Treatment of human immunodeficiency virus/acquired immunodeficiency syndrome

Abstract

Methods of treating a patient with human immunodeficiency virus are disclosed. The method includes a providing intradermal and intravenous doses of a aTh1 composition that can increase the CD4+ cells in a patient that are resistant to HIV. The description includes a method for viral load reduction and a viral purge method. The regimen leads to a spike in the viral load and a then a return to baseline or lower levels of the virus and can lead to reduction and/or elimination of the latent viral reservoirs. Kits configured to provide intradermal doses and intravenous doses according to the regimen are also included.

Claims

1. A kit comprising components of a therapeutic human immunodeficiency virus (HIV) immunogenic composition, wherein the kit comprises: an intradermal aTh1 composition comprising at least two intradermal doses; an intravenous aTh1 composition comprising at least two intravenous doses, the intravenous doses packaged separately with escalating doses of the aTh1 composition in each package, wherein the package with the smallest amount of the intravenous, aTh1 composition is the first intravenous dose and each subsequent intravenous dose has an escalated amount of the aTh1 composition than the preceding intravenous dose; components of highly active antiretroviral therapy (HAART); and one or more HIV antigens.

2. The kit of claim 1 wherein the intradermal composition is divided into single dose packages with the same amount of the intradermal composition in each package.

3. The kit of claim 1 wherein the intravenous composition comprises at least three doses divided into single dose packages, wherein the second dose of the intravenous aTh1 composition has an escalated does relative to the first dose of the intravenous aTh1 composition and the third dose of the intravenous aTh1 composition has an escalated dose relative to the second dose of the intravenous aTh1 composition.

4. The composition of claim 1 wherein the aTh1 composition comprises activated CD4+ T-cells and increases CD4+ cell counts in an individual by creating activated CD4+ Th1 memory cells in circulation in the individual.

5. A composition comprising: an aTh1 immunogenic composition comprising an alloantigen, a molecule that interacts with CD40 surface receptor and Type I cytokines; at least one or more anti-retroviral drugs; and at least one or more HIV-antigens, wherein administration of the aTh1 immunogenic composition to a patient leads to activation of latently infected memory cells in the patient.

6. An AVI composition comprising: an aTh1 immunogenic composition comprising an alloantigen, a molecule that interacts with CD40 surface receptor and Type I cytokines; and at least one or more anti-retroviral drugs, wherein administration of the aTh1 immunogenic composition to a patient leads to activation of latently infected memory cells in the patient.

7. The composition of claim 6 further comprising one or more HIV antigens.

8. The composition of claim 5 wherein the aTh1 composition comprises activated CD4+ T-cells and increases CD4+ cell counts in an individual by creating activated CD4+ Th1 memory cells in circulation in the individual.

9. The composition of claim 6 wherein the aTh1 composition comprises activated CD4+ T-cells and increases CD4+ cell counts in an individual by creating activated CD4+ Th1 memory cells in circulation in the individual.

Description

DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

(1) This present description includes compositions for patients infected with retroviruses, especially Human Immunodeficiency Virus (HIV). The composition can include the aTh1 composition for eliciting an immunological response by the patient. The present description also includes an anti-viral immunotherapy drug composition (aTh1+ anti-viral medication) and methods for using the anti-viral composition for treatment of patients infected with HIV. Methods are described wherein the compositions can be used to: (1) treat immunological failure by increasing the CD4 count (CD4 Enhancement Method); (2) treat virological failure by restoring immune control of viral load (Therapeutic Vaccine Method); and (3) purge virus from the latent viral pool (Viral Purge Method). The combination of all these methods or the combination of the CD4 Enhancement Method and the Viral Purge Method can eliminate the need for daily HAART medication for an extended period of time (HAART Holiday Method).

(2) The anti-viral immunotherapy composition comprising aTh1 and anti-viral medication may be referred to herein as AVI composition.

(3) Patients infected with HIV can be treated with the compositions and methods described herein. The patients may be treated while experiencing immunological or viral failure while on HAART. The patient may be treated with or without simultaneous HAART medication. A biomarker for successful treatment by the composition and methods described herein can be characterized by enhanced serum levels of IL-12 in plasma of the HIV patients. IL-12 can enhance HIV-specific cellular immunity. The methods of the description generally can cause the appearance of IL-12 in the serum by at least 120 days of administering the aTh1 composition, preferably by 90 days, more preferably by 30 days and even more preferably by 7 days. IL-12 can serve as an early biomarker indicating success of the methods in creating anti-HIV immunity.

(4) The aTh1 composition can include i) living cells, or components thereof, containing at least one highly immunogenic antigen, ii) a molecule that delivers a signal through binding to surface CD40 receptor and iii) one or more inflammatory type 1 cytokines and/or chemokines. All of these components of the aTh1 composition can be delivered together or individually at the same time or separately in time.

(5) The highly immunogenic antigen component of the aTh1 composition can be natural, synthetic or recombinant proteins or peptides that have some foreign component that can make them recognizable to the human immune system. The immunogenic antigens can be, for example, allogeneic or xenogenic protein antigens. Self proteins that are altered to be recognized as foreign are also within the scope of the description. The alteration of the self-protein can be by recombinant or chemical means or by mixing the self-protein with an adjuvant. In a preferred embodiment, the highly immunogenic antigen is part of a living cell, preferably an allogeneic living cell, more preferably a living allogeneic immune cell, most preferably an allogeneic living Th1 immune cell. Alloantigens are a preferred highly immunogenic antigen included in the aTh1 composition.

(6) The highly immunogenic antigen(s) of the composition can be capable of being processed by professional antigen presenting cells (APC) for presentation on MHCI and/or MHCII molecules. Examples of highly immunogenic antigens may include also KLH, viral proteins, bacterial protein, yeast proteins, fungal proteins or combinations thereof.

(7) Examples of adjuvants that can increase the immunogenicity of a protein, such as a self protein, include agents which cause immature dendritic cells to mature to IL-12+ DC1 cells. Examples include adjuvant danger signals such as LPS, BCG and Toll-Like receptor agonists (e.g., TLR4 and TLR7). All highly immunogenic peptides and proteins are within the scope of this description.

(8) The aTh1 composition can also include type I cytokines and/or chemokines. Preferred Type 1 cytokines for the aTh1 composition can include interferon-gamma, IL-2, TNF-alpha, TNF-beta, GM-CSF, IL-1, IL-7, IL-15, IL-23 and IL-12 individually or in combinations thereof. Preferred chemokines for the aTh1 composition can include RANTES, MIP-1alpha, MIP-1beta and MCP-1 individually or in combinations thereof. These type I cytokines can either be part of the aTh1 composition or can be induced in the patient by the aTh1 composition.

(9) The aTh1 composition can also include a molecule that delivers a signal through surface CD40 receptor. One preferred molecule in the aTh1 composition that delivers a signal through CD40 is immobilized CD40L (CD154). CD40L (also known as CD154) is a member of the TNF superfamily. CD40L can act as a co-stimulatory molecule that interacts with CD40 expressed on dendritic cells (DC) to support their maturation to a IL-12+ phenotype. CD40L is preferably immobilized by expression on a cell surface so that it provides a positive signal through CD40. Alternatively, an agonist to CD40 can be used to deliver a CD40 signal, such as a fusion protein or an anti-CD40 antibody. The components of the aTh1 composition can be delivered together or separately and in various sequences and at various points in time and are within the scope of this description.

(10) In preferred embodiments, the aTh1 composition can include activated allogeneic CD4+ T-cells, and in more preferred embodiments, allogeneic activated memory CD4+ T-cells with high surface expression of CD40L and which produce interferon-gamma, are used.

(11) IL-12 production and CD40L expression in HIV-infected (HIV+) individuals can be severely impaired. CD40-CD40L interactions are the major mechanism involved in the T cell-dependent activation of antigen-presenting cells (APC), such as DC, to produce IL-12. While CD40, the counter-receptor for CD40L, is expressed on monocytes from HIV+ individuals, IL-12 production can still be suppressed. The appearance of IL-12 in the plasma after administration of the aTh1 composition can indicate successful initiation of the immunological mechanism of the methods.

(12) Different forms of CD40L can also signal through CD40. For example, soluble trimeric CD40L agonist protein (CD40LT), soluble CD40L and CD40L inserted into HIV virus can also provide the same signal and the same effect. All forms of CD40 agonist are within the scope of this description.

(13) In some preferred embodiments, the aTh1 composition can be AlloStim?. AlloStim? are bioengineered CD4 immune cells derived from the blood of normal donors. AlloStim? has an activated Th1 memory phenotype: CD4+, CD45RO+, CD62L.sup.lo, CD40L.sup.hi, CD25+, interferon-gamma+ and IL-4?. AlloStim? can be maintained in an activated state by continuous attachment to CD3/CD28-monoclonal antibody-coated microparticles. The key effector molecules of AlloStim? are the high surface expression of CD40L and the production of high amounts of inflammatory cytokines, such as interferon-gamma, tumor necrosis factor-alpha and granulocyte-macrophage colony stimulating factor (GM-CSF). AlloStim? and methods of making AlloStim? are described, for example, in U.S. Pat. No. 7,435,592, U.S. Pat. No. 7,678,572 and U.S. Pat. No. 7,402,431, all incorporated herein by reference. Other allogeneic or xenogeneic immune cells can also be used as components in the aTh2 composition. Some of the methods of the present disclosure are described with reference to AlloStim? but this is not meant to limit the methods to the use of AlloStim? only and other compositions may be used in the described methods.

(14) The aTh1 compositions described herein may also include anti-viral or anti-retroviral medication (AVI composition). Compositions such as AlloStim? which contain the necessary components of the aTh1 composition have been previously disclosed. In some embodiments, the use of AlloStim? alone or aTh1 composition may not be sufficient for treating HIV infection. The AVI composition includes anti-retroviral medication together with the aTh1 composition.

(15) While the aTh1 composition can be beneficial for cancer treatment, this composition may be detrimental to an HIV patient. This is due to the unique nature of the HIV life cycle. For example, when AlloStim? is used as the aTh1 composition, intradermal injections of AlloStim? can increase the titer of memory CD4+ cells specific for alloantigens. In HIV infection, this increase in CD4+ memory cells alone would only increase the number of CD4+ targets for the virus to infect. If the patient was not viral suppressed to have viral load below the limit of detection, circulating virions would infect the newly formed CD4 cells increasing the pool of latent virus. Thus, intradermal AlloStim? injections alone will lead to increase latent viral pool. The feature of the present method that can protect these newly formed CD4+ cells from viral entry is a step that can activate these cells using intravenous infusions. Activated memory cells can be resistant to viral entry due to up-regulation of CCR5 agonist cytokines and down-regulation of the CCR5 receptor. However, the mass activation of memory cells can awaken the viral production of latently infected cells. Intravenous infusions of AlloStim? can cause activation of T-cells and monocytes that can cause any latently infected cells to begin viral production. This can result in an increase in plasma viral load and can eventually lead to a decrease in CD4+ cell counts. Further, the activation of latent viral pools after intravenous AlloStim? infusion and the subsequent increase in viral replication may lead to increased risks in the development of viral escape mutants that become resistant to HAART drug cocktails. Carefully sequenced administration by dose and route can be required in order to treat HIV using the aTh1 compositions such as AlloStim?, when combined with anti-retroviral medications such as used in HAART. This can slow the production of virus and can allow for establishment of immune control of the virus. Frequent monitoring of CD4 counts and HIV RNA viral load can be performed to assure the proper balance is maintained. The latent viral load can be monitored by monitoring both cellular and plasma viral DNA levels.

(16) AlloStim? or other aTh1 compositions can be used initially together with viral suppressing drugs to slow down the spread of virus to healthy cells and prevent viral mutation.

(17) A variety of anti-retroviral drugs or medications can be included in the AVI composition. The AVI composition can include, for example, one or more drugs from any of the classes of antiretroviral drugs. The anti-retroviral drugs, for example, can include drugs from the following classes. Drugs from other classes are also within the scope of this description.

(18) Nucleoside/Nucleotide Reverse Transcriptase Inhibitors (NRTIs): Sometimes called nukes. These anti-HIV drugs can work to block HIV's ability to use reverse transcriptase to correctly change viral RNA into DNA. Host cells can use DNA to produce the proteins that the virus needs to make copies of itself.

(19) Non-Nucleoside Reverse Transcriptase Inhibitors (NNRTIs): These are called non-nukes. They can work in a very similar way to nukes. Non-nukes also can block the enzyme, reverse transcriptase, and can also prevent HIV from making copies of its own DNA. But unlike the nukes (which work on the genetic material), non-nukes can act directly on the enzyme itself to prevent it from functioning correctly.

(20) Protease Inhibitors (PIs): When HIV replicates inside cells, it can create long strands of its own RNA genetic material. These long strands have to be cut into shorter strands in order for HIV to create more copies of itself. The enzyme that acts to cut up these long strands is called protease. Protease inhibitors can block this enzyme and prevent those long strands of genetic material from being cut up into functional pieces.

(21) Entry/Fusion Inhibitors: These medications can work to block the virus from entering cells. HIV attaches and bonds to CD4 cells through receptor sites. Receptor sites are found on both HIV and CD4 cells (they are found on other types of cells too). Fusion inhibitors can target those sites on either HIV or CD4 cells and can prevent HIV from docking into healthy cells. CCR5 is an example of a receptor site for HIV.

(22) Examples of anti-HIV drugs that can be included in the AVI composition may include the following multi-class combinations: Atripla (efavirenz+tenofovir DF+emtricitabine); Complera (Eviplera, rilpivirine+tenofovir DF+emtricitabine); Stribild (formerly Quad) (elvitegravir+cobicistat+tenofovir DF+emtricitabine); Triumeq (formerly Trii) (dolutegravir+abacavir+lamivudine).

(23) Examples of anti-HIV drugs include the following NNRTs: Edurant (rilpivirine, RPV, TMC-278); Intelence (etravirine, ETR, TMC-125); Rescriptor (delavirdine, DLV); Sustiva (Stocrin, efavirenz, EFV); Viramune and Viramune XR (nevirapine, NVP); Lersivirine (UK-453061).

(24) Examples of anti-HIV drugs include the following NRTIs: Combivir (zidovudine+lamivudine, AZT+3TC); Emtriva (emtricitabine, FTC); Epivir (lamivudine, 3TC); Epzicom (Kivexa, abacavir+lamivudine, ABC+3TC); Retrovir (zidovudine, AZT, ZDV); Trizivir (abacavir+zidovudine+lamivudine, ABC+AZT+3TC); Truvada (tenofovir DF+emtricitabine, TDF+FTC); Videx EC and Videx (didanosine, ddI); Viread (tenofovir disoproxil fumarate, TDF); Zerit (stavudine, d4T); Ziagen (abacavir, ABC); Amdoxovir (AMDX, DAPD); Tenofovir alafenamide fumarate, TAF.

(25) Examples of anti-HIV drugs include the following protease inhibitors: Aptivus (tipranavir, TPV); Crixivan (indinavir, IDV); Invirase (saquinavir, SQV); Kaletra (Aluvia, lopinavir/ritonavir, LPV/r); Lexiva (Telzir, fosamprenavir, FPV); Norvir (ritonavir, RTV); Prezista (darunavir, DRV); Reyataz (atazanavir, ATV); Viracept (nelfinavir, NFV); Prezcobix (Rezolsta, darunavir/cobicistat); Atazanavir+Cobicistat.

(26) Examples of anti-HIV drugs include the following integrase inhibitors: Isentress (raltegravir, MK-0518); Tivicay (dolutegravir, S/GSK-72); Vitekta (elvitegravir, GS-9137).

(27) Examples of anti-HIV drugs include the following fusion inhibitors: Fuzeon (enfuvirtide, ENF, T-20); Selzentry (Celsentri, maraviroc, UK-427,857)

(28) The above stated anti-HIV drugs are exemplary and other anti-HIV drugs are within the scope of this description.

(29) The aTh1 compositions and/or the AVI compositions described herein can be used in methods to reduce and or eliminate HIV from patients. The methods described herein can enhance the CD4+ cells in the patient. The methods can also reduce the viral load and/or purge the virus from the patient.

(30) CD4 Enhancement Method

(31) The methods included in the present description can include the CD4 Enhancement Method. The CD4 Enhancement Method can use the aTh1 composition in a HIV patient taking anti-viral medication. This can increase the CD4+ cell counts of HIV patients, preferably the Th1 memory (CD4+CD45RO+) cells. The new CD4+ cells created by the method can be resistant to viral replication and viral entry. The CD4 Enhancement Method can be used in patients that have immunological failure on HAART medication.

(32) The CD4 Enhancement Method can increase CD4+ cell counts by creating activated CD4+ Th1 memory cells in circulation. Activated CD4+ Th1 memory cells can be resistant to HIV replication. This HIV resistant state can be due to an increase in the production of chemokines that are released from activated memory cells which in turn interact with the CCR5 receptor (i.e., RANTES, MIP-1alpha and MIP-1beta) and due to the down-regulation of CCR5 expression on CD4 memory cells that are activated.

(33) The creation of viral resistant CD4+ cells can be an important aspect of the CD4 Enhancement Method. Methods that would increase numbers of na?ve CD4 cells, Th2 cells, Th0 cells or resting CD4 memory cells would only add fuel to the fire. These undesirable CD4 subtypes are permissive for viral replication. Fuel for the fire means that there would be more CD4 targets for viral entry and thus more cells may be producing virions into plasma, which can lead to more cells with latent infection, increasing viral load and eventually resulting in increased CD4 cell death. Eventually the loss of CD4 cells would reduce the CD4 counts below the original baseline, causing the patient to be worse off than prior to the therapy.

(34) The CD4 Enhancement Method can create high titers of activated Th1 memory cells that are resistant to viral entry and replication due to activation with CD28 co-stimulation (through the co-stimulatory ligands CD80 and CD86 ligands up-regulated on APC) that can cause an increase in expression of native CCR5 ligands and the concomitant down regulation of surface CCR5 expression. To create these HIV resistant cells, the method can include multiple injections of the aTh1 composition (priming doses) and activation of APC to express CD80 and CD86 co-stimulatory molecules. The priming doses of the aTh1 composition can be administered intradermally, subcutaneously, intramuscularly or intravenously. The aTh1 composition could also be administered by a combination of these routes.

(35) In one embodiment, the aTh1 composition priming doses are administered multiple times intradermally. A minimum of two intradermal injections or doses can be required in order to cause memory cells to develop, such as about 4 or more doses are administered. The doses can generally be frequent. The doses may be administered up to about 2 weeks apart, or about 1 week apart, and even about 3-4 days apart. Doses less than about 2 days apart are integrated and still considered a single dose. Once CD4+ memory cells can be detected in the circulation, the patient can be said to be primed (i.e., immune to the antigen(s) in the aTh1 composition).

(36) The CD4 Enhancement Method can result in the increase in absolute CD4+ cell counts. The CD4/CD8 ratio may increase or remain near the same as baseline due to a concomitant increase in CD8 cells. In addition, the method can result in a shift in the Th1/Th2 balance to favor Th1. HIV infection causes a loss of Th1 cells resulting in Th2-dominated immune cells in circulation. The methods described herein can correct this imbalance by increasing the Th1 cell component.

(37) In one embodiment, at least two doses of the aTh1 compositions are administrated in the same location. After at least two doses in the same location, a new location may be selected for administration of subsequent doses. Alternatively, all doses can be administered in the same location. If a new location is selected, at least two doses should be administered at each new location. This cycle of administering doses of aTh1 composition can continue until the desired CD4+ cell count is obtained.

(38) Intradermal doses of the aTh1 composition in the same location can be administered to assure that professional antigen presenting cells (APC), such as Langerhan's cells (LC), macrophages (M) and immature dendritic cells (DC) that traffic to the injection site are exposed to the type 1 cytokines and CD40L in the aTh1 composition at the time they engulf the highly immunogenic antigen(s). It may take 2-3 days before these APC traffic to the administration site. After intradermal administration, LC of the skin can engulf and process the antigens from the aTh1 composition resulting in the activation and priming of antigen-specific T cells.

(39) Type 1 cytokines and CD40L in the aTh1 composition can cause the professional APC that process the aTh1 antigen(s) to mature and express MHCI/II, CD80/86 and IL-12. These mature APC may then traffic to the draining lymph nodes to interact with na?ve T-cells causing the activation, differentiation and proliferation of new effector CD4+ Th1 cells and CD8+ CTL (Tc1) killer cells specific for the antigen(s) in the aTh1 composition. Multiple administrations can convert the effector Th1/Tc1 cells to memory cells. In the presence of anti-retroviral drugs, as the administration of the number of doses of aTh1 composition increase eventually a new, higher CD4 set-point can be achieved. During the course of the aTh1 injections, CD4 counts and viral load can be monitored.

(40) This CD4 Enhancement Method can result in the patient being primed and immune to the antigen(s) in the aTh1 composition. This can result in an increase in memory CD4 cells that are resistant to viral entry upon activation. Multiple priming injections of the aTh1 composition are preferable. Such a pulsed introduction of antigen to the immune system can cause an enhanced delayed-type hypersensitivity (DTH) response at the injection site. The DTH reactions are mediated by memory Th1 cells and the appearance of a DTH reaction at the injection site can confirm presence of CD4 memory cells specific for the aTh1 antigen(s). Increased DTH skin reaction can also correlate with increased titers of CD4 memory cells in the circulation of HIV+ patients.

(41) The CD40L and type 1 cytokines in the aTh1 composition can non-specifically (polyclonally) activate memory T-cells. When memory Th1 cells are polyclonally activated, they can expand and maintain a HIV resistant memory, CCR5-phenotype. The expansion of HIV-resistant, CD4 memory cells can cause a beneficial sustained increase in CD4 counts. In order to polyclonally activate circulating CD4 memory cells, the aTh1 composition may be infused intravenously.

(42) Intravenous infusion of aTh1 composition may also activate latently infected memory cells. These activated cells can begin to produce virus upon polyclonal activation. The method described herein can create a pool of memory CD4 cells resistant to virus, these cells can provide help for HIV-specific CTL killer cells to eliminate cells that are actively producing virus. If the patient remains on HAART medication, the viral production can be slowed so that the CD4 count can be maintained high enough to support the anti-HIV immune response. In this manner, the resident anti-HIV immune response can identify and kill the activated memory cells producing virus while new viral resistant memory cells are replacing these cells. This balance between immune elimination of activated cells producing virus and increase in viral-resistant memory cells eventually leads to an increase in absolute CD4 counts and a decrease in the latent viral burden. Fluctuations in CD4 counts may occur prior to reaching the higher CD4+ cells set point level.

(43) After a patient is primed and the CD4 count has increased, the CD4 counts can be further increased and the memory cells can be continuously protected from HIV elimination by the simultaneous intradermal injection of the aTh1 composition and the intravenous infusion of the aTh1 composition. The polyclonal activation of Th1 memory cells in circulation can cause the establishment of a sustained type 1 cytokine storm. The intravenous infusion can cause activation of memory CD4 cells in the blood of HIV patients, which in turn can cause an increase in the production of type 1 inflammatory cytokines, creating a type 1 cytokine storm. Type 1 cytokines can polyclonally activate by-stander memory cells thus creating a positive feed-back loop for the maintenance of activated memory cells.

(44) Activated memory cells can expand in the presence of type 1 cytokines, thus accelerating the increase in the circulating CD4 counts. A sudden and violent immune reaction is known to occur with a cytokine storm containing type 1 cytokines such as TNF-alpha and IFN-gamma. Such a cytokine storm can be beneficial to HIV patients. Also type I cytokines such as IFN-gamma and IL-12 can enhance the memory cell function and innate immune activity.

(45) In preferred embodiments where the aTh1 composition used is AlloStim?, the intravenous infusion further enhances CD4 counts of HIV resistant memory cells due to the CD3/CD28 coated microbeads attached to the cells in this composition. These microbeads can also interact and activate host memory cells causing them to proliferate. Memory cells activated with CD3/CD28-coated microbeads can resist HIV infection.

(46) In one embodiment, AlloStim? cells are used as the aTh1 composition. The AlloStim? cells are injected intradermally at a dose of between about 0.2?10.sup.6 cells to about 2?10.sup.6 cells, preferably about 1?10.sup.6 cells. An intravenous preferred dose for accelerating CD4 counts is between about 1?10.sup.7 and about 3?10.sup.7 cells (low dose). AlloStim? cells are suspended in buffer solution (e.g., PlasmaLyteA with 1% human serum albumin) at a concentration of about 1?10.sup.7 cells/ml.

(47) One method for accelerated CD4 count enhancement can include one or more low dose intravenous AlloStim? infusions during the intradermal priming. The low dose intravenous infusions may occur within 7 days of the last intradermal injection, or within 24 hours, or at the same time as an intradermal injection. The intravenous dosing does not start until at least two intradermal priming doses have been administered, or after 4 intradermal priming doses or after more than 4 intradermal priming doses.

(48) Variations on the timing, amounts and routes of administration can vary and all are within the scope of the present description.

(49) Viral Load Reduction Method

(50) The Viral Load Reduction Method can reduce viral load through enhancement of cellular immune control of the virus. This method can be useful in patients that are virological failures on HAART medication. The CD4 Enhancement Method and the acceleration of this method can also accomplish a reduction in viral load. However, the formed methods can require a resident anti-HIV immune response to exist that can be awakened by the increased CD4 counts. Some patients may not have an effective, resident anti-HIV immune response and thus are unable to mediate the immune elimination of cells that have been activated to produce virus. In this circumstance, the Viral Load Reduction Method can be helpful as it imprints the missing anti-HIV immune response so that the CD4 counts can be increased and the viral load decreased.

(51) The Viral Load Reduction Method can include one or more HIV antigen components that are administered together with the aTh1 composition. The HIV antigen components can include, for example, whole attenuated virus, as well as natural or recombinant HIV viral proteins. These HIV antigens are administered together with the aTh1 composition at the same route and frequency of administration.

(52) The HIV antigens and aTh1 composition are administered intradermally together or immediately following each other in a patient that has been previously primed. The aTh1 antigens can attract a vigorous memory response due to the prior priming. The viral antigens and the aTh1 antigens can then be engulfed by scavenger APC, such as LC or DC. These cells can process and present the antigens to activate HIV antigen-specific T-cells. By this method, the aTh1 composition together with the Th1 memory cells that arrive at the injection site due to the prior priming, can both act as an adjuvant to steer the development of Th1/Tc1 anti-HIV immunity.

(53) The Viral Load Reduction Method generally includes HIV antigens for use together with the aTh1 composition. These HIV antigens can be natural or recombinant viral proteins, including tat, env and gp120. Whole attenuated virus or virus attenuated by nef substitution can also be used. The proteins can be expressed in a carrier such as pox virus. In a preferred embodiment, the HIV viral protein is the gag protein. The repeated administration of HIV antigens together with the aTh1 composition can establish high titers of CD4 Th1 memory cells and CD8 memory CTL specific for HIV. These memory cells can be maintained in an activated state by infusing the aTh1 composition intravenously.

(54) Viral Purge Method

(55) The Viral Purge Method can include escalating intravenous doses of the aTh1 composition in patients on anti-viral medication. This method is used in patients that have been first subjected to the CD4 Enhancement Method and/or the Viral Load Reduction Method. The Viral Purge Method is administered to patients that have achieved an increased CD4 set-point consisting of viral-resistant memory cells. If the patient has a high latent viral load, activation of these cells by intravenous infusion may cause a burst of viral release and may result in an immediate drop in CD4 counts. Thus it is safer to start the method from as high a CD4 set-point as possible. As an example, the patient is at a CD4 set-point >300 cells/ml, or at a set-point >500 cells/ml or at a set-point of >700 cells/ml.

(56) In certain embodiments, patients that have been previously primed and have at least a 6 month history of viral load below the limit of detection are subjected to increasing intravenous doses of the aTh1 composition while maintaining active anti-viral suppression. The intravenous infusions can occur at least about 3 days apart. After each infusion, the viral load can be assayed to determine if a viral spike has occurred. A spike is any reading over the limit of detection. The doses of aTh1 can be increased at each infusion until a viral spike occurs. The appearance of a viral spike can be indicative that cells from the latent pool have been activated. After a viral spike occurs, the CD4 counts and viral load can be followed until the viral load returns to the undetectable level. When the viral load is undetectable, another IV infusion at the same dose as caused the viral spike can be administered. If again a viral spike is detected, the patient is followed until the viral load returns to baseline and the process can be repeated until no viral spike occurs after the intravenous infusion. At any time no viral spike is detected, the intravenous dose can be again escalated. If the escalated dose causes a viral spike, the process is repeated until no spike is produced. At the point that an escalated intravenous dose does not cause a viral spike, the intravenous dosing can be halted.

(57) Once the intravenous doses are halted, the patient CD4 and viral load can be continued to be monitored. When the CD4 stabilizes with at least two counts a week apart above the baseline CD4 value and the viral load is undetectable, the patient can be taken off the anti-viral medication. While on a holiday from anti-viral medication the patient should be monitored for CD4 count and viral load. The patient should remain without anti-viral drugs until the viral load spikes. When the viral load spike occurs, the anti-viral medication should be started again immediately.

(58) After the spike of viral load on anti-viral drug holiday, the process of escalating intravenous doses can be re-initiated. Each time the patient is placed on holiday from anti-viral medication, the time it takes for a viral spike to occur should be increased.

(59) In embodiments where AlloStim? is used as the aTh1 composition, the escalating intravenous dosing can start at about 3?10.sup.7 cells and can escalate to about 5?10.sup.7 cells to about 10?10.sup.7 cells to about 15?10.sup.7 cells to about 20?10.sup.7 cells. Dose escalation can continue at intervals of 5?10.sup.7 cells to a maximum of 100?10.sup.7 cells.

(60) As discussed above, combinations of the CD4 enhancement method, the viral load reduction method and the viral purge method can be administered. In some embodiments, the patient is administered HAART as appropriate in conjunction with the aTh1 composition.

EXAMPLES

Example 1

(61) During the initial protocol treatment phase, patients are kept on HAART therapy. After detecting a spike in viral burden, indicating successful activation of latent virus, followed by a decrease in viral burden to baseline, indicating immune control patients can be eligible for the HAART interruption phase.

(62) To minimize the risk of treatment interruption, patients are closely monitored and resume treatment should virus replication be detected.

(63) The protocol alters between intradermal and escalating intravenous dosing of AlloStim? in patients on HAART. The intradermal dosing is designed to increase the titer of circulating CD4+ Th1 memory cells that are resistant to HIV infection. The intravenous infusion is designed to provide an inflammatory cytokine storm and activate memory CD4 cells and macrophages (through CD40-CD40L). Activation should stimulate latent virus replication within these reservoirs. In addition, the intravenous infusion should activate NK cells which will target and kill viral replicating cells providing a source of viral antigens. Dendritic cells will process the shed viral antigens and in the inflammatory environment stimulate anti-HIV-specific immunity. The continuous inflammatory storm will disable viral immunoavoidance mechanisms permitting clearance of cells with replicating virus The cycling between intradermal injections to increase CD4 cells and intravenous infusions to activate latent virus and stimulate anti-HIV immunity is expected to clear latent virus. Each IV infusion should cause a spike in viral load and subsequent immune control should then gradually decrease viral load. If there is difficulty in returning viral loads to baseline viral blocker drugs will be added (such as Maraviroc and/or Fuzeon).

(64) Regimen

(65) The initial protocol is 28 days.

(66) Day 0: Intradermal AlloStim?

(67) Day 3: Intradermal AlloStim?

(68) Day 7: Intravenous AlloStim? (1 ml)

(69) Day 10: Intradermal AlloStim?

(70) Day 14: Intradermal AlloStim?

(71) Day 17: Intravenous AlloStim? (3 ml)

(72) Day 21: Intradermal AlloStim?

(73) Day 24: Intradermal AlloStim?

(74) Day 28: Intravenous AlloStim? (5 ml)

(75) Viral load and CD4/CD8 ratio is measured at baseline (Day 0) and Days 10, 21 and 29) and every 28-32 days thereafter for 6 months.

(76) Research blood (45 ml) is drawn at or before baseline (Day 0) and Day 7, 17, 27 before IV infusions. PBMC and plasma is stored frozen until analyzed for Th1/Th2 balance (ELISPOT), HIV-specific immunity (ELISPOT), cytokine bead array.

(77) Phenotype analysis is conducted prior to baseline and Day 60 (+/?2 days) including:

(78) CD3, CD4, CD8, CD45RA, CD45RO, CD62L, CD25

(79) CD14, HLA-DR, CD80, CD86, CD16, CD38, CD117

(80) CBC, CMP, CRP laboratory tests for safety taken at baseline, Day 7, Day 14, Day 21 and Day 28.

(81) HAART Interuption

(82) Access to lymphoid tissue or most anatomic compartments in otherwise healthy HIV subjects in order to determine level of latent infection is difficult. Further, even if such studies fail to detect an infected reservoir, they cannot prove latent virus eradication. The ultimate test of efficacy can only be the withdrawal of HAART.

(83) Patients that experience a spike in viral load and then recover to baseline or lower and remain at base line or lower for at least 60 days are provided the option of entering into a HAART interruption phase of the protocol. In this phase, all viral suppressive drugs are withdrawn and viral load will be measured daily for the first 7 days. If an increase in viral load is detected, HAART will be re-started. If no rise in viral load is detected, the HAART interruption is continued with viral load being measured weekly for 7 weeks. If no viral load increase is detected, monthly viral burden tests are conducted until the 1 year anniversary of the HAART interruption. At any time an increase in viral burden is detected, HAART is reinitiated.

(84) Primary Outcome Measures:

(85) Changes in steady state viremia (so-called viral set point) at baseline and monthly for 6 months after completion of 28 day protocol while remaining on HAART.

(86) Safety and tolerability

(87) Changes from baseline and absolute counts and activation status of CD4 and CD8 na?ve and memory T cells

(88) Changes in absolute counts and activation state of monocyte/macrophages

(89) Changes in the number of interferon (IFN)-gamma generating (in response to HIV antigens) CD4 T cells/million peripheral blood mononuclear cells (PBMCs) as measured by intracellular cytokine staining (ICS) or ELISPOT.

(90) Secondary Outcome Measure:

(91) Time to viral burden increase from baseline after HAART interruption

(92) Inclusion Criteria:

(93) HIV-1-infected

(94) On a stable HAART regimen without changes or interruptions for at least 12 weeks. prior to study entry. Patients must be currently taking regimens containing drugs of at least two different classes.

(95) Two readings of plasma HIV-1 viral load of less than 50 copies/ml within 30 days prior to study entry.

(96) CD4 count greater than 350 cells/mm^3 within 12 weeks prior to study entry.

(97) Lowest CD4 count greater than 250 cells/mm^3 at any time prior to study entry.

(98) Willing to use acceptable forms of contraception.

(99) Karnofsky performance score 90 or higher obtained within 30 days prior to study entry.

(100) Exclusion Criteria:

(101) Age<18 years old.

(102) Patients with failure to HAART.

(103) HIV-1 viral load greater than 500 copies/ml within the 24 weeks prior to study entry.

(104) History of any chronic autoimmune disease (e.g., Graves' disease). Excessive exposure to the sun (e.g., sunbathing, tanning bed) within 2 weeks prior to study entry.

(105) Previous CDC Category B or C event.

(106) Use of immunomodulating therapy, including cyclosporine, IgG-containing products, interleukins, interferons, or systemic glucocorticosteroids (including those inhaled) within 6 months prior to study entry.

(107) Exposure to an experimental HIV vaccine.

(108) Any vaccine within 30 days prior to study entry.

(109) Investigational products within 12 weeks prior to study entry.

(110) Current drug or alcohol use or dependence that, in the opinion of the investigator, would interfere with the study.

(111) Serious illness requiring systemic treatment and/or hospitalization. Participants who complete therapy or are clinically stable on therapy for at least 14 days prior to study entry are not excluded.

(112) Positive hepatitis B surface antigen or positive anti-hepatitis C antibody at screening.

(113) Pregnant or breastfeeding.

(114) Adequate organ function including:

(115) Marrow:

(116) Platelets>100,000/mm.sup.3.

(117) Absolute neutrophil count?1,500/mm.sup.3.

(118) Hemoglobin?10.0 g/dL (transfusion allowed)

(119) Hepatic:

(120) Serum Total bilirubin<1.5?ULN mg/dL,

(121) ALT (SGPT)/AST (SGOT)?1?upper limit of normal (ULN).

(122) Renal:

(123) Serum creatinine (SCR)<1.0?ULN, or

(124) Creatinine clearance (CCR)>30 mL/min.

(125) History of cardiac, pulmonary, gastrointestinal, hepatic, renal, pancreatic, or neurologic disease which, in the opinion of the study official, will compromise study participation

Example #1

(126) A HIV+ man on HAART medication for 19 years with viral load always below detectable limits was entered into the Viral Enhancement protocol.

(127) The patient had absolute CD4 cell counts of 250-350 at baseline.

(128) He was administered 1?10.sup.7 AlloStim? intradermally while on his HAART medication on Day 0, Day 3 in same location. Then again on Day 7 and Day 10 in another location. Over this period of time, his absolute CD4 count increased from 350 cells to 450 cells.

(129) Beginning on day 14 escalating intravenous doses of AlloStim? were administered. On day 14, 1?10.sup.7 cells were infused. There was no detectable viral load. On day 17, 5?10.sup.7 cells were infused. There was no detectable viral load. On Day 21, 10?10.sup.7 cells were infused. The viral load spiked to 66 and remained above detection for 10 days when it again returned to undetectable. CD4 count increased to over 500 during this period and continued to rise over the next 60 days stabilizing at over 600.

Example #1

(130) A HIV positive man on HAART medication for at least six years with undetectable viral load. His absolute CD4 counts ranged from 100-230 over a period of 2 years.

(131) The patient had a 250 CD4 count at baseline.

(132) He was administered 1?10.sup.7 AlloStim? intradermally on Day 0, Day 3, Day 10 and Day 14. His CD4 counts increased to 293. On Day 17 he received a 1?10.sup.7 intradermal injection and a 3?10.sup.7 intravenous infusion. On day 21 he received a 1?10.sup.7 intradermal injection and a 10?10.sup.7 intravenous infusion. On Day 24 he received a 10?10.sup.7 intravenous infusion. On Day 28 and day 31 he received a 10?10.sup.7 intravenous infusion. His viral load spiked at 300 on day 31 and returned to baseline by day 42. During this time his CD4 counts slowly declined to below 200 by day 42.

(133) Beginning on Day 49 until Day 63, he received 1?10.sup.7 intradermal injection of AlloStim every 3-4 days. His CD4 count gradually increased from below 200 to above 300. His viral remained undetectable.

(134) On Day 84, Day 87, Day 91 and Day 94 he received 10?10.sup.7 intravenous AlloStim? infusions. On Day 97 his viral load spiked to 86. By Day 101, his viral load returned to baseline and his CD4 counts remained over 300. He was removed from his HAART medication.

(135) He remained with undetectable viral load for 31 days without HAART medication. On Day 32 the viral load was 300 and CD4 230. HAART was restarted and the viral came back to undetectable and CD4 stabilized at around 250.

(136) Although the present description has been described with reference to preferred embodiments, workers skilled in the art will recognize that changes may be made in form and detail without departing from the spirit and scope of the invention.