Immune Cells Killing Stem Cells and Stem Cells Killing Immune Cells

Knight et al. J Neurol Sci.
Several studies have demonstrated that stem cells are useful in the treatment of multiple sclerosis. The Cellmedicine clinic published previously in collaboration with the University of California San Diego that 3 patients treated with their own fat derived stem cells entered remission. Other studies are ongoing, including a study at the Cleveland Clinic in which bone marrow stem cells differentiated into mesenchymal stem cells are being administered into patients with multiple sclerosis. Unfortunately the mechanisms by which therapeutic effects occur are still largely unknown. One general school of thought believes that stem cells are capable of differentiating into damaged brain cells. The other school of thought believes that stem cells are capable of producing numerous growth factors, called trophic factors, that mediate therapeutic activity of the stem cells. Yet another school of thought propagates the notion that stem cells are merely immune modulatory cells. Before continuing, it is important to point out that stem cell therapy for multiple sclerosis involving autologous hematopoietic transplants is different than what we are discussing here. Autologous (your own) hematopoietic stem cell therapy is not based on regenerating new tissues, but to achieve the objective of extracting cells from a patients, purifying blood making (hematopoietic) stem cells, destroying the immune system of the recipient so as to wipe out the multiple sclerosis causing T cells, and subsequently readministering the patient’s own cells in order to regenerate the immune system. This approach, which was made popular by Dr. Richard Burt from Northwestern University.
In order to assess mechanisms of how stem cells work in multiple sclerosis it is necessary to induce the disease in animals. The most widely used animal model of multiple sclerosis is the experimental allergic encephalomyelitis model. This disease is induced in female mice that are genetically bred to have a predisposition to autoimmunity. These animals are immunized with myelin basic protein or myelin oligodendrocyte protein. Both of these proteins are components of the myelin sheath that protects the axons. In multiple sclerosis immune attack occurs against components of the myelin sheath. Therefore immunizing predisposed animals to components of the myelin sheath induces a disease similar to multiple sclerosis. The EAE model has been critical in development of some of the currently used treatments for multiple sclerosis such as copaxone and interferon.
Original studies have demonstrated that administration of bone marrow derived mesenchymal stem cells protects mice from development of EAE. This protection was associated with regeneration on oligodendrocytes as well as shifts in immune response. Unfortunately these studies did not decipher whether the protective effects of the stem cells were mediated by immune modulation, regeneration, or a combination of both. Other studies have shown that MSC derived from adipose tissue had a similar effect. One interesting point of these studies was that the stem cell source used was of human origin and the recipient mice were immune competent. One would imagine that administration of human cells into a mouse would result in rapid rejection. This did not appear to be the case since the human cells were found to persist and also to differentiated into human neural tissues in the mouse. One mechanism for this “immune privilege” of MSC is believed to be their low expression of immune stimulatory molecules such as HLA antigens, costimulatory molecules (CD80/86) and cytokines capable of stimulating inflammatory responses such as IL-12. Besides not being seen by the immune system, it appears that MSC are involved in actively suppressing the immune system. In one study MSC were demonstrated to naturally home into lymph nodes subsequent to intravenous administration and “reprogram” T cells so as to suppress delayed type hypersensitive reactions. In those experiments scientists found that the mechanism of MSC-mediated immune inhibition was via secretion of nitric oxide. Other molecules that MSC use to suppress the immune system include soluble HLA-G, Leukemia Inhibitor Factor (LIF), IL-10, interleukin-1 receptor antagonist, and TGF-beta. MSC also indirectly suppress the immune system by secreting VEGF which blocks dendritic cell maturation and thus prevents activation of mature T cells.
While a lot of work has been performed investigating how MSC suppress the immune system, relatively little is known regarding if other types of stem cells, or immature cells, inhibit the immune system. This is very relevant because there are companies such as Stem Cells Inc that are using fetally-derived progenitor cells therapeutically in a universal donor fashion. There was a paper from an Israeli group demonstrating that neural progenitors administered into the EAE model have a therapeutic effect that is mediated through immune modulation, however, relatively little work has been performed identifying the cell-to-cell interactions that are associated with such immune modulation.
Recently a paper by Knight et al. Cross-talk between CD4(+) T-cells and neural stem/progenitor cells. Knight et al. J Neurol Sci. 2011 Apr 12 attempted to investigate the interaction between immune cells and neural stem cells and vice versa. The investigators developed an in vitro system in which neural stem cells were incubated with CD 4 cells of the Th1 (stimulators of cell mediated immunity), Th2 (stimulators of antibody mediated immunity) and Th17 (stimulators of inflammatory responses) subsets. In order to elucidate the impact of the death receptor (Fas) and its ligand (FasL), the mouse strains lpr and gld, respectively, were used.
The investigators showed that Th1 type CD4 cells were capable of directly killing neural stem cells in vitro. Killing appeared to be independent of Fas activation on the stem cells since gld derived T cells or lpr derived neural stem cells still participated in killing. Interestingly, neural stem cells were capable of stimulating cell death in Th1 and Th17 cells but not in the Th2 cells. Killing was contact dependent and appeared to be mediated by FasL expressed on the neural stem cells. This is interesting because some other studies have demonstrated that FasL found on hematopoietic stem cells appears to kill activated T cells. In the context of hematopoietic stem cells this phenomena may be used to explain clinical findings that transplanting high numbers of CD34 cells results in a higher engraftment, mediated in part by killing of recipient origin T cells.
The finding that neural stem cells express FasL and selectively kill inflammatory cells (Th1 and Th17) while sparing anti-inflammatory cells (Th2) indicates that the stem cells themselves may be therapeutic by exerting an immune modulatory effect. One thing that the study did not do is to see if differentiated neural stem cells would mediate the same effect. In other words, it is essentially to know if the general state of cell immaturity is associated with inhibition of inflammatory responses, or whether this is an activity specific to neurons. As mentioned above, previous studies have demonstrated that mesenchymal stem cells (MSC) are capable of eliciting immune modulation through a similar means. Specifically, MSC have been demonstrated to stimulate selective generation of T regulatory cells. This cell type was not evaluated in the current study, however some activities of Th2 cells are shared with Treg cells in that both are capable of suppressing T cytotoxic cell activation. In the context of explaining biological activities of stem cell therapy studies such as this one stimulate the believe that stem cells do not necessarily mediate their effects by replacing damaged cells, but by acting on the immune system. Theoretically, one of the reasons why immature cells are immune modulatory in the anti-inflammatory sense may be because inflammation is associated with oxidative stress. Oxidative stress is associated with mutations. Conceptually, the body would want to preferentially protect the genome of immature cells given that the more immature the cells are, the more potential they have for stimulation of cancer. Mature cells have a limited self renewal ability, whereas immature cells, given they have a higher potential for replication are more likely to accumulate genomic damage and neoplastically transform.

Medistem Reports Rheumatoid Arthritis Patient Success Using Adult Stem Cell Protocol

SAN DIEGO, CA – (Marketwire – June 21, 2010) – Medistem Inc. Medistem Inc. together with the Stem Cell Institute reported today publication in the peer reviewed journal Cellular Immunology its paper titled “Autologous stromal vascular fraction cells: A tool for facilitating tolerance in rheumatic disease,” which describes the first use of a patient’s own fat stem cells for treatment of rheumatoid arthritis.

How Fat Stem Cells May Work on Autoimmune Diseases

In collaboration with the company Vet-Stem Inc, the University of Western Ontario, and The University of California San Diego, Medistem scientists detailed the scientific rationale for use of patient’s own fat derived stem cells for “reprogramming” the immune system of patients with autoimmune diseases such as rheumatoid arthritis. A case report of a 67-year-old American woman who recovered from rheumatoid arthritis after intravenous treatment with adult stem cells is provided.

“We have been treating companion animals for osteoarthritis and rheumatoid arthritis for over five years, achieving and publishing excellent efficacy data,” said Robert Harman, CEO of Vet-Stem. “Medistem’s identification of potential mechanisms of action, as well as translation of this technology into the clinic, supports the importance of our findings.”
Medistem previously identified and filed intellectual property covering the co-purification of high concentrations of T regulatory cells using protocols that enrich for adipose derived stem cells, a finding that was later confirmed and published by Diane Mathis’s group from Harvard University (Feuerer et al. Nat Med. 2009 Aug;15(8):930-9). T regulatory cells are used by the body to control autoimmunity, which is explained in this video by Thomas Ichim, the CEO of Medistem.

“We are very excited that Medistem’s protocol for isolation of a patient’s own fat derived stem cells and T regulatory cells, which produced promising results in multiple sclerosis, appears to be useful in rheumatoid arthritis, another autoimmune disease,” said Neil Riordan, Chairman of Medistem.

In a 2009 paper Medistem together with Vet-Stem and University of California San Diego reported substantial clinical improvement in a small group of multiple sclerosis patients treated using a similar protocol. The paper is available at www.translational-medicine.com.

About Medistem Inc.

Medistem Inc. is a biotechnology company developing technologies related to adult stem cell extraction, manipulation, and use for treating inflammatory and degenerative diseases. The company’s lead product, the endometrial regenerative cell (ERC), is a “universal donor” stem cell being developed for critical limb ischemia. A publication describing the support for use of ERC for this condition may be found at www.translational-medicine.com.

Cautionary Statement

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Hope Through Stem Cell Therapy

Mary Posta suffers from multiple sclerosis, a debilitating
disease that progressively degenerates the nervous system of its victims through
stripping away the insulator proteins surrounding the nerves called myelin.

In January of this year Mary Posta completed raising funds
to be treated by Cellmedicine in Central America using stem cells and returned
from treatment feeling "really good".  Specifically, after a month spent at
Cellmedicine, she stated "I can walk and talk better, and there are other
things." She adds "My memory seems to be better. I’m moving faster on thinking
and talking, and I have a lot more energy. I used to have to take sleeping pills
but have not had to start taking them again."

The stem cell therapy comprises of an intensive four-week
program of stem-cell and physical therapies.  The stem cells used are from adult
sources and therefore are not subjected to the ethical controversy associated
with other types of stem cells such as fetal or embryonic stem cells.

Cellmedicine has previously published results of the first
three multiple sclerosis patients in a peer reviewed medical journal which can
be found at this link

http://www.translational-medicine.com/content/pdf/1479-5876-7-29.pdf

The approach used involves administration of the cells
purified from the fat of the patients.  These cells contain two types of stem
cells, one called mesenchymal and the other called hematopoietic.  Additionally,
cells extracted from fat include alternatively activated macrophages and T
regulatory cells.  At a theoretical level these cells may be mediating their
effects as follows: 

Mesenchymal stem cells are known to inhibit multiple
sclerosis when administered in animal models of the disease, as seen in this
video

http://www.youtube.com/watch?v=D2RIuCc5h0A
.  The video discusses one
mechanism by which mesenchymal stem cells achieve this effect, particularly
through induction of an enzyme called indolamine 2,3 deoxygenase, which is
responsible for shutting down autoreactive T cells.  Since multiple sclerosis is
a disease in which T cells are mediating destruction of the myelin sheath,
suppression of autoreactive T cells is theoretically beneficial.  Additionally,
mesenchymal stem cells are known to produce various growth factors that increase
ability of the body’s own cells to repair themselves.  Furthermore, some studies
have suggested that mesenchymal stem cells themselves are capable of
differentiating into oligodendrocytes and Schwann cells, which produce myelin,
as well as into brand new nervous system tissue. 

Hematopoietic stem cells are conventionally known as
the cells that are responsible for the therapeutic effect of bone marrow
transplantation.  That is, these are the cells that produce all the blood cells
in the body.  More recent studies have shown that hematopoietic stem cells, such
as CD34 positive cells,  are capable of producing growth factors such as IGF-1,
these are capable of protecting various cells in the body from premature cell
death.  Additionally, there are some studies that suggest CD34 cells are capable
of regenerating injured nervous system tissue.

Alternatively activated macrophages comprise a
subset of the immune system cell classically known as the "big eater".  While
conventional macrophages are involved in protecting the body from disease by
eating pathogens, as well as producing inflammatory stimuli, alternatively
activated macrophages are involved in healing of damaged tissue.  It is known
that alternatively activated macrophages generate substances such as
interleukin-10 that shut down ongoing immunological/inflammatory reactions, as
well as assist in tissue healing.

T regulatory cells resemble the "anti-matter" of T
cells.  The body has two parallel universes of T cells.  The conventional T
cells are responsible for attacking everything that does not belong to the
body.  That is, conventional T cells recognize and kill bacteria, viruses, and
other pathogens.  On the other hand, T regulatory cells recognize everything
that "belongs" to the body.  For example, there are T regulatory cells in the
body that recognize myelin.  The difference between T regulatory cells and
conventional T cells is that T regulatory cells do not "kill" but instead
prevent what is being recognized by conventional T cells from being killed.  In
other words the T regulatory cells serve as a backup mechanism for the immune
system so that in situations such as multiple sclerosis, where the conventional
T cells are attacking something that they should not be attacking, the T
regulatory cells try to inhibit that attack.  Unfortunately in multiple
sclerosis, by the time the disease is clinically detected, the T regulatory
cells are not exerting their effects for reasons some known and some unknown. 
Adipose tissue contains high numbers of T regulatory cells, which are more
potent than T regulatory cells found from other tissues in the body.  This is
explained in this video, which discusses a publication from Harvard Medical
School
http://www.youtube.com/watch?v=rEJfGu29Rg8.

Given the potent combination of stem cells, and other
therapeutic cells, found in fat tissue, it is interesting that the company
Vet-Stem has already commercialized the procedure of using fat-derived cells for
treatment of companion animals.  Here is a video discussing some of Vet-Stem’s
technologies
http://www.youtube.com/watch?v=hEkSJo3CmPc .

Use of fat stem cells in patients with multiple sclerosis
has been previously reported in numerous other media venues:

CBS News:

http://www.youtube.com/user/cellmedicine#p/u/24/wIcUaKZWOSE

Fox 4 News:
http://www.youtube.com/user/cellmedicine#p/u/25/1j1F57olCdI

Texas Channel 8 News:
http://www.youtube.com/user/cellmedicine#p/u/21/r_mOKM5__00

CBS 4 News:
http://www.youtube.com/user/cellmedicine#p/u/19/mxd6t3izxtw