Immunologic Cell Teeter-Totter*
“Ask Immunologic questions give molecular answers” Andy Blidy
Regenerative Medicine cell replacement with Immunotherapy, Harnessing the Immune Systems to kill or promote life saving responses
https://www.youtube.com/watch?v=-9q4c-QRdes Cancer Immunotherapy - medical animation
https://www.youtube.com/watch?v=r1Fp6C1xx6Q&t=2163s
https://www.youtube.com/watch?v=PphTL6vY4W4 History of Immunotherapy by James Allison at PMWC 2017 Silicon Valley
1-26-2017
Andy Blidy
Version 3.1
Why is the immune response like a Teeter totter? B/c if you fall off the teeter totter b/c your immune system is damaged/ aged due to many factors, some environmental and some genetically, you end up in a disease state…..This can result in disease state of aging and you can get an auto immune disease on the bottom end of the teeter totter or cancer on the top side of the teeter totter….. But how does one balance the immune teeter totter with a replacement cellular therapy and genetic expression from possible gene therapy that results in homeostasis of the immune system? …Balance is the key in immunology, tolerance control by both the (Major Histocompatibility Complex ) MHC I and MHC II pathways….If you are “healthy” or young, your gamma delta /alpha beta cells will keep you homeostasis or balance on the teeter totter. Think of these cells as the fulcrum on the teeter totter. Do you want to suppress the immune system or activate it? A high level process of the immune system is illustrated below in a summary ……Notice balance is needed in this quantum immunologic system.
It is now known that TCR gamma delta show suppression of the immune system while TCR alpha beta are the fulcrum for proliferation during an immune response or in attack mode. Knowing the cellular process and their checkpoints are the key. I have outlined 5 major checkpoints on the immune teeter totter as:
1.) Antigen presentation, start of the innate immunity system
2.) Activated T Cells, the aggressive protection expansion of the human immune system
3.) T Cell receptor, the integrators that discriminate self vs. non self
4.) “The CTLA-4 (cytotoxic T-lymphocyte-associated protein the brakes of the immune response and finally
5.) PD-1 (Programmed death -1 ) killing of foreign invaders or aging cells or cancerous cells of apoptotic gone wrong
Immune checkpoint pathways down regulate T-cell activation to maintain peripheral tolerance, and can be exploited by tumors to induce an immunosuppressive state that allows the tumors to grow and develop instead of being eliminated by the immune system. The differential patterns of the CTLA-4 and PD-1 ligand expression—found primarily in lymphoid tissue and in peripheral tissues, respectively—are central to the hypothesis that CTLA-4 acts early in tolerance induction and PD-1 acts late to maintain long-term tolerance. “ One needs to reboot the immune with the correct stem cells this paper will outline the repair of chronic inflammation or cancer …….The immune system needs to be rebooted so rescue and repair can take place. Notice the Mesenchymal MSC is the central of this biological system …..
Immunosuppression by mesenchymal stem cells. MSCs suppress innate and adaptive immune responses by enhancing regulatory immune cells with tolerogenic properties. MSCs suppress macrophages by favoring monocyte polarization to anti-inflammatory M2 macrophages, increasing the production of IL-10, and decreasing the production TNF-a and IL-12. MSCs can also regulate DCs by downregulating the expression of MHC, CD40, CD80, CD83 and CD86, thus, diminishing their antigen presenting ability, while upregulating the expression of IL-10. MSCs can reduce the NK cell cytotoxicity and decrease their production of TNF-a and IFN-?. Treg and Breg cells can be induced by MSCs, further increase the production of anti-inflammatory cytokines (IL-10 and TGF-ß1). However, the mechanisms of how Breg cells are induced by MSCs are still not clear. MSCs: Mesenchymal stem cells; TNF: Tumor necrosis factor; IL: Interleukin; NK: Natural killer; DCs: Dendritic cells; IFN-?: Interferon-?; Treg: Regulatory T; Breg: Regulatory B; TGF: Transforming growth factor; PGE2: Prostaglandin E2; IDO: Indoleamine 2,3-dioxygenase.
World J Stem Cells. 2016 Sep 26; 8(9): 268–278.
Published online 2016 Sep 26. doi: 10.4252/wjsc.v8.i9.268
1. Immunotherapy Checkpoint “Antigen-presenting cells (APCs)
APCs are a heterogeneous group of immune cells that mediate the cellular immune response by processing and presenting antigens for recognition by certain lymphocytes such as T cells CD4 helper cells and CD8 cytotoxic with the furculum at the TCR T cell receptor which have two alpha beta units to one gamma delta structure. Classical APCs include dendritic cells, mesenchymal and epithelial (EMT), macrophages, monocytes Langerhans cells and B cells”. This is why antigen presentation is key to treating a trauma cell repair, cancer or autoimmune diseases at first line of defense to the individual? These cells present the T cell with an antigen from say a cancer cell with a co- stimulator protein…”Antigen presentation stimulates T cells to become either "cytotoxic" CD8+ cells or "helper" CD4+ cells”.….”This process begins to hunt down diseases by NK cells and CD8 killers cells that are covered with the same antigens or the repair by CD 4 cells. Each type of T-cell is slated to deal with different chronic inflammation, cancers, or pathogens, which may be a bacteria, toxin or virus. This is the least understood of the Immune check points “almost every cell in the body is technically an APC because they can present antigens to T-cells”. “One such example of this could be bacterium internalize and digested by macrophage and expresses MHC II on it cell surface to recognize a T cells…… In order for the immune system to recognize and bind foreign antigen or invader, cancer cell the immune system must be able to tolerate self from foreign.”
http://prohealthinsight.com/wp-content/uploads/2014/10/3-Macrophage-Angigen-Presenting-Cell.jpg
2. Immunotherapy Checkpoint Activated T cells
“A T cell or T lymphocyte is a type of lymphocyte (a subtype of white blood cell) that plays a central role in cell-mediated immunity. T cells can be distinguished from other lymphocytes, such as B cells and natural killer cells, by the presence of a T-cell receptor on the cell surface.” “Antitumor immunity can be improved by ICOS-targeting therapies. Once activated you will alter the microbiome to respond to cancers or any foreign antigen or suppress the immune system in normal functions, the cause of auto immune disease such as Parkinson , MS, lupus. Key subset is Tregs CD4 CD25 FOXp3, GARP, LAP , SATB1 cells and the subsetting of all TH1, TH2 and TH17. “Foxp3 is shown to be the master regulator of Tregs at the transcription level with functional gene regulation by SATB1(special
AT-rich binding protein), LAP (latency-associated peptide),GARP that tethers to TGF-beta”…..And the CD8 pathways “THE CYTOTOXIC T CELL RESPONSE.
The next step is the activation of an antigen-specific cytotoxic T cell response:
o activated APCs present their antigen to the specific cytotoxic T CD8 cell receptor within an MHC I, along with a variety of second signals, including B7 + CD28 and/or 4-IBB + 4-IBBL
o this process is helped along by the secretion of IL-2 – a potent T cell growth factor – by TH1 cells and the cytotoxic CD8 T cells themselves
Once activated, the cytotoxic T cells are very keen to get out and start hunting and killing things. They identify infected cells by recognizing the antigen displayed within MHC I on their surfaces. They then destroy these cells using one of several mechanisms:
o they classically form an immunological synapse with their target cell – this just means the cell membranes touch – and release a substance called perforin to make a hole in the cell wall. They then use this hole to release granzymes and granulysin into the cell, which induce apoptosis and DNA fragmentation.
o Fas ligand interactions between the cell surfaces can also produce apoptosis of the infected cells via the aptly named death-inducing signaling complex (DISC)
o cytotoxic T cells can also release interferon gamma (IFN?), which has an interesting role in viral infections, as it is able to block intracellular viral replication without killing the cell itself. This effect is very useful, as killing and lysing virally infected cells would simply let all the baby viruses out and cause the infection to spread itself even further, which is clearly suboptimal.”
“Once activated, they divide rapidly and secrete small proteins called cytokines that regulate or assist in the active immune response.….This is the marriage of molecular biology with immunology at the cell surface and gene expression levels”…… How does EMT function as a microbiome translation with ACT T cells ? Same is true for CD8 activated cell but less studied over the years B/C of sub populations . Is cancer antigen presentation different than auto immune diseases ? IMO yes “There are myriad molecules involved in the complex biochemical process (called trans-membrane signaling) by which T-cell activation occurs. The signal transduction mechanism by which a T cell elicits this response upon contact with its unique antigen is termed T-cell activation. ” next we look at TCR and their role in Act T cells See diagram below for one example of such Activated T cells
(International Journal of Biological Sciences 10: 0119 image No. 002 http://www.ijbs.com/v10p0119.htm
3. Immunotherapy Checkpoint Alpha beta /’gamma delta T cells
“Alpha beta /’gamma delta T cells what are they? “The seesaw (also known as a teeter-totter or teeterboard) is a long, narrow board supported by a single pivot point, most commonly located at the midpoint between both "ends"; as one end goes up, the other goes down.” In the middle of the immune system at the fulcrum are either your gamma delta or alpha beta expression. In humans, in 95% of T cells the TCR consists of an alpha (a) chain and a beta (ß) chain (encoded by TRA and TRB, respectively), whereas in 5% of T cells the TCR consists of gamma and delta (?/d) chains (encoded by TRG and TRD, respectively).” When the TCR engages with antigenic peptide and MHC (peptide/MHC), the T lymphocyte is activated through signal transduction…..Now my understanding is TCR consisting of gamma and delta is associated with suppression of act T cells and push towards neither CD4 or CD8 T cells pathways. “An alpha (a) chain and a beta (ß) chain On CD4 helper T cells and regulatory T cells, this co-receptor is CD4 that is specific for MHC class II. On cytotoxic T cells, this co-receptor is CD8 that is specific for MHC class I.” Lastly “Human ?d T cells can be divided into three main populations based on d chain expression. ?d T cells expressing Vd1 chains are prominent in the intraepithelial layer of mucosal surfaces, where they are involved in maintaining epithelial tissue integrity when facing damage, infection, or transformation”.
Gamma delta from the TCR
Figure 1
“Ligand recognition by aß and ?d TCRs.
• Full size figure and legend
“(“a) The aß T cells recognize peptide bound to MHC molecules through the interaction of both TCR chains with peptide-bound MHC molecules. ß2m, ß2-microglobulin. (b) In contrast, the NKT cell TCR contacts CD1d-bound glycolipid through its invariant TCRa chain. (c,d) The recognition of ligand by ?d T cells is less well understood. (c) The G8 ?d TCR inserts its CDR3d loop into the hydrophobic groove of T22, with the ?-chain making only a minor contribution to binding. (d) Willcox et al. now show a unique mode of binding for another ?d TCR, LES, to its ligand, EPCR/ The CDR3 loop of the LES TCR ?-chain binds to EPCR independently of lipid antigens through residues on the EPCR ß-sheet.”
EPCR: a stress trigger for ?d T cells
• Deborah A Witherden
• & Wendy L Havran
Nature Immunology13, 812–814(2012)
doi:10.1038/ni.2398
Published online
21 ugust 2012
https://www.youtube.com/watch?v=K09xzIQ8zsg PDL-1 and T cell science
Now Let,s look at CLTA4 VS PD_1 checkpoint different’s relayed to the immune system
Anti-CTLA4 anti PD-1
a. Targets CD28 pathways Targets TCR pathways
b. Hard wired T cells Induce Resistance cells
c. Expands Clonal diversity Does not expand Clonal only cell in tumor
d. .Moves T cell into tumor Does not move T cells into tumor
e. Disease recurrence are rare Disease recurrence are higher
4. Immunotherapy Checkpoint CTLA-4 “Brakes of the Immune System “
CTLA-4 Protein what is its function? “CTLA-4 (cytotoxic T-lymphocyte-associated protein 4), also known as CD152 (cluster of differentiation 152), is a protein receptor that, functioning as an immune checkpoint, down regulates immune responses. The complexity of the CD28/CTLA-4 receptor interactions stems from the fact that there are two natural ligands CD80 (B7-1) and CD86 (B7-2) for these receptors” The protein is the brakes of the cellular immune response. …..cancer cell immune checkpoints that shut down T cell response and allow the cancer stem cells to grow undisturbed…..Checkpoint inhibitors will physically block the checkpoint , which free Act T and other T cells to kill cancer cells. ´ T-cell activation is a critical event in the organization of effective cellular and humoral immune responses. Activated T cells are essential for provision of T-cell help, promoting the development of high-affinity antibody production and the generation of cytotoxic T-cell responses. Accordingly, defects in proteins required for T-cell activation give rise to significant infectious pathology and malignancies. However, the decision to allow T-cell activation also has potentially dangerous consequences for the host and must therefore also be tightly controlled. Defects in proteins involved in regulating activated T-cell behavior therefore tend to lead to autoimmunity. Thus, the major challenge faced in regulating T-cell responses is how to maintain a sufficiently large immune repertoire capable of recognizing all possible foreign antigens, whilst at the same time maintaining T cells in an unresponsive state towards self-antigens.”
The concept of using anti-CTLA4 antibodies to treat cancer was first developed by James P. Allison while he was Director of the Cancer Research Laboratory at the University of California, Berkeley. Clinical development of anti-CTLA4 was initiated by Medarex, which was later acquired by Bristol-Myers Squibb. Bristol-Myers Squibb’s comb therapy is OPDIVO® (nivolumab) (PD-L1) and YERVOY® (ipilimumab) (CTLA4). All human Monoclonal antibodies.
Jim Allison’s talk at Berkeley Ca history of CTLA4 and PD-1
https://www.youtube.com/watch?v=r1Fp6C1xx6Q
Fig. 1
“Indirect interactions between mesenchymal stroma/stem cells and cancer cells. a Cytokines, chemokines, growth factors: MSC secrete a plethora of soluble factors that can bind as substrates to appropriate receptors on the cell surface of cancer cells and vice versa for mutual activation of signaling pathways. b Metabolites: Likewise, MSC-released metabolites such as prostaglandin E2, kynurenine or galectin-1 can act in a paracrine manner on cancer cells altering their properties and functions [14]. c Exosomes: Both, MSC and cancer cells, secrete exosomes for the exchange of small molecules including protein, mRNAs and microRNAs. d Microvesicles: Besides exosomes, microvesicles represent a different type of microparticles for the exchange of small molecules such as mRNAs or microRNAs affecting tumor cells and MSC in mutual ways”
11. World J Stem Cells. 2016 Sep 26; 8(9): 268–278.
Published online 2016 Sep 26. doi: 10.4252/wjsc.v8.i9.268
5. Immunotherapy Checkpoint PD-L1
Last line of defense of self recognizing of good vs. bad before apoptosis cell death involving TCR pathways…… “ Programmed death (PD)-1 Ligand1, PD-1 Ligand2,of the immune cells CD279 (cluster of differentiation 279), is a protein that in humans is encoded by the PDCD1 gene. “PD-1, functioning as an immune checkpoint, plays an important role in down regulating the immune system by preventing the activation of T-cells, which in turn reduces autoimmunity and promotes self-tolerance. The inhibitory effect of PD-1 is accomplished through a dual mechanism of promoting apoptosis (programmed cell death) in antigen specific T-cells in lymph nodes while simultaneously reducing apoptosis in regulatory T cells(suppressor T cells”
This is the MAB that saved Jimmy Carter’s life, KeyTrude
http://www.nbcnews.com/health/cancer/cancer-drug-keytruda-keeps-some-patients-alive-3-years-n576376
5a. Immunotherapy Checkpoint Co stimulator pathway IDO-1and CD4+ pathways
IDO-1 with PD-L1 Inhibitor Combo Could Change Standard of Care in Cancer
“One way is the IDO-1 “This gene encodes indoleamine 2,3-dioxygenase (IDO) This enzyme is thought to play a role in a variety of pathophysiological processes such as antimicrobial and antitumor defense, neuropathology, immunoregulation, and antioxidant activity. Through its expression in dendritic cells, monocytes, and macrophages this enzyme modulates T-cell behavior by its peri-cellular catabolization of the essential amino acid tryptophan . IDO is an immune checkpoint molecule in the sense that it is an immunomodulatory enzyme produced by some alternatively activated macrophages and other immunoregulatory cells (also used as an immune subversion strategy by many tumors). Interferon-gamma has an antiproliferative effect on many tumor cells and inhibits intracellular pathogens such as Toxoplasma and Chlamydia, at least partly because of the induction of indoleamine 2,3-dioxygenase” causing depletion of tryptophan which can cause halted growth of microbes as well as T cells.[7] PGE2 is able to elevate the expression of indoleamine 2,3-dioxygenase in CD11C(+) dendritic cells and promotes the development of functional Treg cells” Normally, tryptophan metabolism by IDO keeps T-cell immune responses under control.18Tumor cells hijack this immunosuppressive process. They have evolved to upregulate IDO activity, in order to suppress T-cell function and help themselves survive.1”
This IDO-1 pathway MUST be combine with say PD-1 protein blocking pathway or other cellular s such as mesenchymal or plasmacytoid dendritic cells (pDCs) and antigen presentation to call for more killer cells.
Learning from PD-1 Resistance: New Combinations Strategy Gordon J. Freeman
“When they analyzed immune infiltrates found:
• Anti-CTLA-4 treatment expands the presence of CD4 effector T cells that are positive for ICOS, an immune-stimulating protein, and that these cells were strongly associated with smaller tumors in the mice.
• Both anti-PD-1 and anti-CTLA-4 treatment greatly expand the presence of CD8 T cells, the most powerful killers in the T cell family, and this expansion was associated with smaller tumors in the mice.
• These PD-1 positive CD8 T cells had what scientists call an exhausted-like phenotype. They have markers of inactivity, including the presence of other immune checkpoints, but are not necessarily known to be inactive and likely still have significant functional activity.” Jim Allison…… www.sciencedaily.com/releases/2017/08/170810124952.htm
"The mechanisms these two therapies use mostly do not overlap, which provides a reason why combining them works better than either alone," said Jim Allison,
• •Anti-PD-1 and anti-CTLA-4 utilize distinct cellular mechanisms
• •T cell responses to different tumor models are fundamentally similar
• •Anti-PD-1 and anti-CTLA-4 both target subsets of exhausted-like CD8 T cells
• •CTLA-4 blockade induces expansion of ICOS+ Th1-like CD4 T cells
DOI: http://dx.doi.org/10.1016/j.cell.2017.07.024
“Analyses reveal a spectrum of tumor-infiltrating T cell populations that are highly similar between tumor models and indicate that checkpoint blockade targets only specific subsets of tumor-infiltrating T cell populations. Anti-PD-1 predominantly induces the expansion of specific tumor-infiltrating exhausted-like CD8 T cell subsets. In contrast, anti-CTLA-4 induces the expansion of an ICOS+ Th1-like CD4 effector population in addition to engaging specific subsets of exhausted-like CD8 T cells. “
What controls the down regulation of the immune system at the microbiome Epithelial to Mesenchymal Cell Transition (EMT) and therefore sends the human body in the reboot, rescue and repair modes by helper CD4 T cells ? Mesenchymal cells
“Epithelial to Mesenchymal Transition (EMT) describes a mechanism by which cells lose their epithelial characteristics and acquire more migratory mesenchymal properties. This transient and reversible process is classified into three subtypes that are dependent on the biological and functional setting in which it occurs. This illustration represents general pathways in the scientific literature and is not to be considered comprehensive nor definitive. TIL tumor-infiltrating-lymphocytes Transcriptomes of T helper 1 (Th1), Th17. Treg cells have early cell surface markers such as programmed death (PD)-1 and CCR8. Therefore major histocompatibility complex (MHC) molecules play a major function in immunotherapy.
Type 1 EMT during development is essential for gastrulation, neural crest cell migration, and organ development. EMT generates fibroblasts following tissue injury that assist in local wound healing.
Type 2 Persistent EMT following attenuation of inflammation can result in organ fibrosis.
Type 3 EMT results in the transformation of epithelial cells into the invasive metastatic mesenchymal cells that underlie cancer progression.”
“Loss of Tight Junctions, Adherens Junctions, and Desmosomes > Disassembly of specialized cell-cell contacts leads to redistribution of cytoskeletal proteins and disruption of the apical-basal cell polarity of epithelial cells. > Key Molecules: Actin, a-Actinin, a-Catenin, ß-Catenin, Claudins, E-Cadherin, Desmogleins, Desmocollin, JAM, Occludin, Plakoglobin, Plakophilin, Vinculin, Zona Occludens
Cytoskeletal Changes Formation of actin stress fibers that anchor to focal adhesion complexes to begin to promote cell migration. > Key Molecules: Actin, Cytokeratins, S100A4, a-Smooth Muscle Actin, Vimentin
Transcriptional Shift Suppression of epithelial genes and activation of mesenchymal genes is mediated by Snail, ZEB, and bHLH family transcription factors. Vimentin is upregulated and extracellular deposition of Fibronectin is increased. > Key Molecules: FoxC2, Goosecoid, LEF-1, Snail 1, Snail 2 (Slug), Twist-1, ZEB1, ZEB2
Increased Migration and Motility > Up regulation of N-Cadherin, secretion of matrix metalloproteases, and stimulation of integrins by extracellular matrix proteins facilitates cell motility. > Key Molecules: N-Cadherin, FAK, Fibronectin, a5ß6 Integrin, Laminin-5, SPARC, Syndecan-1, Vitronectin these are your cancer causing molecules”
Embryonic DC cells ..Why? …..homogeneous population easy of manufacturing and they are the most potent …..Do they migrate like hMSC mesenchymal cells? ….unknown
“Epithelial to Mesenchymal Cell Transition – loss of cell adhesion leads to constriction and extrusion of newly mesenchymal cell.”
How about the proliferation of say TIL (Tumor infiltrated lymphocytes ) cells and how they get exhausted ? “Lack of Dendritic cells maybe at the microbiome effecting the dendritic cells are usually divided into two main groups: the myeloid dendritic cells (mDCs) and the plasmacytoid dendritic cells (pDCs) and antigen presentation to call for more killer cells.”
Developed an efficient method of pre- embryonic generation
eDendritc are Immunomodulatory/ no need for IMMUNOSUPPRESSION
Cells Persist transiently/ minimal risk of TUMORIGENICITY
Platform technology ( cancer )
Why? are pre-embryonic Dendritic superior to other clinically used adult stem cells
greater therapeutic potency
Better MIGRATORY properties
UNLIMITED ( and non variable ) cell source form naive blastomere pluripotent stem cell from
non embryonic destructive ( NED cell lines ) process
Full steam ahead with this Immunotherapy treatment and cellular replacements …We are innovating a new paradigm shift with stem cells at the Mesenchymal and dendritic levels with new cells such as hESCs/iPSC or simple new cell replacement be it gene therapy . Early the cell the better and they migrated to the site of need to alter the microbiome environment.
We need to complete studies ASAP to increase demand of immune therapy and cellular response. We have started the development phase of setting up the manufacturing process for release of approved cellular products. When I started this thread, I was thinking what is a way to short cut the drug product approval cycle, since this is regenerative medicine and we are using cells to treat and halt these diseases? We are trying to release a product for the #1 unmet medical need with a new technology with limited resources (conservative money, no JV yet and no revenues) and the FDA/NIH and world regulatory bodies are on a learning curve.
The patient’s needs are for treatment, yesterday, time lost/ life lost. Who would not want to improve their eye sight with a safe effective treatment or cure cancer like Jim Allison with his CTLA-4 Monoclonal antibodies or using new replacement cells to save lives of your love ones?
My questions are when will the patients’ unmet needs Trump the need of process? And where can the drug process be short circuit for the benefits of the patients? Federal FDA/NIH funding or CIRM funding in California.
After all folks we are tied to our patient’s diagnosis, prognosis, and cellular treatment of the FDA in the USA. Can other countries such as Japan move this process more quickly since they are more focused on the immune process? Time will tell……In the meantime many will suffer and die today…..Why is autophagy important to cancer cell microbiome? What’s the feedback loop to antigen presentation? How to boast the immune response for TIL tumor attacking cells for cancer and or auto immune diseases?
Part 2
How does Cell Replacement / Regenerative Medicine work with the immune system? What happens when you inject stem cells that can migrate to site of need? What are the right stem cells ? How do you define potency of cells and protein cytokines effects on Microbiome? Do you know? , let’s talk ….. Think about what a cancer stem cell is and early immune monitoring is the key for potency, migration and paracrine cytokines . Where scientist are mislead is they deal in the adult system without regards of cell replacement with the correct state …….If you have stage IV cancer you better understand PD-1 biology with CTLA_4????
Three key terms antigen presentation (be it protein or cell–cell interaction) potency (naive blastomere pluripotent stem cells are the gold standard) and migration (or in cancer know as metastasis) and a possible answer to a question of Cancer Stem Cells and what are they? You are more that welcome to share this white paper … Why are the T cells and NK cells considered the police and fire departments? And is Cancer the burning building ? Who do you call when you have a burning building? The plumber , no( T cells )!!!!!!!! Can T and Natural Killer (NK) cells kill cancer? Answer is yes we have know that for 30 years ....Lewis L. Lanier (NK s) and Jim Allison ( CTLA-4)or Gordon Freeman and Arlene Sharpe (PD-1) and Lieping Chen was independently discovering PD-L1, - See more at: http://www.cancerresearch.org/news-publications/our-blog/september-2014/cri-names-winners-of-top-scientific-prize#sthash.9qVBJx7K.dpuf…
.What controls T cells at the microbiome (site of the cancer )? MSC cells, down regulation!!!!! What controls NK cells at the microbiome? Dendritic cells, up regulation!!!!!! More on the up and down regulation of the immune system with these cell types?
“Mesenchymal stromal cells (MSCs) are adult multipotent stem cells residing as pericytes in various tissues and organs where they can differentiate into specialized cells to replace dying cells and damaged tissues. These cells are commonly found at injury sites and in tumors that are known to behave like “wounds that do not heal.” In this article, we discuss the mechanisms of MSCs in migrating, homing, and repairing injured tissues. Reference also reviews a number of reports showing that tumor microenvironment triggers plasticity mechanisms in MSCs to induce malignant neoplastic tissue formation, maintenance, and chemoresistance, as well as tumor growth” Reference # 12
“MSC role in tumor microenvironment. MSCs can induce cancer cell survival, stemness, and chemoresistance by differentiating
into cancer-associated fibroblasts (CAFs) using a tumor growth factor ß type 1 (TGFß1)-dependent mechanism, and by releasing soluble
factors that favor angiogenesis and immunosuppression in the tumor microenvironment, such as prostaglandin E2 (PGE2) and vascular
endothelial growth factor (VEGF). MSCs can mediate anti-cancer effects by releasing anti-cancer factors, such as tumor necrosis factorrelated
apoptosis inducing ligand (TRAIL), via mechanisms that are not well understood.”
Cancer Biol Med 2017. doi: 10.20892/j.issn.2095-3941.2016.0033
So here is the phenotyping and genotyping back to early embryonic development in what is lost of cell surface expression is key to understand what is lost and gain in the life time of the of immune system Highly tumorigenic subpopulation of cancer cells expressing or lacking the cell surface markers CD10, CD24, CD44, CD133. Stro-1 and epithelial-specific antigen (ESA) CAMs are the key to understanding the immune of Cancer. You can substitute any CAM for any organ to be more comprehensive or inclusive tumor specific ……
Why is CD24 key to mesenchymal stem cells …..Why are they very high in blastomere pluripotent stem cells 24% vs. adult MSC 0-1 % ….Robert Lanza MSC papers on Astellas.com ….. see reference #2
1. What is CD 24 “Signal transducer CD24 also known as cluster of differentiation 24 or heat stable antigen CD24 (HSA) is a protein that in humans is encoded by the CD24gene.[1] CD24 is a cell adhesion molecule.” Very important and “CD24 is a glycoprotein expressed at the surface of most B lymphocytes and differentiating neuroblasts. This gene encodes a sialoglycoprotein that is expressed on mature granulocytes and in many B cells. “ And we now know on early embryonic MSC from blastomere pluripotent stem cells –Lanza MSC papers
2. What is CD44 homing receptor know a lot about this ….. “CD44 antigen is a cell-surface glycoprotein involved in cell–cell interactions, cell adhesion and migration. In humans, the CD44 antigen is encoded by the CD44 gene on Chromosome 11.[1] CD44 has been referred to as HCAM (homing cell adhesion molecule), Pgp-1 (phagocyticglycoprotein-1), Hermes antigen, lymphocyte homing receptor, ECM-III, and HUTCH-1.” what is metastases you got it ….migration
3. CD10 this is the blast state of the immature cells “Neprilysin, also known as membrane metallo-endopeptidase (MME), neutral endopeptidase (NEP),cluster of differentiation 10 (CD10), and common acute lymphoblastic leukemia antigen (CALLA) is an enzyme that in humans is encoded by the MME gene. Neprilysin is a zinc-dependent metalloprotease that cleaves peptides at the amino side of hydrophobic residues and inactivates several peptide hormones including glucagon, enkephalins, substance P, neurotensin, oxytocin, and bradykinin.[1] It also degrades the amyloid beta peptide whose abnormal misfolding and aggregation in neural tissue has been implicated as a cause of Alzheimer's disease.” And we now know on early eMSC from blastomere pluripotent stem cells –Lanza MSC papers
4. Stro-1 What is Stro-1? ” Stro-1 is the best-known mesenchymal stem cell marker. However, despite its bone marrow origin, its localization in bone marrow has never been demonstrated. By immunofluorescence staining, it is shown that ~ 0.74% of nucleated bone marrow cells expressed Stro-1. Also found that ~ 8.7% of CD34-expressing cells expressed Stro-1, and more than 20% of Stro-1-expressing cells did not express CD34. In adipose tissue Stro-1 expression was identified in the endothelium of arterioles and capillaries. Stro-1 was also localized in the endothelium of some but not all adipose tissue veins. Endothelial expression of Stro-1 was also identified in blood vessels in penis and in leg muscles, but not in other tested tissues. In these other tissues, Stro-1 was scantly expressed near but not in blood vessels. These variable and endothelial expression patterns of Stro-1 point to a need to re-examine published data that relied on Stro-1 as a mesenchymal stem cell marker.”
Robert Lanza and Etc. found that Stro-1 was negative for his hMSC blastomere pluripotent stem cells vs. very positive for adult MSC ….so if you have Stro-1 in high expression your potency will be varied and low…An err of many MSC researchers because of the maturity of MSC from EMT…..
5. “ESA Or CD326 Ep-CAM consists of two glycoproteins, 34 and 39 kDa, sometimes designated epithelial antigen, epithelial specific antigen, and epithelial glycoprotein. In paraffin sections, the protein is detected with mAbs like Ber-EP4 and MOC-31. The glycoproteins are located on the cell membrane surface and in the cytoplasm of virtually all epithelial cells with the exception of most squamous epithelia, hepatocytes, renal proximal tubular cells, gastric parietal cells and myoepithelial cells.”
6. CD133 “CD133 antigen also known as prominin-1 is a glycoprotein that in humans is encoded by the PROM1gene.[1][2] It is a member of pentaspan transmembrane glycoproteins (5-transmembrane, 5-TM), which specifically localize to cellular protrusions. While the precise function of CD133 remains unknown, it has been proposed to act as an organizer of cell membrane topology” “Tissue distribution[edit]
CD133 is expressed in hematopoietic stem cells,[4] endothelial progenitor cells,[5] glioblastoma, neuronal andglial stem cells,[6] various pediatric brain tumors,[7] as well as adult kidney, mammary glands, trachea, salivary glands, placenta, digestive tract, testes, and some other cell types.[8][9]”
7. All these CAM reagents are important because of EMT
Take in concretion the dendritic cells. What are DC ? “Dendritic cells (DCs) are antigen-presenting cells (also known as accessory cells) of the mammalian immune system. Their main function is to process antigen material and present it on the cell surface to the T cells of the immune system.”
And “The innate immune system is an evolutionarily older defense strategy, and is the dominant immune system found in plants,fungi, insects, and primitive multicellular organisms.[3]
The major functions of the vertebrate innate immune system include:
• Recruiting immune cells to sites of infection, through the production of chemical factors, including specialized chemical mediators, called cytokines
• Activation of the complement cascade to identify bacteria, activate cells, and promote clearance of antibody complexes or dead cells”
• The identification and removal of foreign substances present in organs, tissues, the blood and lymph, by specialised white blood cells
• Activation of the adaptive immune system through a process known as antigen presentation”
Acting as a physical and chemical barrier to infectious agents. “Dendritic cells (DC) are phagocytic cells present in tissues that are in contact with the external environment, mainly the skin (where they are often called Langerhans cells), and the inner mucosal lining of the nose, lungs, stomach, and intestines.[2] They are named for their resemblance to neuronal dendrites, but dendritic cells are not connected to the nervous system. Dendritic cells maturation process.
Dendritic cell life history can be subdivided into a number of phases, all with discrete cellular functionality. Transition between phases is mediated by diverse signals and is accompanied by changes in expression patterns of many surface markers and secreted factors. MDC, myeloid dendritic cell; SCF, stem cell factor; GM-CSF, granulocyte–macrophage colony stimulating factor; TGF, transforming growth factor; HSP, heat shock protein.
Immunology and Cell Biology (2000) 78, 91–102; doi:10.1046/j.1440-1711.2000.00888.x
More work to be done with immunotherapy and cell replacements
References:
1. Identification of Pancreatic Cancer Stem Cells Chenwei Li,1 David G. Heidt,1 Piero Dalerba,4 Charles F. Burant,2,3 Lanjing Zhang,3 Volkan Adsay,4 Max Wicha,3 Michael F. Clarke,5 and Diane M. Simeone1,2 Cancer Res 2007; 67: (3). February 1, 2007
2. Link to Robert Lanza ‘s papers… Mesenchymal stem cell population derived from human pluripotent stem cells displays potent immunomodulatory and therapeutic properties.
Kimbrel EA1, Kouris NA, Yavanian GJ, Chu J, Qin Y, Chan A, Singh RP, McCurdy D, Gordon L, Levinson RD, Lanza R. Stem Cells Dev. 2014 Jul 15;23(14):1611-24. doi: 10.1089/scd.2013.0554. Epub 2014 May 2.
Addendum
Robert Lanza http://www.robertlanza.com/
abstract from reference #2
“ Mesenchymal stem cells (MSCs) are being tested in a wide range of human diseases; however, loss of potency and inconsistent quality severely limit their use. To overcome these issues, we have utilized a developmental precursor called the hemangioblast as an intermediate cell type in the derivation of a highly potent and replenishable population of MSCs from human embryonic stem cells (hESCs). This method circumvents the need for labor-intensive hand-picking, scraping, and sorting that other hESC-MSC derivation methods require. Moreover, unlike previous reports on hESC-MSCs, we have systematically evaluated their immunomodulatory properties and in vivo potency. As expected, they dynamically secrete a range of bioactive factors, display enzymatic activity, and suppress T-cell proliferation that is induced by either allogeneic cells or mitogenic stimuli. However, they also display unique immunophenotypic properties, as well as a smaller size and > 30,000-fold proliferative capacity than bone marrow-derived MSCs. In addition, this is the first report which demonstrates that hESC-MSCs can inhibit CD83 up-regulation and IL-12p70 secretion from dendritic cells and enhance regulatory T-cell populations induced by interleukin 2 (IL-2). This is also the first report which shows that hESC-MSCs have therapeutic efficacy in two different autoimmune disorder models, including a marked increase in survival of lupus-prone mice and a reduction of symptoms in an autoimmune model of uveitis. Our data suggest that this novel and therapeutically active population of MSCs could overcome many of the obstacles that plague the use of MSCs in regenerative medicine and serve as a scalable alternative to current MSC sources.”
3. Human ESC-Derived MSCs Outperform Bone Marrow MSCs in the Treatment of an EAE Model of Multiple Sclerosis
Xiaofang Wang Erin A. Kimbrel7 Kumiko Ijichi Debayon Paul Adam S. Lazorchak Jianlin Chu Nicholas A. Kouris Gregory J. Yavanian Shi-Jiang Lu Joel S. Pachter Stephen J. Crocker Robert Lanza stem cell reports DOI: http://dx.doi.org/10.1016/j.stemcr.2014.04.020
4. Stem Cells Dev. 2011 Oct;20(10):1747-52. doi: 10.1089/scd.2010.0564. Epub 2011 Feb 24.
Tissue distribution of mesenchymal stem cell marker Stro-1.
Lin G1, Liu G, Banie L, Wang G, Ning H, Lue TF, Lin CS.
5. Products for EMT Research Species are at R and D Systems https://www.rndsystems.com/products/epithelial-mesenchymal-transition-products
6.Wikipedia
7.CCL21/CCR7 Axis Contributed to CD133+ Pancreatic Cancer Stem-Like Cell Metastasis via EMT and Erk/NF-?B Pathway
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0158529&elqTrackId=6906a59297b74aacb93a1e5f39d66123&elq=31dd791262264b54b7c53df36192ff03&elqaid=16390&elqat=1&elqCampaignId=11
8. Cancer cells enter dormancy after cannibalizing mesenchymal stem/stromal cells (MSCs) http://www.pnas.org/content/113/42/E6447.long
Thomas J. Bartosha,b,1, Mujib Ullaha, Suzanne Zeitounia, Joshua Beavera,b, and Darwin J. Prockopa,1Thomas J. Bartosh, E6447–E6456, doi: 10.1073/pnas.16122901138.
Abstract
Patients with breast cancer often develop malignant regrowth of residual drug-resistant dormant tumor cells years after primary treatment, a process defined as cancer relapse. Deciphering the causal basis of tumor dormancy therefore has obvious therapeutic significance. Because cancer cell behavior is strongly influenced by stromal cells, particularly the mesenchymal stem/stromal cells (MSCs) that are actively recruited into tumor-associated stroma, we assessed the impact of MSCs on breast cancer cell (BCC) dormancy. Using 3D cocultures to mimic the cellular interactions of an emerging tumor niche, we observed that MSCs sequentially surrounded the BCCs, promoted formation of cancer spheroids, and then were internalized/degraded through a process resembling the well-documented yet ill-defined clinical phenomenon of cancer cell cannibalism. This suspected feeding behavior was less appreciable in the presence of a rho kinase inhibitor and in 2D monolayer cocultures. Notably, cannibalism of MSCs enhanced survival of BCCs deprived of nutrients but suppressed their tumorigenicity, together suggesting the cancer cells entered dormancy. Transcriptome profiles revealed that the resulting BCCs acquired a unique molecular signature enriched in prosurvival factors and tumor suppressors, as well as inflammatory mediators that demarcate the secretome of senescent cells, also referred to as the senescence-associated secretory phenotype. Overall, our results provide intriguing evidence that cancer cells under duress enter dormancy after cannibalizing MSCs. Importantly, our practical 3D coculture model could provide a valuable tool to understand the antitumor activity of MSCs and cell cannibalism further, and therefore open new therapeutic avenues for the prevention of cancer recurrence.
9. Transcriptional Landscape of Human Tissue Lymphocytes Unveils Uniqueness of Tumor-Infiltrating T Regulatory Cells
http://dx.doi.org/10.1016/j.immuni.2016.10.021
Marco De Simone13Alberto Arrigoni13Grazisa Rossetti1Paola Gruarin
Summary
Tumor-infiltrating regulatory T lymphocytes (Treg) can suppress effector T cells specific for tumor antigens. Deeper molecular definitions of tumor-infiltrating-lymphocytes could thus offer therapeutic opportunities. Transcriptomes of T helper 1 (Th1), Th17, and Treg cells infiltrating colorectal or non-small-cell lung cancers were compared to transcriptomes of the same subsets from normal tissues and validated at the single-cell level. We found that tumor-infiltrating Treg cells were highly suppressive, upregulated several immune-checkpoints, and expressed on the cell surfaces specific signature molecules such as interleukin-1 receptor 2 (IL1R2), programmed death (PD)-1 Ligand1, PD-1 Ligand2, and CCR8 chemokine, which were not previously described on Treg cells. Remarkably, high expression in whole-tumor samples of Treg cell signature genes, such as LAYN,MAGEH1, or CCR8, correlated with poor prognosis. Our findings provide insights into the molecular identity and functions of human tumor-infiltrating Treg cells and define potential targets for tumor immunotherapy.
10. Interaction of MSC with tumor cells
• Catharina Melzer,
• Yuanyuan Yang and
• Ralf HassEmail authorView ORCID ID profile
Cell Communication and Signaling201614:20
DOI: 10.1186/s12964-016-0143-0
Abstract
Tumor development and tumor progression is not only determined by the corresponding tumor cells but also by the tumor microenvironment. This includes an orchestrated network of interacting cell types (e.g. immune cells, endothelial cells, fibroblasts, and mesenchymal stroma/stem cells (MSC)) via the extracellular matrix and soluble factors such as cytokines, chemokines, growth factors and various metabolites. Cell populations of the tumor microenvironment can interact directly and indirectly with cancer cells by mutually altering properties and functions of the involved partners. Particularly, mesenchymal stroma/stem cells (MSC) play an important role during carcinogenesis exhibiting different types of intercellular communication. Accordingly, this work focusses on diverse mechanisms of interaction between MSC and cancer cells. Moreover, some functional changes and consequences for both cell types are summarized which can eventually result in the establishment of a carcinoma stem cell niche (CSCN) or the generation of new tumor cell populations by MSC-tumor cell fusion.
Keywords
MSC Mesenchymal stroma/stem cells Tumor cell signaling Tumor microenvironment Cellular interaction Cell fusion
Fig. 1
Indirect interactions between mesenchymal stroma/stem cells and cancer cells. a Cytokines, chemokines, growth factors: MSC secrete a plethora of soluble factors that can bind as substrates to appropriate receptors on the cell surface of cancer cells and vice versa for mutual activation of signaling pathways. b Metabolites: Likewise, MSC-released metabolites such as prostaglandin E2, kynurenine or galectin-1 can act in a paracrine manner on cancer cells altering their properties and functions [14]. c Exosomes: Both, MSC and cancer cells, secrete exosomes for the exchange of small molecules including protein, mRNAs and microRNAs. d Microvesicles: Besides exosomes, microvesicles represent a different type of microparticles for the exchange of small molecules such as mRNAs or microRNAs affecting tumor cells and MSC in mutual ways
11. World J Stem Cells. 2016 Sep 26; 8(9): 268–278.
Published online 2016 Sep 26. doi: 10.4252/wjsc.v8.i9.268
Immunomodulation by mesenchymal stem cells: Interplay between mesenchymal stem cells and regulatory lymphocytes
Oscar Ka-Fai Ma and Koon Ho Chan
Mesenchymal stem cells (MSCs) possess immunomodulatory properties, which confer enormous potential for clinical application. Considerable evidence revealed their efficacy on various animal models of autoimmune diseases, such as multiple sclerosis, systemic lupus erythematosus and uveitis. MSCs elicit their immunomodulatory effects by inhibiting lymphocyte activation and proliferation, forbidding the secretion of proinflammatory cytokines, limiting the function of antigen presenting cells, and inducing regulatory T (Treg) and B (Breg) cells. The induction of Treg and Breg cells is of particular interest since Treg and Breg cells have significant roles in maintaining immune tolerance. Several mechanisms have been proposed regarding to the MSCs-mediated induction of Treg and Breg cells. Accordingly, MSCs induce regulatory lymphocytes through secretion of multiple pleiotropic cytokines, cell-to-cell contact with target cells and modulation of antigen-presenting cells. Here, we summarized how MSCs induce Treg and Breg cells to provoke immunosuppression.
Key Words: Mesenchymal stem cells, Regulatory T cells, Regulatory B cells, Immunomodulation,Autoimmunity
Core tip: In this review, we summarized the mechanisms involved in regulatory T (Treg) and B (Breg) cell induction by mesenchymal stem cells (MSCs). In an inflammatory environment, MSCs secrete various anti-inflammatory cytokines, actively interact with immune cells and modulate them to acquire regulatory properties, thus, generate a tolerogenic environment. Particularly, by inducing Treg and Breg cells, the immunomodulation of MSCs is amplified. Therefore, genetic engineered MSCs to enhance their ability to induce Treg and Breg cells may increase their therapeutic efficacy.
12. Mesenchymal stromal cells’ role in tumor microenvironment: involvement of signaling pathways Armel Herve Nwabo Kamdje1, Paul Takam Kamga2, Richard Tagne Simo1, Lorella Vecchio3, Paul Faustin Seke Etet3, Jean Marc Muller4, Giulio Bassi2, Erique Lukong5, Raghuveera Kumar Goel5, Jeremie Mbo Amvene1, Mauro Krampera2 Cancer Biol Med. 2017 May; 14(2): 129–141.
doi: 10.20892/j.issn.2095-3941.2016.0033
ABSTRACT Mesenchymal stromal cells (MSCs) are adult multipotent stem cells residing as pericytes in various tissues and organs where they can differentiate into specialized cells to replace dying cells and damaged tissues. These cells are commonly found at injury sites and in tumors that are known to behave like “wounds that do not heal.” In this article, we discuss the mechanisms of MSCs in migrating, homing, and repairing injured tissues. We also review a number of reports showing that tumor microenvironment triggers plasticity mechanisms in MSCs to induce malignant neoplastic tissue formation, maintenance, and chemoresistance, as well as tumor growth. The antitumor properties and therapeutic potential of MSCs are also discussed. KEYWORDS Mesenchymal stromal cells; systemic circulation; migration; homing; tumor modulation; signaling pathways; chemoresistance
13. Distinct Cellular Mechanisms Underlie Anti-CTLA-4 and Anti-PD-1 Checkpoint Blockade , Cell Volume 170, Issue 6, p1120–1133.e17, 7 September 2017 Et el Spencer C. Wei, Jim Allison
Highlights
•Anti-PD-1 and anti-CTLA-4 utilize distinct cellular mechanisms
•T cell responses to different tumor models are fundamentally similar
•Anti-PD-1 and anti-CTLA-4 both target subsets of exhausted-like CD8 T cells
•CTLA-4 blockade induces expansion of ICOS+ Th1-like CD4 T cells
“Ask Immunologic questions give molecular answers” Andy Blidy
• http://www.1001freedownloads.com/free-clipart/seesaw
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