THE FAUCI FILES, 3( 86): Does Fauci's IL-2 Patent Prevail Over The Nuremberg Code?

Re: NIAID News Release: B CELLS MAY HELP MAINTAIN HIV INFECTION

August 29, 2000

Does Fauci's IL-2 Patent Prevail Over The Nuremberg Code?

Today's NIAID News announcement confirms the detrimental
effects of overactive antibody responses (non-Tcl) in
HIV disease and cancer. But what about the research
which has shown recombinant human IL-2 (rhIL-2) to
be a B-cell growth factor that utilizes IL-2 surface
receptors similar to those on activated T cells?
(Emmrich et al., 1985; Mingari et al., 1984)

NIAID IL-2 Trials

After more than a decade of dozens of NIAID-sponsored trials of
interleukin-2 as a treatment for HIV/AIDS, it was discovered
in the early trials that people with AIDS progressed faster
to disease and death. Undaunted by any nagging suspicions
of how IL-2 may have been a cause of AIDS progression,
the IL-2 NIAID trials took a different approach, choosing to
test IL-2 on healthy HIV+ individuals who did not have
symptomatic disease (AIDS). Despite no proof of efficacy
or improved survival, the Phase III IL-2 trials continue today.

IL-2 Favors B-cells Not Tcl T-cells

In HIV disease and cancer vaccines, IFN-gamma-producing memory/effector
CD8 CTLs (Tc1) are considered to have beneficial effects, while
overactive antibody responses (non-Tc1) are considered to have
detrimental effects. Yet IL-2 favors the B-cells, not the Tcl
which use IFN-gamma and not IL-2 (Brinchmann et al., 1998;
Caruso et al., 1998).

NIAID: B Cells Are Bad

In the NIAID News release "B CELLS MAY HELP MAINTAIN HIV INFECTION",
Aug 29, 2000, the following comments by NIAID Director Dr.
Anthony Fauci directly conflict with Fauci's patent for IL-2
"intermittent therapy", which, according to the most recent
reports, has moved to Phase III HIV trials at NIAID.

Here's what Fauci says today, on August 29, 2000:

  "This study enhances our understanding of how HIV
   persists in the body and might partly explain the
   abnormalities seen in B-cell function in people
   with HIV infection,"

  "Identifying this pool of HIV-carrying cells also
   opens new avenues for treating the infection."

      Anthony S. Fauci, M.D.
      NIAID director and chief of the laboratory
      where the research took place

Given Fauci's "enhanced understanding" about B-cells now,
let's take a look at what Fauci published 15 years ago
as senior author (Bich-Thuy et al., 1985):

   "Taken together, these data clearly demonstrate that IL-2
   can induce human B-cell proliferation in vitro. This
   proliferation is not due to T cells themselves nor to
   T-cell help or T-cell helper factors, although its magnitude
   can be increased in the presence of a sufficient proportion
   of T cells. Therefore, it is concluded that IL-2 can directly
   trigger human peripheral blood B cells to proliferate. Such
   findings have important potential implications in our
   appreciation of the broad spectrum of the effects of
   T-cell-derived factors on B-cell function."

In 1985, IL-2 patent co-inventors Fauci and Lane, were fully
aware of the harm resulting from polyclonal B-cell activation:

   "This polyclonal B-cell activation has been extensively
   characterized and is manifested by elevated levels of Ig and
   circulating immune complexes, an inability to mount an
   antigen-specific antibody response following immunization,
   elevated numbers of spontaneous Ig-secreting cells in the
   peripheral blood, enhanced responsiveness to B-cell growth
   factors, and refractoriness to the normal in vitro signals
   for B-cell activation."


If IL-2 Worsens B-Cell Harm, IL-2 = Food for Virus

In repeating Fauci's words on August 29, 2000, doesn't the notion
of "treating the infection" include doing no harm, as in avoiding
treatments like IL-2 which can only favor the disease
and worsen the infection, indeed as "food for virus"?

  "Identifying this pool of HIV-carrying cells also
   opens new avenues for treating the infection."


Will Fauci call an end to the IL-2 NIAID trials, IL-2 vaccines,
or IL-2 therapy for disease states requiring the vital increase in
Tcl responses and decreases in antibody responses?

Or does the Nuremberg Code take a backseat to an IL-2 patent?



W. Fred Shaw, Editor
THE FAUCI FILES

----------------------------------------------------------------

U.S. Patent Office

5696079 :    Immunologic enhancement with intermittent
             interleukin-2 therapy

INVENTORS:   Lane; H. Clifford, Bethesda, MD
             Kovacs; Joseph A., Potomac, MD
             Fauci; Anthony S., Washington, DC

ASSIGNEES:   The United States of America as represented by
             the Department of Health and Human Services,
             Washington, DC

ISSUED:      Dec. 9 , 1997
FILED:       May 26, 1995

SERIAL NUMBER: 452440

----------------

On Tue, 29 Aug 2000 09:59:28 -0400, in <8ogfnc$f81@icet.net.nih.gov>
"Greg " <gfolkers@nih.gov>    in misc.health.aids wrote:

>NIH NEWS
>******************************************************
>National Institute of Allergy and Infectious Diseases
>National Institutes of Health
>
>FOR IMMEDIATE RELEASE
>Tuesday, Aug. 29, 2000
>
>Contact: Sam Perdue
>(301) 402-1663
>sp189u@nih.gov
>
>B CELLS MAY HELP MAINTAIN HIV INFECTION
>
>The human immunodeficiency virus (HIV) devastates the body's ability to
>fight off infection by destroying a key class of T cells essential for
>maintaining a vigorous immune response.  Now, scientists from the National
>Institute of Allergy and Infectious Diseases (NIAID) report for the first
>time that B cells-the antibody-producing cells of the immune system-help
>ferry HIV throughout the blood and can likely deliver the virus to nearby T
>cells.  This discovery, reported in the September 4 issue of the Journal of
>Experimental Medicine, helps explain several phenomena associated with HIV
>infection and paves the way for new approaches to eliminating the virus from
>the blood.
>
>"This study enhances our understanding of how HIV persists in the body and
>might partly explain the abnormalities seen in B-cell function in people
>with HIV infection," says Anthony S. Fauci, M.D., NIAID director and chief
>of the laboratory where the research took place. "Identifying this pool of
>HIV-carrying cells also opens new avenues for treating the infection."
>
>The research, conducted by Susan Moir, Ph.D., Angela Malaspina, Ph.D., and
>colleagues from NIAID's Laboratory of Immunoregulation, follows reports from
>several laboratories that HIV can infect B cells in the test tube and that
>low levels of HIV genetic material can exist in B cells of HIV-infected
>individuals.  None of these reports, however, have identified viable HIV
>associated with the B cells of infected individuals.
>
>To address this concern, the researchers isolated B cells from the blood of
>people with chronic HIV infection.  When they examined these cells for the
>presence of HIV, they found significant levels of the virus attached to the
>surface of the cells.  The scientists then used test-tube experiments to
>show the virus could infect T cells under laboratory conditions.
>
>Because of the close interaction between B and T cells in the immune system,
>this discovery casts new light on how HIV can interact with T cells.  The
>two cell types constantly form temporary attachments with each other to
>exchange information and coordinate the immune response.  This gives the B
>cells ample opportunity to pass HIV to previously uninfected T cells.  Dr.
>Moir explains that B cells are not a hidden reservoir of HIV, however,
>because they do not house internalized, replicating virus, and the amount of
>B-cell-bound virus decreases as HIV levels decline in the blood.  "HIV does
>not appear to reproduce inside B cells, but rather hitches a ride on the
>cell surface so it is free to jump to nearby T cells."
>
>The studies also might explain why B-cell-mediated immunity in individuals
>with chronic HIV infections can go awry.  "People with high levels of HIV in
>the blood often have malfunctions in their B cell responses, such as
>uncontrolled activation of antibody production," explains Dr. Moir.   "In the
>past, scientists have thought this was caused largely by indirect effects of
>HIV infection, but now we have evidence that some B cell abnormalities might
>be due to direct viral interference."
>
>Dr. Moir and co-workers looked at the B cells carefully to determine how the
>virus attached to the cells.  They identified the docking point for HIV, a
>molecule called CD21.  This protein appears on the surface of B cells and
>normally binds complement, a molecule that tags invading microbes and
>targets them for destruction.  The researchers discovered that HIV, when
>attached to complement, could use CD21 as a binding site on the B cell.
>Because complement normally "tickles" CD21, thereby signaling B cells to
>produce antibodies, the scientists believe the uncontrolled production of
>multiple antibodies in people with HIV might be caused by the repeated
>stimulation of the B cell as the virus binds CD21.
>
>Now that the researchers have shown how B cells might play a role in HIV
>infection, they are testing to see if the HIV in infected T cells is
>genetically related to that on the B cells.  They are also pursuing more
>studies on how HIV might directly cause deficiencies in B-cell function.
>"Scientists haven't looked at B cells much during HIV infection," says Dr.
>Moir, "This research opens a new opportunity for better understanding the
>complex nature of the disease."
>
>Scientists from the National Cancer Institute and Advanced BioScience
>Laboratories, Inc., Kensington, Maryland, also participated in this study.
>
>NIAID is a component of the National Institutes of Health (NIH).  NIAID
>conducts and supports research to prevent, diagnose and treat illnesses such
>as HIV disease and other sexually transmitted diseases, tuberculosis,
>malaria, asthma and allergies.  NIH is an agency of the U.S. Department of
>Health and Human Services.
>                                         ###
>Reference:
>S Moir, et al. B cells of HIV-1-infected patients bind virions through
>CD21-complement interactions and transmit infectious virus to activated T
>cells. J Exp Med 192(5):637-45 (2000).  This article is available online at
>http://www.jem.org/cgi/content/full/192/5/637.
>
>Press releases, fact sheets and other NIAID-related materials are available
>on the NIAID Web site at http://www.niaid.nih.gov.

===================================

Bich-Thuy LT, Lane HC, Fauci AS. Human B-cell proliferation in
response to recombinant interleukin 2 is not due to T-cell help. Cell
Immunol 1985 Sep;94(2):353-9. 

Abstract: Positively selected human B-cell suspensions with no
detectable T cells and containing more than 99.5% B cells both at the
initiation and termination of culture were shown to proliferate in
response to interleukin 2 (IL-2) in a dose-dependent fashion. The lack
of influence of residual T cells on this proliferative response was
demonstrated in experiments where T cells were added back in increasing
numbers to B-cell suspensions. No detectable enhancing effect on B-cell
proliferation was noted when 2.5% T cells were purposely added back to
culture, a proportion far in excess of that which might be expected to
contaminate B-cell suspensions under the present methodology. In
contrast, when 10% T cells were added back to B-cell cultures, an
enhanced proliferation of B cells was observed suggesting that the lack
of effect of lower numbers of T cells was due to their inefficiency in
helping B-cell proliferation in response to IL-2. Therefore, it is
concluded that highly purified IL-2 is capable of triggering human
peripheral blood B cells to proliferate and that this proliferation is
not due to T-cell help.

===================================

Brinchmann JE, Rosok BI, Spurkland A. Activation and proliferation of
CD8+ T cells in lymphoid tissues of HIV-1-infected individuals in the
absence of the high-affinity IL-2 receptor. J Acquir Immune Defic Syndr
Hum Retrovirol 1998 Dec 1;19(4):332-8

Institute of Transplantation Immunology, Rikshospitalet, Oslo, Norway.
j.e.brinchmann@rh.uio.no

Abstract: Activation of CD8+ T cells may have important pathogenic
implications in HIV-1 infection. Studies of this process have so far
been confined to cells from the peripheral blood. In the present study,
we have examined molecules involved in activation and proliferation of
CD8+ T cells in lymphoid tissues from HIV-1-infected patients. Tonsillar
tissue and blood samples from 13 HIV-1-infected patients and 6
seronegative controls were examined for cell surface expression and the
presence of mRNA for CD69, CD25, and HLA-DR. Intonsillar tissue, the
number of CD8+ T cells was increased several fold in HIV-1-infected
patients compared with controls. The majority of these cells expressed
CD69 and HLA-DR, but virtually no tonsillar CD8+ T cells were found to
express CD25 on the cell surface or at the mRNA level. Following in
vitro activation, however, almost all activated CD8+ T cells were found
to express CD25. Tonsillar CD4+ T cell numbers were maintained or
reduced compared with controls, and a considerable proportion expressed
CD25. The data suggest that CD8+, but not CD4+ T cells proliferate
extensively in lymphoid tissues in HIV-1-infected patients in the
absence of the high-affinity interleukin-2 (IL-2) receptor.

===================================

Caruso A, Licenziati S, Morelli D, Fiorentini S, Ricotta D, Malacarne F,
Sfondrini L, Balsari A. Segregation of type 1 cytokine production in
human peripheral blood lymphocytes: phenotypic differences between
IFN-gamma and IL-2-producing cells in the CD8+ T cell subset. Eur J
Immunol 1998 Nov;28(11):3630-8. 

Institute of Microbiology, University of Brescia, Italy.

Abstract: T cell clones are classified as type 0, 1 or 2 depending on
the lymphokines they produce. However, it has remained unclear whether
single cells of a given type produce one or several cytokine species.
Flow cytometric analysis of peripheral blood lymphocytes (PBL) obtained
from 20 healthy donors for the production of the type 1 cytokines
IFN-gamma and IL-2 revealed very few cells that co-expressed both
cytokines independently of the mitogenic stimulus used for PBL
activation. Similarly, kinetic studies of cytokine synthesis
indicated a low percentage of IFN-gamma/IL-2 double-positive T
cells at all time points. Reverse transcription-PCR analysis of sorted
IL-2- and IFN-gamma-positive T cells showed the presence of IL-2- or
IFN-gamma-specific mRNA only in those cells expressing the corresponding
cytokine. This segregation of the two type 1 cytokines was lost in
long-term cultured T cells and in T cell clones. A high percentage of
cells expressing only IL-2 or IFN-gamma was observed even when the
production of these cytokines was evaluated on CD4- and CD8+ subsets.
Moreover, in some healthy individuals, IFN-gamma and IL-2 production by
CD8+ T cells was related to CD8+ expression levels and cell size, i. e.
IL-2-expressing cells were generally smaller with more intense CD8+
staining as compared with IFN-gamma-producing T cells. These data
indicate that activated T lymphocytes are strongly committed in vivo to
produce IFN-gamma or IL-2 and emphasizes the independent regulation of
the two cytokine genes.

===================================

Emmrich F, Moll H, Simon MM. Recombinant human interleukin 2 acts as a B
cell growth and differentiation promoting factor. Immunobiology 1985
Feb;169(1):97-102. 

Abstract: Human B cells appropriately activated by a B cell mitogen are
rendered susceptible to human Interleukin 2 (IL-2) as demonstrated with
recombinant human IL-2 (rec. h IL-2). They show increased proliferation
and drastically enhanced immunoglobulin secretion. Susceptibility to IL-
2 is accompanied with the expression of the IL-2 receptor (Tac antigen)
on B cells. The data suggest that IL-2 is one of the lymphokines
directly involved in the activation of B lymphocytes.

===================================

Lane HC, Fauci AS. Immunologic abnormalities in the acquired
immunodeficiency syndrome. Annu Rev Immunol 1985;3:477-500.

Abstract: The immune systems of patients with AIDS are characterized by
a profound defect in cell-mediated immunity which is predominantly due
to a decrease in the number and function of the helper/inducer T
lymphocytes, particularly the antigen-reactive cells. This defect is
manifested primarily as decreases in delayed-type hypersensitivity
reactions and decreased in vitro proliferation to soluble antigen. A
variety of secondary manifestations of immunologic dysfunction occur,
some of which result from a lack of effective inducer-cell function,
others from the occurrence of opportunistic infections. Among these
secondary phenomena are decreased cytotoxic lymphocyte responses,
polyclonal B-cell activation, decreased monocyte chemotaxis, and a
number of serologic abnormalities. The primary cause of this defect in
the antigen-reactive helper/inducer T lymphocyte is infection with a
class of T-cell tropic retroviruses known as HTLV-III or LAV. This virus
is capable of selectively infecting T4 + lymphocytes and can be isolated
consistently from patients with AIDS or AIDS-related conditions. Despite
substantial knowledge concerning the nature of the immune defect in AIDS
and its causative agent, little progress has been made in developing
effective therapies for this uniformly fatal illness. Because the
incidence of this disease continues to increase, and patients stricken
with this illness have a median survival of two years, additional
investigation in this area is greatly needed. Continued effort aimed at
delineating the precise nature of the immune defect in these patients
should be of value in attempting to enhance our understanding of the
human immune system in both normal and disease states.

===================================

Mingari MC, Gerosa F, Carra G, Accolla RS, Moretta A, Zubler RH,
Waldmann TA, Moretta L. Human interleukin-2 promotes proliferation of
activated B cells via surface receptors similar to those of activated T
cells. Nature 1984 Dec 13-19;312(5995):641-3. 

Abstract: Human interleukin-2 (IL-2) is a glycoprotein of relative
molecular mass (Mr) 15,000, which is released by T lymphocytes on
stimulation with antigen or mitogen and functions as a T-cell growth
factor (TCGF) by inducing proliferation of activated T cells. It is
generally accepted that resting or activated B cells do not respond
directly to IL-2 but require for their proliferation other T-cell-
derived lymphokines usually referred to as B-cell growth factors
(BCGFs). Recently, however, a monoclonal antibody reacting with the IL-2
receptor molecules expressed by activated T cells (anti-Tac) was shown
to react also with certain B tumour cells; in addition, murine B cells
proliferate in response to pure human IL-2. We now show that recombinant
IL-2, derived from Escherichia coli expressing the human gene, is able
to promote strong proliferation of human B cells activated with protein-
A-rich Staphylococcus aureus Cowans strain I. Moreover, we demonstrate
that the anti-Tac antibody also reacts with S. aureus-activated normal B
cells and inhibits sharply the proliferative response of such cells to
IL-2. Finally, immunoprecipitation experiments reveal that anti-Tac
defines similar molecules on activated T and B cells.

=== end