The Grouppe Kurosawa HIV Treatment Protocol
8.8.06
THE NEW REVISED GROUPPE KUROSAWA HIV TREATMENT PROTOCOL, Parts One and Two. August, 2006
This protocol is out of date. It is posted here for informational purposed only.
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INTRODUCTION
There are two parts to this treatment protocol. In part one, we are attempting to inhibit the synthesis of the virus. We want the viral load reduced as low as possible in order to enhance the efficacy of part two. In part two, we will attempt to activate the immune response against virally infected cells in an effort to finally clear the virus from the body. I have learned from bitter experience that you cannot successfully block HIV synthesis and activate the immune system at the same time.
This protocol has only one purpose…to kill off virally infected cells. In brief, we are looking for a cure.
Many of the sections in this protocol are taken directly from our blog essays. Since the current Natural Medicine blog is private, we have pasted most of the text into this document. Some of these essays are directed toward cancer immunotherapy protocols. This is irrelevant. The same immunological principles apply to the destruction of HIV infected cells.
PART ONE
This treatment phase uses niacinamide (nicotinamide), lactoferrin, ibuprofen, vitamin A, anti-oxidants, and cimetidine. This phase of treatment is to be continued for 2 to 3 months. The compounds are listed in order of importance.
1. Niacinamide (Nicotinamide)
Nicotinamide is indeed the perfect anti-HIV compound. First, it inhibits the ability of the viral TAT protein to induce activation of the HIV gene. Second, it blocks the infection of new cells. What more could you ask for from an inexpensive, safe supplement?
I became interested in the role nicotinamide plays in HIV synthesis when I read the following article. This review article, which can be read online by clicking the University of Chicago box, presents some startling information. The article speaks for itself. For now, it is enough to know that nicotinamide is an effective treatment agent against BOTH TB and HIV. Wow...
The question is why?
As it turns out, there are two reasons. I won't discuss TB in this essay.
First, the SIRT1 deacetylase enzyme, discussed in the last two essays, binds directly to the viral TAT protein, thereby promoting it's activity. TAT is an acetylated protein. The acetylation of TAT inhibits its ability to stimulate HIV synthesis. The activation of the HIV gene absolutely requires a functional TAT protein.
SIRT1 recycles TAT to its non-acetylated form, thereby acting as a cofactor of TAT in the activation of HIV synthesis.
SIRT1 deacetylase activity is inhibited by nicotinamide. This clearly means that nicotinamide inhibits the TAT activation of the HIV gene.
There is more.
In order for the HIV gene to be integrated into the DNA of a cell, the DNA must be slightly damaged. Under non-stress conditions, the viral protein integrase cuts the DNA, thereby initiating the complex process of HIV gene insertion into the cellular DNA. However, if the cellular DNA is damaged by oxygen radicals, this process is tremendously accelerated. This is one of the reasons anti-oxidants inhibit HIV synthesis.
When DNA is damaged, the free ends of the DNA initiate a stress response that activates a protein called Poly ADP-ribose polymerase-1 or PARP-1 for short. PARP-1 promotes DNA repair. It is also necessary for the integration of different viruses into cellular DNA. In brief, PARP-1 is absolutely necessary for the integration of the HIV gene into the DNA of newly infected cells. If PARP-1 is inhibited, the infection of new cells is blocked.
In addition, PARP-1 inhibits the UV light induction of HIV synthesis. In this case, the cells are already stably infected with the HIV virus. UV light generates free oxygen radicals which in turn activate the HIV gene. This process is PARP-1 dependent, and inhibited by nicotinamide.
It is very likely that this PARP-1 dependent process involves the activation of the NF-kappaB genetic factor, an established activator of HIV viral synthesis.
Nicotinamide inhibits HIV synthesis in both acute and chronic laboratory infections.
In addition, nicotinamide inhibits DNA fragmentation in lymphocytes of HIV infected persons. DNA fragmentation correlates well with the apoptosis of HIV infected and non-infected cells.
Nicotinamide, aka niacinamide, is indeed a perfect HIV treatment molecule. And this is how it works.
All NAD dependent biochemical reactions work this way. The ADP ribose part of the NAD molecule is donated to a target protein. Nicotinamide is the OTHER part of the NAD molecule. Normally, nicotinamide is recycled back to NAD. However, if excessive amounts of nicotinamide build up, there is a feedback inhibition of the NAD biochemical reaction. In our case, SIRT1 and PARP-1 are both NAD requiring enzymes. They serve different functions, but that is completely irrelevant. Any NAD dependent reaction can be inhibited by nicotinamide.
2. Lactoferrin
This subject area is completely new to me. I tripped over it by accident while doing literature searches on HIV and dendritic cell function.
Background.
Lactoferrin is an iron binding protein found in milk. Its immunological claim to fame is that it stimulates the synthesis of the powerful immune hormone IL-18 in the gastric immune system. This means that lactoferrin really is a powerful immune adjuvant. Lactoferrin's adjuvant properties are not the topic of this essay.
The immune system in the intestine, the GI tract mucosa, is the major site of HIV synthesis in the body. Since the major sites of HIV entry into the body are the vaginal and anal mucosa, the virus eventually ends up in the GI tract.
Immature dendritic cells play the primary role in detecting and responding to foreign antigens in the gastric mucosa. These cells have many ways of detecting pathogens, and one of them is via a lectin (carbohydrate binding protein) expressed on their membranes called DC-SIGN. DC-SIGN binds a variety of pathogen related molecules, resulting in their uptake into the dendritic cells. The dendritic cells then process these pathogens into fragments that can associate with class 2 histocompatibility antigens for the activation of naive CD4 T cells.
The HIV virus binds DC-SIGN via its membrane protein gp120/160. The dendritic cell takes up the virus, but the virus is not killed. Quite to the contrary, the virus remains alive. When the dendritic cells bind naive T cells, the HIV virus is directly transmitted to these new cells. This type of cell-cell transmission is the major method of virus dissemination in the body. The following abstract is a good review of this topic.
Although cell free virus can certainly infect CD4 T cells, the process is very inefficient. The half life of HIV infected or activated CD4 T cells in the body is less than 2 days. In the face of such a rapid turnover of HIV infected cells, the state of viral persistence can ONLY be maintained if infected dendritic cells transfer live virus to non-infected CD4 T cells. The following article is an excellent review of this most important topic.
The viral particles released from the dendritic cells in the face of CD4 T cells are TEN TIMES more infectious than free viral particles.
Lactoferrin binds strongly to DC-SIGN thereby blocking the gp120--DC-SIGN interaction. This prevents the uptake of virus into the dendritic cells and the subsequent infection of new CD4 T cells. Interestingly, bovine or cow lactoferrin, which is extremely plentiful, has a higher affinity for DC-SIGN than human lactoferrin.
When immature dendritic cells pick up HIV particles through its DC-SIGN receptor, these cells migrate to lymph nodes to activate CD4 T cells. Unfortunately, the HIV virus they carry is NOT dead. As a result, these dendritic cells infect all the naive CD4 T cells that surround them in the lymph nodes. The lymph nodes are the primary site of HIV synthesis.
Bovine lactoferrin is plentiful, easy to purify and totally non-toxic. It is truly a powerful natural medicine for the treatment of HIV infections. Again, keep in mind that HIV infected CD4 cells turn over rapidly. This means that CD4 T cell counts can rebound if the dendritic--CD4 T cell transmission of virus is blocked.
3.Ibuprofen
Over activation of the Cox-2 enzyme results in the production of large amounts of prostaglandins. One prostaglandin, PGE2, is BOTH a major immunosuppressant AND a stimulator of cancer cell growth, metastasis and angiogenesis. There are hundreds and hundreds of scientific articles published on the role of Cox-2 prostaglandins in cancer cell development, yet oncologists never prescribe indomethacin, a powerful cox-2 inhibitor (primarily used to treat gout), to treat cancer or leukemia. Why?
The following article is a good general introduction of the role of Cox-2 enzyme activity and breast cancer. This is just one of many, many such articles.
Prostaglandins such as PGE2 induce immunosuppression in many different ways. Lets cite a few.
1. PGE2, via cyclic AMP stimulation, induces immunological tolerance against tumor antigens. This means the immune system is being taught to ignore the presence of the tumors. Not a good idea.
2. PGE2 induces the synthesis of IL-10, an immunosuppressive hormone.
3. PGE2 inhibits the maturation of dendritic cells, antigen processing cells absolutely necessary for activating CD8 and CD4 T cells against tumors and other foreign substances.
4. PGE2 inhibits the gene for the powerful immune hormone IL-2. IL-2, along with IL-12 and IL-18, is absolutely necessary for tumor cell rejection.
5. PGE2 inhibits the ability of IL-12 and IL-18 to activate natural killer cells, the most important anti-cancer cells in our innate immune response.
6. PGE2 and other cyclic AMP elevating agents inhibit the synthesis of alpha and beta interferon. The implications of this inhibition of alpha/beta interferon synthesis is nothing short of horrific.
At least 10,000 scientific papers have been published on the use of alpha and beta interferon as a treatment for cancers of all kinds, leukemias, fungal infections, multiple sclerosis, hepatitis C, HIV, etc. The database on these two critical immune hormones is massive.
In addition to their use as chemotherapeutic agents, alpha and beta interferon control the maturation of the immune system. A particular form of dendritic cell, called pDC, is now known to be a "professional" alpha/beta interferon secreting cell. This dendritic cell, via type I interferon release, controls the interaction between the innate and genetic immune systems. The activation of the gamma delta T cells, now known to be a mainstay of the innate immune response against pathogens, is controlled by pDC cells and interferons.
Any agent that increases cyclic AMP levels in lymphoid tissues blocks alpha and beta interferon synthesis. This list includes Cox-2 derived PGE2 prostaglandins, and stress hormones such as epinephrine (adrenaline) and norepinephrine.
In reference three, note that alpha interferon induces its own resistance by inducing Cox-2 enzyme synthesis. This is a normal feedback mechanism of many other immune hormones, including the powerful IL-18
In review, cox-2 enzyme expression produces prostaglandins which are immunosuppressive. If cox-2 inhibitors are combined with beta interferon, human tumors grafted into mice are either killed outright or fail to grow.
"Beta interferon combined with cox-2 inhibition was associated with an increased number of T cells witin tumors and resulted in cures of small tumors, significant inhibition of the growth of large established tumors, and inhibition of the growth of metastatic tumor foci..."
PGE2 serves as an immportant feedback inhibitor of many different immune responses. Without it, an immune response might continue without constraint. Unfortunately, many different diseases are characterized by overexpression of this important enzyme. This results in inhibition of immune responsiveness and increased cellular growth in cancer.
The use of ibuprofen or indomethacin is clearly necessary to enhance the immune response against cancers and leukemias. In addition, general beta blockers, such as propranolol or pindolol, should also be used to prevent an increase in cyclic AMP induced by psychological or physical stress.
4.Vitamin A
When I was researching the affects of retinoic acid, vitamin A, on cancer growth, I stumbled onto some papers citing the utility of vitamin A in the treatment of HIV infections. I ignored them...for the moment anyway. After my recent protocol failure, I reinvestigated the role of vitamin A in HIV infections.
Apparently, vitamin A truly is a natural inhibitor of both new HIV infections and the reactivation of HIV from latently infected cells.
Two billion people, primarily in SE Asia and Sub Sahara Africa, are at risk for vitamin A and iron deficiencies.
Low concentrations of vitamin A are associated with accelerated HIV infections and increased mortality.
However, vitamin A supplementation has generally not proven effective in controlling HIV infections. There are reasons for this, which I will address later.
How does vitamin A control HIV infections?
The follow study shows that the enzyme PI-3K controls HIV infections in previously non-infected (primary) macrophages and CD4 T cells. In brief, PI-3K activated enzymes promote viral integration into cellular DNA. If the PI-3K pathway is blocked, the HIV virus is unable to stably infect new cells.
When retinol binds its intracellular storage protein CRBP-1, this complex interacts with and inhibits the activation of the PI-3K enzyme. It is clear that vitamin A is a natural inhibitor of this most critical cellular activation pathway.
We can conclude that vitamin A's ability to block PI-3K activation translates to an ability to block HIV infections in fresh macrophages and CD4 T cells.
But this is only part of the story.
Once the viral DNA is integrated into the cellular genome, PI-3K specific inhibitors have no affect on HIV synthesis. But vitamin A still plays a role in inhibiting the activation of the HIV gene.
This is exciting stuff.
Vitamin A inhibits the expression of the HIV virus in infected monocytes/macrophages. This means that the concentrations of vitamin A found "normally" in serum are able to transcriptionally repress the HIV gene in monocytes/macrophages, major long term reservoirs for the HIV virus. Apparently, this repression does not apply to CD4 T cells.
Apparently, retinoic acid blocks activation of the HIV gene by inhibiting chromatin remodeling at the HIV promotor. This paper is a bit complex, but it clearly shows that retinoic acid affects the chromatin in the area of the HIV gene. This prevents activation of the HIV promotor. Interesting, histone deacetylase inhibitors, anti-cancer agents, activate the HIV gene.
It would be wonderful if we could use common vitamin A to control any and all HIV infections. Well, we can in the asymptomatic phase of the infection. However, once diarrhea occurs the absorption of vitamin A from the diet becomes seriously impaired. Of course, the vitamin A could always be introduced via injection.
There is another problem. As we learned from the essays on vitamin A and cancer, frequently vitamin A receptor genes become dormant or silent. We can reactivate these receptors with histone deacetylase inhibitors, but these inhibitors will also reactivate the HIV gene. This is a temporary problem. We can live with a temporary increase in HIV synthesis if we can enhance retinoic acid responsiveness in infected cells.
5.Anti-oxidants
It is well established that ROS or reactive oxygen species promote both NF-kappaB synthesis and HIV gene activation. The use of high concentrations of anti-oxidants is therefore a viable treatment protocol for the treatment of HIV. However, at step one you have to decide what you are trying to accomplish. If your goal is to reduce HIV viral titers, the use of large amounts of anti-oxidants is fine. BUT, this protocol cannot be used if your goal is to reactivate the immune system. The proper functioning of the immune system depends on NF-kappaB activation.
Our New HIV treatment protocol is divided into two parts. In phase one, we are attempting to reduce viral titers. Therefore, the use of high amounts of vitamin C, vitamin E, alpha lipoic acid ( ALA) and N-acetylcysteine (NAC) is recommended.
Review
Very low concentrations of hydrogen peroxide, an extremely reactive ROS, promotes HIV synthesis via activation of NF-kappaB. The anti-oxidant NAC inhibits this response.
HIV infection, per se, is well known to increase the generation of ROS species. These oxygen radicals promote further HIV synthesis.
The following article is a nice review on the use of anti-oxidants to treat HIV infections. It can be read online.
ALA is a particular important anti-oxidant because it increases the level of glutathione in HIV infected cells. In addition, ALA directly inhibits the activation of NF-kappaB.
In addition, ALA can regenerate both vitamin C and E back to their anti-oxidant form.
In addition to anti-oxidant supplements, the mineral selenium is necessary to reduce HIV titers. Selenium is a mineral cofactor for all glutathione synthesis enzymes. Glutathione reduces ROS activity which activates HIV synthesis. Selenium is commonly deficient in people with HIV infections.
In summary, ALA and NAC are the most important anti-oxidants necessary to reduce HIV synthesis. Vitamins C and E are important, but substantially less so than the former. Selenium supplementation is critically important in the activation of glutathione synthesizing enzymes.
6.Cimetidine
I am intrigued by the idea that cimetidine, Tagamet, has a biological affect that transcends its ability to inhibit the histamine H2 receptor.
To my knowledge, cimetidine is known to stimulate the following pathways that have little or nothing to do with the inhibition of the histamine H2 receptor.
1. Blocks the infectivity of the HIV virus.
2. Blocks the expression of the E-selectin molecule on blood vessel cells.
3. Promotes antigen presentation.
4. Promotes IL-18 release via caspase-1 activation.
I believe cimetidine activates deacetylase enzyme activity. These enzymes remove acetyl groups from cellular and nuclear proteins. I know this info means next to nothing to you, the reader, but its a big deal if correct. Many cellular proteins undergo reversible acetylation reactions. These reactions modulate their biological activity. Cimetidine is a cheap drug that has shown to be active at VERY low concentrations. It might turn out to be a blockbuster treatment for a host of different diseases.
Consider the following...
1. HIV Infectivity.
When cimetidine is added along with virus to NON-infected cells, it blocks viral infectivity. No one knows why. What is remarkable is that cimetidine blocks viral infectivity, 100%, at the low dose of 10 microMolar. Further, cimetidine is completely non-toxic at concentrations 100 times this effective dose. The affect of cimetidine on HIV infectivity is NOT due to its role as a H2 receptor antagonist.
When viral membrane protein gp120 binds the CD4 protein on lymphocytes, it activates the acetylation of the protein alpha-tubulin, the major component of microtubules. This acetylation reaction enables the virus to enter the cell. If the acetylation reaction is inhibited by HDAC6, a deacetylase enzyme bound to microtubules, the virus cannot fuse into the membrane of the target cell. This makes infection impossible. For this and other reasons, I believe cimetidine is activating HDAC6 and possibly other deacetylase enzymes. This is SO easy to test experimentally.
If this is true, cimetidine could be used to inhibit the infection of most if not all enveloped viruses.
2. Downregulation of E-selectin
E-selectin is an adhesion protein expressed on the membranes of blood vessel cells. It has many normal roles in the body. In cancer, highly malignant cells which carry the Lewis carbohydrate antigen leave the blood and metastasize into other tissues by binding E-selectin. Cimetidine inhibits the expression of E-selectin thereby impairing the metastasis of these cancer cells. The affect of cimetidine on E-selectin is not due the blockage of the histamine H2 receptor, nor is it due to an inhibition of E-selectin synthesis.
Butyrate, a histone deacetylase inhibitor, promotes the expression of E-selectin on blood vessel cells. This implies that deacetylase enzymes, such as HDAC6, would down regulate the expression of this important adhesion protein.
Interestingly, tubulin, microtubules, are responsible for the down regulation of E-selectin.
3. Antigen presentation.
Cimetidine enhances antigen presentation in dendritic cells. Again, this response has nothing to do with histamine H2 receptors.
The interaction site that forms when dendritic cell class 2 histocompatibility antigens bind the T cell receptor is called an immune synapse. This is a dynamic structure that is linked to the tubulin cytoskeleton. HDAC6 mediated deacetylation of alpha tubulin plays a major role in the cytoskeletal rearrangements necessary for organization of the immune synapse.
It is conceivable that cimetidine enhances antigen presentation by activating the deacetylation of microtubules.
4. IL-18 release.
Cimetidine stimulates IL-18 release by activating the caspase-1 enzyme. This enzyme cleaves inactive IL-18 into an active form.
It is possible that caspase-1 is inactivated by acetylated cytoskeletal proteins, and that cimetidine reverses this inactivation.
DOSAGE
Niacinamide, 6-10 grams a day based on body weight. This molecule is water soluble and quite bioavailable.
Lactoferrin, 2 grams a day, 1 gram twice a day, preferably dissolved in warm cream or ½ and ½. This improves bioavailability.
Ibuprofen, 1.2 grams a day, 200 mg individual doses 6 times a day.
Vitamin A, 15000IU a day.
Anti-oxidants. 600 mgs ALA, 2 grams NAC, 2 grams vitamin C ester or ascorbyl palmitate. Vitamin E as desired. Selenium, 250 micrograms a day. No NOT use yeast derived selenium. It is poorly bioavailable. Purchase sodium selenite only.
Cimetidine, 800 mgs a day, 400 mgs twice a day.
PART TWO
This treatment phase has no time limit. The objective is to activate the immune system against virally infected cells.
These essays were originally written for our cancer immunotherapy protocol. The same information applies to HIV infections.
1.Lactoferrin
Lactoferrin is an iron binding protein found commonly in mammalian milk, tears, saliva, and lungs. It is naturally made in most cells of the body. Lactoferrin is known to be a powerful natural antibiotic when degraded into peptide form. As an intact protein, it is also known to enhance the synthesis/release of many immune hormones, including IL-18, the subject of our previous blog essay.
Lactoferrin is the newest addition to our immunotherapy protocol.
Cow's milk lactoferrin powerfully activates the innate immune system in humans. As a protein, lactoferrin is largely degraded in the stomach into peptides which have biological activity when they penetrate the gastric mucosa. The activation of the gastric immune system provides systemic protection against solid tumors and their metastatic cells. The ability of lactoferrin/lactoferrin peptides to stimulate IL-18 release is considered fundamental to its ability to active innate immunity.
If the information in these articles doesn't impress you, nothing will. I am a cynical immunologist and I am absolutely stunned by the potent immune enhancing properties of lactoferrin.
The efficacy of lactoferrin can be enhanced by its incorporation into liposomes, soap bubbles really. In our case, the lactoferrin is removed from its capsules and added to warm coconut milk. Coconut milk is an emulsion of water and fat. It's perfect for delivering the lactoferrin directly to the walls of the intestinal mucosa. No one knows why this is important, but apparently it is.
The second article is particularly significant because it shows that fat encapsulated lactoferrin increases alpha interferon secretion in normal human volunteers. Alpha interferon is a MAJOR activator of the immune system. This is great stuff folks. Non-lipid encapsulated lactoferrin does not induce alpha interferon synthesis in human volunteers.
At an oral dose of 2 grams of bovine lactoferrin a day, the immune system is significantly activated in human volunteers.
Protocol.
Two grams a day of cow's milk lactoferrin is the minimum dose per day. You can and should take as much as you want, especially if you have cancer or leukemia. Lactoferrin is TOTALLY non-toxic...except to cancer/leukemia cells.
Heat your coconut milk to a quick boil, let it cool to warm and mix in your lactoferrin. You can make a stock solution of this mixture and store it in the refrigerator. If you intend to use 2 grams a day, open 8x 250 mg capsules for a day's dose. A weeks dose is 56 capsules in as much coconut milk as you want. Consume 1/7 of the stock solution per day.
Lactoferrin Induced IL-18 and IL-12 Synergy
The immune hormone IL-12 activates and promotes the development of both genetic (adaptive) and innate immune cells. It is absolutely necessary for our defense against microbial agents and cancers.
What about IL-18? We talked about IL-18 in previous essays. Remember, it is powerfully activated by lactoferrin, the iron binding protein found in milk. What, exactly, does IL-18 do to the immune system to make it so important?
IL-12 stimulates CD4 and CD8 T cells to proliferate, but these cells are immature and incapable of killing targets. IL-18 stimulates these growing cells to differentiate into "effector cells", lymphocytes that express high levels of the IL-2 receptor and acquire the ability to secrete gamma interferon. Effector cells are "fully loaded" to kill their targets. IL-12 stimulates growth, while IL-18 follows behind to promote differentiation of these cells into fully functional cells. It is not surprising that IL-12 stimulates the expression of the IL-18 receptor on stimulated cells.
In addition to stimulating gamma interferon synthesis, IL-18 also activates the CD134 pathway. This is a big deal. IL-18 stimulates the synthesis of the OX40 (aka CD134) LIGAND on macrophages, dendritic cells and propably CD4 T cells as well. The OX40 (not to be confused with CD40) Ligand stimulates T and B lymphocytes that express the OX40 receptor on their membranes to become activated. This activation promotes the migration of T and B cells in the body, while prolonging their lifespan.
The lifespan of activated CD4 and CD8 cells is short unless activated by the OX40/OX40 Ligand system. CD8 cytotoxic T cells, those that directly attack tumor cells in the genetic immune response, will not accumulate in the body without activation by the OX40 Ligand. It is rather obvious that immune surveillance by memory CD8 and CD4 T cells is useless if these cells are allowed to die off after a first round of activation. IL-18, a well documented immune adjuvant or activator, stimulates OX40 Ligand synthesis in dendritic cells, which in turns binds the OX40 receptor on lymphocytes, thereby stimulating their long term survival.
One of the fascinating roles of IL-18 is its ability to activate memory T cells in the ABSENCE of recall antigen. Ever had a TB test? They scratch your skin with a common TB protein to see if your immune system reacts. This is a recall immune response. It means that you have previously been exposed to TB. I was taught that recall immune responses do NOT occur unless you have been reexposed to a partticular antigen. Apparently, this is wrong.
IL-18 can stimulate memory T cells to become reactivated, or recalled all by itself. The immune cells do not need exposure to actual antigen.
On the surface, this sounds great, but an overactivated immune system can cause autoimmune disease. IL-18 has been linked to various autoimmune diseases, such as Lupus. Moderation is the key to a long life.
In summary, it is clear that there is a highly defined relationship between IL-12 and IL-18 that transcends the ability of IL-12 to activate the IL-18 receptor.
In the absence of blocking molecules such as the prostaglandin PGE 2, the activation of IL-12 by antigen is not a problem. However, the factors that stimulate IL-18 synthesis have been less clear.
Lactoferrin is clearly an important component of our HIV Immunotherapy protocol.
2.Indomethacin
This important anti-inflammatory drug is critically important in the activation of the immune system. First, it inhibits the immunosuppressive Cox-2 enzyme, Second, it does not inhibit NF-kappaB like ibuprofen. Third, it activates the genetic factor EGR -1 which stimulates the synthesis of CD40 LIGAND, a critically important activator of both genetic and innate immunity.
Some background information on the role dendritic cells play in immune reactivity.
Think of circulating dendritic cells as guard dogs. They move all around the property looking for trouble.
Immature dendritic cells, the ones that circulate, have some interesting properties.
First, they have a tremendous ability to ingest cellular debris. Some of this debris is processed as peptide antigens which form a complex with class II histocompatibility antigens on the membranes of the dendritic cells.
Second, these cells possess a high concentration of TOLL receptors on their membranes. TOLL receptors bind a host of different bacterial, viral, and fungal derived products. The activation of TOLL receptors literally tells the dendritic cells that they are under attack by pathogens.
Three, these cells, once initially activated, migrate out of the tissues, blood, etc. to the nearest lymph node.
The immune response against newly discovered antigens, such as those shed from cancer cells, is initiated ONLY in the lymph nodes. Activating T cells against a specific antigen is a very complex process. Once the dendritic cell arrives at the lymph node, it binds naive CD4 T cells that can recognize the tumor antigen via the CD4 cell's T cell receptor ( TCR ). This recognition process is purely genetic This sets off a chain reaction of events.
When the CD4 TCR binds the class II histocompatibility antigen complex on the membrane of the immature dendritic cell, a signal is sent to the CD4 cell to make a genetic factor called EGR -1. EGR -1 stimulates the synthesis of a molecule called CD40 LIGAND which is then expressed on the membrane of the CD4 T cells. This LIGAND then binds a membrane protein called CD40 on the membrane of the dendritic cell (CD40 is found on many other cells including macrophages, B cells, endothelial cells and epithelial cells).
Once CD40 is stimulated, the immature dendritic cell begins to develop into a fully mature antigen processing cells. These mature cells now secrete a host of immune hormones including IL-12, which is absolutely necessary for the expansion of newly activated CD4 T cells. Also, the maturation of dendritic cells induces the expression of class I histocompatibility antigens on its membrane. This now allows the dendritic cell to activate CD8 cells which are specifically cytotoxic to various tumors, etc.
The CD40/CD40 ligand interaction is ABSOLUTELY necessary for the activation of both the genetic and innate immune response. IL-12 activates NK cells, and other innate immune cells. IL-12 is NOT made in dendritic cells unless the CD40 molecule on the surface is activated by CD40 ligand on the CD4 T cell.
Naturally, the CD40/CD40 ligand interaction is defective in HIV infections. I now know why this happens...the topic of another essay.
NSAID inhibitors such as sulindac and indomethacin powerfully stimulate the synthesis of EGR -1. Aspirin and ibuprofen are much less effective activators.
Since EGR -1 activates the synthesis of CD40 ligand, and this molecule literally controls the proper functioning of the immune system, sulindac and indomethacin are perfect activators of the immune system against cancers, HIV...whatever. Right?
Not exactly.
Sulindac is a histone deacetylase inhibitor and a direct inhibitor of NF-kappaB activation. HDAC inhibitors are immunosuppressive (and stimulate HIV synthesis) and NF-kappaB MUST be activated for proper immune functioning. However, we can certainly use sulindac in phase one of the cancer treatment protocol. There is nothing wrong with activating EGR -1, which is also a tumor suppressor, in phase one. Stimulating the expression of CD40 ligand on CD4 cells in preparation for phase 2, the immunotherapy phase, sounds good to me.
However, in phase 2 we CAN use indomethacin. Indo is not an HDAC inhibitor and it doesn't inhibit the activation of NF-kappaB. Since it is also a powerful cox1/2 inhibitor, indomethacin is a perfect activator of the immune system in phase 2.
The two primary immune defects caused by HIV viral proteins are the excessive production of immunosuppressive prostaglandins via the COX -2 enzyme, and an inability to synthesize the CD40 LIGAND.
For a very long time, I have argued that activation of the Cox-2 enzyme, and the ensuing immunosuppression, was fundamental to the development of chronic HIV infections.
Prostaglandin PGE 2, which increases cyclic AMP concentrations in HIV infected and non-infected cells, is extremely immunosuppressive. But I never knew HOW HIV viral particles increased the expression of the Cox-2 enzyme.
Now I do. But there is more to the story.
Antigen presenting (activating) cells ( APC ) such as dendritic cells, macrophages, and endothelial cells posses an molecule called CD40 on their membranes. When a naive, non-activated, CD4 T cell binds the class 2 histocompatibility antigens on these cells via the T cell receptor ( TCR ) on the CD4 cells, a signal is sent to the CD4 cell to stimulate the synthesis and expression of a molecule called CD40 LIGAND. When CD40 LIGAND is expressed on the membrane of the CD4 T cell, it binds the CD40 molecule on the APC and induces maturation, i.e. activation, of these cells. In point of fact, these immature CD4 T cells and immature dendritic cells activate one another.
Immature dendritic cells circulate throughout the body in a methodical search for antigen, such as HIV proteins. Once these cells pick up antigens, they migrate to the lymph nodes to begin the activation of naive CD4 T cells. Upon further activation by the newly expressed CD40 LIGAND on CD4 cells, the dendritic cells begin a maturation process that results in the expression of class 1 histocompatibility antigens and the synthesis of the critical TH1 (cell mediated immunity) hormone IL-12. These class 1 antigens are necessary for the activation of CD8 cytotoxic T cells, the cells that attack and kill HIV virus producing cells. IL-12 is a growth factor that is absolutely necessary for the synthesis of gamma interferon, an immune hormone that controls both innate and genetic (adaptive) immunity.
The following are good reviews on this topic.
CD40 LIGAND and CD154 are the same molecule...different names.
One of the major recognized defects in immune responsiveness in HIV infections has been linked to an inability to induce CD40 LIGAND (CD154) expression on CD4 cells. Since CD40 T cells and dendritic cells really activate one another, this means that dendritic cells will never reach a state of maturity necessary to mount an effective immune response against the HIV virus. The defect in CD40 LIGAND expression has been linked to the ability of viral gp120 proteins to cross link the CD4 molecule.
The inability of non-infected CD4 T cells, removed from HIV infected persons, to respond to pathogens other than HIV has been linked to the defective CD40 LIGAND expression induced by the viral membrane protein gp120/gp160. This contributes to the development of opportunistic infections.
The lack of CD40 LIGAND expression on CD4 T cells prevents dendritic cell maturation and the secretion of IL-12 and subsequently gamma interferon. Since free gp120 proteins and dead/live viral particles freely circulate throughout the blood and lymph, they have the capacity to bind non-infected CD4 T cells, thereby preventing both their activation and their ability to promote the maturation of dendritic cells. Antibodies targeted to CD40 (which mimic the activating effects of CD40 LIGAND) reactivate the previously defective immunological response found in HIV infections.
We now know that viral gp120/160 proteins, either free or bound to dead/live virus, can cross link the CD4 receptor on T cells. This results in an inability of these cells to synthesize and express CD40 LIGAND on their membranes. For all intents and purposes, the lack of CD40 LIGAND expression on T cells inhibits both TH1 (cell mediated) and TH2 (antibody mediated) immune responses to ANY pathogen...viral, bacterial, fungal or cancer.
During normal immune reactions, the CD4 molecule and the TCR (genetically specific T cell receptor) on T cells bind the class 2 histocompatibility antigens on antigen presenting cells. During this process, the CD4 molecule is NOT cross linked.
When the CD4 molecule on T cells is cross linked by HIV particles or free gp120/160 proteins, it initiates a biochemical response that is completely inappropriate for normal immune functioning. In fact, it initates a response that promotes cancer cell development.
How?
Cross linking CD4 by HIV particles promotes the chronic activation of RAS , a major tumor gene. One of the most important roles of activated RAS is the activation of the PI-3K pathway.
Activated RAS inhibits the activation of the EGR -1 transcription factor via the PI-3K signaling pathway.
The EGR -1 transcription factor activates the gene for CD40 LIGAND.
Clearly, viral particles activate RAS , which activates PI-3K signaling and the down regulation of EGR -1. The down regulation of EGR -1 inhibits the expression of CD40 LIGAND and the activation of the immune response to ALL pathogens.
The PI-3K pathway activates an enzyme called AKT or protein kinase B. This enzyme activates Cox-2 gene activity.
Since HIV viral particles or gp120/160 proteins can activated RAS and subsequently PI-3K signaling in CD4 membrane expressing cells, it is clear that the HIV virus, dead or alive, can induce a massive immunosuppression to its own existence by DOWN regulating EGR -1, the activator of CD40 LIGAND expression and general immunity, while simultaneously UP regulating the expression of the Cox-2 gene, an activator of cyclic AMP mediated immunosuppression and viral expression.
Indomethacin is a potential HIV wonder drug because it can correct both of these defects.
And everything is linked to the "simple" process of CD4 molecule cross linking by viral particles.
3.Theanine
Theanine is an amino acid made only in tea plants. A breakdown product of theanine activates a major population of gamma delta T cells, a population of immune cells critically important in our innate immune system.
Some gamma delta T cells are now known to be antigen processing cells like dendritic cells and macrophages. This is new information, right off the press.
Gamma delta T cells are known to be defective in HIV infections.
The defect associated with these cells may be attributed to a general lack of type 1 interferon synthesis during HIV infections. Gamma delta T cells require a direct interaction with immature dendritic cells in order to become activated.
The lack of CD40 LIGAND synthesis, and the corresponding defect in dendritic cell maturation, may be responsible for the lack of gamma delta T cells in HIV infections.
4.Melatonin
Melatonin, the sleep hormone, also activates the cell mediated immune response. Melatonin synthesis is defective in HIV infections. By an unknown mechanism, melatonin does stimulate IL-12 synthesis, suggesting that its primary target are dendritic cells.
5.Cimetidine
Cimetidine is a component of our immunotherapy protocol for cancer. In the last few days, I found some additional articles on cimetidine and the immune response that are worth mentioning.
Histamine acts on immature dendritic cells to shift their differentiation into a pathway that promotes the differentiation and activation of CD4 T cells in the TH2 and/or TH3 direction. When naive CD4 T cells encounter dendritic cells, they can develop into the following:
TH1 cells which promote cell mediated immunity.
TH2 cells which promote antibody formation.
TH3 cells which promote tolerance.
In order to stimulate the production of CD4 T cells that promote cell mediated immunity, dendritic cells MUST secrete IL-12. If they secrete IL-4, IL-5 and IL-13 instead, the CD4 T cells will promote the secretion of antibody. On the other hand, if the dendritic cells secrete IL-10, tolerance or immune non-responsiveness will occur.
Histamine plays a non-appreciated role on dendritic cell maturation. It inhibits IL-12 synthesis, while promoting the synthesis of IL-10 and IL-4, etc. If you want activate the immune system against cancer cells or virally infected cells, this is BAD news. Antibody based immune responses will not control these pathogens.
Histamine, via H2 receptors, can also directly stimulate CD4 T cells to secrete specific immune hormones.
It is well established that immune functioning decreases as we age. During aging, a TH2 dominant immune response becomes established. A TH1 immune response dominates at an earlier age. A TH2 dominant immune response cannot protect us from most infections and cancer.
Histamine related problems are well known to increase as we age. Histamine, acting via its H2 receptor on dendritic cells and macrophages, is responsible for much of the altered immune response during aging.
Histamine H2 antagonists, such as Tagamet (cimetidine), block these histamine mediated effects on immune responsiveness.
Cimetidine is a weird molecule. It is not a true H2 antagonist, because some of its biological effects cannot be attributed to a pure H2 antagonism. For example, cimetidine can stimulate the release of IL-18, a powerful immune hormone that along with IL-12 TREMENDOUSLY promotes cellular immunity. The cimetidine mediated release of IL-18 involves the activation of caspase-1, an enzyme that cleaves inactive, PRO-IL-18, into the active IL-18 molecule. Only the active IL-18 molecule is secreted into the blood.
Ten years ago, a study was published showing that histamine H2 antagonists, cimetidine in particular, blocked HIV synthesis in a culture dish. Many compounds inhibit HIV expression, but these compounds are useless clinically. Cimetidine, on the other hand, substantially blocked HIV synthesis at VERY small doses, doses that are attainable clinically. I haven't read this article yet. Yet, the abstract seems to imply that cimetidine blocked the infection of new cells. Now, that is interesting.
A year later, a human study, using 400 mg doses of cimetidine, refuted the use of cimetidine in the treatment of HIV infections.
400 mgs doses of cimetidine are probably too low to affect immune responsiveness OR HIV synthesis. Also, I seriously doubt that cimetidine alone can be used to treat HIV infections.
Cimetidine is a complex molecule. It inhibits both histamine H2 activity and other biochemical pathways that have NOTHING to do with the histamine H2 receptor. It would be nice if we could figure out HOW cimetidine inhibited HIV infectivity. I am working on it.
For now, we know that cimetidine promotes cell mediated immunity by blocking histamine H2 receptor activity. This makes cimetidine very useful as an immune adjuvant for both cancer and viral infections such as HIV.
DOSAGE
Indomethacin, 100 mgs a day, 2 times 50 mgs. This is a minimum dose. Double it if necessary.
Lactoferrin, 2 grams a day, 2 times 1 gram a day in warm cream or ½ and ½.
Theanine, 600 mgs a day, 3 times 200 mgs. This compound will make you tired.
Melatonin, 6 mgs a night before bed.
Cimetidine, 800 mgs, 2 times 400 mgs a day. Cimetidine is quite non-toxic so you can take more.
SUMMARY
This is a ton of information to intellectually digest so take your time. HIV is a complicated virus and AIDS is an even more complicated syndrome of physiological defects that transcends the immune system.
We are looking for volunteers, Kurosawa MICE, to test this protocol. You cannot be using HIV protease inhibitor drugs. Other drugs are OK. Of course, the optimal MOUSE is someone who has never been treated with any HIV drug.
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KEEP THE FAITH!!!
END
Copyright © 2002, Stephen Martin, Ph.D
Chief Scientist, Grouppe Kurosawa
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