Sue, can we agree that these trials will not proceed until
the "automated assay" is completed?
Did Faustman have this automated assay during the mouse trials?
I am guessing the answer is no.
So, why the need for automation for a small human trial?









--- In nathanfaustmantrials@yahoogroups.com, Sue root
<susan_root@...> wrote:
>
> What will be different in Dr. Faustman and Dr. Nathan's human
clinical trial is that they will be using multiple dosing of BCG in
humans at an increasing scale. Most importantly, they will beable to
measure the effect of the dosing of BCG by her developed human blood
assay which they have been working to automate for the trial.
Another words, as the dosing of BCG increases, they will measure its
effect by being able to count the number of targeted T cells dying.
What is hoped to be expected is a decrease in the number of only the
targeted T cells as the dosing of BCG increases until ultimately
they are depleted. In past human trials using BCG, only one
vaccinated dose of BCG was given without measuring any effect. A
good anology would be using one dose of antibiotics to heal an
infection and dismissing the use of antibiotics because the
infection did not clear up with this single dose. We all know that
it usually requires 10 days worth of antiobiotic dosing to do the
job.
>
> BCG is an immune stimulant. It is known to stimulate the
production of TNF alpha in the body. In the presence of TNF alpha,
the specific defected autoreactive T cells die (see below for
supportive research). This was shown with Dr. Faustman's mice
experiments using multiple doses of a similar stimulant called
Complete Freud's Adjuvant or CFA. Note: Dr. Faustman's lab also used
BCG in the mice and had the same effect. This past year, multiple
independent labs successfully replicated her mice experiments.
> Dr. Faustman is attempting to use BCG in the human clinical
trial to target and eliminate one of the 2 similar defects found in
NOD mice and humans with diabetes.
> Hope this helps.
>
>
> Here are some experiments to support the effect and use of BCG:
>
> In vivo apoptosis of diabetogenic T cells in NOD mice by IFN-c/TNF-
a
>
> International Immunology 2004; 1 of 10
>
> Hui-Yu Qin, Pratibha Chaturvedi and Bhagirath Singh
>
> Department of Microbiology and Immunology, University of Western
Ontario, Robarts Research Institute and Canadian
>
> Institutes of Health Research, Institute of Infection & Immunity,
London, Ontario, Canada
> Our results suggest that BCG down-regulates destructive
autoimmunity by TNF-a/IFN-c-induced apoptosis of diabetogenic T
cells through both Fas and TNF pathways. These studies provide a
novel mechanism for blocking disease recurrence and immune
modulating effect of BCG immunization in type 1 diabetes.
>
> Apoptosis of autoreactive CD8 lymphocytes as a potential
mechanism for the abrogation of type 1 diabetes by islet-specific
TNF-alpha expression at a time when the autoimmune process is
already ongoing.
> Ann N Y Acad Sci 2002 Apr;958:166-9 (ISSN: 0077-8923)
> Christen U; Von Herrath MG Departments of Neuropharmacology and
Immunology, The Scripps Research Institute, La Jolla, California
92037, USA. christen@...
> Most interestingly, late expression of TNF-alpha resulted in a
reversion of mice that were already diabetic to a nondiabetic state.
Here we provide a model of how experienced autoaggressive CD8
lymphocytes are dying by apoptosis as a result of beta cell-specific
TNF-alpha expression at a time when the autoimmune process is
already ongoing.
> Repeated BCG vaccination is more effective than a single dose in
preventing diabetes in non-obese diabetic (NOD) mice.
> Isr J Med Sci 1997 Nov;33(11):711-5
> Shehadeh N; Etzioni A; Cahana A; Teninboum G; Gorodetsky B;
Barzilai D; Karnieli E
> Juvenile Diabetes Unit, Rambam Medical Center, Haifa, Israel.
glass@...
> Our report demonstrates that repeated BCG vaccination is safe
and more effective than a single dose in preventing type I diabetes
in NOD mice. This data should be considered in planning new human
trials with BCG.
> Effect of adjuvant therapy on development of diabetes in mouse
and man Lancet 1994 Mar 19;343(8899):706-7
> Shehadeh N; Calcinaro F; Bradley BJ; Bruchlim I; Vardi P;
Lafferty KJ
> Rambam Medical Centre, Juvenile Diabetes Centre, Halfa, Israel. In
a preliminary trial in 17 newly diagnosed, type 1 diabetic patients,
intracutaneous administration of 0.1 mL of BCG 1 mg/mL led to
clinical remission in 11 (65%)--by week 4 in 6. Remission has been
sustained in 3 for 6-10 months. No side-effects were reported. A
double-blind trial of BCG is warranted.
>
>
> Scott Strumello <sstrumello@...> wrote: In the most
recent edition of the New England Journal of Medicine
> (NEJM), a letter from Dr. Faustman written in response to a letter
to
> the NEJM from Douglas Melton in July 2006 was published. The text
of
> both these letters is included at the end of this message for
> reference.
>
> I was hoping someone could respond with a better explanation than
we
> have received so far. While Dr. Faustman's response addresses most
> of the elements Mr. Melton's letter raised, I did find one
citation
> troubling, mainly because Dr. Faustman has never completely
explained
> why she and her team believes that the use of bacille Calmette-
Guérin
> (BCG) will have the desired effect of arresting the autoimmune
> attack. She has written and stated that earlier trials (the one
> cited was published in 2005) occurred before they knew exactly how
> BCG worked, but I'm afraid that explanation still does not clarify
> what will be different about how they are using this drug, or why
> they are convinced it will have the same effect as Freund's
adjuvant
> had in the NOD mouse. Will they be doing something different in
> terms of dosage, frequency of dosing, etc.?
>
> The quote I am referring to from Mr. Melton's letter is as follows
> (the full article follows):
>
> (Unfortunately, controlled studies have shown that neither the
> complete Freund's adjuvant nor the bacille Calmette-Guérin
adjuvant
> appears to have this effect in humans.8)
>
> Citation:
> 8. Huppmann M, Baumgarten A, Ziegler A, Bonifacio E. Neonatal
bacille
> Calmette-Guerin vaccination and type 1 diabetes. Diabetes Care
> 2005;28:1204-6.
>
> There may certainly be convincing evidence to support the reason
they
> believe that BCG will accomplish this, but I'd like for the
Faustman
> lab to explain in more detail why they believe it will do so!!
>
> Regards!
>
> Reversal of Type 1 Diabetes in Mice
> [Correspondence]
> Faustman, Denise L.
>
> Harvard Medical School, Boston, MA 02115
>
> To the Editor: Dr. Melton (July 6 issue) [1] misrepresents my
study
> on the reversal of type 1 diabetes in mice and its implications
for
> new treatment strategies in humans. [2] As in a previous study,
[3]
> my colleagues and I discovered that an immune therapy triggered a
> permanent reversal of end-stage type 1 diabetes in mice. The
> treatment involved two components: injecting the mice with an
immune
> adjuvant (to induce the production of tumor necrosis factor, which
> destroys autoreactive T cells) and injecting splenocytes from a
donor
> mouse.
>
> Dr. Melton writes that we ascribed the reversal of type 1 diabetes
in
> a mouse model solely to the transplantation of spleen cells and
that
> we claimed to have identified a stem cell among the donor
splenocytes
> that contributed to regrowth of the islets in the recipient. It is
> true that we observed adult stem cells and that these cells can
> contribute, in part, to the regrowth of the islets. However, Dr.
> Melton does not acknowledge that we also observed regeneration of
> pancreatic islets and complete reversal of type 1 diabetes without
> the introduction of any live donor splenocytes. [2] Infusion of
live
> splenic cells hastened the development of permanent normoglycemia
in
> the mice but did not enhance the rate of cure. We did not claim
that
> the regenerative process required a stem cell, and we did not rule
> out other mechanisms, such as regrowth or rescue of host islets.
Our
> research simply found regeneration of the pancreas once the
> autoimmune process was removed.
>
> Instead of cheering the fact that our laboratory's
immunomodulatory
> approach was replicated successfully by three recent studies, [4-
6]
> Dr. Melton places emphasis on the failure of these cited studies
to
> identify a splenocyte contribution to the observed regeneration of
> the pancreas. It is possible that methodologic differences between
> our protocol and theirs precluded finding a contribution of
splenic
> stem cells to pancreatic regeneration in these studies. But since
> then, the optional splenic contribution has been replicated. [7]
>
> From a clinical perspective, the existence of an adult stem cell
in
> the spleen seems to be beside the point. Many studies have since
> shown that the regenerative process in the pancreas is likely to
be
> intact and that targeted immune intervention may unleash the
> spontaneous regeneration of the pancreas. It seems reasonable to
test
> the hypothesis that for end-stage diabetes, an immune intervention
> that destroys autoreactive T cells in the mouse can also work in
the
> clinic.
>
> Dr. Faustman reports owning stock in General Electric, Pfizer,
> Microsoft, IBM, Keel, and Johnson & Johnson, all of which have
> research programs or products involving stem cells. Dr. Faustman's
> employer, Massachusetts General Hospital, owns patent applications
on
> the nuclear factor-(kappa)B-tumor necrosis factor pathway for the
> treatment of autoimmunity. Should the hospital receive income from
> those applications, Dr. Faustman or her laboratory could receive
> income.
>
> Denise L. Faustman, M.D., Ph.D.
>
> Harvard Medical School, Boston, MA 02115
>
> REFERENCES
>
> 1. Melton DA. Reversal of type 1 diabetes in mice. N Engl J Med
> 2006;355:89-90.
>
> 2. Kodama S, Kuhtreiber W, Fujimura S, Dale EA, Faustman DL. Islet
> regeneration during the reversal of autoimmune diabetes in NOD
mice.
> Science 2003;302:1223-7.
>
> 3. Ryu S, Kodama S, Ryu K, Schoenfeld DA, Faustman DL. Reversal of
> established autoimmune diabetes by restoration of endogenous beta
> cell function. J Clin Invest 2001;108:63-72.
>
> 4. Nishio J, Gaglia JL, Turvey SE, Campbell C, Benoist C, Mathis
D.
> Islet recovery and reversal of murine type 1 diabetes in the
absence
> of any infused spleen cell contribution. Science 2006;311:1775-8.
>
> 5. Suri A, Calderon B, Esparza TJ, Frederick K, Bittner P, Unanue
ER.
> Immunological reversal of autoimmune diabetes without
hematopoietic
> replacement of beta cells. Science 2006;311:1778-80.
>
> 6. Chong AS, Shen J, Tao J, et al. Reversal of diabetes in non-
obese
> diabetic mice without spleen cell-derived beta cell regeneration.
> Science 2006;311:1774-5.
>
> 7. Faustman DL, Tran SD, Kodama S, et al. Comment on papers by
Chong
> et al., Nishio et al., and Suri et al. on diabetes reversal in NOD
> mice. Science 2006;314:1243.
>
> Reversal of Type 1 Diabetes in Mice
> Douglas A. Melton, Ph.D.
>
> In 2003, the field of diabetes was shaken up by an article
published
> in Science,1 stating that established autoimmune diabetes in mice
> could be permanently reversed with the injection of spleen cells.
> Kodama et al. reported that a known immune modulator, complete
> Freund's adjuvant, and the temporary transplantation of islets to
the
> kidney capsule could be coupled with the injection of splenocytes
to
> cure diabetes in mice. The idea was that the adjuvant modulates
the
> immune attack and the transplanted islets maintain normal
> blood glucose levels long enough for the spleen cells to
regenerate
> the insulin-producing beta cells. The authors described the
approach
> as follows:
>
> "Stem cells of the spleen have been demonstrated to home to the
> pancreas where they mature into fully functional islet cells
> responsible for restoring normoglycemia."2 This article was
important
> because of its claim that readily available cells, spleen cells,
> could be used to produce pancreatic
> beta cells.
>
> The report attracted considerable attention because it pointed to
new
> clinical strategies for treating type 1 diabetes. In addition, the
> use of spleen cells avoided the ethical questions surrounding the
use
> of embryonic stem cells as a
> possible route to the production of beta cells. At the same time,
> some scientists were skeptical of the claim that spleen cells
could
> form pancreatic beta cells. To their credit, Kodama et al.
provided
> detailed protocols so that others could repeat and independently
> verify their conclusions.
>
> Recently, the results of three independent efforts to replicate
the
> experiment were reported, again in Science.3-5 The results were
> remarkably consistent: no splenocyte contribution to the islets
was
> observed, and no evidence was found to support the principal
> conclusion of Kodama et al.
> All three studies showed, as others had previously,6,7 that
diabetes
> can be reversed in this mouse model. However, the recovered host
beta
> cells, rather than spleen cells, were responsible for this
reversal.
> Each of the three studies supports the conclusion that the
adjuvant-
> dependent dampening of the autoimmune attack, coupled with the
> recovery of residual host islets, underlies the cure in mice.
>
> What does all this mean for possible clinical treatment? It is
> noteworthy that in these studies, the progression of autoimmune
> diabetes was reversed in control mice receiving the adjuvant.
> (Unfortunately, controlled studies have shown that neither the
> complete Freund's adjuvant nor the bacille Calmette-
> Guérin adjuvant appears to have this effect in humans.8) Although
a
> mechanistic understanding of the autoimmune reversal in mice due
to
> the adjuvant is still lacking, the finding emphasizes the
importance
> of identifying the initiating antigen or antigens and the
subsequent
> cascade of immune T and B cells responsible for the autoimmune
> attack. This approach is the best way to find interventions that
> can be effectively designed and applied. The value of this
approach
> is supported by the promising results of early-stage clinical
trials
> wherein other modulators of the immune system - for example, the
anti-
> CD3 antibody9 - are administered during the "honeymoon" period of
> type 1 diabetes, when there is still considerable betacell mass.
>
> The three recent studies also illustrate the dramatic response of
> beta cells to environmental signals. In each study, the beta cells
> recovered from an immune attack and proliferated to restore beta-
cell
> mass. Beta-cell mass has been shown to increase in several other
> circumstances, including pregnancy, obesity, and in some patients,
> after gastric bypass surgery. Mice in which the insulin receptor
in
> the liver cells has been knocked out respond by increasing beta-
cell
> mass by a factor of 10.10 If we can harness this endogenous
capacity
> of beta cells to proliferate and can combine this ability with a
more
> effective blunting of the autoimmune attack in humans, it may well
be
> possible to devise important new treatments for type 1 diabetes.
>
> No potential conflict of interest relevant to this article was
> reported.
>
> From the Harvard Stem Cell Institute, Department of Molecular
> and Cellular Biology, Harvard University, Boston.
>
> REFERENCES
>
> 1. Kodama S, Kuhtreiber W, Fujimura S, Dale EA, Faustman DL.
> Islet regeneration during the reversal of autoimmune diabetes in
NOD
> mice. Science 2003;302:1223-7.
>
> 2. Kodama S, Faustman DL. Routes to regenerating islet cells:
> stem cells and other biological therapies for type 1 diabetes.
> Pediatr Diabetes 2004;5:Suppl 2:38-44.
>
> 3. Chong AS, Shen J, Tao J, et al. Reversal of diabetes in
nonobese
> diabetic mice without spleen cell-derived beta cell regeneration.
> Science 2006;311:1774-5.
>
> 4. Nishio J, Gaglia JL, Turvey SE, Campbell C, Benoist C, Mathis
D.
> Islet recovery and reversal of murine type 1 diabetes in the
absence
> of any infused spleen cell contribution. Science 2006;311:1775-8.
>
> 5. Suri A, Calderon B, Esparza TJ, Frederick K, Bittner P, Unanue
ER.
> Immunological reversal of autoimmune diabetes without
hematopoietic
> replacement of beta cells. Science 2006;311:1778-80.
>
> 6. Sadelain MW, Qin HY, Lauzon J, Singh B. Prevention of type I
> diabetes in NOD mice by adjuvant immunotherapy. Diabetes
> 1990;39:583-9.
>
> 7. Wang T, Singh B, Warnock GL, Rajotte RV. Prevention of
> recurrence of IDDM in islet-transplanted diabetic NOD mice by
> adjuvant immunotherapy. Diabetes 1992;41:114-7.
>
> 8. Huppmann M, Baumgarten A, Ziegler A, Bonifacio E. Neonatal
> bacille Calmette-Guerin vaccination and type 1 diabetes.
> Diabetes Care 2005;28:1204-6.
>
> 9. Herold KC, Hagopian W, Auger JA, et al. Anti-CD3 monoclonal
> antibody in new-onset type 1 diabetes mellitus. N Engl J Med
> 2002;346:1692-8.
>
> 10. Kulkarni RN, Jhala US, Winnay JN, Krajewski S, Montminy
> M, Kahn CR. PDX-1 haploinsufficiency limits the compensatory
> islet hyperplasia that occurs in response to insulin resistance. J
> Clin Invest 2004;114:828-36.
>