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Are NK cells lost when Mesenchymal, Dendritic Cells Immunologic Cell Teeter-Totter controls the Immune System?
“Ask Immunologic questions give molecular answers” Andy Blidy
https://www.youtube.com/watch?v=PphTL6vY4W4 History of Immunotherapy by James Allison at PMWC 2017 Silicon Valley

https://www.youtube.com/watch?=vyMxegkW9p-k Lewis L. Lanier Immunotherapy: Unleashing the Body’s Natural Defense Systems to Fight Cancer



Version 1.3
10-5-2017

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 immunologic Teeter Totter or cancer on the top side of the Teeter Totter….. But how does one balance the immune Teeter Totter as one age’s a replacement cellular therapy will be employed in the near future and genetic expression from possible gene therapy ( CRISPR-CAS9 technology) 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 certain cells as the fulcrum on the Teeter Totter. A question can be asked, Do you want to suppress the immune system or activate it? But what cells control the microenvironment to do this? A high level process of the immune system is illustrated below in several illustration ……Notice balance is needed in these quantum immunologic systems with the Mesenchymal (MSC) and Dendritic cells (DC) in the middle of balance immunity acting as fulcrum points of checkpoints associated with biomarkers …….Now let’s look at the microbiome and the immunomodulatory effects of Mesenchymal and Dendritic cells at the local tissue level related to immune system and how these systems function together, keeping in mind the effects on auto immune diseases and cancers in an inflammation or tumor microenvironment or tissue organ disease….If NK cells are first to fight a cancer cell and if they lose the battle b/c of environmental controls or have their activity blocked by local MSC immunomodulatory activate then the cancer grows and possibly spreads….

Mesenchymal / Pericytes are contractile cells that wrap around the endothelial cells. What is epithelial transition to mesenchymal, EMT? Answer is in last illustration….(MSC) are near such tissue as the skin and mucous membranes, providing the first line of defense for innate immunity. Many non-myeloid cells contribute to this defense strategy, including fibroblasts and epithelial/mesenchymal cells, producing immunomodulatory and antimicrobial factors. MSCs and IMO are centrally located within the epithelial microbiome associated with both of the innate cellular CD8 and humor B cell of immune system. Expression of toll-like ligand receptors (TLRs) on the cell surface of mesenchymal stromal cells (MSCs) is another key role suggesting their inherent role in recognition of antigen presentation

MSCs effects many different cells and are a major component of the innate immunity trigging the cascade of the complement system. The central step in complement activation is the cleavage of C3 into C3a and C3b. These proteins become bound to the immune cells and express the complement receptors, such as CR1, CR2, CR3, and CR4. Binding of MSC to C3 has been shown to suppress the proliferation of Peripheral Blood Mononuclear Cells. Therefore, this inhibits Activated T cells that are required for rescue and repair by CD4 pathways. This suppressive activity of MSCs may be a major role to regulating immune control of the regeneration and repair of tissue. Notice the Mesenchymal MSC is the central cell of this biological system …..



“The multi-faceted anti-inflammatory actions of MSCs. In response to pro-inflammatory cytokines or TLR3 stimuli, MSCs will develop an anti-inflammatory profile. Through the secretion of soluble factors these licensed cells can act on numerous innate immune cells affecting both effector function and phenotype. Individual effects are discussed further within the main text.https://doi.org/10.1016/j.imlet.2015.05.004 Mesenchymal stromal cells and the innate immune response” reference #1


Other cells affected by MSCs are:
1. Neutrophils (PMNs) are major part of innate immune cell, responding to foreign challenge by homing to the wound site within several minutes of injury. These non-proliferative, phagocytic cells respond to such foreign antigen as bacterial invasion, these are control in the microbiome by MSCs such as in sepsis to help in microbial defense. Myeloid-derived suppressor cells (MDSCs) implicated in Autoimmune Diseases and Cancer to maintain a cancer microenvironment or chronic inflammation microbiome.
The immunology and molecular biology has come a long ways from the Aids Crisis in the early 1980’s to the embryonic age in 1998 and the field has not developed a bio-process manufacturing of MSC cell in large volume for therapy by cell replacement. In fact the MSC are non-standard for potency, migration and Paracrine production in most clinical trial studies. The uncharacterized cells use in most therapy (adult stem cells are the least potent and do not migrate to the site of injury ( chronic inflammation) where they product proteins for control of the immunity be it up or downing the immune response).

2. Mast cells (MCs) are also innate responder cells during MSC homing levels of IL-6 IL-1a secretions aid in the prevention of pro-apoptotic activity on neutrophils. MCs are related to IgE mediated MC degranulation that can call for the recruitment of PMNs a Dendritic cells. Most scientists relate MCs to allergy pathways. Including myself (Tufts School Allergy Department with Ross E. Rocklin and Lanny Rosenwasser graduate advisors.) We studied histamine response on WBC

3. Dendritic Cells (DCs) provide a link between the innate and adaptive immune systems ( Cartoon below) Main function of Dendritic cells is go between of the NK cells and T cells or the control of innate and cellular immunity .



“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


4. Natural killer (Lewis Lanier, expert in NKs cells , my boss at BD Monoclonal center ) cells have are activated by exposure to IL-2 or IL-15 Causing the expression of IFN? and TNFa pathways for cancer tumor and viral infection….. Both IDO and PGE2 offer multiple mechanisms for dampening NK responsiveness to the MSCs.

5. Monocytes ( IL-1 Monocytes discovered by Charles Dinarello at Tufts Medical ) represent approximately 10% of circulating leukocytes detecting microbial pathogens and enhance the recruitment of monocytes and macrophages into inflamed tissues to promote wound repair through the secretion of the chemokine.

6. Macrophages are characterized into two phenotypes, the M1 pro-inflammatory macrophage with antimicrobial activity and the M2 anti-inflammatory macrophage





What is the allogenic effect of MSC on the response related to immune T cells? In vivo Robert Lanza and group have shown that human embryonic MSC does alter the secretion of cytokines to Dendritic Cells, Naive and effector T cells both TH1 and TH2 and NK s causing an anti-inflammatory response along with phenotype change. tumor necrosis factor (TNF- alpha) secretion and mature DC to increase interleukin-10 (IL-10) and interferon gamma (IFN-g) this will cause an increase in Treg CD4 cells by IL-4 …Leading to prostaglandin E2 (PGE2), and inhibitors of PGE2 production mitigated hMSC-mediated immune modulation in the microenvironment that controls inflammation and may lead to lower GVHD disease from transplantation of cells or organs. Recently, McGuirk (Reference #3 ) has shown MSCs secrete an array of cytokines, chemokines, and soluble receptors that act locally in microbiome. MSCs regulate immunity by interacting with innate immune cells (including macrophages, natural killer (NK) cells, and dendritic cells), and adaptive immune cells (including B and T cells)


“Immune profile of mesenchymal stem cells (MSCs). Graphic summary of the interactions between MSC and the immune system. MSCs can suppress proliferation of both T helper (TH) and cytotoxic T cells (Tc) through multiple pathways. Differentiation of MSCs to TH2 and regulatory T-cells (Treg) is triggered, resulting in an anti-inflammatory environment. Interleukin (IL)-6 blocks the maturation of dendritic cells (DC) by inhibiting upregulation of CD40, CD80, and CD86, which subsequently reduces T-cell activation. Monocytes are stimulated by MSCs to preferentially differentiate towards the M2 phenotype. IL-10, produced by M2 macrophages, can boost the formation of Treg, and simultaneously reduces neutrophil tissue migration. Neutrophils (polymorphonuclear granulocytes; PMN) have a longer life span; however, production of reactive oxygen species (ROS) is decreased. Natural killer (NK) cell proliferation and cytotoxic activity are both suppressed. B-cell proliferation is inhibited, and production of antibodies is reduced. HGF, hepatocyte growth factor; IDO, indoleamine-pyrrole-2-3-dioxygenase; PGE2, prostaglandin E2; and TGF-ß, transforming growth factor-ß. (Adapted from van den Akker F, de Jager SC, Sluijter JP. Mesenchymal stem cell therapy for cardiac inflammation: iummunomodulatory properties and the influence of toll-like receptors.” Mediators Inflamm2013: 181020, 2013.)


“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.” 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 #5



“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 factor related
apoptosis inducing ligand (TRAIL), via mechanisms that are not well understood.”
Cancer Biol Med 2017. doi: 10.20892/j.issn.2095-3941.2016.0033









In Conclusion:
It is now known that TCR gamma delta show suppression of the immune system while TCR alpha beta is 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
An example of Tumor resistance that may be alter by the correct MSC cell population. Indoleamine 2,3-dioxygenase (IDO) is the rate-limiting enzyme in the catalysis of tryptophan. The mechanism by which IDO promotes tolerance is still an area of active investigation and hot clinical trials for Cancer with PDL-1 and other biomarkers, such as CTLA-4.


Immune checkpoint pathways down regulate or up regulation of T-cell activation to maintain peripheral tolerance 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. But MSCs will be the key in stem cell replacement in Regenerative Medicine in the near future. The major question is what MSC type of Replacement cell, adult vs embryonic vs IPSC ? IMO, Naïve blastomeric pluripotent MSC ( google Robert Lanza and MSC) derived from NED cell lines are the most potent, migrate the best and produce the immunomodulatories need for reboot, rescue and repair, therefore will be the best cells for therapy , but how long will the bioprocess take and the scientific field to realize what are the best cells ?

Major problem of Tumor infiltrated Lymphocytes, TIL Cells in Cancer……What is the solution? Can MSC control fibroblast and be the magic cell to help cure cancer? IMO, yes


Learning from PD-1 Resistance: New Combinations Strategy Gordon J. Freeman
Where is Immunotherapy now August 2017
“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. “

In the next puzzle piece is for understanding microbiome and early microenvironments of cancer development and proliferation.


Highlights from http://dx.doi.org/10.1016/j.celrep.2017.08.089

“Lung-tumor-derived MSCs (T-MSCs) reduce NK cell function and modulate NK phenotype
T-MSCs are more immunosuppressive than their non-tumor associated counterparts
CD56 dim/bright and functional NK cell subsets are differentially modulated by MSCs
Modulation of NK cell function and phenotype by MSCs occurs mainly through PGE2”


Does the loss of NK function push the immune system to tumor infiltrated lymphocytes TIL CD3/CD8 cytotoxic cells and exhaustion in late tumors? :: Therefore it is possible that the loss of NK functioning is also related to cancer growth in late stages and metastases? This is the same process in HIV+ AIDs patients as CD4 helper cells are lost in HIV+ patients in late stage when the helper /suppressor ratios are inverted with increased CD 8 cells ….. “Tumor-Derived Mesenchymal Stem Cells Use Distinct Mechanisms to Block the Activity of Natural Killer subsets “ So replacement of fresh MSC especially the most potent cell such as naïve blastomere pluripotent MSC stem cells that Robert Lanza( Astellas Pharma http://www.the-scientist.com/?articles.view/articleNo/39575/title/Making-Better-Mesenchymal-Stem-Cells/ ) is working with might change the microbiome to attach all tumors … Since Lewis L. Lanier (the grandfather of NK cells and my former boss ad BD Monoclonal Center) calls the NK cells the “Marines” first responders on the scene to kill foreign/ caners cells …… Most pathologist miss these NK cells and MSC when diagnosing a cancer with biopsy Immunopathology because they are not there or in low numbers since the process is destroy by the cancer environment such as “compare natural killer (NK) cell immunosuppression by mesenchymal stem cells (MSCs) from primary human squamous cell carcinomas and adjacent normal lung tissue. Tumor-associated MSCs exert stronger immunosuppression than normal-tissue-derived MSCs and modulate different NK functions by distinct mechanisms.” But understanding the microenvironment is necessary to understanding the possible way to treat and attack cancer by cellular therapy using both dendritic and mesenchymal stem cells.

Will simple regenerative medicine in the near future with cancer microbiome contaminated by MSC alter this paradigm shift with pure potent mobile embryonic MSC and reboot the immune teeter-totter?…..time will tell.


References:

1. Mesenchymal stromal cells and the innate immune response, Volume 168, Issue 2, December 2015, Pages 140-146, Immunology letters Katarina Le Banc and Linsay C. Davies
2. Human mesenchymal stem cells modulate allogeneic immune cell response Blood 2005 ,105 1815 -1822 Sudeepta Aggarwal and Mark F. Pittenger
3. Review: Mesenchymal Stromal Cells: What Is the Mechanism in Acute Graft-Versus-Host Disease? Neil Dunavin , Ajoy Dias , Meizhang Li and Joseph McGuirk , biomedicine 1 July 2017 5, 39 doi:10,3390
4. Rebuilding the Damaged Heart: Mesenchymal Stem Cells, Cell-Based Therapy, and Engineered Heart Tissue Samuel Golpanian, Ariel Wolf, Konstantinos E. Hatzistergos, Joshua M. Hare Physiological Reviews Published 22 June 2016 Vol. 96 no. 3, 1127-1168 DOI: 10.1152/physrev.00019.2015
5. 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
6. 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
7. Tumor-Derived Mesenchymal Stem Cells Use Distinct Mechanisms to Block the Activity of Natural Killer Cell Subsets Sabine Galland, Joanna Vuille, Patricia Martin, Igor Letovanec, Anne Caignard, Giulia Fregni, Ivan Stamenkovic, Cell Reports 20, 2891–2905, September 19, 2017