steere/Yale
University, and the Barbour/Connaught ImuLyme vaccines.
Steere's standard became the CDC's standard for Lyme serodiagnosis:
http://www.cdc.gov/epo/mmwr/preview/mmwrhtml/00038469.htm
References 
   1.Engstrom SM, Shoop E, Johnson RC. Immunoblot interpretation
criteria for serodiagnosis of early Lyme disease. J Clin Microbiol
1995;33:419-22.
   2.Dressler F, Whelan JA, Reinhart BN, Steere AC. Western blotting
in the serodiagnosis of Lyme disease. J Infect Dis 1993;167:392-400.
The IgM criteria of Engstrom and Johnson, was also doubtful, since any
IgM, which was specific for burgdorferi borreliosis, should have been
accepted.  The specificities of Borrelia burgdorferi antigens range in
the 90s percentiles, meaning, the presence of any one specific IgM
antibody, was as sure a sign of this infection, as the percent
specificity.  For hypothetical example, if OspA has 97% specificity,
and if an IgM antibody was detected, as bound to that antigen, one
could be 97% sure, the infection was burgdorferi.  If IgM antibodies
persist, one can assume the infection persists.  If the range of
detectable IgM antibody expands over time, as Steere explained earlier
(1986), one could fairly surely assume the infection persisted.
This same unusual immune-evading infectious process occurs in
Trypanosomes:
http://www.cdc.gov/ncidod/eid/vol6no5/barbour.htm
Synopsis 
Antigenic Variation in Vector-Borne Pathogens
Alan G. Barbour* and Blanca I. Restrepo?
*University of California Irvine, Irvine, California; and ?Corporaci?n
para Investigaciones Biol?gicas, Medell?n, Colombia
Several pathogens of humans and domestic animals depend on
hematophagous arthropods to transmit them from one vertebrate
reservoir host to another and maintain them in an environment. These
pathogens use antigenic variation to prolong their circulation in the
blood and thus increase the likelihood of transmission. By convergent
evolution, bacterial and protozoal vector-borne pathogens have
acquired similar genetic mechanisms for successful antigenic
variation. Borrelia spp. and Anaplasma marginale (among bacteria) and
African trypanosomes, Plasmodium falciparum, and Babesia bovis (among
parasites) are examples of pathogens using these mechanisms. Antigenic
variation poses a challenge in the development of vaccines against
vector-borne pathogens.
 
Res Microbiol 1991 Jul-Aug;142(6):711-7	 
Antigenic variation in Borrelia.

Saint Girons I, Barbour AG.

Unite des Leptospires, Institut Pasteur, Paris.

Antigenic variation was demonstrated for the agent of relapsing fever,
Borrelia hermsii. The phenomenon is correlated with changes in major
surface proteins called Vmp. The genes encoding these antigens are
located on linear plasmids. Expression occurs by transposition of
genes encoding Vmp to a telomeric expression site located on another
linear plasmid. Activation of a vmp gene occurs by placing it
downstream from a promoter. Resemblance to the antigenic variation of
trypanosomes is discussed. PMID: 1961981 [PubMed - indexed for
MEDLINE]
Absurdly, the criteria for Dressler/Steere diagnosis of borreliosis,
was that 5 of 10 of these antibodies must be present, although the
specificity of one, flagellin (fla or 41 kilodaltons) is less than
90%, rather than just one specific antigen of higher than 90%.
DEARBORN: 
MMWR WEEKLY.    August 11, 1995 / 44(31);590-591 
Notice to Readers Recommendations for Test Performance and
Interpretation from the Second National Conference on Serologic
Diagnosis of Lyme Disease
"It was recommended that an IgM immunoblot be considered positive if
two of the following three bands are present: 24 kDa (OspC) * , 39 kDa
(BmpA), and 41 kDa (Fla) (1). It was further recommended that an that
IgG immunoblot be considered positive if five of the following 10
bands are present: 18 kDa, 21 kDa (OspC) *, 28 kDa, 30 kDa, 39 kDa
(BmpA), 41 kDa (Fla), 45 kDa, 58 kDa (not GroEL), 66 kDa, and 93 kDa
(2)."
Even more absurdly, the primary antigens were left out-- OspA and
OspB, which are encoded on the same plasmid, which when this plasmid
is dropped, as it does naturally, would render the borrelia not quite
a burgdorferi.
Later, Steere reported that indeed antibodies were detectable early to
OspA and B.  A Mayo Clinic researcher named David Persing (now at
CORIXA Corporation, State of Washington, USA) patented an OspA/B
plasmidless borrelia, and issued the rights to use this organism to a
biotechnology "spinoff" firm of Yale University, named "L2
Diagnostics", and to a immunology laboratory based in Norwood,
Massachusetts, USA, "Imugen".
From a Yale Bulletin and Calendar Release:
http://www.yale.edu/opa/ybc/v26.n26.news.04.html
"L2 Diagnostics, which was established with investment funds from the
School of Medicine as a spin-off of its Lyme/Lupus Diagnostics Lab.
The firm's new diagnostic methods will be essential when Yale's Lyme
disease vaccine is approved for public use. The tests will distinguish
patients infected with the Lyme disease from those who have been
vaccinated. "
In Summary, regarding diagnosis: While it was earlier recognized that
antigenic variation of these persisting spirochetal infections could
be detected by variable IgM, once vaccines were patented and under
development, the serodiagnosis criteria changed, and this was accepted
not only by the CDC, but by the United States Food and Drug
Administration, as criteria for assessment of vaccine safety and
efficacy outcomes.
Advisors to the SmithKline in 1998, regarding the Yale LYMErix vaccine
were:
Sponsor's Presentation 
  Dr. Robert Pietrusko           25
  Dr. Robert Schoen    32
  Dr. Vijay Sikand        38
  Dr. Yves Lobet    48
  Dr. Allen Steere         67
  Dr. Dennis Parenti               79
  Dr. Robert Pietrusko           122
http://www.fda.gov/ohrms/dockets/ac/98/transcpt/3422t1.rtf
 
At the 1994 FDA Lyme vaccines meeting, Raymond Dattwyler said (pages
43 ? 44)
In discussing how to assess Lyme diagnosis and vaccine response:
Paragraph 4 of the meeting minutes transcript:
"Now, what serologic assays could one use in a study such as a vaccine
trial.  Would a single ELISA be adequate.  Would a single Western Blot
be adequate.  Or, should one do serial ELISAs and serial Western
Blots.
It is my opinion that the best way to assess most infectious diseases
is to get acute and convalescent serology.  If one thought that the
person was acutely infected, I think that is a classic way of
assessing.
We know that, if you immunize someone with a vaccine and get an
appropriate immune response, that they should have some antibody and
perhaps be positive in a single ELISA.  So, a single ELISA under those
circumstances, I don't think, wuold be terribly useful.
A single Western Blot, since we are immunizing ? at least in this
discussion ? with OspA, would that be useful.  The answer is, I think,
yes, and I will get back to that in a minute.
A serial ELISA certainly could be helpful if one did it in an acute
and convalescent.  A rising serologic response would suggest an
infection. And the same, I think would be true about serial Western
Blots, where one would see an increase in repertoire of immune
response against various antigens to the bacteria?."
 
Page 45, Paragraph 3 (regarding the specificities of the early,
although "common" antigens?Common means common to other bacteria, such
as flagella and heat shock proteins, and means, these are less
specific to burgdorferi.  One would be less sure the infection is
burgdorferi), Dattwyler continues:
"The difficulty with OspC, though, is that it is a plasmid encoded
antigens [sic-transcription].  And most strains of this bacteria that
express OspC, as yo passage them repetitively over time and in
cultures, they will lose the plasmid that encodes for this, and that
is a problem.
And there are a number of commercial laboratories right now that have
organisms which are simply not expressing this any longer."
 
It was well-recognized that this problem of difficulty in truly
recognizing whether or not the vaccines had prevented Lyme disease
infection, and would require serological follow-up; repeat testing for
the expansion of antigens that arise, and in IgM antibodies due to
antigenic variation.  The CDC, upon the advice of its EIS officers,
Steere and Barbour, and a few others involved in facilitation of the
Dearborn Conference (e.g., Arthur Weinstein and Edward McSweegan)
adopted this strange criteria of Dressler and Steere, which reflected
largely arthritis serology.
In the field, in practice, this Dressler/Steere standard was only 14 %
(Imugen's submission to Dearborn) to 25% (Aguero-Rosenfeld) accurate. 
When Dressler and Steere themselves used this criteria in a
prospective study, it detected Lyme arthritis in 39/54 patients and
less so the meningitis/neurological presentations.
There would be no other reason to drop what was encoded on the OspA/B
plasmid, when OspA and B are primary antigens, except to qualify these
vaccines, because one could not distinguish OspA from vaccination,
from natural OspA.
The harm to the potential victim population, was in that once this CDC
standard was adopted, OspA and B could not be used to diagnose Lyme
disease in non-vaccinated individuals.
Additionally, it made no sense to drop the expanding development of
antibodies to different antigens expressed by the bacteria, as a
method to assess infection in IgM type antibodies, other than as it
appears now from post-vaccine adverse event results, the Asymptomatic
population were at risk.  How do you find Asymptomatically but
currently infected?
J Clin Invest 1986 Oct;78(4):934-9	 

Antigens of Borrelia burgdorferi recognized during Lyme disease.
Appearance of a new immunoglobulin M response and expansion of the
immunoglobulin G response late in the illness.

Craft JE, Fischer DK, Shimamoto GT, Steere AC.

Using immunoblots, we identified proteins of Borrelia burgdorferi
bound by IgM and IgG antibodies during Lyme disease. In 12 patients
with early disease alone, both the IgM and IgG responses were
restricted primarily to a 41-kD antigen. This limited response
disappeared within several months. In contrast, among six patients
with prolonged illness, the IgM response to the 41-kD protein
sometimes persisted for months to years, and late in the illness
during arthritis, a new IgM response sometimes developed to a 34-kD
component of the organism. The IgG response in these patients appeared
in a characteristic sequential pattern over months to years to as many
as 11 spirochetal antigens. The appearance of a new IgM response and
the expansion of the IgG response late in the illness, and the lack of
such responses in patients with early disease alone, suggest that B.
burgdorferi remains alive throughout the illness.

PMID: 3531237 [PubMed - indexed for MEDLINE]
 
The vaccine trial results:
http://www.fda.gov/ohrms/dockets/ac/01/slides/3680s2_11.pdf
Page 4 of that .pdf relates what were the previous criteria for
diagnosis.
THE PRE-DEARBORN DIAGNOSTIC STANDARD
     [Slide 7 - page 29 Dearborn Conference Summary]
     [Slide 8- Zoom]
Changing bands over time was formerly the criteria for determining
later stage Lyme disease, in place before the Dearborn conference, as
reported by David Dennis of the CDC:
 
"1) Isolation of Bb from Clinical specimens
2) Demonstration of diagnostic levels of IgM or IgG antibodies to the
spirochete in the serum or the CSF, or
3) Significant change in IgM or IgG antibody response to Bb in paired
acute-phase and convalescent sera phase
 
Although potentially useful in confirming active Lyme disease, neither
cultural isolation
nor paired serum specimen testing has been much used for validating
cases in routine Lyme testing, since the procedures are not often
performed in the general medical setting."
--------------------------------------------------------------------------------
-------------------------
HOW DOES DEARBORN APPLY TO THE VACCINE TRIAL?  
If few people have Lyme disease - and this Dressler/Dearborn criteria
will exclude most Lyme patients - the vaccine will not be shown to be
a failure or cause adverse events.
We believe this is exactly what happened in the trial.  
[Slide 9 Table 2 of NEJM SKB Vaccine Results]
Only 22 people got Lyme disease the first year in the vaccine group,
while there were 515 unconfirmed cases ? compared to in the placebo
group of 468.
There 10% more unconfirmed  cases than in the placebo group in the
first year of the trial.
There were ~1750 Unconfirmed Lyme disease cases reported during the
SKB trial of ~11,000 over two years.
The Western Blot serology from these unconfirmed Lyme cases will need
to be reviewed for evidence of other Bb specific bands and compared to
the placebo group by an  independent group of analysts.   If there are
any other specific bands besides OspA, the case must be counted as a
Lyme disease case, in the presence of symptoms.
Note that there were only 2 asymptomatic cases the first year in the
vaccine group vs 13 in the placebo group.  In the second year, there
were 0 (zero) in the vaccine group and 15 in the placebo group.
We believe these results do not show that the vaccine is effective at
preventing asymptomatic Lyme, which SKB  reports, but rather, that it
is turning asymptomatic Lyme cases into symptomatic ones.
As a support group leader in Southeastern CT, I have met ~10 people,
who found my name on the internet, who had adverse events and were
ill, looking for help.  After learning more about these patients, I
found that all but one of these cases had previous Lyme,  and that one
got the Erythema Migrans rash during the series of vaccination.  NOT
ONE SINGLE PERSON DID NOT HAVE OTHER BANDS ON FOLLOW UP WESTERN BLOT.
It is because I have gotten so many calls from patients looking for
help because of their illness, that I am here today.
Continued follow up on these Unconfirmed patients should have been
with further Western blotting from one of the CDC recommended strains
(B31, 297, 2591) and  the original case definition, to look for
changing bands, and/or one of the newer antigen-decomplexing methods,
like that of Len Sigal's of RWJ or Steven Schutzer's, for IgM or IgG.
In the re-tabulated results, which we insist be performed, cases where
active infection is not found by these follow up methods, should be
resummarized as the "Uncomfirmed Lyme/Possible Seronegative Lyme".
VACCINE FAILURE AND ADVERSE EVENT
[Slide 10 Persing's Patent]
Dr. David Persing, formerly of Mayo, now with CORIXA recorded in his
US patent 6,045,804:
  "Additional uncertainty may arise if the vaccines are not completely
protective; vaccinated patients with multisystem complaints
characteristic of later presentations of Lyme disease may be difficult
to distinguish from patients with vaccine failure. Vaccine failures
have been occasionally noted in animal models (E. Fikrig et al.,
Science, 250, 553-6 (1990)),..."
Vaccine failure and vaccine adverse event cannot be distinguished from
each other.  An  asymptomatic Bb infected adverse LYMErix event case
may never be detected until the patient is vaccinated and symptoms
occur, which we  think explains the majority of the adverse events
reported to FDA re:  LYMErix.  Many previously infected Lyme cases
report systemic symptoms after vaccination.  Many find out they had
Lyme after being vaccinated, becoming ill, being tested for Lyme and
finding other specific antibodies.
FDA should therefore not be looking for only arthritis as a potential
adverse event, to the exclusion of systemic illness.
FREQUENCY OF ASYMPTOMATIC INFECTION
[Slide ? 10]
According to Allen Steere's 1986 report, it is possible that, for
every one Bb-infected person with symptoms, there is one walking
around without symptoms.
SUMMARY
Vaccine failure and exacerbation of asymptomatic infection are
identical, according to the patient data collected, and on the online
VAERS database.
Dearborn/Dressler is not a valid criteria for assessing Lyme, the
former CDC criteria of changing bands is valid.
Until there is an independent review of the WB data from the trial, we
have no idea how safe this OspA vaccine is.
[Slide- SBK Results Table] 
 
OUTLOOK
By what mechanism vaccination of the asymptomatic Bb infected patients
is causing the Lyme like illness, we do not know exactly.
Previous infection could be "priming" the immune system, as Denise
Huber of Tufts has suggested, in "Identification of LFA-1 as a
Candidate Autoantigen in Treatment-Resistent Lyme Arthritis"  July 31,
1998, Science, Vol 281, p 703.
or the vaccine is activating a dormant infection by the immune
dysregulation it causes, as demonstrated by the effect of Bb infection
and Osp A alone, on NK cells population, T cells, neutrophils, and the
effects on the various inflammatory regulating biomoleclues, such as
IL-10.
We simply don't know all the variables, at present, that effect
systemic illness from immune dysregulation caused by Bb infection, and
especially the effect of a sucha a large dose of a known immune
irritant, Osp A upon  this system, the asymptomatic Lyme patient.
-----------------------------------------------------------end except
from 2001 FDA testimony.
The standard was changed to suit the intended false positive vaccines
outcomes.
 
THE CONTROVERSY
Knowing the vaccines were being developed, and knowing Lyme
borreliosis is an incurable infection, the disease was spun:
 
Ann Intern Med 1992 Aug 15;117(4):281-5	 

Lyme disease associated with fibromyalgia.

Dinerman H, Steere AC.

Tufts University School of Medicine, Boston, Massachusetts.

OBJECTIVE: To describe the clinical and laboratory findings as well as
results of treatment in patients with Lyme disease associated with
fibromyalgia. DESIGN: Observational cohort study. The mean duration of
observation was 2.5 years (range, 1 to 4 years). SETTING: Diagnostic
Lyme disease clinic in a university hospital. PATIENTS: Of 287
patients seen with Lyme disease during a 3.5-year period, 22 (8%) had
fibromyalgia associated with this illness, and 15 (5%) participated in
the observational study. MEASUREMENTS: Symptoms and signs of
fibromyalgia, immunodiagnostic tests for Lyme disease, and tests of
neurologic function. RESULTS: Of the 15 patients, 9 developed
widespread musculoskeletal pain, tender points, dysesthesias, memory
difficulties, and debilitating fatigue a mean duration of 1.7 months
after early Lyme disease; the remaining six patients developed those
symptoms during the course of Lyme arthritis. At the time of our
evaluation, late in the course of their illness, 11 patients had
positive immunoglobulin (Ig) G antibody responses to Borrelia
burgdorferi by enzyme-linked immunosorbent assay (ELISA), one had a
positive Western blot, and the three seronegative patients had
positive cellular immune responses to borrelial antigens. Four
patients had abnormal cerebrospinal fluid analyses that showed an
elevated protein level, a slight pleocytosis, or intrathecal antibody
production to the spirochete. The signs of Lyme disease resolved with
antibiotic therapy, usually intravenous ceftriaxone, 2 g/d for 2 to 4
weeks, except in one patient with persistent knee swelling. However,
14 of the 15 patients continued to have symptoms of fibromyalgia.
Currently, only one patient is completely asymptomatic. CONCLUSIONS:
Lyme disease may trigger fibromyalgia, but antibiotics do not seem to
be effective in the treatment of the fibromyalgia.
PMID: 1637022 [PubMed - indexed for MEDLINE] 
JAMA 1993 Apr 14;269(14):1812-6	 

The overdiagnosis of Lyme disease.

Steere AC, Taylor E, McHugh GL, Logigian EL.

Division of Rheumatology/Immunology, New England Medical Center,
Boston, MA 02111.

OBJECTIVE--To analyze the diagnoses, serological test results, and
treatment results of the patients evaluated in a Lyme disease clinic,
both prior to referral and from current evaluation.
DESIGN--Retrospective case survey of prescreened patients.
SETTING--Research and diagnostic Lyme disease clinic in a university
hospital. PATIENTS--All 788 patients referred to the clinic during a
4.5-year period who were thought by the referring physician or the
patient to have a diagnosis of Lyme disease. MAIN OUTCOME
MEASUREMENTS--Symptoms and signs of disease, immunodiagnostic tests of
Lyme disease, and tests of neurological function. RESULTS--Of the 788
patients, 180 (23%) had active Lyme disease, usually arthritis,
encephalopathy, or polyneuropathy. One hundred fifty-six patients
(20%) had previous Lyme disease and another current illness, most
commonly chronic fatigue syndrome or fibromyalgia; and in 49 patients,
these symptoms began soon after objective manifestations of Lyme
disease. The remaining 452 patients (57%) did not have Lyme disease.
The majority of these patients also had the chronic fatigue syndrome
or fibromyalgia; the others usually had rheumatic or neurological
diseases. Of the patients who did not have Lyme disease, 45% had had
positive serological test results for Lyme disease in other
laboratories, but all were seronegative in our laboratory. Prior to
referral, 409 of the 788 patients had been treated with antibiotic
therapy. In 322 (79%) of these patients, the reason for lack of
response was incorrect diagnosis. CONCLUSIONS--Only a minority of the
patients referred to the clinic met diagnostic criteria for Lyme
disease. The most common reason for lack of response to antibiotic
therapy was misdiagnosis. PMID: 8459513 [PubMed - indexed for MEDLINE]
 
Steere used his own standard in these two determinations (that Lyme
becomes something else), which later was the Dearborn IgG standard,
knowing that treatment would render patients serologically suppressed
in antibodies.
Thus, a CONTROVERSY was spawned.
 
OTHER TESTIMONY AT THE 2001 FDA MEETING
http://www.fda.gov/ohrms/dockets/ac/01/slides/3680s2.htm
 
Combined, 1) the adverse events reported to the FDA through VAERS and
as described in the FDA testimony during the January 31, 2001 FDA
vaccine meeting, 2) the Asymptomatic infection results as reported by
SmithKline Beecham, 3) the "Unconfirmed Lyme", the fact that
Asymptomatic cases disappeared from the SmithKline data table, and 4)
that these later manifestations of Lyme disease (which amazingly
transformed themselves from a known illness, into one of unknown
origin), present one picture:  Asymptomatic infection is a risk for
vaccination with OspA.  Someone who has Fibromyalgia or Chronic
Fatigue Syndrome, does not have Lyme disease, yet these patients were
left out of the vaccine trial (neurologic presentations remaining
after Lyme infection, according to SmithKline).  If someone has Lyme
arthritis, there is not even any need to do bloodwork, because as
Steere says in Rahn and Evans, 1998 "Lyme Disease", ACP Key Diseases
Series, page 114
"It should be stressed that seronegative Lyme is a contradiction in
terms if a patient has marked arthritis, particularly chronic
arthritis, because the immune response seems to be involved in the
pathogenesis of joint inflammation."
What is the % of the population who are infected with this permanent
infection and don't know it?  For every person who is ill, and knows
it, there is another, who is not ill, but may find out, once they have
been vaccinated with OspA.
Still, they won't know it is Lyme, if by Steere's criteria, unless
they have the genetic background for arthritis.  They may end up with
Fibromyalgia after vaccination.  Robert Schoen of Yale also advised in
Rahn and Evans, pages238-239, not to do serologic testing of patients
with fatigue and pain who have received the vaccine.  Why?
The recognition of patients without the Steere/Dressler Lyme disease
would be the recognition of the invalidity of the standard, and the
invalidity of the vaccine trial outcomes.
 
Lobet, of SmithKline argued that persisting infection was the reason
for the LYMErix adverse events:
http://www.fda.gov/ohrms/dockets/ac/01/slides/3680s2_02_lobet.pdf
 
PERSISTING INFECTION:
United States Patent   4,721,617
Johnson January 26, 1988
Vaccine against lyme disease
Abstract
A vaccine for the immunization of mammals against Lyme borreliosis
(Lyme disease) is disclosed which contains an effective amount of
inactivated Borrelia burgdorferi spirochetes dispersed in a
physiologically-acceptable, non-toxic liquid vehicle.
"Spirochetes are introduced into the host at the site of the tick bite
and this is also the location of the initial characteristic skin
lesion, erythema chronicum migrans (ECM). A systemic illness ensues
due to the lymphatic and hematogenous spread of B. burgdorferi. The
early phase of the illness often consists of the ECM, headache,
fatigue, muscle and joint aches, stiff neck and chills and fever. This
phase of the disease may be followed by neurologic, joint or cardiac
abnormalities. The chronic forms of the disease such as arthritis
(joint involvement), acrodermatitis chronica atrophicans (skin
involvement), and Bannwart's syndrome (neurological involvement) may
last for months to years and are associated with the persistence of
the spirochete. A case of maternal-fetal transmission of B.
burgdorferi resulting in neonatal death has been reported. Domestic
animals such as the dog also develop arthritis and lameness to this
tick-borne infection. For every symptomatic infection, there is at
least one asymptomatic infection.  Lyme disease is presently the most
commonly reported tick-borne disease in the United States.
 
The infection may be treated at any time with antibiotics such as
penicillin, erythromycin, tetracycline, and ceftriaxone. ***Once
infection has occurred, however, the drugs may not purge the host of
the spirochete but may only act to control the chronic forms of the
disease.*** Complications such as arthritis and fatigue may continue
for several years after diagnosis and treatment."
DOES Osp A cause Immune Suppression?
"Thus, IL-10 could potentially down-regulate inflammatory and
microbicidal effector mechanisms of the innate immune response to a B.
burgdorferi infection, facilitating the establishment of the
spirochete."
 From: Infect Immun 2002 Apr;70(4):1881-8	 
Autocrine and exocrine regulation of interleukin-10 production in
THP-1 cells stimulated with Borrelia burgdorferi lipoproteins.

Giambartolomei GH, Dennis VA, Lasater BL, Murthy PK, Philipp MT.
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=1
1895951&dopt=Abstract
 
"Purified lipidated outer surface protein A (OspA), but not its
unlipidated form, induced the production of high levels of IL-10 in
uninfected human PBMC. Thus, the lipid moiety is essential in the
induction of IL-10 in these PBMC. B. burgdorferi M297, a mutant strain
that lacks the plasmid that encodes OspA and OspB, also induced IL-10
gene transcription in PBMC, indicating that this phenomenon is not
causally linked exclusively to OspA and its lipid moiety. These
results demonstrate that B. burgdorferi can stimulate the production
of an antiinflammatory, immunosuppressive cytokine in naive cells and
suggest that IL-10 may play a role both in avoidance by the spirochete
of deleterious immune responses and in limiting the inflammation that
the spirochete is able to induce."
 
From:
Infect Immun 1998 Jun;66(6):2691-7	 
  
Borrelia burgdorferi stimulates the production of interleukin-10 in
peripheral blood mononuclear cells from uninfected humans and rhesus
monkeys.

Giambartolomei GH, Dennis VA, Philipp MT.
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9
596735&dopt=Abstract
 
 
BARBOUR AND PERMANENT CNS BORRELIAL INFECTIONS (the focus of his
current USA Federal grants)
  
In vivo activities of ceftriaxone and vancomycin against Borrelia spp.
in the mouse brain and other sites.

Kazragis RJ, Dever LL, Jorgensen JH, Barbour AG.

Department of Medicine (Infectious Diseases), University of Texas
Health Science Center at San Antonio 78284, USA.

Borrelia burgdorferi, the agent of Lyme disease, and B. turicatae, a
neurotropic agent of relapsing fever, are susceptible to vancomycin in
vitro, with an MIC of 0.5 microgram/ml. To determine the activity of
vancomycin in vivo, particularly in the brain, we infected adult
immunocompetent BALB/c and immunodeficient CB-17 scid mice with B.
burgdorferi or B. turicatae. The mice were then treated with
vancomycin, ceftriaxone as a positive control, or normal saline as a
negative control. The effectiveness of treatment was assessed by
cultures of blood and brain and other tissues. Ceftriaxone at a dose
of 25 mg/kg of body weight administered every 12 h for 7 to 10 days
eliminated cultivable B. burgdorferi or B. turicatae from all BALB/c
or scid mice in the study. Vancomycin at 30 mg/kg administered every
12 h was effective in eliminating infection from immunodeficient mice
if treatment was started within 3 days of the onset of infection. If
treatment with vancomycin was delayed for 7 days or more, vancomycin
failed to eradicate infection with B. burgdorferi or B. turicatae from
immunodeficient mice. The failure of vancomycin in eradicating
established infections in immunodeficient mice was associated with the
persistence of viable spirochetes in the brain during antibiotic
treatment.  PMID: 8913478 [PubMed - indexed for MEDLINE]
 
 
Andrew Pachner, Neurology, was at Yale, when this occurred
 
http://groups.google.com/groups?q=bicycle+boy+group:sci.med.diseases.lyme&hl=en&
lr=&ie=UTF-8&oe=UTF-8&selm=20020322203622.14030.00002745%40mb-ct.aol.com&rnum=1
 
 "His Behavior Was Compulsive , It's Origins Unknown;  Then a Good
Doctor
Seemed to Make a Miracle Happen"
 
 
Russell Johnson, in addition to the patent above, edited the book, 
"The Biology of Parasitic Spirochetes", 1976, in which it says, tick
borne borrelia persist in the eye and brain past antibiotic treatment,
and that this is well-known (page 391).
 
 
ACTIVATION OF OTHER LATENT INFECTIONS:
 
Folia Biol (Praha) 2003;49(1):40-8 
Interaction of Borrelia burgdorferi sensu lato with Epstein-Barr virus
in Lymphoblastoid cells.
 
    Hulinska D, Roubalova K, Schramlova J.
 
    National Reference Laboratory on Borreliosis, Electron Microscopy,
National Institute of Public Health, Prague, Czech Republic.
 
    Since the possibility of interruption of latent EBV infection has
been suggested by the induction of the lytic virus cycle with chemical
substances, other viruses, and by immunosuppression, we hypothesized
that the same effect might happen in B. burgdorferi sensu lato
infection as happens in Lyme disease patients with positive serology
for both agents. We have observed EBV replication in lymphoblastoid
cells after superinfection with B. garinii and B. afzelii strains
after 1 and 4 h of their interaction. We found that viral and
borrelial antigens persisted in the lymphoblasts for 3 and 4 days.
Morphological and functional transformation of both agents facilitate
their transfer to daughter cells. Association with lymphoblasts and
internalization of B. garinii
by tube phagocytosis increased replication of viruses more
successfully than B. afzelii and chemical inductors. Demonstration of
such findings must be interpreted cautiously, but may prove a mixed
borrelial and viral cause of severe neurological disease.PMID:
12630667 [PubMed ? in process]
 
 
Paul Duray, Pathologist, in "Lyme Disease, Molecular and Immunologic
Approaches":
Page 12:
 
            "Atypical precursor B and T immunoblastoid cells have been
observed in our work on target organs [Duray and Steere, 1988].  On
occasion, these atypical-appearing lymphocytes have been
misinterpreted in biopsy by several laboratories as cells of malignant
lymphoma or leukemia.  Bb antigens, then, may stimulate growth of
immature lymphocytic subsets in large organs, as well as in the
cerebrospinal fluid [Szyfelbein and Ross 1988].  Unusual bacterial
infections do not produce such lymphocyte infiltrates in tissue. 
These immunoblastoid cells in Bb infections sometimes resemble those
found in Epstein ?Barr virus infections.  Does Bb reactivate latent
virus infections in tissues?  Do siome tick inocula deliver
simultaneous infectious agents [ixodid ticks can harbor Rickettsiae,
Babesia microti, and Ehrlichia bacteria, in addition to Bb], producing
multi-agent infections in some hosts?  Further studies can clarify
these issues by means of tissue-based molecular probe analysis."
 
===========
 
*** Rev Infect Dis 1989 Sep-Oct;11 Suppl 6:S1460-9 ***
 
    Epidemiology and clinical similarities of human spirochetal
diseases.
 
    Schmid GP.
 
    Division of Sexually Transmitted Diseases, Centers for Disease
Control, Atlanta, Georgia 30333.
 
    Lyme disease, first identified in 1975, is the most recently
recognized of the seven human spirochetal diseases; the evolving
clinical picture of Lyme disease indicates it shares many features
with the other diseases. These similarities are striking in view of
the
diverse epidemiology of the seven diseases, which are caused by
Treponema species (spread by human-to-human contact) or Leptospira or
Borrelia species (zoonoses). These similarities include the following:
(1) skin or mucous membrane as portal of entry; (2) spirochetemia
early in the course of disease, with wide dissemination through tissue
and
body fluid; and (3) one or more subsequent stages of disease, often
with intervening latent periods. Lyme disease shares with many
spirochetal diseases a tropism for skin and neurologic and
cardiovascular manifestations, whereas chronic arthritis is unique to
Lyme disease.  These similarities and dissimilarities offer
opportunities to discover
which properties unique to the pathogenic spirochetes are responsible
for clinical manifestations and suggest that certain clinical features
of patients with spirochetal diseases other than Lyme disease may
someday be recognized in patients with Lyme disease.  PMID: 2682958
[PubMed - indexed for MEDLINE]
 
Schmid writes, "the brain is a favored target in animals", of
borrelia.  Page S1466.
 
 
Dattwyler and Luft say, in the same supplement:
 
Rev Infect Dis 1989 Sep-Oct;11 Suppl 6:S1518-25 
 
    A perspective on the treatment of Lyme borreliosis.
 
    Luft BJ, Gorevic PD, Halperin JJ, Volkman DJ, Dattwyler RJ.
 
    Department of Medicine, University of New York, Stony Brook
11794-8153.
 
    Lyme borreliosis has become the most common tick-borne infection
in the United States. Although both beta-lactam and tetracycline
antibiotics have been shown to be effective in the treatment of this
spirochetosis, the development of optimal therapeutic modalities has
been hampered by the lack of reliable microbiologic or immunologic
criteria for the diagnosis or cure of this infection. In vitro
sensitivity studies have been performed by several laboratories, but
there has been no standardization of the methodology for measuring
either inhibitory or bactericidal levels. ***Clinical studies have
documented the efficacy of antibiotics, but therapy has failed in as
many as 50% of cases of chronic infection. Although new antibiotic
regimens appear promising, the optimal treatment of this infectious
disease remains to be determined. In this report we review the
clinical and experimental rationale for the antibiotic regimens that
we currently use and the need for a more standardized approach to
treatment trials.*** PMID: 2682965 [PubMed - indexed for MEDLINE]
 
 
***  We never got past this, because then Yale and Barbour had vaccine
patents.  Instead of Lyme disease, a patient with persisting symptoms,
now suddenly had Fibromyalgia.  Rather than continue with the
decades-old acknowledgement that spirochetal infections persist,
knowing that half of all those infected aren't aware of it, and stand
to be potential vaccine recipients, rather than prescreening vaccine
test subjects with the original valid method (IgM), these patients
were thrown out ("waste-basket" diagnoses), as were the non-arthritis
patients in the vaccine trial.
 
 
Andrew Pachner says, in the same supplement:
 
Rev Infect Dis 1989 Sep-Oct;11 Suppl 6:S1482-6 
 
    Neurologic manifestations of Lyme disease, the new "great
imitator".
 
    Pachner AR.
 
    Department of Neurology, University Hospital, Georgetown
University
Medical School, Washington, D.C. 20007.
 
    The causative agent of Lyme disease, Borrelia burgdorferi, is a
highly neurotropic organism that not only can produce symptomatic
neurologic disease but also can exist dormant within the central
nervous system (CNS) for long periods. Two distinct types of
neuroborreliosis occur at different stages of Lyme disease.
Second-stage Lyme meningitis resembles aseptic meningitis and is often
associated with facial palsies, peripheral nerve involvement, and/or
radiculopathies. Lyme meningitis may be the first evidence of Lyme
disease, occurring without a history of erythema chronicum migrans or
flu-like illness. ***Third-stage parenchymal involvement causes a
multitude of nonspecific CNS manifestations that can be confused with
conditions such as multiple
sclerosis, brain tumor, and psychiatric derangements.***
Manifestations of CNS parenchymal involvement in Lyme disease are
generally associated, however, with a history of erythema chronicum
migrans, meningitis, or carditis. Both second- and third-stage Lyme
neuroborrelioses are commonly misdiagnosed because they are relatively
uncommon and because they mimic many better-known disorders.  PMID:
2682960 [PubMed - indexed for MEDLINE]
 
Eur Neurol 1995;35(2):113-7	 

Seronegative chronic relapsing neuroborreliosis.

Lawrence C, Lipton RB, Lowy FD, Coyle PK.

Department of Medicine, Albert Einstein College of Medicine, New York,
N.Y., USA.

We report an unusual patient with evidence of Borrelia burgdorferi
infection who experienced repeated neurologic relapses despite
aggressive antibiotic therapy. Each course of therapy was associated
with a Jarisch-Herxheimer-like reaction. Although the patient never
had detectable free antibodies to B. burgdorferi in serum or spinal
fluid, the CSF was positive on multiple occasions for complexed
anti-B. burgdorferi antibodies, B. burgdorferi nucleic acids and free
antigen.

PMID: 7796837 [PubMed - indexed for MEDLINE]
 
http://www.acponline.org/journals/annals/15oct94/lyme.htm
 
Annals of Internal Medicine 
The Long-Term Clinical Outcomes of Lyme Disease
A Population-based Retrospective Cohort Study
Annals of Internal Medicine, 15 October 1994; 121: 560-567.
Nancy A. Sha____, MD, MPH; Charlotte B. Phillips, MPH; Eric L.
Logigian, MD; Allen C. Steere, MD; Richard F. Kaplan, PhD; Victor P.
Berardi, AB; Paul H. Duray, MD; Martin G. Larson, ScD; Elizabeth A.
Wright, PhD; Katherine S. Ginsburg, MD, MPH?; Jeffrey N. Katz, MD, MS;
and Matthew H. Liang, MD, MPH
Patient 12 had had high fever, meningeal symptoms, and subsequent
arthritis in 1982. She was noted to have a positive serologic test
result for Lyme disease 4 years later and was treated with 2 weeks of
parenteral penicillin. She later developed a progressive speech
disorder, bradykinesia, and abnormal ocular motor function. Magnetic
resonance imaging of the brain showed scattered white matter lesions
in the hemispheres and pons, and she was diagnosed with supranuclear
palsy. Lumbar puncture showed no selective concentration of antibody
in the spinal fluid. Nevertheless, she was re-treated with 2 weeks of
parenteral ceftriaxone in 1989 that had no effect on her neurologic
symptoms. During the time of observation, this patient died. At
autopsy, lymphoid mononuclear cells were observed surrounding the
intracerebral vessels in one section. Using Dieterle silver stain, a
spirochete was present in the cortex and another was exterior to a
leptomeningeal vessel.
 
PERSISTENCE, KLEMPNER:
J Infect Dis 1992 Aug;166(2):440-4	 

Fibroblasts protect the Lyme disease spirochete, Borrelia burgdorferi,
from ceftriaxone in vitro.

Georgilis K, Pea____e M, Klempner MS.

Department of Medicine, New England Medical Center, Boston,
Massachusetts.

The Lyme disease spirochete, Borrelia burgdorferi, can be recovered
long after initial infection, even from antibiotic-treated patients,
indicating that it resists eradication by host defense mechanisms and
antibiotics. Since B. burgdorferi first infects skin, the possible
protective effect of skin fibroblasts from an antibiotic commonly used
to treat Lyme disease, ceftriaxone, was examined. Human foreskin
fibroblasts protected B. burgdorferi from the lethal action of a 2-day
exposure to ceftriaxone at 1 microgram/mL, 10-20 x MBC. In the absence
of fibroblasts, organisms did not survive. Spirochetes were not
protected from ceftriaxone by glutaraldehyde-fixed fibroblasts or
fibroblast lysate, suggesting that a living cell was required. The
ability of the organism to survive in the presence of fibroblasts was
not related to its infectivity. Fibroblasts protected B. burgdorferi
for at least 14 days of exposure to ceftriaxone. Mouse keratinocytes,
HEp-2 cells, and Vero cells but not Caco-2 cells showed the same
protective effect. Thus, several eukaryotic cell types provide the
Lyme disease spirochete with a protective environment contributing to
its long-term survival.
PMID: 1634816 [PubMed - indexed for MEDLINE]
Later, when Klempner studied the effects of ceftriaxone treatment in
Chronic Lyme patients, he could find only 78 out of about 1700
patients who had still met the Dressler/Steere standard.  The study
was incomplete and invalid for a number of reasons. The only real
result, was not published:  His seronegative patient were very highly
overrepresented in one of the two most frequently observed Multiple
Sclerosis haplotypes, HLA-DQB1*-0602.  Klempner then presented this
data at the NIH Rare diseases Conference on Neuroborreliosis, hosted
by the representatives of the NINDS Multiple Sclerosis group,
including Roland Martin, who found the first MS haplotype in a
neuroborreliosis patient.
Not yet has it been discovered a treatment for borreliosis, but we
have had 12-15 years of CONTROVERSY, which is not a controversy at
all, since Steere himself has published about persisting infection
past treatment on more than one occasion.   It was shown to be a
hazard after all, to be infected with borrelia and then be vaccinated
with the immune suppressing recombinant OspA.  Not necessarily, the
data suggests, does OspA ractivate latent Lyme infection, but perhaps
other latent infections, such as Epstein-Barr.
The way to determine persisting infection, if the method is to look
for spirochetal DNA, is to use code from the CHROMOSOME, because
plasmids are dropped, and not all borrelia are burgdorferi.
 And finally, as part of this CONTROVERSY, it was published two years
after the vaccines trials were concluded, that the method for
assessing the vaccine results, was useless:
http://www.journals.uchicago.edu/CID/journal/issues/v31n1/991200/991200.html
"Since specimen quantities were available, representative study
subjects were also tested by FDA-licensed ELISA and WB kits for
descriptive and comparative purposes, ***making no claim or attempt to
formally validate or invalidate these tests' performances for
vaccinated study subjects.***"
The manufacturer of the only currently FDA-approved (and released)
recombinant OspA Lyme disease vaccine has suggested that vaccination
does not interfere with serological evaluation of Lyme disease in
vaccine recipients [?] ***a statement that is not supported by the
data presented here.***
 From:
Clinical Infectious Diseases    2000;31:42-47
? 2000 by the Infectious Diseases Society of America. All rights
reserved.
1058-4838/2000/3101-0010$03.00
Detection of Multiple Reactive Protein Species by Immunoblotting after
Recombinant Outer Surface Protein A Lyme Disease Vaccination
Philip J. Molloy,1,2 Victor P. Berardi,2 David H. Persing,2,3,a and
Leonard H. Sigal4
1Rheumatology Associates of Southeastern Massachusetts, Plymouth, and
2IMUGEN, Norwood, Massachusetts; 3Department of Laboratory Medicine
and Pathology, Mayo Clinic, Rochester, Minnesota; and 4Division of
Rheumatology, Robert Wood Johnson Medical School, University of
Medicine and Dentistry of New Jersey, New Brunswick
Received 31 August 1999; revised 3 December 1999; electronically
published 17 July 2000.
 
They could not read their results.  They could not read the blots. 
They could not tell if someone who was vaccinated, was infected, or
not.
 Who validated methods?
 Arthur Weinstein:
 For evaluation of Klempner's alleged long term treatment results:
 
Arthritis Care Res 1999 Feb;12(1):42-7	 

The Fibromyalgia Impact Questionnaire: a useful tool in evaluating
patients with post-Lyme disease syndrome.

Fallon J, Bujak DI, Guardino S, Weinstein A.

Leinhard School of Nursing, Pace University, Pleasantville, New York,
USA.

OBJECTIVE: To determine the reliability and validity of a modified
version of the Fibromyalgia Impact Questionnaire (FIQ) in evaluating
patients with post-Lyme disease syndrome (PLDS). ?.The findings
suggest that there may be some differences in the etiopathology of the
symptoms experienced by PLDS and FM patients.  PMID: 10513489 [PubMed
- indexed for MEDLINE]
 THEREFORE, THIS METHOD IS NOT VALID.  It's not valid if it finds a
distinction.
  Arthritis Rheum 1994 Aug;37(8):1206-11	 

Western blot band intensity analysis. Application to the diagnosis of
Lyme arthritis.

Kowal K, Weinstein A.

New York Medical College, Valhalla 10595.

OBJECTIVE. To determine the usefulness of quantitative band-intensity
analysis of Western blots for the diagnosis of Lyme arthritis.
METHODS. IgG Western blots for antibodies to Borrelia burgdorferi were
performed on sera from 39 patients with Lyme arthritis, 30 patients
with syphilis, 50 patients with connective tissue diseases, and 10
healthy individuals. Band positions and band intensities were
calculated using a computerized image analysis system. RESULTS. Lyme
arthritis patients had more bands and higher-intensity bands than did
non-Lyme patients. The presence of at least 2 bands of moderate to
high intensity (> 40 optical units) or at least 5 bands of lower
intensity (> 20 optical units) was over 90% sensitive and 100%
specific for the diagnosis of Lyme arthritis. A 60-kd band was present
in all Lyme arthritis patients. The presence of an 83-, 39-, 21-, or
18-kd band was highly specific for Lyme arthritis. CONCLUSION. Band
intensity analysis increases the objectivity and accuracy