MXPA99012105A - Herbal extract composition and method with immune-boosting capability - Google Patents

Abstract

A herbal extract composition comprising extract of ARUM, extract of POMEGRANATE, extract of HIBISCUS and extract of TEA. The herbal extract composition of the invention demonstrates in vitro stimulation of lymphocyte transformation and cytokine production, and in vitro inhibition of gp120 binding, and provides a potential candidate for therapies and treatments for immune disorders and HIV infection.

Description

COMPOSITION OF HERBAL EXTRACTS AND METHOD WITH IMMUNE REINFORCEMENT CAPACITY DESCRIPTION OF THE INVENTION This invention pertains generally to herbal extracts, methods and treatments, and more particularly to a composition derived from vegetable or herbal extract and the method having therapeutic immunity booster and antiviral effects to reinforce or stimulate the immune response in humans. The history of herbology is intertwined intrinsically with modern medicine. Many of the modern family medicines have been developed from ancient healing traditions associated with specific plants. The medical properties of many plants have been identified with specific chemical compounds that have been isolated, purified and, in many cases, reproduced synthetically. Many of the well-known drugs were originally derived from plants. Salicylic acid, the precursor of aspirin, was originally isolated from the white willow bark and the ulmaria plant. Quinine, which was used to treat malaria, was derived from Quinine bark. Vrincristine, which is used in cancer treatment, comes from vincafervinca. The cancer drug taxol was originally isolated from the bark and spikes of the Pacific Tejo tree. Perhaps the most famous are morphine and codeine, which were derived from the opium poppy. Morphine is still the standard against which new synthetic pain-releasing drugs are measured. The use of plants for medical purposes precedes recorded history. The root of acalia, hyacinth and millennium has been found carefully placed around the bones of a man of the stone age in Iraq. The acalia root is a soothing herb with anti-inflammatory properties and is used to treat inflamed or irritated mucous membranes. Hyacinth is used as a diuretic to encourage tissues to dislodge excess water. The mielenrrama is a remedy against the cold and fever that could have been used once as widely as aspirin is now. Modern physicians, particularly in the United States, tend to rely on treatments that use drugs manufactured in synthetic or chemical form. Instead of using whole plants or plant extracts for treatment, pharmacologists tend to identify, isolate, extract and synthesize active compounds from plants for use in treatment. This approach however has disadvantages. In addition to the individual physiologically active compounds present in the plant, there are also minerals, vitamins, glycosides, oils, alkaloids, bioflavinoids and other substances that may be important in supporting the medical properties of a particular plant. These additional substances can provide a synergistic effect that is absent when the purified physiological or synthetic compounds are used alone. Additionally, the toxicity of the purified physiologically active compounds is generally higher than when the physiologically active compounds are present with other plant substances. The effectiveness of various herbal remedies, extracts, potions and treatments is well known and therapeutic herbal products are increasingly recognized as desirable alternatives to synthetic drugs. For example, U.S. Patent No. 4,446,130 discloses the use of a ginseng extract that has stimulatory and diuretic effects. U.S. Patent No. 4,886,665 teaches the use of a pharmaceutical preparation of oat and nettle extracts. U.S. Patent No. 4,671,959 describes the use of blends of natural oils for stress reduction. U.S. Patent No. 5,064,675 relates to an herbal extract composition that provides a calming effect. U.S. Patent No. 5, 407,675 describes an herbal extract used for treatment of the scalp. U.S. Patent No. 5,178,865 discloses a herbal extract mixture that inhibits infection of the human immunodeficiency virus or HIV in vitro. U.S. Patent No. 5,500,340 and U.S. Patent No. 5,294,443 describe the use of herbal extracts for immunosuppression and the treatment of autoimmune disorders. The human immune response is extremely complex and requires a constant interaction of the cells involved in the immune system. Intracellular communication is facilitated by the secretion of chemical messenger proteins known as cytokines, which act to increase cell growth, promoting cellular activation, directing cellular traffic, stimulating macrophage function, destroying antigens and other functions. The cytokines generally comprise lymphokines, which are secreted by the T lymphocytes and the lymphokines secreted by the monocytes. Interferons which are used for defense against viral infection and tumor cell growth are an important class of cytokines. Cytokines also include interleukins, which are also involved in cell differentiation and tumor necrosis factors, and transformation growth factors that are involved in mediating inflammation and cytotoxic reactions. Another important set of proteins in the human immune response are immunoglobulins, which are secreted by B lymphocytes. Immunoglobulins serve as antibodies that counteract viruses, bacteria and other antigens. People suffering from immunosuppressive conditions or conditions such as human immunodeficiency virus (HIV), cancer, hepatitis, kidney disease, diabetes, asthma, arthritis and the like often experience decreased levels of lymphocytes and correspondingly decreased levels of cytokines. For example, when individuals have been infected by HIV lymphocytes that have a cell surface antigen known as CD4 they are present in low numbers in an uncharacteristic manner. While in healthy individuals those CD4 lymphocyte cells are present at concentrations of approximately 800 cells per milliliter of serum, individuals infected with HIV exhibit something as low as 200 CD4 cells per milliliter of serum when opportunistic infections develop. The specific types of known CD4 lymphocytes are TH1 and TH2 cells that appear to be particularly important in the measured response by cell for HIV infection. TH1 cells produce interleukin-2 (IL-2) and gamma interferon (IFN?). TH2 cells produce interleukins-4, -5 and -10 (IL-4, IL-5 and IL-10). The cytokines secreted by the THl and TH2 cells are considered to have an opposite effect on each other with the cytokines secreted by THL cells that act to regulate the cytokine production of TH2 cells and vice versa. At the beginning of the course of HIV infection the THl response dominates and the secretion of IL-2 by THL cells increases the activity of CD8 lymphocytes The naturally occurring interferons, interleukins and immunoglobulins secreted by lymphocytes are well structured to regulate the human immune response and the treatment of immunodeficient or immunosuppressed patients with interferons, interleukins and immunoglobulins that can be effective. The industrial production of interferons, interleukins and immunoglobulins by genetic engineering techniques is well known, where the genes responsible for producing these proteins are introduced into the bacteria, which are grown and harvested. The interferons, interleukins and immunoglobulins secreted by the genetically modified bacteria are then purified and delivered to patients through syringe or intravenous methods. An oral delivery method to provide interferons, interleukins and immunoglobulins to patients has not been achieved, since stomach acids and enzymes tend to separate these proteins before they can be delivered to the bloodstream. Several herbs are considered to have a beneficial effect on the human immune system. For example, it is considered that Echinacea Purpurea and Echinacea Angustifolia are believed to stimulate the activity of the T cell. Mo ery, D. B.; The Scientific Validation of Herbal Medicine, Keats Publishing, Inc., New Canaan Press. 1986, P. 118-119. Membranaceous Astragulas is considered to be capable of stimulating the production of interferon and immunoglobulins A and G in mice (IgA and IgG) Kaiser, J. D.: Immune Power A Comprehensive Treatment Program for HIV. St. Martin 's Press. New York, 1993. p. 59-60. However, a plant or herbal derivative that effectively provides a stimulating or reinforcing effect to the human immune system is not known until now. Accordingly, there is a need for an herbal extract composition and a composition and method that provide a health supplement and for the general treatment of people and animals having diseases or conditions that suppress the immune response. The present invention satisfies those needs as well as others and generally overcomes the deficiencies found in the prior art.

The present invention is a herbal extract composition and the method for making and using same to stimulate or enhance the immune response in humans and animals. The names, classifications and geographical origins of the plants or herbs used with the invention are given below in Table 1. TABLE I Typically, the fresh or dehydrated leaves, seeds, bark, fruits, husk, flowers and / or roots of each of the above plants or herbs can be used in the preparation of the herbal extract composition of the invention as described in detail then. However, the preferred portions of ARUM, POMEGRANATE, HIBISCUS and TE used for the extraction are shown in Table 1. In general, the herbs are extracted with hot water, although the aqueous ethanolic solutions and the ethanol can alternatively be used for the extraction. extraction. In general terms, the herbal extract composition of the composition comprises the ARUM extract. Another embodiment of the invention is an herbal extract composition comprising about ten percent and about 90 percent extract of ARUM, and between about ten percent and about ninety percent extract of POMEGRANATE. The herbal extract composition of the invention may further comprise between about ten percent and about ninety percent extract of HIBISCUS and / or between about ten percent and about ninety percent extract of TE. Preferably, the herbal extract composition of the invention comprises generally equal portions of ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extract. The herbal extract composition of the invention provides a healthy and nutritious complement to the human diet as it contains Vitamin C, Vitamin E, Vitamin D2, Vitamin D3, Vitamin K, solubilized minerals that include phosphorus, sodium, potassium, zinc, magnesium and copper and numerous beneficial proteins. The proteins include various cytokine and cytokine proteins and similar proteins and immunoglobulin or immunoglobulin which is considered to provide a stimulating or reinforcing effect to the immune system. As a nutritional supplement the herbal extract composition of the invention is ingested orally or can be applied topically. The experimental results (described below) indicate that the herbal extract composition of the invention is effective in boosting the human immune response. In vitro experiments show that the herbal extract composition of the composition stimulates cell-mediated immunity by stimulating blastogenesis or lymphocyte blasts formation by stimulating cytokine production in the lymphocyte suspension from normal persons , from leucine patients and patients suffering from renal failure. Initial experimental results also indicate that the herbal extract composition of the invention is effective in inhibiting HIV infection in vitro. These experiments and the resulting data demonstrate that the herbal extract composition of the invention is a useful source or candidate for potential immune disorder therapies.

The method for the preparation of the herbal extract composition of the invention preferably comprises the steps of preparing an ARUM extract, preparing an extract of POMEGRANATE, preparing a HIBISCUS extract, preparing a TE extract, and combining the ARUM extracts. , POMEGRANATE, HIBISCUS and TE. The method of use of the invention for stimulating cell-mediated immunity in vitro comprises contacting or exposing the cells to an efficient amount of a preparation consisting essentially of ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extract. . The method of use of the invention for stimulating lymphocyte blastogenesis in vitro comprises contacting or exposing the lymphocyte suspensions to an effective amount of a preparation consisting essentially of an ARUM extract., POMEGRANATE extract, HIBISCUS extract and TE extract. The method of use of the invention for stimulating cytokine production in vitro in peripheral blood mononuclear cells comprises contacting or exposing such cells to an effective amount of a preparation consisting essentially of extract of ARUM, extract of POMEGRANATE, extract of HIBISCUS and extract of An object of the invention is to provide an herbal extract composition and method that is a useful or candidate source for therapies and treatments of humans and animals suffering from immunosuppressive and autoimmune diseases, conditions, infections or conditions of this type. Another object of the invention is to provide a herbal extract composition and the method that provides a nutritious dietary supplement for humans and animals. Another object of the invention is to provide an herbal extract composition and the method that stimulates or reinforces the immune response in humans and animals. Another object of the invention is to provide a herbal extract composition and the method that stimulates immunity by in vitro cell. Another object of the invention is to provide an herbal extract composition and method that stimulate cell-mediated immunity in humans and animals. Another object of the invention is to provide a herbal extract composition and method that stimulate in vitro blasto transformation or lymphocyte blastogenesis. Another object of the invention is to provide a herbal extract composition and method that stimulate the transformation of lymphocyte blasto or blastogenesis in humans and animals. Another object of the invention is to provide a herbal extract composition and method that stimulate cytokine production in vitro in peripheral blood mononuclear cells. Another object of the invention is to provide herbal extract and method that stimulates the production of cytokine in humans and animals. Another object of the invention is to provide an herbal extract composition and the method for in vitro inhibition of the gpl20 binding to MT4 cells. Another object of the invention is to provide an herbal extract composition and the method for in vitro inhibition of HIV infection of MT-4 cells. The extract composition comprising the ARUM extract, POMEGRANATE extract, HIBISCUS extract, and TE extract to inhibit HIV infection in humans. Another object of the invention is to provide a herbal extract composition and method for the treatment of immunosuppressant conditions and conditions of this type that are relatively non-toxic. Another object of the invention is to provide an herbal extract composition that includes various cytokine and / or cytokine-like proteins and immunoglobulin and / or immunoglobulin-like proteins. Another object of the invention is to provide a herbal extract composition containing Vitamin C, Vitamin E, Vitamin D2, Vitamin D3, Vitamin K, solubilized minerals, including phosphorus, sodium, potassium, zinc, magnesium and copper and numerous beneficial proteins. Additional objects and advantages of the invention will be described in the following portions of the specification, wherein the detailed description is for the purpose of fully describing the preferred embodiment of the invention without placing limitations thereto. BRIEF DESCRIPTION OF THE DRAWINGS The present invention will be understood more fully by reference to the following drawings, which are for illustrative purposes only. Figure 1 is a graphical representation of the effect of the composition of the herbal extract of the invention on the blastogenesis of normal lymphocytes. Figure 2 is a graphic representation of the effect of the herbal extract of the infection on the blastogenesis of lymphocytes from patients with leukemia. Figure 3 is a graphical representation of the effect of the herbal extract of the invention on the blastogenesis of lymphocytes from patients having renal failure. The present invention generally belongs to the use of ARUM herbal extracts, POMEGRANATE, HIBISCUS and TE, and different combinations thereof to reinforce or stimulate the immune response in humans and animals for the treatment of people and animals suffering from immunosuppressive and autoimmune diseases or conditions, and to inhibit HIV infection in humans. humans. Example 1 illustrates the preparation process and the composition of the herbal extracts of ARUM, POMEGRANATE, HIBISCUS and TE. Example 2 relates to the use of an herbal extract composition comprising ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extract for the in vitro blasto transformation of normal lymphocyte suspensions. Example 3 relates to the use of the herbal extract composition comprising the ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extract for the in vitro blastogenesis of peripheral blood lymphocytes from patients with acute leukemia. Example 4 relates to the use of the herbal extract composition comprising the ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extract for the in vitro blastogenesis of peripheral blood lymphocytes from patients with renal failure.

Example 5 relates to the use of the herbal extract composition comprising the ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extract for the in vitro stimulation of cytokine production in peripheral blood mononuclear cells (PMBC) a from normal patients. Example 6 describes the use of the herbal extract composition comprising the ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extract for the in vitro inhibition of the gpl20 binding to MT-4 cells as measured with the OKT link. -4A mAb. Example 7 describes the use of the herbal extract composition comprising ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extract for in vitro inhibition of gpl20 binding to MT-4 cells, as measured with an anti gpl20 mAb. Example 8 describes the analytical examination of the combined extracts of ARUM, POMEGRANATE, HIBISCUS and TE. EXAMPLE 1 Preparation of Extract This Example describes the preparation and combination of extracts of ARUM, POMEGRANATE, HIBISCUS and TE. Reference is made to Table 1 above for the preferred plant parts of ARUM, POMEGRANATE, HIBISCUS and TE used for the extraction processes. The processes described below can be improved to produce larger quantities of the extracts. The details provided for the preparation of the following extracts reflect the currently preferred method for the preparation of extracts and should not be considered as limiting. The amounts and times described below can be substantially varied to provide suitable extracts of ARUM, POMEGRANATE, HIBISCUS and TE according to the invention. 1. Preparation of ARUM extract Approximately 20 grams of finely ground and dehydrated ARUM and one liter of distilled water were added to a stainless steel pressure cooking vessel and sealed therein with a stainless steel lid. The mixture therein was heated to boiling (about 100 ° C) and allowed to boil for about 45 minutes and then heated to about 130 ° C and allowed to boil for about 15 minutes, then allowed to cool below. 100 ° C to provide a liquid extract from ARUM. The ARUM extract was used while it was warm or hot in the manner described below. 2. Preparation of POMEGRANATE Extract Approximately 60 grams of finely ground dehydrated POMEGRANATE and one liter of distilled water were added to a stainless steel pressure cooking vessel and sealed therein with a stainless steel lid. The mixture in it was heated to boiling (about 100 ° C) and allowed to boil for about 45 minutes and then heated to about 130 ° C and heated for about 15 minutes, then allowed to cool to below 100 ° C. to provide a liquid effect of POMEGRANATE. The extract of POMEGRANATE was used while it was warm or hot in the manner described below. 3. Preparation of HIBISCUS extract Approximately 15 grams of finely ground and dehydrated HIBISCUS and one liter of distilled water were added to a stainless steel pressure cooking vessel and sealed therein with a stainless steel lid. The mixture therein was heated to boiling (about 100 ° C) and allowed to boil for about 45 minutes and then heated to about 130 ° C and boiled for about 15 minutes, allowing to cool to below 100 ° C to provide a liquid extract of HIBISCUS. The extract of HIBISCUS was used while it was warm or hot in the manner described below. 4. Preparation of TE Extract Approximately 40 grams of finely ground and dehydrated TE and one liter of distilled water were added to a stainless steel pressure cooker and sealed therein with a stainless steel lid. The mixture was heated to boiling (about 100 ° C) and allowed to boil for about 45 minutes and then heated to about 130 ° C and allowed to boil for about 15 minutes, then allowing to cool to below 100 ° C to provide a liquid extract of TE. The TE extract was used while it was warm or hot in the manner described below. 5. Preparation of Combined Extracts of ARUM and POMEGRANATE Approximately 50 ml of each of the extracts of ARUM and extract of POMEGRANATE were transferred while they were warm to hot (between approximately 40 ° C and 100 ° C) to a pressure cooker of stainless steel and were sealed therewith with a stainless steel lid and heated together with mixing or continuous stirring for about 1.5 hours at about 100 ° C. The mixture thus obtained could be cooled and then stored under refrigeration or used when it was warm to hot as described below to prepare the additional extract combinations. The combined extracts of ARUM and HIBISCUS, ARUM and TE, POMEGRANATE and HIBISCUS and POMEGRANATE and TE were also prepared by a generally identical procedure. 6. Preparation of Combined Extracts of ARUM, POMEGRANATE, HIBISCUS AND TE. Approximately 100 ml of combined extracts of ARUM and POMEGRANATE as prepared above, together with about 50 ml of each HIBISCUS extract as prepared above and about 50 ml of TE extract as prepared above, were transferred from warm to warm (between about 40 ° C and 100 ° C) to a stainless steel pressure cooking vessel and sealed therewith with a stainless steel lid and heated together with mixing or continuous stirring for about 1.5 hours at about 100 ° C. C. The mixture thus obtained generally had equal portions of each of the extracts of ARUM, POMEGRANATE, HIBISCUS and TE described above. The combined mixture of the extracts of ARUM, POMEGRANATE, HIBISCUS and TE was allowed to cool to room temperature. During cooling, the pH buffer (Na2HP04 NaH2P04) was added to the blended mixture while the mixture was approximately 30 ° C to 60 ° C to provide a slightly acidic pH to the mixture. Preferably, the pH of the combined extracts was adjusted to about 4.0 and more preferably to about 3.8 although a pH below 7 which is adequate was provided. After the above procedure, the combined extracts of ARUM, POMEGRANATE, HIBISCUS and TE were stored under refrigeration at approximately 4 ° C. A trace of sodium benzoate can be added to the combined extracts as a preservative and a flavoring or food coloring can be added if desired. The results of the chemical analysis for the combined mixture of extracts of ARUM, POMEGRANATE, HIBISCUS, TE are described below in Example 8. The above procedure was used to prepare a combined mixture of extracts of ARUM, POMEGRANATE and HIBISCUS but omitting the extract of TE.- The above procedure was used to prepare a combined mixture of extracts of ARUM, POMEGRANATE and TE but omitting the HIBISCUS extract. The above procedure was also used to prepare the combined mixture of extracts of POMEGRANATE, HIBISCUS and TE, but omitting the ARUM extract. The above procedure was also used to prepare the combined mixture of extracts of POMEGRANATE and HIBISCUS, but omitting the extracts of ARUM and TE. Example 2 In Vitro Blasto Transformation of Lymphocytes from Normal Patients This Example describes the effect of the herbal extract composition comprising the ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extract described in Example 1 in the cell-mediated immunity, which can be measured by blasto in vi tro transformation or blastogenesis of normal lymphocytes. Blastogenesis in the initial stage in the induction of cell-mediated immunity and is associated with the release or secretion of several interleukins that are essential for the intercellular interaction of the immune system. Cell-mediated immunity is "involved with the defense of the human body against malignancies, certain viral infections that include HIV, bacterial-intercellular infection as well as the regulation of different cellular and immunological humoral interactions and the prevention of autoimmune reactions. of blasto was measured by means of a conventional lymphocyte stimulation test in which 3H (tritiated) thymidine is added to the lymphocyte suspensions, followed by incubation, cell culture and radioactivity measurement of the harvested cells. They indicate that the lymphocytes have undergone transformation and conform to 3H thymidine.The lymphocytes from the whole blood of three normal patients were obtained by density gradient separation in Ficoll Isopaque.The multiple well microtiter plates were prepared containing lymphocyte suspensions in solution of Hanks supplemented with 10% serum - veal fetal, penicillin and streptomycin, 5 μl, 10 μl and 20 μl of the combined mixture of the extracts of ARUM, POMEGRANATE, HIBISCUS, TE was added to the wells in each of the plates. The final volume of the lymphocyte suspension in each well was 0.2 ml. Microtiter plates were then incubated for 72 hours at 37 ° C in a C02 incubator. Each well of 0.05 ml of H (tritiated) thymidine was then added, followed by an additional 24 hour incubation at 37 ° C, after which the wells were harvested with an automatic multiple harvester and the radioactivity was measured with a counter of scintillation. A stimulation index was calculated from the average counts for each concentration of combined extract. The results are shown in Table 2 and Figure 1. TABLE 2 Average Count (Stimulation Index Speed 3H Intake) Non-Combined Extracts 284 Minimum Counts (Background) 20 μl Combined Extracts 815 Minimum Counts 2.86 10 μl Combined Extracts 723 Counts Minimums 2.54 5 μl Combined Extracts 463 Minimum Counts 1.63 The results illustrated in Table 2 and in FIGURE 1 show that the addition of increased amounts of the combined extracts to lymphocyte suspensions results in increased amounts or degrees of lymphocyte or blastogenesis transformation as they are shown by the increased radiation counts due to the increased 3H intake. These results indicate that the herbal extract composition comprising the ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extract reinforces or stimulates cell-mediated immunity by stimulating blastogenesis and the resulting secretion of cytokines in normal persons. These results further demonstrate that the herbal extract composition of the invention is a useful or candidate source for the therapies and treatments of potential immune conditions. EXAMPLE 3 In Vitro Blastogenesis of Peripheral Blood Lymphocytes from Patients with Acute Leukemia This Example describes the effect of the herbal extract composition comprising the ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extract described in Example 1 on cell-mediated immunity, which can also be measured by transforming blasto in vi tro or blastogenesis of lymphocytes from five patients with leukemia. PHA (fithomatoglutinin) a known powerful mitogen that stimulates lymphocyte transformation was used as a control to calibrate or measure the efficiency of the combined extracts. Multiple well microtiter plates were prepared containing lymphocyte suspensions from five patients with leukemia in Hanks' solution supplemented with 10% fetal calf serum, penicillin and streptomycin. Five μl, 10 μl and 20 μl, quantities of the combined mixture of the extracts of ARUM, POMEGRANATE, HIBISCUS, TE were added to the wells in each of the plates. Additionally, PHA (Phytohemaglutamine) in concentrations of 1:50 and 1: 100 were added to the wells as controls. The final volume of the lymphocyte suspension in each well was 0.2 ml. The microtiter plates were incubated for 72 hours at 37 ° C in a C02 incubator, after which 0.05 ml of 3H tritiated thymidine were added to each well, followed by an additional 24 hour incubation at 37 ° C. The wells were harvested with an automatic multiple harvester and the radioactivity was measured with a scintillation counter and a stimulation index was calculated for each concentration of combined extracts. The results are shown in Table 3 and FIGURE 2.

TABLE 3 The results illustrated in Table 3 and in FIGURE 2 illustrate that the addition of increasing amounts of the combined extracts to lymphocyte suspension from patients with leukemia resulted in increased amounts of lymphocyte transformation or blastogenesis, as shown by the counts of increased radiation due to the increased intake of 3H. The herbal extract composition comprising ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extract exhibits an ability to stimulate lymphocyte transformation which is at least as large as PHA. The stimulation and transformation of the blast was increased by 306% at the highest concentration (20 μl). These results indicate that the herbal extract compositions of the ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extract reinforce or stimulate the cell-mediated immunity in vitro by stimulating the lymphocyte blastogenesis and thus the resulting secretion of cytokines in lymphocytes from patients suffering from leukemia. These results further demonstrate that the herbal extract composition of the invention is a useful source or candidate for therapies and potential treatments of immune disorders. EXAMPLE 4 In Vitro Blastogenesis of Peripheral Blood Lymphocytes from Patients with Renal Failure This Example describes the effect of the herbal extract composition comprising ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extract described in Example 1 on cell-mediated immunity, which was further measured by in vitro blasto transformation or lymphocyte blastogenesis from fifteen patients suffering from renal failure. PHA (phytohemagglutinin) in two concentrations was used as a control to calibrate or measure the efficiency of the combined extracts. Multiple well microtiter plates were prepared containing lymphocyte suspensions from five patients with leukemia in Hanks' solution supplemented with 10% fetal calf serum, penicillin and streptomycin. The combined mixture of the extracts of ARUM, POMEGRANATE, HIBISCUS, TE was added to the wells of each of the plates in quantities of 5 μl, 10 μl and 20 μl. PHA at a concentration of 1:50 and 1: 100 was added to the wells as controls. The final volume of the lymphocyte suspension in each well was 0.2 ml. The microtiter plates were incubated for 72 hours at 37 ° C in a C02 incubator, after which 0.05 ml of 3H tritiated thymidine were added to each well followed by an additional 24 hour incubation at 37 ° C. The wells were harvested with an automatic multiple harvester and the radioactivity was measured with a scintillation counter and the stimulation index was calculated for each concentration of the combined extracts. The results are shown in Table 4 and FIGURE 3.

The results illustrated in Table 4 and FIGURE 3 show that the addition of increasing amounts of the combined extracts to the suspensions of lymphocytes from patients with renal failure results in increasing amounts of lymphocyte transformation or blastogenesis, as shown by the increased radiation counts due to increased 3H intake. The combined extracts of ARUM, POMEGRANATE, HIBISCUS and TE exhibit an ability to stimulate lymphocyte transformation which is at least as large as PHA. The stimulation of the blasto transformation is increased by 306% in the highest concentration (20 μl). The results indicate that the combined extracts of ARUM, POMEGRANATE, HIBISCUS and TE reinforce or stimulate cell-mediated immunity in vitro by stimulating lymphocyte blastogenesis and the resulting secretion of cytokines in lymphocytes from patients suffering from renal failure. These results further demonstrate that the herbal extract composition of the invention is a useful source or candidate for treatments or potential therapies of immune disorders. EXAMPLE 5 Stimulation of Cytokine Production in Normal People The effect of the herbal extract composition comprising ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extract on cytokine production by human peripheral blood mononuclear cells (PMBC) was measured using the method described by D. Schols and E. De Clencq in Human Immunodeficiency Virus Type gpl20 Induces Anergy in Human Peripheral Blood Lymphocytes By Inducing Interleukin Production. J. Virol. Vol. 70. p. 4953-4960 (1996), the contents of which are incorporated by reference. The average results for normal patients are shown in Table 5.

TABLE 5 As can be seen in Table 5, the presence of the herbal extract composition comprising the ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extract induces the production of considerable amounts of interleukin 10 (IL-10) and ? -interferon (IFN-?) on levels secreted in wells without herbal extracts. Interleukin 2 (IL-2) is detectable in wells where the combined extracts are present in a concentration of 10%. No toxicity was observed in cell cultures at 10% concentration of the combined extracts as measured by trypan blue. These results indicate that the combined extracts of ARUM, POMEGRANATE, HIBISCUS and TE reinforce or stimulate the cytokine production in vitro in PMBC from normal persons. These results further demonstrate that the herbal extract composition of the invention is a useful source or candidate for potential therapies and treatments of immune disorder. EXAMPLE 6 In Vitro Inhibition of gpl20 Link to MT-4 cells Measured with Monoclonal Antibody OKT-4A. The effect of the herbal extract composition comprising extract of ARUM, extract of POMEGRANATE, extract of HIBISCUS and extract of TE ("combined extracts") on the binding of rpgl20 to MT-4 as verified with OKT-4A monoclonal antibodies (mAb ) was determined using the method described by J. Neyts, D. Reymen, D. Letoumeur, J. Josefonvicz, D. Schols, J. Este, G. Andrei, P. McKenna, M. itvrouw, S. Ikeda. J. Clements and E. De Clercq in Differential Antiviral Activity of Derva tized Dextrans. Biochem. Pharmacol. Vol. 50 p. 743-751 (1995), the contents of which are incorporated herein by reference. The HIV coat protein gpl20 that binds to the OKT-4A blocks of the competitive binding of OKT-4A mAb. The combined extracts of ARUM, POMEGRANATE, HIBISCUS and TE of the gpl20 link blocks are as shown in Table 6. TABLE 6 As can be seen from Table 6, when the increasing amounts of herbal extract composition comprising the ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extract results in the increased inhibition of the gpl20 binding to the cells MT-4. These results demonstrated that the herbal extract composition comprising an extract of A-RUM, extract of POMEGRANATE, extract of HIBISCUS and extract of TE is a useful source or candidate for treatments and potential anti-HIV therapies in infected hosts. EXAMPLE 7 In Vitro Inhibition of the gpl20 Link to MT-4 Cells. Measured with Monoclonal Antibody Anti-gpl20 The effect of the herbal extract composition comprising the ARUM extract, the POMEGRANATE extract, the HIBISCUS extract, and the TE extract ("combined extracts") on the link from rpgl20 to MT- 4 as labeled with anti-gpl20 monoclonal antibodies (mAb) was determined using the method described by D. Schols, M. Baba, R. Pauwels, and E. De Clercq in Fl ow Cytometri c Method To Demonstrate For Anti-HIV- 1 Agen ts Inhibit Virion binding To T4 + Cells. the contents of which are incorporated into it by reference. The combined extracts of ARUM, POMEGRANT, HIBISCUS and TE in blocks block the binding of gpl20 as shown in Table 7.

TABLE 7 As can be seen from the Table, the herbal extract composition comprising ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extract, inhibit the binding of gpl20 to MT-4 cells. These results show that the composition of herbal extract containing ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extract is a useful source or candidate for treatment and potential anti-HIV therapies in vivo in infected hosts. EXAMPLE 8 Chemical Analysis of Combined Extracts Conventional analytical techniques were used to investigate the chemical composition of the herbal extract composition comprising ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extract for various vitamins, fats, dyes, proteins , minerals and other compounds. This analytical results are summarized in Table 8. TABLE 8 As can be seen from the results of Table 8, the herbal extract composition comprising the ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extract include numerous essential vitamins including Vitamin C, Vitamin E, Vitamin K , Vitamin D2, Vitamin D3 as well as several essential minerals different from beneficial materials and therefore provides a useful and nutritious dietary supplement when ingested orally. The analytical results shown in Table 7 also indicate that various cytokines or cytokine-like proteins are present in the herbal extract composition comprising the ARUM extract, the POMEGRANATE extract, the HIBISCUS extract and the TE extract. Particularly, detectable amounts of IL-4, IL-5, IL-6, IL-10, IL-12, IFN-? and TNFa are present in the combined extracts of ARUM, POMEGRANATE, HIBISCUS and TE. Additionally, traces of IgG or an immunoglobulin-like protein appear to be present in the herbal extract composition comprising the ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extract. The presence of human IL-4, IL-5, IL-6, IL-10, IL-12, IFN ~ ?, TNFa and IgG is in some way incompatible with the plant origin of the herbal extract composition. However, plant antiviral factors (AVF) are known to have a close sequence homology to human β-interferon (IFNβ) and are considered to exert an antiviral activity in a manner similar to that of human interferons. Therefore, the positive results for IL-4, IL-5, IL-6, IL-10, IL-12, IFN-? TNFa and IgG described above may be due to interleukin-like proteins, interferon-like proteins , TNF-like proteins, and immunoglobulin-like proteins that are present in the herbal extract composition comprising ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extract. Therefore, the term "herbal extract composition comprising ARUM extract POMEGRANATE extract, HIBISCUS extract and TE extract" as used herein which also includes or comprises IL-4, IL-5, IL-6 , IL-10, IL-12, IFN- ?, TNFa and IgG, as well as interleukin-like proteins, inteferon-like proteins, TNF-like proteins and immunoglobulin-like proteins. DISCUSSION The above in vitro experiments and the resulting data demonstrate that the herbal extract compositions of the invention are a potential useful or candidate source for therapies and immune booster treatments for humans and animals suffering from disorders, diseases, infections or conditions that include immunosuppressed conditions, due to leukemia, renal failure, various types of cancer and tumors, viral infections, bacterial infections, and parasitic infections. The precise composition or chemical compositions and pharmacological mechanisms that result in the stimulation of in vitro blasto transformation and cytokine production have not been determined. The herbal extract composition of the invention may contain an individual pharmacological active ingredient, component or agent acting alone or a combination of such ingredients. components or agents and / or biological metabolites or derivatives thereof acting separately or synergistically. In the in vivo therapies and treatments using the herbal extract composition of the invention which are similarly based on orally ingested doses on the oral extract composition or in liquid or solid form. Rectal, parenteral, intravenous, topical, aerosol, inhalation or subcutaneous routes for in vivo administration of the herbal extract composition of the invention are also possible. The herbal extract composition can also be administered in vivo in a mixture or combination with excipients, carriers, antiviral agents, immune modulators, chemotherapeutic agents, antibodies or combinations thereof. The pharmacological preparations of the invention may be in unit dosage forms, such as tablets, capsules, suppositories, ampoules or metered doses in liquid or aerosol. Preferably, it is considered "that the pharmacologically active component or components of the invention are present in ARUM and / or POMEGRANATE." Initial experiments indicate that the ARUM extract alone, the POMEGRANATE extract alone and the combined extracts of ARUM and POMEGRANATE exhibit some effects of immunological reinforcement However, the preferred embodiment of the invention comprises combined extracts of ARUM and POMEGRANATE as well as HIBISCUS and TE It is considered that HIBISCUS and TE provide other additional components or agents that act as excipients or carriers or that otherwise have a synergistic effect together with the components provided by ARUM and POMEGRANATE Additionally, the preferred method of preparing herbal extract composition comprising ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extract, wherein the combined extracts they are heated together in stages as described in Example 1 is considered to be more advantageous than the pure mixture or the combination of ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extracts without further heating. The above in vitro experiments and the resulting data also demonstrate that the herbal extract composition of the invention is a potential useful source or candidate for anti-HIV and other antiviral therapies and treatments for humans and animals suffering from HIV, other viral infections or complications or conditions associated with such viral infections. The herbal extract of the invention can potentially inhibit or prevent the infection of cells by a variety of different viruses, including but not limited to, herpes simplex virus, human papilloma virus (HPV), vaccinia virus, vesicular stomatitis virus, coxsaquie virus, syncytial virus, cytomegalovirus, and varicella zoster virus. The anti-HIV activity of the herbal extract composition of the invention may contain a single pharmacologically active ingredient, component or agent acting alone, or in combination of such ingredients, components or agents and / or biological metabolites or derivatives thereof. In vivo therapies and treatments using the herbal extract composition of the invention for HIV and antiviral treatments will likewise be based on topically applied or orally ingested doses of the herbal extract composition in liquid or solid form. Rectal, parenteral, intravenous, or aerosol or subcutaneous inhalation routes for in vivo administration of the herbal extract composition of the invention is also possible. As for the treatment of immunosuppressive treatments, the herbal extract composition can also be administered in vivo to antiviral treatments in admixture or combination with the excipients, carriers, antiviral agents, immune modulators, chemotherapeutic agents, antibodies, antiviral agents or preferred combinations of the same. Chemotherapeutic preparations may be in the form of unit doses such as tablets, capsules, suppositories, lotions, ampoules or metered doses of liquid or aerosol. Accordingly, it will be noted that this invention provides an herbal extract composition that provides a nutritional supplement that stimulates cell-mediated immunity in vitro, which stimulates in vitro lymphocyte blastogenesis, which stimulates in vitro cytokine production by lymphocytes, which inhibits HIV infection in vitro, and which provides a potential candidate for treatments and therapies to strengthen and stimulate the immune response in humans and animals. Although the above description contains many specifications, these should not be considered as limiting the scope of the invention but only that they provide an illustration of the currently preferred embodiment of the invention. Therefore, the scope of the invention should be determined by the appended claims and their legal equivalents.

Claims (38)

1. CLAIMS 1. An herbal extract composition to stimulate cell-mediated immunity, characterized in that it comprises ARUM extract.
2. 2. The herbal extract composition according to claim 1, characterized in that it also comprises extract of POMEGRANATE.
3. 3. The herbal extract composition according to claim 1, characterized in that it also comprises extract of HIBISCUS.
4. 4. The herbal extract composition according to claim 1, characterized in that it also comprises TE extract.
5. 5. The herbal extract composition according to claim 2, characterized in that it also comprises extract of HIBISCUS.
6. 6. The herbal extract composition according to claim 2, characterized in that it also comprises TE extract.
7. 7. The herbal extract composition according to claim 5, characterized in that it also comprises extract of TE.
8. 8. The herbal extract composition to stimulate cell-mediated immunity, characterized in that it comprises POMEGRANATE extract.
9. 9. The herbal extract composition according to claim 8, characterized in that it also comprises extract of HIBISCUS.
10. 10. The herbal extract composition according to claim 8, characterized in that it also comprises TE extract.
11. 11. The herbal extract composition according to claim 9, characterized in that it also comprises TE extract.
12. 12. A method of preparing an herbal extract composition for stimulating cell-mediated immunity, characterized in that it comprises the steps of: (a) preparing ARUM extract by boiling finely ground and dehydrated ARUM in water; and (b) preparing POMEGRANATE extract by boiling, of finely ground POMEGRANATE and dehydrated in water; and (c) combining the extract of ARUM and the extract of POMEGRANATE and boiling such combined extracts of ARUM and
13. POMEGRANATE The method of preparing an herbal extract according to claim 12, characterized in that it further comprises the steps of: (a) preparing the HIBISCUS extract by boiling HIBISCUS finely ground and dehydrated in water; (b) preparing TE extract by boiling finely ground TE dehydrated in water; and (f) combining the HIBISCUS extract and the TE extract with the combined extracts of ARUM and POMEGRANATE and boiling such combined extracts of ARUM, POMEGRANATE, HIBISCUS, TE.
14. 14. A method for stimulating in vitro cell-mediated immunity characterized in that it comprises contacting the cells with an efficient amount of a preparation consisting essentially of ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extract.
15. 15. The method for stimulating the cell-mediated immunity in vitro according to claim 14, characterized in that the cells comprise cells from normal patients.
16. 16. The method for stimulating in vitro cell-mediated immunity according to claim 14, characterized in that the cells comprise cells from patients having leukemia.
17. 17. The method for stimulating the cell-mediated immunity in vitro according to claim 14, characterized in that the cells comprise cells from patients having renal failure.
18. 18. A method for stimulating in vitro lymphocyte blastogenesis characterized in that it comprises contacting the lymphocyte suspensions with an effective amount of a preparation consisting essentially of ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extract.
19. 19. The method for stimulating the cell-mediated immunity in vitro according to claim 18, characterized in that the lymphocyte suspensions comprise lymphocytes from normal patients.
20. 20. The method for stimulating in vitro cell-mediated immunity in accordance with claim 18, characterized in that the lymphocyte suspensions comprise lymphocytes from patients having leukemia.
21. 21. The method for stimulating the cell-mediated immunity in vitro according to claim 18, characterized in that the lymphocyte suspensions comprise lymphocytes from patients having renal failure.
22. 22. A method for stimulating in vitro cytokine production, comprising contacting the peripheral mononuclear blood cells with an effective amount of a preparation consisting essentially of ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extract.
23. 23. The method for stimulating the cell-mediated immunity in vitro according to claim 22, characterized in that peripheral blood mononuclear cells are obtained from normal patients.
24. 24. The method for stimulating the cell-mediated immunity in vitro according to claim 22, characterized in that peripheral blood mononuclear cells are obtained from patients having leukemia.
25. 25. The method for stimulating the cell-mediated immunity in vitro according to claim 22, characterized in that peripheral blood mononuclear cells are obtained from patients having renal failure.
26. 26. A method for inhibiting HIV infection in vitro characterized in that it comprises contacting the cells with an efficient amount of a preparation consisting essentially of -ARUM extract, POMEGRANATE extract, HIBISCUS extract and TE extract.
27. 27. A method for inhibiting the in vitro binding of gpl20 to MT-4 cells characterized in that it comprises contacting the MT-4 cells with an effective amount of a preparation consisting essentially of extract of ARUM, extract of POMEGRANATE, extract of HIBISCUS and TE extract.
28. 28. An herbal extract composition for stimulating cell-mediated immunity characterized in that it comprises a mixture of an aqueous extract of ARUM, an aqueous extract of POMEGRANATE, an aqueous extract of HIBISCUS and an aqueous extract of TE.
29. 29. The herbal extract composition according to claim 28, characterized in that the composition further comprises at least one cytokine.
30. 30. The herbal extract composition according to claim 29, characterized in that the cytokine is an interleukin.
31. 31. The herbal extract composition according to claim 29, characterized in that the cytokine is an interferon.
32. 32. The herbal extract composition according to claim 28, characterized in that the composition further comprises at least one tumor necrosis factor.
33. 33. The herbal extract composition according to claim 28, characterized in that the composition further comprises at least one immunoglobulin.
34. 34. The herbal extract composition according to claim 28, characterized in that the composition further comprises polysaccharide.
35. 35. The herbal extract composition according to claim 28, characterized in that the composition further comprises proteins.
36. 36. The herbal extract composition according to claim 28, characterized in that the composition further comprises RNA.
37. 37. The herbal extract composition according to claim 28, characterized in that the composition further comprises DNA.
38. 38. The herbal extract composition according to claim 28, characterized in that the composition contains approximately equal portions of such aqueous extracts.