WO1998010792A1

WO1998010792A1 - Immune direction therapy

Info

Publication number: WO1998010792A1
Application number: PCT/IB1996/000945
Authority: WO
Inventors: Patrick T. Prendergast
Original assignee: Individual
Current assignee: Individual
Priority date: 1996-09-11
Filing date: 1996-09-13
Publication date: 1998-03-19
Anticipated expiration: 1999-03-11
Also published as: JP2001503613A; WO1998010787A3; WO1998010787A2; SE9900812L; SE9900812D0; AU6887096A; EP0929568A2; IL128806A0; NZ335039A; CN1230195A; CA2265885A1; AU4132097A

Abstract

Herein is described a specific amino acid sequence which exhibits specific Ion (bridge) pair arrays enclosed on at least one side by non polar hydrophobic transmembrane segments, as a mechanism used by many infectious agents and a number of cytokine inhibitory factors, such as Interleukin 10 and Prolactin Inhibitory factor and alfa-fetoprotein, to not only undermine the hosts immune defences but to also allow for the infection of target lymphoid tissue. It has been demonstrated that certain vaccines, when inoculated into a host, produced a range of neutralising antibodies but failed to prevent infection when that host is later challenged with live infectious organism. This present patent illustrates that when such vaccine inoculation is coupled with passive immunisation with mono or polyclonal antibodies to these specific amino acid sequences as specificied herein that the host is then capable of overcoming the infectious challenge. Herein is described the therapeutic use of mono or polyclonal antibodies to these said specific sequences as a treatment for Acquired Immune Deficiency Syndrome (AIDS) and other disease states that persist due to the presence of a cytokine inhibitory factor of viral, fungal, bacterial or host origin such as Chronic Fatigue Syndrome where Interleukin 10 mimic molecules are responsible for a multitude of disease symptoms identified as indicative of Myalgic Encephalitis. Herein is described the therapeutic use of mono or polyclonal antibodies to these specific amino acid sequences as a combination therapy with vaccines and anti-viral agents to prevent side effects from certain immune modulation and anti-viral agents (e.g. DHEA and IL-12) which cause enhanced production of Interleukin 10 or AFP mimic molecules during therapy. Also herein is described the therapeutic use of these specific sequences either isolated from the organism source or produced by direct synthesis or recombinant protein synthesis. These peptides when administered to a patient suffering from an auto-immune disease, such as Multiple Sclerosis (MS), Lupus (systemic Lupus erythematoses) or diabetes or rheumatoid arthritis as limited examples or to transplant organ recipients, will allow the patient's immune state to be shifted to a Th2 antibody dependent immune response and curtail the Th1 (T cell dependent) immune attack which is evident in such immune malfunctions as MS and graft versus host disease. Certain dermatological conditions which are today treated by the use of corticosteroid creams and ointment may also be successfully treated by replacing the corticosteroid with these mimic immunosuppressive AFP/Interleukin 10 sequences outlined in this patent.

Description

IMMUNE DIRECTION THERAPY

BACKGROUND TO THE INVENTION It is identified in this patent that a very specific amino acid sequence which exhibits specific Ion bridge pair arrays enclosed on at least one side by a non-polar hydrophobic transmembrane segment of at least one amino acid can, if presented to a cell membrane, induce endocytosis and cause activation of specific intracellular and extracellular events which would normally only result from the interaction of an Antigen MHC-II complex with both the T cell receptor (TCR) and the CD3 membrane complexes. These specific amino acid Ion bridge pair sequences when presence within the cell membrane normally results from the proper functioning of the immune system presenting antigen and activating T cell clone expansion, however, if this specific molecular signal of a specific Ion bridge pair enclosed on at least one side by hydrophobic segments consisting of at least one amino acid is presented to the cell from a non immune source. Increased cytoplasmic circulation from cell membrane of specific marker molecules occurs and this removes the normal immune functions of the cell types impacted by these specific peptide sequences. A proportion of amino acid sequences according to the patent have demonstrated in a dose dependent manner the ability to down regulate the expression of la molecules on human macrophages . Some in-vitro experiments suggest that direct T cell antigen interactions without the mediation of la bearing macrophages may result in the generation of antigen specific suppressor T cells. All experimental evidence indicates that the development of antigen-reactive clones of helper T cells requires the presence of la bearing cells in the tissue. This inhibition of expression on the membrane surface of these class II molecules (la) as produced with alfa-fetoprotein and/or cytokine inhibitory factor (IL-10) , signals the immune system to accept the appearance of new antigens as self to the immune system. Hence the often reported observation that an immune activation on polyclonal B cell activation producing auto-antibodies follow certain viral, bacterial and parasitic infections (e.g. HIV, Malaria). The effects of these specific peptide ion-bridge pairs attached to a hydrophobic amino acid sequence demonstrate that they are the component peptide segments within both the alpha-fetoprotein and Interleukin 10 molecules that are involved in the process of inducing tolerance and maintaining the tolerant state to infectious organisms presenting these sequences.

In identifying this specific type of sequence and its ability to generate specific immune signals together with its ability to enhance or trigger endocytosis of attached peptides or glycopeptides we have been able to identify these specific amino acid sequences as a mechanism used by many infectious agents to not only undermine the hosts immune defences but to also allow for the penetration or infection of target ly phoid tissue.

The large scale efforts to produce a broad spectrum vaccine candidate for Influenza Virus has proved impossible due to the rapid rate of mutation of the outer coat of this virus, however, without the ability- to attach and fuse and signal host immune cells in the body using those specific polar single Ion bridge pair arrays of amino acid specific sequences, outlined herein, it is non-infectious. If these amino acid sequences are altered in the Influenza Virus, this virus then cannot undermine the hosts immune system and achieve cell entry or create immune dysfunction, this ability is restricted to a specific number of sequences all of which must present to the cell membrane the charge distribution shown to activate endocytosis and TCR/CD3 cell activation and neutralise T cell immune surveillance as it relates to MHC-Class I and II. Therefore, if the infected host, human or animal, already posses neutralising antibodies to these amino acid changed depole sequences as specified herein it will not be possible for infection to be established because these neutralising antibodies perform a duel function (A) they prevent anchorage and endocytosis of the infecting organism into the host cell thus preventing productive infection and (B) they prevent the circulation in the plasma of these mimic Interleukin 10 or AFP type molecules which are released by infecting organisms. These specific said sequences confuse the normal signalling system involved in immune T and B cell activation since when applied to cells they trigger an intracellular Biochemical signal similar to when a T cell receptor (TCR) molecule coupled to a CD3 molecule interact with a MHC-II antigen complex, together with the fact that these sequences also cause increased turnover of surface receptor molecules such as the Interleukin I receptor molecule, thus leading to an increase of Interleukin I levels and cause a shift in T cell performance due to the shifting Th-_^/T^ cytokine balance. It has been demonstrated with non-protective vaccine candidate antigens which when previously inoculated into a host produced a range of neutralising antibodies but failed to prevent infection being established when that host was later challenged with live infectious organism. When the initial vaccine inoculation is coupled with passive immunisation with mono or polyclonal antibodies to these said specific sequences of this present patent that an immune response to the vaccine antigen from both T cell and B cell immune components results which includes antibodies to these hitherto unchallenged sequences results, the host is then capable of overcoming an infectious challenge without becoming infected or producing the usual antibody and autoantibody peak and subsequent immunosuppression normally associated with infections caused by organisms who utilise these specific amino acid sequences to direct the hosts immune signalling system towards a more pronounced B cell or Th₂ cytokine profiled response.

Malaria is one of the most important infectious diseases in the World, each year there are 270 million new infections resulting in over 100 million episodes of illness and approximately 2 million deaths. World-wide the malaria problem is getting worse each year. The reason for this worsening situation include (A) increased levels of drug resistance on the part of the parasites, (B) increased levels of insecticide resistance on the part of the vectors . No vaccine has yet been produced which can successfully induce a protective antibody response and the reason for this is that although antibodies which cross react with many epitopes of the P. vivax circumsporozoite are produced in abundance by the current unprotective vaccine candidates, because of the immune blind spot or immunologically privileged sites offered by these specific sequences identified herein, like Interleukin 10 and AFP are not visible to the host immune system which both allow the parasite to gain access to the host cell and to cause the non-specific polyclonal B cell activation and immunosuppressive (Interleukin 10 and/or AFP) like effects which are so universal for people suffering from parasites such as malaria, and Leishmania, the host cannot gain enough immune reactive monocytes to overcome the infection initially because these Interleukin 10/AFP mimic molecules carried by the infecting organism shuts down the vital Th_x T cell response needed to clear intra-cellular infections. We have identified the specific polar array sequence on the coat protein of malaria which this organism uses like Influenza Virus to attach and activate endocytosis together with activating a Th₂ (B cell) response and subsequently undermining the host ' s immune response and allowing infection to take hold while still producing an array of neutralising antibodies which creates mutational pressure for the generation of more virulent strains of the organism within the host . Our studies clearly demonstrated in mouse models that polyclonal or monoclonal antibodies generated to the above polar sequence arrays to these specific amino acid sequences either taken from the sequences present in human alfa-fetoprotein or human Interleukin 10 resulted in protection of mice from challenge by malaria sporozoites. Therefore a vaccine for malaria which will enable a human to raise a protective antibody titre against malaria sufficient to prevent infection may be manufactured using amino acid sequences displaying these specific polar arrays. Another method expected to be more successful as a vaccine combination (because the immune system of primates including man appear to be blinded to these specific signal sequences) for protection is to use passive immunisation with either polyclonal or monoclonal antibodies to these said specific sequences generated either in animal human and/or tissue culture given either before or simultaneously with any of the current malaria vaccine candidates which previously could not produce a protective immune response. When these mono or polyclonal antibodies are given to the host in conjunction with the antigen the hosts immune system does not produce the well documented polyclonal B cell activation of the host immune system and the immune system of the host so challenged will produce a protective antibody and T cell immune response which allows it to deal effectively with any later malaria infection challenge.

In malaria, as in other infections the said specific sequence when embedded in the cell membrane of the host activate the phosphatidylinositol pathway, which causes the release of Ca++, the phosphosylation of cell proteins and the activation or enhanced activity of certain enzymes related to metabolism. This does not occur in the presence of antibodies to the disclosed specific sequences and the organism like malaria, Mycobacterium Tuberculosis, Leishmania, HIV and others are not able to cause metabolic and immune Th₂ activation and exhaustion. It is an important coincidence that in certain malaria endemic areas that genetic mutations that have caused the deletion of the metabolic activity control enzyme glucose- 6 -phosphate dehydrogenase has conferred on the host immunity to malaria. By intervening at an early stage of infection and neutralising certain properties of the malaria parasite to alter cellular reactions by interfering with these specific membrane signal transduction sequences as defined herein it is possible to confer protective immunity to this organism.

The present invention utilises the novel discovery that certain amino acid sequences which exhibit specific Ion (bridge) pair arrays enclosed on at least one side by non-polar hydrophobic transmembrane segments can be utilised to enhance the humoral antibody response and down-regulate the T cell or delayed-type hypersensitivity (DTH) response of humans and animals. These CD3/TCR mimic membrane interaction molecules which present as hydrophobic Ion bridge pairs are utilised by both the organism itself as specific peptides and by cytokines and also by infectious agents to modulate immune response (A) during periods of reproductive foetal gestation as with the alpha-fetoprotein molecule to prevent foetal rejection by the maternal immune system and (B) during cytokine control of immune functions as with cytokine synthesis inhibitory factor (Interleukin 10) when a Th₂ cytokine profile is required or to curtail the uncontrolled Th^ T4 cell immune response. This cytokine (IL-10) is particularly evident following vaccination to enhance humoral immunity and secure antibody formation, and often causes the temporary disappearance of the Tuberculin reaction which is associated with Th_: (DTH) response in patients following vaccination. (C) Infectious agents such as viruses (RNA & DNA) mycoplasma, bacteria, malaria and a wide array of human and animal parasites also carry these specific charged array of amino acid sequences which cause the down regulation of the T4 cell response and enhance the humoral (antibody mediated) immune response of their infected host, see sample listing enclosed.

Now that these specific control sequences have been identified and verified we herewith outline a number of therapeutic modalities that result from this new found ability to intervene therapeutically to control, neutralise or enhance specific immune type reactions dependent upon the nature of the patient's or animal's own immune system status, infection or disease state. Examples : - 1

Anti-serum generated to these specific sequences as presented in AFP, Interleukin 10, EBV-BCRF1 and other peptides and as specified in amino acid sequence, listing enclosed, with this patent can be used to remove Interleukin 10 mimic molecules from the circulation of immunosuppressed patients suffering from viral and/or bacterial and/or fungal, mycoplasmic or parasitic infections, which infection's principle method of defence against the host is to stimulate a Th2 cytokine response and curtail or abolish the Thl cell mediated immune attack.

This invention relates to methods of treatment of persons and animals with indications of immunodeficiency, wherein the said indication is resultant from viral and/or retroviral infection and/or infectious parasites, bacteria and/or mycoplasma. The invention further relates to treatment with the above antiserum either poly or monoclonal in nature for establishing improved immuno response for persons and prophylactic treatment for persons where immuno-malfunction due to genetic pre-disposition or infection is considered a future risk.

The invention further relates to a screening method for vaccines, manufactured by the use of coat or other peptides from viral, bacterial, parasitic or mycoplasma, to determine and remove and/or neutralise inherent immune suppressive properties - such suppressive potential properties is determined by the manufactured vaccine's reactivity with the said specific amino acid sequences as outlined herein, be they synthetic or natural in origin, e.g. AFP, Interleukin 10, viral or bacterial coat peptides. In one embodiment, the host organism (man or animal) is treated with mono or polyclonal antibodies to any one or combination of the specific amino acid sequences as defined herein. This will result in the removal of Interleukin 10 and AFP type mimic immunosuppressive peptides and initiate a Th_x cell response, allowing Interleukin 2 and gamma interferon synthesis to occur. Treatments used according to this invention employing the poly or monoclonal antiserum to these specific cytokine inhibitory sequences are administered as treatments against viral, bacterial and mycoplasma and parasitic infections which cause immunosuppression by any suitable route including enteric, parenteral, topical, oral, rectal, nasal or vaginal routes. Parenteral routes include subcutaneous, intramuscular, intravenous and sublingual administration. The preferred route of administration would be an intravenous one.

The present invention further provides pharmaceutical formulations, for use in treatments against HIV/HTLV- 1 , II, III and other viral diseases and diseases caused by mycoplasma, bacteria or parasites.

The present invention also relates to a method comprising inoculating into a patient a human, animal, synthetic or recombinant amino acid sequence with or without adjuvant, to produce an antibody response, the antibodies, mono or polyclonal will cause the binding of the immunosuppressive CD3/TCR mimic interaction molecules already present in the plasma of the infected host will be removed from the circulation of the infected host and normal immune function demonstrating a Th_x cytokine profile, i.e. Interleukin 2 and gamma interferon, capable of resisting the infection will be re-established.

Vaccines manufactured by the use of coat or other peptides from viral, bacterial, parasitic or mycoplasma may be screened to determine whether they posses these specific amino acid sequences which exhibit these specific Ion bridge pair arrays capable of mimicking the actions of AFP or Interleukin 10 and there inherent immune suppressive properties - such suppressive potential properties is determined by the manufactured vaccine's reactivity with any of the said specific amino acid sequences listed herein which may be removed or neutralised by the antiserum specified in this patent .

Suitable dosages in accordance with the present invention depend on many factors, e.g. the patient's weight, the mode of administration, the frequency of administration, the type of affliction being treated or prevented, whether the infection presently exists, and if so, to what degree. Suitable dosages for given situations can readily be determined by those skilled in the art without undue experimentation.

The total treatment time according to the present invention will vary from patient to patient based on sound medical judgement and factors particular to the patient being treated, such as, for example, the age and physical condition of the patient. Those skilled in the art can easily determine suitable total treatment time on a patient by patient basis.

The following is a description of a suitable protocol in accordance with the present invention.

PROTOCOL FOR ADMINISTRATION OF AN IMMUNOGLOBULIN IgM MONOCLONAL ANTIBODY AGAINST HUMAN INTERLEUKIN 10. 1.0 INTRODUCTION The Human Immunodeficiency Virus Type 1 (HIV-1) is the etiological agent of Acquired Immune Deficiency Syndrome (AIDS) (1,2). AIDS is characterised as a profound breakdown in host's cellular and humoral immunity and increased susceptibility to a wide range of opportunistic infections. One of the consequences of this immune dysfunction is a marked depletion in absolute CD4+ cells in HIV- infected individuals. Studies over the past years have demonstrated that the destruction of the immune system by HIV-1 is a chronic process, starting at the moment of infection. The results indicate that strategies for effective therapeutic intervention using antibodies to these specific mimic CD3/TCR peptide interaction molecules should start early in infection to prevent irreversible damage occurring to the immune system. Since it has been demonstrated in HIV that an early loss of CD3/TCR mediated T cell activation is evident. This imbalance in turn effects monocyte and B cell function.

Recent studies have established the functional binding and immunosuppressive similarities between specific amino acid charged sequences present on the alfa-fetoprotein molecule and on Interleukin 10 and certain HIV envelope amino acid sequences, see fig. Laboratory data demonstrates that i munoglobulin G (IgG) or IgM to the said specific amino acid sequence inhibits syncytial formation and prevents HIV-1 laboratory strains MN, RF, and IIIB replication in C8166-45 cells (lymphocyte cell-line) in-vitro. In addition, IgG to the said amino acid sequence inhibits replication of HIV-1_BAL in fresh macrophage culture in a dose-dependent manner. 1.2 RATIONALE

The basic rationale for using this therapy is the understanding that there exists a functional binding and immunosuppression similarity between certain peptides containing specific ion pair arrangements of amino acids enclosed within two hydrophobic amino acids present within the AFP molecule Interleukin 10 and specific external HIV glycoproteins together with other specific viral coat peptides and glycopeptides . This discovery shows that as the body defends itself against the HIV virus by producing antibodies to specific viral coat proteins, these antibodies, while restricting in a normal antibody fashion the HIV virus, are themselves together with certain viral glycopeptides sequences identified herein and produced by the infecting virus are inherently immunosuppressive in that they perform a similar task as AFP or Interleukin 10 in that they selectively down regulate the T cell dependent immune system in favour of a humoural, B cell response which although it produces neutralising antibodies to the infectious agent (e.g. malaria, HIV, Tuberculoses) also allows the infective agent to persist and reproduce within the host cells and to ultimately undermine its immune status.

The major histocompatibility complex (MHC) is a collection of 40-50 genes arrayed within a long continuous stretch of DNA on chromosome 6 in humans . The MHC is referred to as the HLA complex in humans. The MHC genes are organised into regions encoding three classes of molecules: Class I, Class II and Class III. The Class I genes encode glycoproteins expressed on the surface of nearly all nucleated cells, where they present peptide antigens of altered self-cells necessary for the activation of T_c cells. The Class II genes encode glycoproteins expressed primarily on antigen-presenting cells (macrophages, dendritic cells, and B cells), where they present processed antigenic peptides to Th cells. The Class III genes encode somewhat different products that are also associated with the immune process. These include a number of soluble serum proteins (including components of the complement system) , steroid 21-hydroxylase enzymes, and tumour necrosis factors.

The administration of antibodies poly or mono clonal to these specific CD3/TCR mimic molecules will cause an immediate antibody-dependent cell-mediated cytotoxicity (ADCC) stimulated reduction in Viral Load as measured by the culturing of peripheral blood mononuclear cells and following the removal of the mimic Interleukin 10/AFP like viral peptide molecules and in the patient's blood we should see a re-awakening of a CD8 cytotoxic T cell reaction directed against HIV infected cells and this will coincide with a second HIV Viral Load reduction. Also in patient's who have received this antibody therapy we should see the generation of Interleukin 2 and gamma interferon and a dramatic increase in T4 cell number, together with a decrease in PCR and Quantitative Viral culture levels.

A number of white blood cells have cytotoxic potential and express membrane receptors for the Fc region of the antibody IgG molecule. When this antibody is specifically bound to a target cell which occurs when these specific poly or monoclonal antibodies to these sequences present on AFP, Interleukin 10 bind to HIV infected cells or free viral peptides causing immune Th2 shift. These cytotoxic Fc receptor-bearing cells can bind to the antibodies' Fc region, and thus to the infected HIV cells, and subsequently cause lysis of these cells. Although the cytotoxic cells involved are non-specific, the specificity of the antibody to a common immunosuppressive mimic peptide of Interleukin 10/AFP present on a large number of infecting organisms directs them to HIV infected target cells. This type of cytotoxicity is referred to as antibody-dependent cell-mediated cytotoxicity (ADCC) . The variety of cells that have been shown to exhibit ADCC include NK cells. Macrophages, monocytes, neutrophils, and eosinophils. 2.0 OBJECTIVES 2.1 To provide for an administration of monoclonal antibodies to these specified sequences present on AFP and Interleukin 10 and other cytokines and infectious organisms to HIV+ patients. 2.2 To monitor immune system functioning before and after the administration of these mono or polyclonal antibodies.

2.3 To monitor the effect of these type of antibody on cutanous lesions in those study participants who have Kaposi's Sarcoma. 2.4 To monitor viral load in patient's peripheral blood mononuclear cells prior to beginning, during and post this type of antibody infusion therapy.

2.5 To monitor the course or incidence of opportunistic infections in the study participants. 2.6 To determine the safety of these type of antibody administration in persons with HIV disease.

3.0 CLINICAL ENDPOINTS

To determine if these antibodies either poly or monoclonal may be of therapeutic benefit for widespread use in HIV disease based on the following criteria. -

3.1 Changes in T-cell phenotyping and cytokine profile.

3.2 Changes in the size, colour intensity, and palpable skin characteristics of cutaneous Kaposi's sarcoma lesions . 3.3 Changes in HIV load burden as indicated by endpoint - dilution culture quantitation in peripheral - blood mononuclear cells.

3.4 Changes in p24 antigen level.

3.5 Changes in Beta-2-microglobulin level 3.6. Appearance of new or improvement of active opportunistic infections.

3.7 Changes in system functioning (liver, kidney, haematology) .

Claims

I CLAIM

1. A method of treatment of a patent, either animal or human against any one or more of the following indications or infections listed below. (a) Immunodeficiency resultant from a viral infection.

(b) Immunodeficiency resultant from one or more of the following, bacterial, mycoplasmic, fungal and/or parasitic infections. (c) Immunodeficiency resultant from the growth of neoplastic tissue.

(d) Immunodeficiency resultant from any cytokine or hormone imbalance or imbalance of any natural product within the patient. (e) Myalgic Encephalo yelitis (ME) .

(f) Post inoculation or viral infection fatigue syndrome .

(g) Tuberculosis infection, (h) Malarial infection. wherein the treatment comprises administering an effective dosage of a pharmaceutical formulation comprising polyclonal or monoclonal antibodies generated to any one or more sequences as specified in the following list. Amino Acid Sequences as per This Patent Application Table A Sequence I.D. No

A001-1A Ala-His-Asp A002-1A Ala-His-Glu A003-1A Ala-Lys-Asp

A004-1A Ala-Lys-Glu

A005-1A Ala-Arg-Asp

A006-1A Ala-Arg-Glu A007-1I Ile-His-Asp

A008-1I Ile-His-Glu

A009-1I Ile-Lys-Asp

A010-1I Ile-Lys-Glu

A011-1I Ile-Arg-Asp A012-1I Ile-Arg-Glu

A013-1L Leu-His-Asp

A014-1L Leu-His-Glu

A015-1L Leu-Lys-Asp

A016-1L Leu-Lys-Glu A017-1L Leu-Arg-Asp

A018-1L Leu-Arg-Glu

A019-1M Met-His-Asp

A020-1M Met-His-Glu

A021-1M Met-Lys-Asp A022-1M Met-Lys-Glu

A023-1M Met-Arg-Asp

A024-1M Met-Arg-Glu

A025-1F Phe-His-Asp

A026-1F Phe-His-Glu A027-1F Phe-Lys-Asp

A028-1F Phe-Lys-Glu

A029-1F Phe-Arg-Asp

A030-1F Phe-Arg-Glu

A031-1P Pro-His-Asp A032-1P Pro-His-Glu

A033-1P Pro-Lys-Asp

A034-1P Pro-Lys-Glu

A035-1P Pro-Arg-Asp A036-1P Pro-Arg-Glu

A037-1W Trp-His-Asp

A038-1W Trp-His-Glu

A039-1W Trp-Lys-Asp

A040-1W Trp-Lys-Glu A041-1W Trp-Arg-Asp

A042-1W Trp-Arg-Glu

A043-1V Val-His-Asp

A044-1V Val-His-Glu

A045-1V Val-Lys-Asp A046-1V Val-Lys-Glu

A047-1V Val-Arg-Asp

A048-1V Val-Arg-Glu

A049-2A Ala-Asp-His

A050-2A Ala-Glu-His A051-2A Ala-Asp-Lys

A052-2A Ala-Glu-Lys

A053-2A Ala-Asp-Arg

A054-2A Ala-Glu-Arg

A055-2I Ile-Asp-His A056-2I Ile-Glu-His

A057-2I Ile-Asp-Lys

A058-2I Ile-Glu-Lys

A059-2I Ile-Asp-Arg

A060-2I Ile-Glu-Arg A061-2L Leu-Asp-His

A062-2L Leu-Glu-His

A063-2L Leu-Asp-Lys

A064-2L Leu-Glu-Lys A065-2L Leu-Asp-Arg

A066-2L Leu-Glu-Arg

A067-2M Met-Asp-His

A068-2M Met-Glu-His

A069-2M Met-Asp-Lys A070-2M Met-Glu-Lys

A071-2M Met-Asp-Arg

A072-2M Met-Glu-Arg

A073-2F Phe-Asp-His

A074-2F Phe-Glu-His A075-2F Phe-Asp-Lys

A076-2F Phe-Glu-Lys

A077-2F Phe-Asp-Arg

A078-2F Phe-Glu-Arg

A079-2P Pro-Asp-His A080-2P Pro-Glu-His

A081-2P Pro-Asp-Lys

A082-2P Pro-Glu-Lys

A083-2P Pro-Asp-Arg

A084-2P Pro-Glu-Arg A085-2W Trp-Asp-His

A086-2W Trp-Glu-His

A087-2W Trp-Asp-Lys

A088-2W Trp-Glu-Lys

A089-2W Trp-Asp-Arg A090-2W Trp-Glu-Arg A091-2V Val-Asp-His A092-2V Val-Glu-His A093-2V Val-Asp-Lys A094-2V Val-Glu-Lys A095-2V Val-Asp-Arg A096-2V Val-Glu-Arg

2. A method of treatment of a patient, either animal or human against any one or more of the following indications :

(a) Septic Shock

(b) Multiple Sclerosis

(c) Lupus Erythematoses

(d) Auto- immune diseases - myasthema gravis, rheumatoid arthritis, sjogrens disease

(e) Replacement of corticosteroid and hydrocortisteroid in the therapy of auto- immune and dermatological indications where these steroids were used to induce immuno- suppression. (f) Graft v host disease to reduce immune activity in organ and tissue transplant rejection.

The treatment comprises administration of an effective dosage of pharmaceutical formulation wherein the active constituent is one or more of the amino-acid charged ion bridge pairs attached to a hydrophobic amino acid or acids as outlined herein table

3. A method of preparation of a prophylactic vaccine antigen using inactivated coat or capsid peptides. Since vaccine preparations whose antigens contain the specified ion bridge pair hydrophobic amino acid sequences identified in the patient will not be capable of engendering protective immunity this preparation method for a vaccine that will produce both T & B cell memory response requires that when preparing the antigenic peptide it is necessary to delete or otherwise neutralise these specific sequences by the use of antibodies or deletion during synthesis. In live vaccine organism generation or synthesis this may be achieved by using anti-sense RNA and/or DNA strands to prevent synthesis in the organism of these cytokine like messenger signal sequences thus producing a viable infecting organism for use in vaccine preparation but one without the means to effect immunosuppression or avoidance of the T cell defendant immune system deletion chiron corporation malaria vaccine. Antigen Vivax-1 and SmithKline Beecham Malaria Vaccine NSI₈₁ V20 sequence D-R-A-A-D-G-Q-P-A-G both contain the specified sequences which are immunologically privileged and act as cytokine signal molecules similar to AFP and Interleukin 10. If these sequences are deleted and the vaccine antigen for malaria contained only the plasmodium vivax circumsporozoite (CS) protein minus ion bridge pairs associated with hydrophobic amino acid/or acids together with antibodies to these specific sequences as outlined in earlier claims then a proper response by both the T & B cell components of the immune system can be expected which will confer immunity. Another method capable of conferring immunity to infection by organisms which have previously resisted efforts to be good vaccine candidates and this applies to organisms such as Plasmodium which causes human malaria and to the HIV-1 HHV and influenza virus is to culture these organisms in the presence of antisense RNA or DNA to these specific sequences and then use the inactivated organisms produced to act as vaccine antigen. Also it is possible to make deletions to the infectious organisms genetic material so preventing it from generating these specific sequences, such genetically modified organisms could be used because they would infect, replicate and generate an immune system attack which would completely remove the infection since it would have been disarmed by not having these sequences to allow it shift the balance of the hosts immune attack on it and the vaccinated subject would retain a balanced complement of B & T cell memory defences against further infection.

4. A method whereby polyclonal or monoclonal antibodies generated to the specific sequences listed under Claim X can be used as a blood/serum or body fluid assay to determine the levels of these specific peptides since no antibody response would be expected by the affected human since these specific sequences are immunologically privileged and do not present as foreign. Since we have identified elevated levels of these peptides in patients suffering from Myalgic Encephalomyelitis this assay could be used for both diagnosis and for determining the progress of therapy in these and other conditions where elevated levels of these peptide cause disease states.

5. A method of immune treatment in human and/or animal with pharmaceutical formulations containing in whole or in part polyclonal or monoclonal antibodies generated to amino acid sequences which exhibit specific Ion bridge charged pair arrays of a positively charged amino acid and a negatively charged amino acid aligned together enclosed on one or both sides by a hydrophobic transmembrane segment of amino acids. There may be more than one Ion bridge pair separated by polar or non-polar amino acids present within the peptide to which the antibodies are generated.

6. A method of immune treatment in human and/or animal with a pharmaceutical formulation containing in whole or in part polyclonal or monoclonal antibodies generated to the peptide of sequence L-R-D-L-R-D-A which encloses two ion bridge pairs within non-polar amino acids on both sides.

7. A method of immune treatment in human and/or animal with a pharmaceutical formulation containing in whole or in part polyclonal or monoclonal antibodies generated to the specific peptide sequence

V-E-R-Y-L-K-D-Q which encloses two ion bridge pairs within both polar and non-polar amino acids.

8. A method of immune treatment in human and/or animal with a pharmaceutical formulation containing in whole or in part polyclonal or monoclonal antibodies generated to one or a combination of these specific peptide sequences (a) P-K-E-I-A

(b) A-D-K-V-M V-E-K-Y L-E-K-Y Y-D-K-I

L-E-K-I

S-E-R-L

G-E-K-I

L-E-R-G Y-E-H-V

L-E-K-C

G-D-R-A

G-E-K-L

T-E-R-V T-D-R-V

V-E-R-Y

Q-D-K-L

T-E-H-L

L-D-R-L F-E-K-T

S-R-D-L

L-E-K-Y

N-E-R-L

I-E-K-T N-E-K-F

9. A method according to Claims 1-4 wherein said antigenic peptide is selected from the group consisting in whole or in part, of human, animal, synthetic or recombinant alpha-fetoprotein (AFP) and/or cytokine inhibitory factor (Interleukin 10) , Malaria circumsporozite, Viral peptides.

10. A method for treating a patient, comprising administering a pharmaceutical formulation containing polyclonal and/or monoclonal antibodies to sequences as specified in Claims 1-4 as a therapeutic for the binding and removal of peptides generated by the infected host or infecting organism which have been specifically enhanced by the infecting organism to render a down regulation in Th_x cell type dependent immune resistance to infection.