A follow up with Dr. Paul Chapman and PLX4032 .. BRAF Mutation..Melanoma..Jim Breitfeller

Advancements in the treatment of metastatic malignant melanoma
23. September 2009 05:05

Researchers have made significant advances in the treatment of metastatic malignant melanoma - one of the most difficult cancers to treat successfully once it has started to spread - according to a study to be presented at Europe's largest cancer congress, ECCO 15 - ESMO 34, in Berlin on Thursday.

In the phase I extension study, researchers have seen rapid and dramatic shrinking of metastatic tumours in patients treated with a new compound that blocks the activity of the cancer-causing mutation of the BRAF gene, which is implicated in about 50% melanomas and 5% of colorectal cancers. In new results from 31 melanoma patients with the BRAF mutation who were treated with 960mg of PLX4032 twice a day, 64% (14) of the 22 patients who could be evaluated so far met the official criteria for partial response (this involves the diameter of tumours shrinking by at least 30% for at least a month). A further six of the 22 patients also showed a response, but, at the time of the congress presentation, it was too early to say whether the tumours would shrink far enough to meet these criteria.

Dr Paul Chapman, an attending physician on the Melanoma/Sarcoma service at Memorial Sloan-Kettering Cancer Center (New York, USA) and who was one of the leaders of the trial, told a news briefing: "We are very excited about these results. Of the 22 patients we have been able to evaluate so far, 20 have had some objective tumour shrinkage. This is impressive as they all had metastatic disease and most of them had failed several prior therapies. A lot of these patients were pretty sick but many of them had a significant and rapid improvement in the way they function. We've had patients come off oxygen and we've got several patients who have been able to come off narcotic pain medication soon after starting treatment."

The trial is investigating PLX4032 in patients with the BRAF mutation, and results from the first 55 patients were reported at a cancer meeting earlier this year (ASCO 2009). These data had been aimed at finding the best dose of PLX4032 to give to patients. However, the phase I extension data reported at ECCO 15 - ESMO 34 focuses on a subsequent group of an additional 31 patients who were all treated at the maximum tolerated dose of the drug (a 960 mg pill twice a day). All the patients had the BRAF mutation.

Dr Chapman said: "What makes this treatment different from standard chemotherapy is that standard chemotherapy attacks the machinery involved in cell division; so to stop the cancer cells dividing uncontrollably, most standard chemotherapy aims to block the mechanism of division by interfering directly with DNA replication or with microtubules in the dividing cells. PLX4302 is different because it attacks the genetic programme that is causing the cells to divide uncontrollably, and we think the BRAF mutation is driving that programme. The drug is blocking the genetics of the tumour, rather than trying to interfere with the proliferation of the cells and, as a result, there are fewer side effects, although there are some. We are seeing some pretty dramatic and rapid responses, and they are occurring in sites where we rarely see responses to chemotherapy, such as in the bone.

"There are some important caveats. All these patients had failed previous therapies, either chemotherapy or treatment with Interleukin 2, as well as surgery. However, we know that only 10-30% of patients will respond to standard chemotherapy, so it's not surprising that our patients had not responded, or have responded and then the cancer has recurred. In our study 64% of patients have had a partial response, but because we are only treating patients with the BRAF mutation, we are cutting out about 40% of melanoma patients who do not have this mutation and whom we know will not respond to this treatment. That is one reason why we are seeing a much higher response than with conventional treatments.

"Also, we don't know yet how long these responses will last, and we have had patients whose cancer has progressed after initially responding; so we are putting a lot of effort in to studying the patients who do relapse, trying to understand how their tumours have become resistant.

"In addition, one of the main side effects we've seen is that some patients develop early, non-melanoma skin cancers such as squamous cell skin cancer. We are very vigilant about this and although they are very easy to cut out, it's something we are keeping a close eye on."

Dr Chapman and his colleagues are planning a phase II trial of 90 patients starting at the end of this year. In addition, a large phase III randomised controlled trial involving several hundred patients is planned to start either at the end of this year or beginning of next year involving centres in North America, Europe and Australia.

Dr Chapman said it was too early to be talking about a cure for advanced melanoma, but that this drug had potential. "Most of us think that a drug like this would ultimately be part of the regimen, but that we might need additional drugs with it to complete the cure. Right now we are seeing dramatic responses but it's too early to say whether we've actually cured people because most patients still have evidence of some level of tumour on their skin. I think this is a huge step forward; whether or not it will be sufficient by itself really remains to be seen."

http://www.ecco-org.eu/Conferences-and-Events/ECCO-15/


You need to be genetically tested for the BRAF Mutation.



Take Care,

Jimmy B

Experimental Vaccine Improves Response in Melanoma!!!..Jim Breitfeller

Laino, Charlene
Oncology Times:
25 August 2009 - Volume 31 - Issue 16 - pp 39-41
doi: 10.1097/01.COT.0000360410.38201.57


ORLANDO, FL-For the first time, a synthetic peptide vaccine designed to boost immunogenicity has been shown to improve outcomes in patients with metastatic melanoma in a Phase III clinical trial.
Used in conjunction with standard interlekin-2 (IL-2) therapy, the vaccine improved the clinical response rate and prolonged the time to disease progression, compared with IL-2 alone, Patrick Hwu, MD, Professor and Chairman of the Department of Melanoma Medical Oncology at the University of Texas M. D. Anderson Cancer Center, reported here at the ASCO Annual Meeting.

There was also a trend toward improved survival rates among patients given the vaccine, but the patients have to be followed longer before we can see statistical significance, he said.

'A Nice Milestone'

The so-called gp100:209-217(210M) peptide vaccine is a synthetic peptide cancer vaccine consisting of amino acid residues 209 through 217 of the glycoprotein 100 (gp100) melanoma antigen, with a methionine substitution at position 210 designed to improve immunogenicity. It's given along with IL-2, a powerful immune-system stimulant.


Vaccination with gp100:209-217 (210M) peptide stimulates the patient's immune system to mount a cytotoxic T-lymphocyte response against tumor cells expressing gp100, Dr. Hwu explained. The IL-2 multiplies the T cells into a large army that can then attack the tumors to kill them.
In a Phase II trial led by Steven Rosenberg, MD, PhD, 42% of 31 patients with metastatic melanoma receiving high-dose IL-2 plus the vaccine had objective responses, but this is the first Phase III study of the vaccine that is positive, Dr. Hwu said. It's a nice milestone.

21-Center Study

The 21-center, Phase III trial involved 185 patients with Stage IV or locally advanced Stage III cutaneous metastatic melanoma, randomized to receive either the peptide vaccine followed by four days of high-dose IL-2 at 720,000 IU/kg/dose or high-dose IL-2 alone. The cycle was repeated every three weeks until the patients responded or progressed.
The overall clinical response rate was almost double in the vaccine arm-22.1%, compared with 9.7% for the patients who got interleukin-2 alone, Dr. Hwu said.

The median progression-free survival time was 2.9 months in the vaccine arm vs 1.6 months for IL-2 alone, again a significant difference.
The median overall survival time was 17.6 months in the vaccine arm compared with 12.8 in the IL-2 alone arm, but the difference did not reach statistical significance, Dr. Hwu said.

The vaccine was well tolerated. Side effects were swelling and redness at the injection site.

'Rare, Positive Study in Metastatic Melanoma'

This is a rare positive study in metastatic melanoma, said the Discussant, Antoni Ribas, MD, Associate Professor of Medicine (Hematology/Oncology) and Surgery (Surgical Oncology) at UCLA.
Dr. Ribas said he would like to see further study to determine if the vaccine prolongs overall survival times. But since accruement for the current study took seven years, it's uncertain that anyone will want to undertake a confirmatory Phase III trial for overall survival, he said.
Louis M. Weiner, MD, Director of the Lombardi Comprehensive Cancer Center, said he views this as a conceptual advance, teaching us that focusing the attention of the immune system on a specific cancer-related target increases the ability of the immune system to attack tumors, at least for a while.

Many of us believe that a combined approach that includes an immune system attack on cancer cells will ultimately prove to be most useful in controlling cancers such as melanoma, he said.
Dr. Weiner said one reason this vaccine may have worked while others have failed is because it was combined with high-dose IL-2.



We have known for a while that IL-2 therapy is effective in a small percentage of melanoma patients. But when it is effective, it is highly effective, he said.

But IL-2 can be toxic and is not simple to use, Dr. Weiner said.
The moderator of a news conference at the meeting that featured the study, Sonali M. Smith, MD, Associate Professor of Medicine at the University of Chicago Medical Center, called the findings very promising for a very poor-prognosis group of patients, and said that the approach might act as a platform for more potent vaccines.
Dr. Hwu agreed, noting that 80% of patients did not respond. He said he hopes to improve response rates in future trials by combining the vaccine with other immune-stimulating agents such as anti-CTLA-4.

Cytotoxic T lymphocyte antigen 4 (CTLA-4), a key negative regulator of T-cell responses, can restrict the antitumor immune response. By using an anti-CTLA-4 agent, he said, we can take the brakes off the immune system, which should allow the vaccine and IL-2 to work even better.

Source:http://journals.lww.com/oncology-times/Fulltext/2009/08250/Experimental_Vaccine_Improves_Response_in_Melanoma.15.aspx

I believe this aproach will show tremendous progress in Melanoma Therapy






Take Care,

Jimmy B

What We Can Learn From Individual Patients is Often Overlooked.Melanoma..Jim Breitfeller

"What we can learn from individual patients is often overlooked in oncology," he said, adding that many of these remarkable cases have led to the development of new treatment strategies for melanoma such as vaccinations against specific antigens and bone marrow transplantation. "From clinical observation, we can learn a lot from these remarkable cases."

Alan Houghton, M.D., chief of immunology at Memorial Sloan-Kettering Cancer Center, New York



This my Theory to Date as of 6/5/2009.

I will be sending out a Very, Very long list of ACKNOLOGEMENTS.

"In melanoma patients treated with CTLA-4 blockade, not only were more T cells specific for melanoma antigens present, but those CTL were more likely to be polyfunctional — thus more likely to be effective at destroying the tumor — and those patients were much more likely to have regression of their tumors than in people without CTLA-4 blockade.

So the concept that TRegs — or some other inhibitory effect associated with CTLA-4 — suppress anti-tumor immune responses is likely to be correct, and it seems that at least in some cases it’s possible to override that inhibition and drive T cells to once again attack the tumor effectively. When that happens, cancer can be cured. It’s just a question of being able to do this on a consistent basis. Unfortunately, that’s still the hard part."

Unknown

“There are three parts of the equation in a clinical trial. There’s who has control, who gets the reward, and who takes the risk. Patients take all the risk, they have no control, and they get no reward. Patients ought to be the ones driving the process and get the reward out of it and have the control, since they are the ones that take the risk.”

Greg Simons

“Don’t ever give up. Don’t ever give up.” “Cancer can take away all my physical abilities. It can not take away my mind, it can not take away my heart, and it can not take away my soul”.

Jimmy Valvano from his speech during the 1993 ESPN ESPY Awards


I have so many papers that I don't recall where I got the above quote.

My Hope is that this doesn't fall on deaf ears.

Take care
Jimmy B

Cellular Immunotherapy I (Supressive cells in Cancer Patients) Melanoma ..Jim Breitfeller

In my research, I ran across this powerpoint presentation on Cellular Immunotherapy. I gave me a better understanding on what may be happening to Us cancer Patients.

It is from Tumor Genetics, Clinic I Internal Medicine
Center for Molecular Medicine Cologne Germany.

Please take time to Review it.
Source:http://www.zmmk.uni-koeln.de/content/seminare/lmm_08/common/lecture_cellular_immunotherapy_I.pdf

Cellular Immunotherapy I

Take care

Jimmy B

Cutting Edge Technology “Infection-mimicking materials to program dendritic cells in situ”Melanoma .. Jim Breitfeller

Cutting Edge Technology “Infection-mimicking materials to program dendritic cells in situ”

Thanks to Donald Bohlken for bring this to my attention.

“I thought you might be interested in that attached article from the January 25, 2009 issue of the journal "Nature Materials" concerning a Harvard study of a new vaccine methodology which resulted in a 90% survival of mice infected with a melanoma strain which would normally kill them in 25 days. The article speaks of this result as a "cure".

The article synopsis notes: "Cancer vaccines typically depend on cumbersome and expensive manipulation of cells in the laboratory, and subsequent cell transplantation leads to poor lymph-node homing and limited efficacy. We propose that materials mimicking key aspects of bacterial infection may instead be used to directly control immune-cell trafficking and activation in the body. It is demonstrated that polymers can be designed to first release a cytokine to recruit and house host dendritic cells, and subsequently present cancer antigens and danger signals to activate the resident dendritic cells and markedly enhance their homing to lymph nodes. Specific and protective anti-tumour immunity was generated with these materials, as 90% survival was achieved in animals that otherwise die from cancer within 25 days. These materials show promise as cancer vaccines, and more broadly suggest that polymers may be designed to program and control the trafficking of a variety of cell types in the body."
This is Nano and Transdermal Technology at its best.

Implants Mimic Infection To Rally Immune System Against TumorsMain Category:

Melanoma / Skin CancerAlso Included In: Immune System / Vaccines; Medical Devices / Diagnostics; Biology / Biochemistry

Article Date: 25 Jan 2009 - 0:00 PST

“Bioengineers at Harvard University have shown that small plastic disks impregnated with tumor-specific antigens and implanted under the skin can reprogram the mammalian immune system to attack tumors.

The research -- which ridded 90 percent of mice of an aggressive form of melanoma that would usually kill the rodents within 25 days -- represents the most effective demonstration to date of a cancer vaccine.

Harvard's David J. Mooney and colleagues describe the research in the current issue of the journal Nature Materials.

"Our immune systems work by recognizing and attacking foreign invaders, allowing most cancer cells -- which originate inside the body -- to escape detection," says Mooney, Gordon McKay Professor of Bioengineering in Harvard's School of Engineering and Applied Sciences. "This technique, which redirects the immune system from inside the body, appears to be easier and more effective than other approaches to cancer vaccination."

Most previous work on cancer vaccines has focused on removing immune cells from the body and reprogramming them to attack malignant tissues. The altered cells are then reinjected back into the body. While Mooney says ample theoretical work suggests this approach should work, in experiments more than 90 percent of the reinjected cells have died before having any effect.
The implants developed by Mooney and colleagues are slender disks measuring 8.5 millimeters across. Made of an FDA-approved biodegradable polymer, they can be inserted subcutaneously, much like the implantable contraceptives that can be placed in a woman's arm.
The disks are 90 percent air, making them highly permeable to immune cells. They release cytokines, powerful attractants of immune-system messengers called dendritic cells.
These cells enter an implant's pores, where they are exposed to antigens specific to the type of tumor being targeted. The dendritic cells then report to nearby lymph nodes, where they activate the immune system's T cells to hunt down and kill tumor cells throughout the body.”

Source: Http://www.medicalnewstoday.com/articles/136473.php

Implants Mimic Infection To Rally Immune System Against Tumors



Donald Bohlken actually contacted one of the collaborators, Dr. Glenn Dranoff of the Dana Farber Cancer Institute and Harvard Medical School. He indicated: "We are initiating efforts to bring this to clinical testing, but it will take some time to adapt the procedures to patients. A one year time frame is a reasonable guess."

If this crosses over from the mouse model to the human model, we may have a winner on our hands. A 90 % response and if they are complete responses, this technology would surpass any therapy out there to date. The main take away is that they are prompting our immune system to recognize the tumors by activation of the T-cell. Rosenberg and Hwu and other colleagues are doing just that with (ACT) Adoptive Cell Transfer. Kirkwood, Camacho, Webber, Hodi, Maker, O’Day and Wolchok did that with anti-CTLA-4 blockage.

I can see the Light at the end of the tunnel!!!!!!!!!!!!!!!!!!!!!!!!!!!

Thank you Don for advocating for us and your Brother Ron who is a fighting Warrior of Melanoma.

I will post the research paper on Melanoma Missionary for everybody.

Jimmy B

EDITORIAL CTLA-4 Blockade: Unveiling Immune Regulation Melanoma ..Jim Breitfeller

Originally published as JCO Early Release 10.1200/JCO.2005.09.923 on December 21 2004

CTLA-4 Blockade: Unveiling Immune Regulation

Glenn Dranoff

Department of Medical Oncology, Dana-Farber Cancer Institute; Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, MA

To maintain tissue homeostasis under the duress of infection or injury, the immune system is endowed with a diverse repertoire of soluble and cellular effectors. The innate response, composed of granulocytes, macrophages, dendritic cells, natural killer cells, and complement, is rapidly triggered into action, detecting tissue disturbance through a set of germline-encoded pattern recognition receptors. The adaptive reaction, consisting of antibody-producing B cells and T lymphocytes, is slower to develop, but manifests exquisite specificity and memory. These attributes reflect the requirement for expansion of rare clones harboring somatically rearranged immunoglobulin molecules or T-cell receptors specific for foreign proteins or processed peptides presented by major histocompatibility complex (MHC) molecules. These innate and adaptive responses are carefully orchestrated through soluble and membrane-bound regulators, resulting in the deployment of the most suitable effectors for containing the disorder, while minimizing tissue damage.

Cancer cells similarly provoke immune recognition. In one pathway, innate effectors detect tumor cells directly.1 Natural killer cells and phagocytes express NKG2D molecules that function as receptors for stress-related genes such as MICA and MICB, which are induced as a consequence of cellular transformation. Natural killer cells further scan for the loss of MHC class I molecules on the surface of tumor cells. Dendritic cells use a variety of scavenger receptors to result in the phagocytosis of dying tumor cells.

The adaptive response exploits an indirect pathway, termed cross-priming, to recognize cancer cells.2 In this mechanism, dendritic cells capture tumor cell debris, migrate to regional lymph nodes, and stimulate CD4+ and CD8+ T cells with tumor specificity. Primed T cells thereby acquire the capacity to detect tumor cells directly in a MHC-restricted fashion. CD4+ T cells also contribute to B-cell antibody production.

Antitumor innate and adaptive responses are frequently detected in cancer-bearing hosts, but their biologic importance remains incompletely understood. In some cases, endogenous reactions may function to attenuate disease progression. In multiple cancer types, clinicopathologic studies of early-stage lesions demonstrate that dense intratumoral lymphocyte infiltrates are strongly correlated with reduced frequencies of metastasis and improved patient survival.3 Moreover, in advanced ovarian carcinoma, lymphocyte infiltrates also predict for complete responses and prolonged survival following cytotoxic therapy.4 Consistent with this protective role, several strains of immune-deficient mice display enhanced susceptibility to spontaneous and carcinogen-induced tumors.5 Nonetheless, other compelling data indicate that tumor cells sometimes exploit host responses to promote disease progression.6 Unresolved inflammation may facilitate tumor development by modulating tumor cell growth, apoptosis, invasion, and metastasis. Together, these divergent outcomes underscore a dual role for immunity in carcinogenesis.7

Source:http://jco.ascopubs.org/cgi/content/full/23/4/662
CTLA-4 Blockade: Unveiling Immune Regulation




Take care

Jimmy B

Tumors Evading Detection!!!!! Melanoma.. Jim Breitfeller

Tumors Evading Detection
Lack of costimulation:

Many Melanoma tumor cells do not have the B7 protein on their surface so this co-stimulatory second signal cannot take place. Theoretically, they should cause an immune response but they do not stimulate an effective anti-tumor immune response. The first signal originates from the binding of the T cell receptor (TCR) to its antigen-MHC, and provides the specificity of the interaction. Without this signal, the cell enters anergic state and can act as a T reg cell. Expression of B7 on the surface of a cell is the costimulatory signal necessary to allow for the cytolytic CD8+ T cell attack on the tumor. B7 display renders tumor cells capable of effective antigen presentation, leading to their eventual eradication.

Secretion of immunosuppressive cytokines:

Another way tumors evade detection is by secretion of certain cytokines. They are low-molecular weight proteins that use their communication ability to regulate the immune response. Cytokines can act upon either the cells secreting them (autocrine) or on neighboring cells (paracrine) to generate activities in the targeted cells. This means they can act as light switches for on and off immune responses. For example, interleukin-2 activates a cell-mediated immune response, while interleukin-10 suppresses cell-mediated responses. Many types of cancer, including Melanoma, take advantage of this ability to down regulate this appropriate immune response to help extend their survival and proliferation. This causes cancer patients to fail in mounting a successful attack on the tumors. Immunosuppressive cytokines secreted by cancer cells include transforming growth factor-beta (TGF-beta), interleukin-10 (IL-10) and vascular endothelial growth factor (VEGF).

TGF-beta is one of the most potent immunosuppressive cytokines characterized to date. It is capable of affecting the proliferation, activation and differentiation of cells participating in both the innate and acquired immune response.TGF-beta inhibits the profilation T-cells, B cells, Natural killer cells (NK), and macrophages.TGF-beta also converts T-cells, which normally attack cancer with an inflammatory (immune) reaction, into regulatory (suppressor) T-cells, which turn off the inflammatory reaction. Another of TGF-beta's affect is on cytotoxic T lymphocytes (CTLs) This is very important for anti-tumor immunity because of their cytotoxic effects. TGF-beta down-regulates many of the processes necessary for CTL activation. Without this activation, there is no assault on the tumor cells from the CTLs. In addition to suppressing proliferation, TGF-beta has been shown to induce apoptosis (cell death) in B and T cells.

Another immunosuppressive cytokine is IL-10. It is capable of inhibiting the prodction of of pro-inflammatory cytokines like IFN-gamma, IL-2, and GM-CSF made by cells such as macrophages and T helper cells. IL-10 also displays potent abilities to suppress the antigen presentation capacity of antigen presenting cells. Secretion of IL-10 in the vicinity of a tumor can render the tumor totally insensitive to CTL-mediated lysis. It is most likely that the tumor’s microenvironment is altered enough to block or turn off the discharge granules that would lyses the tumor cell. However, it is also stimulatory towards certain T cells, mast cells and B cells. It enhances B cell survival, proliferation, and antibody production. As you can see, IL-10 has many rolls to play when it come to the immune system.

The cytokine VEGF is produced by most tumors.
Vascular endothelial growth factor (VEGF) is a cytokine that is produced by most tumors. This growth factor enables the tumor to expand vascularly when is in its growth phase. VEGF production can be induced in tumor cells that are not receiving enough oxygen.

Regulatory T-cells (Tregs) (suppressor T cells) are a specialized subpopulation of T cells that act to suppress activation of the immune system and thereby maintain immune system homeostasis and tolerance to self-antigens.1

Tumor Growth kinetics

The cell cycle has four stages:

1. G1 phase when the cell increases in size and gets ready to replicate its DNA.
2. S phase when the cell synthesizes or copies its chromosomes
3. G2 phase in which the cell prepares to divide
4. M phase when mitosis occurs.

When the various growth inhibitory proteins and checkpoint controls which regulate this cycle become disabled due to mutations characteristic of cancerous cells, the cell cycle is no longer under tight regulation. Tumor cells are capable of proliferating so quickly that the immune response is not fast enough to keep their growth in check. The growth of the tumor cells outpaces the immune response and escape the detection of the immune system. Lack of cell cycle controls leads to excessive proliferation of tumor cells.





I am getting closer!!!!

Take care

Jimmy B

Hand and Hand Tumor Rejection ..Melanoma ..Jim Breitfeller

This diagram goes with the "Tumor Rejection Piece"


Tumor Rejection!!!!!!

To cause the rejection of the tumors cells, The immune system must orchestrate a chain of events mediated by several types of Leukocytes including Dendtric cells (DC), Natuaral Killer cells (NK), CD4+ and CD8+ lymphocytes and others. This orchestration has many players in it including the T-Regs, sercreted cytokines, and Monoclonal antibodies (mAb’s) and complexes. It is a delicate balance between self and non-self.

At the Lymph Node drainage area, the lymphocytes (CD4+ and CD8+) with their T-cell receptors (TCRs) are able to scan the Dendtric cells (DC’s) for antigen-MHC molecules. (ag-MHC) major histocompatability complex (class I or II). Based on the two signal model, a second signal from CD28 molecule is needed to activate the T-cells (T-lymphocytes). If communcation breaksdown, and the TCR signal only happens, it can lead to tolerance by means of fuctional paralysis of the (APCs) Antigen presenting cells (Anergy) or by the induction of clonal deletion (apoptosis). Anergic cells can act as regulatory T cells by competing at the sites of antigen presentation and adsorbing out stimulatory cytokines such as IL-2. This can halt the activation of the T- lymphocytes and no immune response is initiated.

Once activated fully, the CD4+ T-cells can mobilize to where the event will take place and usually sends out a “danger signal” inflammation. The activated CD4+ T-cells can secrete many different cytokines including IL-4, IL-2 and activate the TH2 cells which are a subset of the CD4+ cells. The TH2 cells stimulate the B cells to mature into plasma cells that secrete antibodies. These antibodies that are produced are the cell-destructive kinds that have anti-tumor behavior. The CD4+ can also cross-prime CD8+ T-cells in the presence of IL-2 and are called (CTLs)Cytotoxic T Lymphocytes. Cross-priming is another name for cross-presentation. The role of the CD8+ T cells is to monitor all the cells of the body, ready to destroy any that express foreign antigen fragments in their class I molecules.

Some CD4+ T cells can develop into CTLs, but they can attack only those cell types (e.g. B cells, macrophages, dendritic cells) that express class II MHC molecules. Virtually every cell in the body expresses class I MHC molecules, so CD8+ CTLs are not limited in the targets they can attack. CTLs have cytoplasmic granules that contain the proteins perforin and granzymes. When the CTL binds to its target, the contents of the granules are discharged. A dozen or more perforin molecules insert themselves into the plasma membrane of target cells forming a pore that enables granzymes to enter the cell. Granzymes are serine proteases. The serine proteases are a family of enzymes that cut certain bonds in other proteins. It is similar to what is in your laundry detergent. They are known as detergent enzymes. They break the bond between the dirt and the fabric. By breaking up these proteins, they start destroying the intracellular workings of the tumor cells.

Next Piece is how the tumor protects itself from the Killer T-cells through a microenviroment and Tregs.


Just plugging away!!

Jimmy B

Failure at the effector phase: immune barriers at the level of the melanoma tumor microenvironment Melanoma Jim Breitfeller

Thomas F. Gajewski
Author's Affiliation: Departments of Pathology and Medicine, University of Chicago, Chicago, Illinois

Requests for reprints: Thomas F. Gajewski, University of Chicago, 5841 South Maryland Avenue, MC2115, Chicago, IL 60637. Phone: 773-702-4601; Fax: 773-702-3163; E-mail: tgajewsk@medicine.bsd.uchicago.edu.



"Metastatic melanoma tumors seem to be deficient in expression of B7-1 and B7-2 (16), which are important costimulatory factors for full T-cell activation (17). Stimulation through the T-cell antigen receptor without B7 costimulation has been shown to induce a hyporesponsive state termed anergy (18, 19). In support of this possibility, we recently have observed in a mouse model that a T-cell hyporesponsive state consistent with anergy occurs in mice bearing B7-negative tumors (20). In this situation, tumor rejection indeed can occur when B7-1 is transfected into the tumor cells, an observation that has been reported by multiple laboratories in other model systems (21, 22)."

In my case we stimulated the B7 receptor with anti-CTLA-4 Blockage, in the present of the antigen presenting cell (APC) that activated the CD4+T-cell it was allowed to propagate for 49 days (43 days gave the maximum propagation. Than we added the IL-2 therapy which we know by experiments the done by Rosenberg that Il-2 is able to pass throught the Tumor's microenvironment.

In 1980, Dr. Steven A. Roesnberg and colleagues discovered novel novel method for killing metastatic cancer cells. They took lymphoid cells and exposed them to interluekin-2 (IL-2).These cells were able to lyse the tumor cells and kill them. The were a different population than the Natural Killer cells. They coined the term “Lymphokine-activated killer cells” (LAK) for short.

Also, When patients do IL-2 therapy, they have side-effect Known as cell leakage. Il-2 makes the cell wall porous.

With the tumor cells walls compromised and with the activated effector T cells, the assualt on the tumors begin.

At Least, that is the way I envision it. There is probably a lot more going on behind the scenes.

Jimmy B

Here Goes!!! I am going to Spill the Beans!!!!!! Melanoma Jim Breitfeller

"The induction of an immunological antitumor response capable of eradicating metastatic tumors is the ultimate goal of immunotherapy. We have recently shown that this can be achieved by interleukin 2 (IL-2) therapy directed to the tumor microenvironment by a recombinant antibody–IL-2 fusion protein. It is not known, however, whether this curative treatment is associated with a predominance of T cells carrying specific T cell receptor variable ? regions (TCRBV) or the presence of clonally expanded T cells."

Activation of preexisting T cell clones by targeted interleukin 2 therapy

Per thor Straten,* Per Guldberg,* Tina Seremet,* Ralph A. Reisfeld,†‡ Jesper Zeuthen,* and Jürgen C. Becker§
*Department of Tumor Cell Biology, Division of Cancer Biology, Danish Cancer Society, DK-2100 Copenhagen, Denmark; †Department of Immunology, Scripps Research Institute, La Jolla, CA 92037; and §Department of Dermatology, School of Medicine, D-97080 Würzburg, Germany
‡To whom reprint requests should be addressed at: Department of Immunology, Scripps Research Institute, IMM 13, R218, 10550 North Torrey Pines Road, La Jolla, CA 92037.
Communicated by Frank J. Dixon, Scripps Research Institute, La Jolla, CA
Received February 19, 1998; Accepted April 30, 1998.

"Melanoma is a highly malignant tumor but several lines of evidence suggest that it is capable of eliciting a specific immune response, i.e., a number of melanoma-associated antigens have been identified and the presence of clonotypic T cells has been demonstrated in melanoma lesions (1–4). Therefore, several immunomodulatory therapeutic approaches were initiated to improve the prognosis of melanoma patients. Interleukin 2 (IL-2) is one of the most potent antitumor cytokines known quote (5), and was recently approved for treatment of metastatic melanoma. However, objective responses induced by systemic IL-2 therapy are still insufficient, and the associated side effects are severe (6). These findings are due to the fact that a systemic application of IL-2 disregards the paracrine nature of this cytokine under physiological conditions (7).

As a means to target IL-2 directly to the tumor site, we have recently shown that human IL-2 can be genetically engineered as a fusion protein with the chimeric mouse–human mAb 14.18 which recognizes the ganglioside GD2, retaining both antigen binding and cytokine activity (8).

Furthermore, we have shown that treatment with this antibody–IL-2 fusion protein can eradicate human hepatic and pulmonary melanoma metastases in severe combined immunodeficient mice (9) as well as autologous murine B16 melanomas (10). Although it was shown in these studies that tumor eradication was dependent on CD8+ T cells, it is not known whether tumor clearance is associated with a clonal expansion of T cells. Furthermore, it remains to be established whether such a clonal expansion would be due to a de novo induction or to the activation and expansion of preexisting T cell clones. Here, we demonstrate both the overexpression of certain T cell receptor variable β regions (TCRBV) as well as the clonal expansion of T cells in melanoma lesions subsequent to targeted IL-2 therapy.

However, clonally expanded T cells were also detectable prior to the
thrapy, suggesting that antibody–IL-2-targeted therapy acts as an activator rather than an inducer of an antitumor T cell response."

Source:http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=21154
Activation of preexisting T cell clones by targeted interleukin 2 therapy


"Melanoma and the Magic Bullet (Monoclonal Antibodies)"

You have read it here First!!!!!!!!!

I still have write the IL-2 part, But I believe the concept is all there.

Now I have to work on the protocol of the "Antibody-IL-2 therapy"

That is why my therapy "Antibody-IL-2 therapy" worked!!!!!!!!

I can smell the sweet Success!!!!!!!



Jimmy B

Delayed Melanoma Response Could Impact Clinical Decisions with Anti-CTLA-4 Agents.. Melanoma..Jim Breitfller

Elsevier Global Medical News. 2008 Nov 7, JS MacNeil

STOCKHOLM (EGMN) - Clinical studies of two experimental agents - tremelimumab and ipilimumab - have shown delayed and mixed responses among patients who gained control of refractory melanomas with these therapies.

Both agents come from a new class of monoclonal antibodies that seek to promote immune response by blocking cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4). In three phase II trials presented at the European Society for Medical Oncology Congress, tremelimumab and ipilimumab ultimately achieved disease control rates of 14%-29% as second-line therapies. No complete responses were reported.

Among patients classified as having progressive disease after ipilimumab treatment, however, were some people who subsequently improved. Likewise, among six patients with "mixed response" to tremelimumab were five patients who initially developed new lesions, but then had slow decreases in targeted lesions. All six were still alive at the time of the presentation.

Investigators suggested that the modified World Health Organization (WHO) criteria used to assess activity of cytotoxic agents may not capture the benefit in some patients classified with progressive disease. They cited four observed patterns of response, which were described in a poster by Dr. Kaan Harmankaya, of the department of dermatology at University of Vienna, and associates:

-Response in baseline lesions.

-Stable disease with a slow, steady decline in tumor volume.

-Response after increase in total tumor volume.

-Response in index and new lesions after the appearance of new lesions.

While delayed response is an issue for researchers, the clinical impact could be more important, according to a discussion of the tremelimumab and ipilimumab studies by Dr. Ulrich Keilholz of the Charité University in Berlin and the European Organisation for Research and Treatment of Cancer melanoma group. In comparing "classical and nonclassical efficacy assessment," he said that nonclassical assessment captures delayed efficacy, but the definitions are complicated and the issues in patient management are distinct from efficacy evaluation in studies.

"Nonclassical assessment does change the response rate, but it does not change the survival rate," Dr. Keilholz said, downplaying the importance of response measures, compared with overall survival in phase III trials.

In clinical practice, however, the physician must decide when to switch a patient from a therapy that is not working. The three trials showed that delayed efficacy does occur with these agents, but the limit appears to be week 20 after treatment, Dr. Keilholz said, adding: "I think that it is important to allow patients a certain amount of time ... but also to put a stop at a certain time point."

Tremelimumab

Dr. John M. Kirkwood, director of the Melanoma Center at the University of Pittsburgh Cancer Institute, presented the tremelimumab data from an open-label phase II trial in 251 patients (nearly all with stage IV disease), of whom 242 were evaluable. The protocol called for a 15 mg/kg dose to be delivered intravenously on the first day of up to four 12-week cycles. Sixteen (7%) patients achieved partial responses and 36 (15%) had stable disease - a clinical benefit rate of 22%.

Dr. Kirkwood said all but 1 of the partial responses lasted at least 170 days, and 11 were ongoing. Median overall survival reached 10.1 months, he said; median progression-free survival reached 2.8 months, with 15.6% of patients progression-free 6 months after treatment. Factors correlating with survival were still being analyzed. The trial was sponsored by Pfizer Inc.

Ipilimumab

The first ipilimumab trial was a multinational, open-label study of 155 patients with advanced disease that had failed previous therapies. Patients received 10 mg/kg of ipilimumab every 3 weeks for four cycles, followed by maintenance therapy at the same dose every 12 weeks from week 12 to week 60.

Dr. Vanna Chiarion Sileni of the Instituto Oncologo Veneto in Padua, Italy, reported 9 patients had partial responses and 33 had stable disease by modified WHO criteria, adding up to a disease control rate of 27% (42/155). The median duration of stable disease was 4.1 months at a median follow-up of 5.7 months, she said; 19 patients were still stable at their last assessment.

Among those classified with progressive disease were patients with the four patterns of response. Small subgroups had a "slow steady decline" in tumor volume after an initial increase in target lesions or the appearance of new lesions, she said.

In the second ipilimumab trial, Dr. Celeste Lebbé of Saint-Louis Hospital in Paris reported on a multinational dose-finding study that randomized patients with unresectable relapsed stage III or IV melanoma to 10 mg/kg, 3 mg/kg, or 0.3 mg/kg of ipilimumab given once every 3 weeks for four cycles followed by maintenance treatment once every 12 weeks.

The 10 mg/kg dose produced the best overall response rate, a composite measure of complete and partial responses, at 11%, and a disease control rate of 29% (the lowest rate, 14%, was at the lowest dose). Nearly half, 48% of 73 patients given the highest dose were alive at 1 year. Their median survival was estimated at 11 months at a median follow-up of 10.4 months.

The four patterns of response were observed in this study as well, Dr. Lebbé reported, and about 35% of patients at the highest dose had a decline in total tumor volume. Patients at this dose also had the most toxicity, she said; about a quarter had grade III adverse events, including gastrointestinal side effects in 16%. In most cases, it was manageable and reversible.

The ipilimumab studies were sponsored by Bristol-Myer Squibb and Medarex Inc, which are jointly developing the agent. Dr. Lebbé was the only investigator to disclose a conflict of interest, having served on two advisory boards for Bristol-Myer Squibb.

Dr. Kirkwood said phase III trials for both agents have been completed and are being analyzed, but applications for approval have not yet been filed. In April 2008, Pfizer halted a phase III study of tremelimumab monotherapy in advanced melanoma when interim data showed it would not be superior to standard therapy.

http://www.oncologystat.com/news-and-viewpoints/news/Delayed_Melanoma_Response_Could_Impact_Clinical_Decisions_with_Anti-CTLA-4_Agents_US.html

Delayed Melanoma Response Could Impact Clinical Decisions with Anti-CTLA-4 Agents


I believe the delayed response was what happen to me.

Jimmy B