Note: Descriptions are shown in the official language in which they were submitted.
~~~'~499
TREATMENT METHODS AND VACCINES FOR
STIMULATING AN IMMUNE RESPONSE
The present invention relates to the field of
immunology, particularly methods for inactivating and/or
attenuating viruses and/or virus infected cells, particularly
virus infected CD4 cells, using photopheresis and employing said
inactivated and/or attenuated viruses and/or virus infected cells
to engender an immune~response. A particular aspect of the
invention relates to the treatment of patients who are infected
with a virus, particularly an HIV retrovirus, and who have an
abnormally low white blood cell count, by using photopheresis in
combination with the administration of a photoactive compound
such as 8-methoxypsoralen. The invention also relates to
vaccines against viruses, particularly HIV retroviruses, and
methods for producing said vaccines.
1
~~C3'~49~
There are many debilitating intectioua diseases which
have been attributed to viral infections. There are also many
different types of viruses including DNA viruses and RNA viruses.
Retroviruse fore a sub-group of RNA viruses which, in
order to replicate, must first employ reverse transcription of
the RNA of their genome into DNA ("transcription" conventionally
describes the synthesis of RNA from DNA). once in the form of
DNA, the viral genome is incorporated into the host cell genome,
allowing it to take full advantage of the host cells
transcription/translation machinery for the purpose of
replication. once incorporated into the host cells DN~1, the
virus may persist for as long a~ the cell live.
Certain retroviruse are known to cause a depression in
an infected patients white blood cell count. This sub-set of
ratroviruses which reduce white blood cell count are known as
Human Immunodeficiency Viruses (HIV).
Particular species of HIV retroviruses have been
isolated from patients who suffer from Acquired Immune Deficiency
Syndrome (AIDS) and have been given the designations HIV 1 and
HIV 2, sometimes collectively referred to as "HTLV III", "LAV" or
simply "HIV". These retroviruses will infect cells expressing
the CD4 marker, such as human T-lymphocytes and monocytes. These
calls are involved in the functioning of the immune system. This
infection, in turn, results in the progressive loss of the CD4 T-
2
i
~~C~'~4J~
cell population and disturbs the function of other CD4 cells,
such as aonoeyt~~ hereby reducing the patients ability to
combat other infections, and predisposing the patient to
opportunistic infections which frequently prove fatal.
There are at least three clinical manifestations of
AIDS infection. In the initial "carries state, the only
indication of infection is either the presence of IiIV antibodies
in the blood-stream or the ability to culture the~virus. The
next stage is known as MAIDS related complex~ (ARC) and the
physical symptom associated therewith may include persistent
general lymphadenopathy, general malaise, increased temperature
and chronic infections. This condition usually progresses to the
final, fatal AIDS condition, when the patient loses the ability
to tight infection.
A particularly troublesome problem associated with
combatting HIV retrovirus infections is that the RNA to DNA
reverse transcription process is fraught with repeated mutation
which makes it extremely difficult for the body~s immune system
to recognize and attack infected cells along with the virus
itself.
It is therefore, an object of this invention to provide
a method of treating virus infections in patients having an
abnormally low white blood cell count.
It is a further object of the invention to provide a
method of treating patients infected with an HIV retrovirus.
A still further object of the invention is to provide a
3
vaccine against virus intoctiona, particularly wh~rt the inl.ction
is causb by a ratrovirua such as an tilt/ r~trovirua.
Others objects o! the invention will b~ apparent from
the d~acription o! the invention which follows.
4
~~~''~4~~
In accordance with the present invention a method has
been found for treating patients who are infected with a virus,
particularly an ~IIV retrovirus, using a photoactive compound
that binds, in the case of a virus infected cell, to the cell
membrane (e. g., by binding to a receptor and/or a nucleic acid
fragment on the cell membrane) and/or to nucleic acid in the cell
nucleus or cell cytoplasm, or, in the case of either free virus
or cell associated virus, that binds to the virus surface (e. g.,
to a receptor and/or to a nucleic acid tragaent on the virus
surface) and/or to nucleic acid (e.g. , DNlI or RNI~) which is
incorporated in the virus, upon activation by exposure to
electromagnetic radiation of a prescribed spectrum, such as
ultraviolet light, for the purpose of inactivating and/or
attenuating the virus and permitting the so treated virus and/or
virus infected cells to be presented to the immune system of the
patient for the purpose of engendering an immune response to the
viral infection. Psoralen compounds are particularly preferred
for this purpose, especially the compound 8-methoxypsoralen -- in
which, case WA radiation is preferred for activating said
compound.
An especially important feature of the present
invention is that the treatment methods described herein are used
in patients having an abnormally low number of circulating
5
~~~'~4~~
lymphocytes or white blood cells. In addition, it has been found
by the imrentors that the treatment methods according to the
invention can ~~ used to reconstitute the immune aystam~s
!unction in such patients. This is of tremendous importance in
the treatment of ARC/AIDS patients who, in addition to having an
HIV infection (which itself has heretofore been extremely
ditticult to treat), also have a depruead immune syetea. such
patients generally cannot tolerate a reduction in the number e!
their~lymphocytes or white blood cells without succumbing to
opportunistic infections.
It has thus been found by the inventors that the
treatment methods in accordance with the present invention err
useful to control HIV and other retrovirus intection~ in patients
having an abnormally low white blood cell level, without causing
a harmful depression of the patients immune systea.
Accordingly, the methods of the invention can be employed in the
treatment of conditions such as ARC/AIDS without subjecting the
treated patient to a risk of opportunistic infection. The
treatment methods in accordance with the invention may also be
used in such patients to improve the functioning of their immune
systems. In addition, HIV retroviruses may also be responsible
for causing diseases other than AIDS. The inventors believe that
the inventive methods should be useful for treating such other
diseases as well.
In accordance with the invention, a photoactive compound
such as 8-methoxypsoralen is administered to the patients
6
~~3U'~4~9
nlood, or aom~ traction thereol, in vitro or in vivo using
conventional administration routes. A portion o! the patient's
bloody is tben treated (preferably, extracorporeally) using
photopheruia, which comprise subjecting the blood to
electromagnetic radiation in a wavelength suitable !or activating
the photoactive compound, such as ultraviolet light, preferably
long wavelength ultraviolet light in the wavelength range of 320
to 400 nm, commonly called WJ1 light. The treated blood, or a
traction thereol, is returned to the patient (in the case of
extracorporeal photopheresis) or remains in the patient
(following in vivo photopheresia).
Vaccines against viruses and methods of making same are
also provided according to the invention. According to they
invention, a photoactive compound (as described herein) is
administered to the blood or some fraction thereof of a donor who
is infected with a virus, such as an HIV retrovirus and/or who
is suffering Eros AIDS or AIDS Related Complex. At least a
portion of the donor's blood is then treated using
photophsreais, as described above. The treated blood or soma
fraction thereof (eg., treated free isolated virus) may be used
as a vaccine. In the case of treating an HIV infection it is
preferred to use treated virus infected cells along with the
treated virus in order to obtain the desired immune response.
Optionally, the treated blood is processed by
conventional techniques to substantially remove the
erythrocytes. The resulting processed fraction is then used as
7
s . 2007499
a vaccines which can be administered to a patient.
A further aspect of the invention is as follows:
A method for producing a vaccine against an HIV
retrovirus infection in an infected donor, said method comprising
the steps o!:
a. administering to at least a portion of the donor's
blood a photoactive compound that binds, in the case of a virus
infected cell, to the cell membrane (e.g., by binding to a
receptor and/or a nucleic acid fragment on the cell membrane)
and/or to nucleic acid in the cell nucleus or cell cytoplasm, or,
in the case o! either free virus or cell associated virus, that
binds to the virus surface (e.g., to a receptor and/or to a
nucleic acid fragment on the virus surface) and/or to nucleic
acid (e. g., DNA or RNA) which is incorporated in the virus, upon
activation by exposure to electromagnetic radiation o! s
prescribed apect~ for the purpose o! inactivating and/or
attenuating the virus and permitting the so treated virus and/or
virus infected cells to be presented to the immune system o! the
patient for the purpose o! engendering an immune response to the
~ira~ inlectiont and
b. then treating the portion o! the donor's blood to
which the photoactive compound has been adainisterad, said
treatment comprising subjecting at least a traction o! said
portion o! blood to photopheresis using said electro-magnetic
radiation, said photoactive compound and said electro-magnetic
A
radiation being administered in amounts which are
pharmaceutically effective for attenuating and/or inactivating at
least some o! the HIV retrovirus which is present in the treated
traction.
Fig. 1 is a block diagram of a photopheresis apparatus which can
be used to
practice the inventive method:=
Fig. is a graph of changes in CD4 cells and GPZ4 and GPiZO
antibody levels of patient No. 3 in Example Z;
Fig. 3 is a graph of changes in GP24 antibody lever of the five
patients treated in Example 2:
Fig. 4 is a graph of changes in GPiZO antibody levels of the five
patients treated in Example Z; and
Fig. 5 is a graph of changes in CD4 helper cell percentages o~
the Live patients treated in Example Z.
30
A
~..
While it is not intended that the scope of the present
invention be limited by any specific theory of operation, it is
believed that viral infections, particularly those which are not
controlled by the normal immunological response of a patient, can
be treated using a photopheresis treatment according to the
invention. It is believed by the inventors that the
photopheresis treatment according to the invention not only
treats the viral infection, but is believed by the inventors to
(i) restore the ability of a treated patient~a immunm systes
(which has been weakened by the viral infection) to coabat othir
infections, including non-viral infections, and (ii) restore the
immune system's anamnestic response to previous infections.
The photopheresis treatment method according to the
invention is of particular value in the treatment of frequently
mutating viral infections, such as ratroviruses, for instance HIv
retroviruses. In accordance with the photopheresis methods of
the invention, treated infected cells as well as killed and/or
attenuated virus, peptides, native sub-units of the virus itself
(which are released upon cell break-up and/or shed into the
blood) and/or pathogenic noninfectious viruses may be used.
Mutation of the viral antigen does not shield it from
attenuation/inactivation during photopheresis and consequent
generation of an immune response to the mutant forma of the viral
9
'~~~''~49~
antigen. Thus, the treatment m~thod~ according to the invention
provide s dyna,ia autogenous vaccine against viral infections.
The inventive methods haw been found by the inventors
to bs useful in the treatment of patients having a virus
infection and who have an abnormally low white blood cell count
and are particullarly useful in treating HIV retrovirus
infections. The inventive methods are also particullarly useful
for treating patients who are AIDS Carriers or who have AIDS or
AIDS Related Complex.
According to the claimed methods, a photoactivs
compound is first administered to the blood of a patient who is
infected with a virus. The photoactivs compound may bs
administered in vivo (..g. orally os intravenously os may ber-
administered in vitro to a portion of the patisnt~s blood which
has been removed from tha patient by~employing conventional blood
withdrawal techniques.
Alternatively, free virus is isolated from infected
cells using conventional virus isolation methods which are known
in the art. The photoactivs compound can be administered to the
infected cells prior to virus isolation or can be administered to
the free isolated virus. In the case of treating HIV infection,
however, it is presently preferred to use both treated virus and
treated virus infected cells in the methods described
hersinbelow.
In accordance with the present invention, the
photoactive compound selected should preferably be one that
' ~~~'~499
rinds, in the case of a virus infected cell, to the cell membrane
(e~g~. ~ biding to a receptor and/or a nucleic acid fragment on
the cell meabrantf and/or to nucleic acid in the cell nucleus or
cell cytoplasts, or, in the case o! either tree.viru~ or cell
associated virus, that binds to the virus surface (e.g., to a
receptor and/or to a nucleic acid fragment on the virus surface)
and/or to nucleic acid (e. g., DNA or RNA) which is incorporated
in the virus, upon activation by exposure to electromagnetic
radiation o! a prescribed spectrum, such as ultraviolet light,
for the purpose of inactivating and/or attenuating the virus and
permitting the so treated virus and/or virus infected cells to be
presented to the immune system o! the patient. Psoralen
compounds are particularly preferred for this purpose especially
the compound 8-methoxypsoralen -- in which case WA radiation is
preferred. for activating said compound.
Next, the portion of the patient's blood, or the free
isolated virus, to which the photoactive compound has been
administered is treated by subjecting the portion of the blood,
or the fre. isolated virus, to photopheresis using said
electromagnetic radiation -- for example, ultraviolet light. The
photopheresis step is preferably carried out in vitro using an
extracorporeal photopheresis apparatus.
The photopheresis step in accordance with the present
invention may also be carried out in vivo.
A presently preferred extracorporeal photopheresis
apparatus for use in the methods according to the invention is
11
.. 2007499
currently manufactured by Therakos, Inc., Westchester,
Pennsylvania under the name UVAR. A description of the Tharakos
WAR photopheresis apparatus may be found in U.S. Patent No.
4,683,889, granted to R.L. Edelson on August 14, 1987,
As illustrated diagramatically in Pig. 1, the apparatus
includes a pump 10 for removing blood from the patient via a
donor needle placed in an appropriate vein of the patient: an
irradiation chamber 2o; a radiation source 30 in close proximity
to the irradiation chamber and a centrifuge 40, preferably of the
continuous type. The various parts of the apparatus, such as
tubing collection bags for the blood and the like, which corns in
contact with the patient's blood or some fraction thereof, are
preferably replaceable so that they may be disposed of after each
use to prevent the possibility of transmitting blood-borne
infections from one patient to others who are subsequently
treated with the apparatus.
The exposure of blood, or free isolated virus, to
ultraviolet light in a photopheresis apparatus is within the
ability of persons having ordinary skill in the art.
When the photopheresis step is carried out in vitro, at
least a fraction of the treated blood, or the treated free
isolated virus, is returned to the patient following the
photopheresis treatment. Preferably, the treatment method
described hereinabove is repeated at an interval of about once
12
A
2007499
par week to about once every four weeks. Most preferably, in the
treatsant~o! MST infection, the treatment methods described
herein arm adsinistered on two successive days and repeated
approximately once per month (ie, the patient preferably receives
two treatments every month).
In view of th~ disclosure contained herein, those
persons who are skilled in the art will be able to adjust the
treatment parameters -- ie, dosage o! the photoactive compound
and electromagnetic radiation, periodicity of treatment (a. g.,
monthly, weekly, etc.) and the number of treatments administered
in each period ( a.g., twice per month on two successive days) -,
depending on the condition of the patient and the patiant~s
response to the treatment.
Preferred photoactive compounds for use in accordance
with the present invention are compounds known as psoralens (or
furocoumarins) which are described in U.S. Patent No. 4,321,919.
The preferred photoactive compounds for use in
accordance with the present invention include the following:
psoralen:
8-methoxypsoralen;
4,58-trimethylpsoralen:
5-methoxypsoralen;
4-msthylpsoralen:
4,4-dimethylpsoralen;
13
A
~Q(~ :49~
4-5~-dimethylpsoralent and
4~,8-mathoxypsoralen
Tha most particularly preterrad photoactive compound
for use in accordance with the invention is 8-methoxypsoralen.
The determination of an elective dosage for in vitro
virus inactivation of free isolated virus is within the ability
o! persons having ordinary skill in the art.
The photoactive compound, when administered to the
patient's blood in vivo is preferably administered orally, but
also can ba administered intravenously and/or by other
conventional administration routes.
The preferred dosage o! the photoactivs compound is in
the range o! about 0.3 to about 0.7 mg/kg of body weight although
larger or smaller doses may be employed. when the photoactiva
compound is administered in vitro to only a portion o! the
patient's blood or fraction thereof, it is within the ability of
those skilled in the art to calculate a dosage which is
equivalent to said range based upon the volume of treated blood
or fraction thereof.
When administered orally, the photoactive compound
should preferably be administered at least about one hour prior
to the photopheresis treatment and no more than about three hours
prior to the photopheresis treatment. The timing of
administration may be adjusted up or down as needed depending on
tha bioavailability of the photoactive compound, its expected
half-life, etc. If administered intravenously, the times would
14
~~f~'74~9
generally be shorter.
The photopheresis treatment in the treatment methods
according to the invention is pr~terably carried out using long
wavelength ultraviolet light (UVA) at a wavelength within the
range of 3a0 to 400 nm. The exposure to ultraviolet light during
the photopheresis treatment preferably has a duration of about
three to tour hours, although shorter or longer treatment periods
may be used it desired.
Whatever the spectrum of electromagnetic radiation, the
exposure of virus infected cells and/or virus thereto, following
administration of the photoactive compound, should be of
sufficient intensity/duration to effectively inactivate and/or
attenuate the virus. The selection of an appropriate wavelength
for photopheresis as wall as the exposure, depending upon the
photoactive compound being employed and the conditions of
treatment (e.g., in vivo exposure or in vitro exposure), is
within the ability of those skilled in the art in view of the
present disclosure.
When the photoactive compound is 8-methoxypsoralen, it
is preferred in accordance with the invention to utilize an
exposure to WA radiation of about 2 Joules/meter2 based upon the
surface area of the virus and virus infected cells undergoing
treatment.
When the photopheresis treatment according to the
invention is carried out in vivo, careful attention should be
paid to controlling the maximum radiant exposure so as to avoid
' ~~~'~49~
,.nnecessary injury to the patient. Methods for calculating
maximum radiant exposure to ultraviolet light are known in the
art and,_ ttterelore, shall not be described herein.
In summary, the invention provides a.novel treatment
for patients who are infected by a virus and who have depressed
immune systems as a result of such infection, as well as for
patients who are infected with an HIV retrovirus or who are AIDS
Carriers or who have AIDS or AIDS Related Complex. Such patients
cannot tolerate a treatment that would depress their immune
system .
The treatment methods according to the invention have
beers loured by the inventors to be sale in this latter regard
while also being ettective in combatting HIV infection fn h~ans.
The invention also provides methods !or making
vaccines. According to the invention, a donor who is infected
with a virus, such as an HIV retrovirus, may be utilized to
produce a vaccine against his infection as follows.
First, a photoactive compound as described hereinabove
is administered to at least a portion of the donor's blood
containing free virus and/or virus infected cells either prior to
removal of the blood, either orally or intravenously, or after
removal from the donor in which case it is administered in
vitro. Optionally, a portion of the donor's blood could first be
processed using known methods to substantially remove the
erythrocytes and the photoactive compound is then administered
to the resulting fraction.
16
~~3C~'~4~~
In any caws, the portion o! blood (eg., an enriched
lsukoayt~ !:'action thsrso!) and/or !ru isolated virus to which
the photoactivs compound has been administered i~ subjected to a
photophersais treatment using slsctromagnstia radiation o! a
prescribed opsctrum, s.g.,ultraviolst light, preferably WA, in
the manner previously described. Ths treated blood, the treated
portion thsrso! or the treated tree isolated virus (as the case
may bs) is then administered back to the donor as an autogsnous
vaccine. It will be understood that in accordance with the
present invention the treated virus can also bs isolated from the
treated blood or portion thsrso! following photophsrsais
traatmsnt !or use as a vaccine.
Additionally, in accordance with the prusnt
invention, the treated blood, which is itssl! a mixture o!
various blood components including peptides or polypsptidss, eg.,
cytokinss, lymphokinss, monokinsa, stc., and/or the treated
portion of blood may bs processed, as is within the ability o!
parsons having ordinary skill in the art, to isolate a particular
component or components which may be used in the treatment of the
virus infection of the donor and/or may be used as a vaccine
against the virus.
A male patient, 39 years of age, weighing approximately
70 kg and who had bean diagnosed as having AIDS Related Complex
was treated in accordance with the present invention as follows:
8-methoxypsoralen was administered orally to the
17
patient during the afternoon of the first day of treatment at a
dosage! o! 3a mg (i.a. about 0.4 ~/kg). Approximately one hour
after the 8-methoxypeoralen had been administered to the pats~nt
h~ was prepared for withdrawal of a portion of his blood for
photopheresis treatment using said Therakos WJ,R photopheresis
machine.
A centrifuge which is integral with the photopheresis
machine was used to spin oft substantially all of the
orythrocytu from the withdrawn blood and these were
subsequently returned to the patient. Next, approximately 30occ
of plasma and 240ca of butty coat (which includes the T-
lymphocytea) were removed in six cycles (40cc o! bu!!y coat pegs
cycle). The buoy coat and plasma were subjected to W1~ light
axposur~ beginning after 40cc of bully coat had been collected in
the first cycle. Tha irradiation was continued through all six
cycles and then for an additional one and one-halt hours for a
total irradiation time of approximately tour hours. The
irradiated bully coat and plasma wire then returned to the
patient. This process was repeated in the morning of the
following day.
The blood parameters of the patient before receiving
the photopheresis treatment according to the invention and five
w~eks after receiving the treatment are set forth in Table 1.
18
~~U~'~4~:~
Hlood Paramatera o! ARC Patient Traatad
~ Photooherae ~ s Mst_hod Accords nc To Th! Twvs.,~. a ,:
Blood Belora Altar Normal
Parameter Treatmg~
HEMOGLOHIZ1 11.7 G/DL 10.8 G/DL --
HEMACRIT 33.9% 33.6% -_
5.2 /UL 4.7 /UL -_
Hands 16% 16~
-.
Sag. Neut. 29%
23% -.
Lymphocytes 43% 46~
-.
Atyp. Lymph. 7% 7; -.
Monocytss 5% 6% --
P~~~TS 208,000 241,000 --
LYMPHOCYTE9s
CD3 (T3) 17% 85% 56-78%
CD4 (T4) 4% 22% 32-50%
CD8 (T8) 26% 62$ 13-38%
T4/T8 RATIO 0.15 0.4 0.8-1.9
In addition, the patient, who had a negative mumps skin
test (which was carried out in the usual manner known in the art)
prior to receiving the treatment in accordance with the present
19
~:~C~'r~~~
invention a: described hereinabove, developed a positive mumps
skin teat after six monthly treatments in accordance with the
present invention-- this positive mumps skin test being
documented by skin biopsy demonstrating the characteristic
histologic changes of delayed hypersensitivity. The above
results demonstrate a reconstitution o! normal immune function as
a result o! the treatment method according to the invention.
This patient continued to receive the described
treatment on two successive days on a monthly basis as part of a
l0 clinical study involving five patients, the results o! which are
discussed in Example 2 below in which thin patient is identified
as patient No. 1.
Five patients with ARC characterized by fatigue,
lymphadenopathy, fever, absent akin-test reactivity, and
decreased CD4 cells volunteered and ent.red into a clinical study
that was conducted in accordance with the provisions of the
Institutional Review Board for the treatment of human subjects at
Morristown Memorial Hospital, Morristown, New Jersey. Three
homosexual males ages 27 to 39, one 42-year-old male reformed
intravenous drug abuser, and one 27-year-old female reformed
intravenous drug abuser comprised the study group. The rangs of
time from the first known HIV antibody positivity until
enrollment for the HIV positive patients was one to four years.
' W~~'~4 ~;~
inclusion criteria included only patients not having undergone
any previous therapy against HIV infection.
Each patient received a monthly regimen o! the
treatment substantially as described in.Example 1 repeated on two
succesive days. The dosage o! 8-methoxypsoralen employed varied
between O.t and 0.6 mg/kg and was administered orally.
Four of the patients received a total o! twelve
treatments each over the six month course o! the study, that is,
on two successive days at monthly intervals for six months. The
to filth patient voluntarily discontinued his treatment alter the
filth month and only received ten treatments. On the basis o!
the physical examination, monthly antibody and antigen studies,
and CDR percentage, all five patients had either a stabilization~
of their disease or a positive response to the treatment. None
of the patients experienced damage to their immune systems during
the course of the treatment.
By strictly clinical criteria, all five patients
demonstrated improvement with respect to lymph node size. Energy
index and general state of ~~well being" improved in four. At the
start of treatment, the increase in energy level lasted for
approximately one week post-treatment. By the third month of
treatment, a more consistent rise in energy level persisted.
Most notably, the first patient treated (the patient in
Example l, above), a homosexual male who before treatment was
able to walk up a flight of stairs only with difficulty, is now
capable of jogging three and one-half miles per day and of
21
lifting weights.
~ onm patient who did not sees to have a persistent
rise in energy level was the feaale reformed intravenous drug
abuser with three small children. Although her fatigue had been
chronic prior to her entry into the study, no persistent change
in energy level was observed after therapy.
Additional clinical observations include increased
appetite and weight stability in four of the patients, the
exception being the female who lost 3.3 kg over the =ix month
treatment period. Lymphadenopathy disappeared in all live
patients by the third month and has remained absent. The four
male patient: also regained delayed cutaneous hypersensitivity
sites six months of therapy. During the six-month period of
treatment all patients were able to manage common community-
acquired upper respiratory tract infections without difficulty.
Other than normal variations, hemoglobin and
hamatocrit, reticulocyte count, and platelet count remained
stable. There were minor variations in the white blood count,
and circulating total lymphocytes remained stable in the four
male patients: however, the single female patient showed a
decrease in white blood count (4,000 prior to treatment to 2,000
after six months of therapy) and a parallel decrease in
lymphocytes. In all cases, ESR, urinalysis, and SMAC-20 remained
unchanged with the exception of minor rises in serum SGOT and
SGPT. EBV titers remained constant in all but one patient in
whom the IgG capsid antigen decreased from a titer of 1:1280 to a
2a
IGr~~ l ~~~
~citsr o! 1:160 paralleling an increse in HIV antibody titers.
CMV titers regained unchanged in all patients. Those patients
who wets PZ4 antigen negative resained negstivet those who were
positive remained positive, and no changes in titer occured~by
the end o! the six-month period. Viral cultures o! blood for IiIV
turned negative in one patient (Patient No. 3) but regained
positive in the others aft~r the reported therapy.
The course of the patient whose HIV culture turned
negative (Patient No. 3) is outlined in Fig. 2. The Gp 24
antibody livels o! all five patients over the course o! treatment
are plotted in Pig. 3. The Gp 120 antibody levels o! all five
patients over the course of treatment are plotted in Fig. 4.
Finally, the CD4 helper cell percentages o! all five patients
over the course o! tr~atment are plotted in Fig. 5.
A summary of the clinical characteristics and
laboratory results is provided in Table 2 below.
23
~~3~1'~4~~
Summary O! Clinical characteristics And Laboratory Resulta
Patient No. 1
Age: 38
sex: Male
Predisposing Factor: Homosexual
Known Duration O! HIV Antibody Poaitivity: 3 years
r~
Weight(kg) 71.3 70.5
CD4 Cells(8) 4 34
GP24 Antibody Level 2.5Z
Z76
(absorbance)
GP120 Antibody Level 2.2Z 1.68
(absorbance)
Delayed Skin Test neg. pos.
Hypersensitivity
Virus Culture (blood) pos. pos.
Patient No. 2
Age: 30
Sex: Male
Predisposing Factor: Homosexual
Known Duration o! HIV Antibody Positivity: 2 years
- PRE-TREATMENT POST-TREATMENT
Weight(kg) 64.5 66.0
CD4 Calls() 29 31
24
GP24 Antibody Level O.iZ 0.74
(ab~orbance)
GPiZO Antibody I,ewel 0.03 0.14
(abaorbance)
Delayed Skin Teat neg. . pos.
Hypersensitivity
Virus Culture (blood) poi, poi.
Patient No. 3
Ages Z7
Seu: Male
Predisposing Factor: Homosexual
Known Duration of H=V Antibody Positivity: 4 years
r
i5 Weight(kg) 63.6 64.7
CD4 Cells(%) 34* 33**
GP24 Antibody Level 0.7Z
1.69
(absorbance)
GP120 Antibody Level 1.04 2.07
(absorbance)
Delayed Skin Test neg. pos.
Hypersensitivity
Virus Culture (blood) pos. neg.
* Data for this patient's pre-treatment CD4 Cells(%) is not
available. The value shown in the table was determined for blood
drawn one month after the patient received his first treatment.
** This patient received only ten treatments over a five month
period at which point he voluntarily opted to discontinue
receiving treatment. Three months after voluntarily
discontinuing treatment this patient's CD4 Cells(%) had risen to
40. However, five months after discontinuing treatment the CD4
Cells(%) fell to 24.
Patient No. a
Age: 4Z
Sax: Mal~
Predisposing Factor: Reformed iVDA
Known Duration of Hiv Antibody Positivity: Z years
-
P
~ - ~T
Weight(kg) TBE~ 108
109
CD4 Calls () 11 33
GPZ4 Antibody Lwel 0.6Z
1.76
(absorbancej _
GP120 Antibody Level 0.21 1. Z3
(absorbanc~)
D~layed Skin Test neg. pos,
Hyp~rs~nsitivity
Virus Culture (blood) pos. poe.
Patient No. 5
Age: 27
Sex: Female
Predisposing Factor: Reformed IVDA
Known Duration Ot HIV Antibody Positivity: 2 years
PRE-TREATMENT POST-TREATMENT
Waight(kgj 61.0 57,7
CD4 C~lls(~) 12 13
GP24 Antibody Level 1.41 2, g9
(absorbance)
26
GP120 l~ntibody Level 0.46 1.95
(absorbancej
Delayed Skin Teat nag. nag.
Hypersensitivity
Virus Culture (blood) pos. pos.
Upon completion of the six month course of therapy described
in Example 2, wherein patient Nos. 1,2,4 and 5 received treatment
on two succusiVe days at monthly intervals, the course of
therapy was modified with each patient receiving only one
treatment per month instead of two. It was found by~th~
inventors that each patients CD4 Cell(%) tell oft markedly. The
original course of therapy, wherein the patients received
treatment on two successive days once per month was reinstituted
with the result that in patients 1 and 2 the CD4 Cell(%) rose.
Data for CD4 Cell(%) in patients 4 and 5 is not yet available.
The individual results were as follows:
At the conclusion of the original six month course of
therapy at two treatments per month, this patient's CD4 Cell(%)
was 34. Attar receiving only one treatment per month for five
months, the CD4 Cell(%) fell to 18. The original course of
therapy was reinstituted and attar two months the CD4 Cell(%)
rose to 39.
27
A~~~ l ~~~
At the conclusion of the original six month course of
therapy at two treatments per month, this patient s CD,~ Call(;)
was 31. Attar receiving only one traataant par month for tour
months, the CD4 Call(%) fell to 10. The original course of
therapy way reinatituted and after two months the CD4 Cell(%)
rose to 27.
Patient No. 4
At the conclusion of the original six month course of
therapy at two treatments per month, this patiant~~ CD4 Cell(%)
was 33. After receiving only one treataant per month for four
months, the CD4 Cell(%) fell to 7. The original course c!
therapy has been reinstituted but data on changes in the CD4
Cell(%) is not yet available.
Patient No. 5
At the conclusion of the original six month course of
therapy at two treatments par month, this patient s CD4 Cell()
was 13. After receiving only one treatment per month for four
months, the CD4 Cell(%) fell to 8. The original course of
therapy has bean reinstituted but data on changes in the CD4
Cell() ii not yet available.
It should be understood that while the foregoing
description has been provided to illustrate the present
inventions, it is not intended to limit the scope of the
inventions as various modifications to the inventions described
28
i~~~ i~~~
herein may be made by p~rson~ having ordinary skill in the art
without departing lroa the spirit and scope thereo! as dslinsd in
the lolloving claims.
29
