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.

AuxoCell Laboratories Licenses Umbilical Cord Tissue Stem Cell Service to PerkinElmer’s ViaCord

Viacord Press Release
Cord blood private banking involves storing your own cord blood mononuclear cells in case you need them later. Cord blood public banking involves banking the cells into a public pool so that if others need them, they have access to them. In some ways it seems like cord blood private banking is based more on hope than on reality. The majority of uses of cord blood are in leukemias. In patients with leukemia you need to use the cord blood of a related or unrelated donor, but rarely if ever do you want to use your own cord blood because it may have the leukemic mutations in it that caused the leukemia to appear in the first place. Therefore, the majority of cord blood banking is based on the belief that in the future the FDA will allow for procedures to take your banked cord blood, manipulate it to generate certain tissues in vitro and then reimplant those tissues back in you if you need them. There are of course exceptions to this. For example, there are clinical trials using your own cord blood for the treatment of cerebral palsy. Specifically, Georgia Health Sciences University is doing a 40 patient cord blood study in patients with cerebral palsy who have stored their own cord blood http://www.clinicaltrials.gov/ct2/show/NCT01072370. Additionally, Joanne Kurtzberg from Duke is performing an 120 patient study in children with cerebral palsy that have stored their own cord blood http://www.clinicaltrials.gov/ct2/show/NCT01147653. Other diseases are also being explored experimentally. Clinical trials are also being performed using patient’s own cord blood for type 1 diabetes. A group in Germany is doing a 10 patient trial http://www.clinicaltrials.gov/ct2/show/NCT00989547 and a group in Florida recently completed a 23 patient trial http://www.clinicaltrials.gov/ct2/show/NCT00305344.
Thus at present the field of private cord blood banking may have some very high future potential. Large companies are realizing this and accordingly are moving into this space. Perkin Elmers announced today that it has licensed technologies patented by AuxoCell Laboratories involving processing and storage of mesenchymal stem cells from the umbilical cord. As we discussed previously on the Cellmedicine website, the umbilical cord possesses mesenchymal stem cells that are in some ways more potent than bone marrow mesenchymal stem cells because they are more immature. The licensing of this technology will allow for Perkin Elmers to deliver to customers the ability to bank not only hematopoietic stem cells but also mesenchymal stem cells. There are many uses for mesenchymal stem cells. In fact numerous clinical trials have been performed using autologous mesenchymal stem cells for conditions ranging from heart failure, to graft versus host, to spinal cord injury.
“AuxoCell is pleased to partner with PerkinElmer’s ViaCord in offering umbilical cord tissue banking and expand our strategic partnerships to bring novel stem cell therapies from the bench to the bedside,” said Kyle Cetrulo, chief operating officer of AuxoCell Laboratories, Inc. “Partnering with ViaCord was an easy decision. They are the first family bank in the United States to freeze treatment-ready cord tissue stem cells upon arrival at the lab, which enables them to be ready for immediate use, if needed.”
“ViaCord is excited to offer another source of stem cells to our customers and believe we have found an excellent partner in AuxoCell. The agreement grants ViaCord’s customers exclusive access, in family banking, to expanding MSCs derived from cord tissue through AuxoCell’s elite patents,” said Morey Kraus, ViaCord’s chief scientific officer. “AuxoCell’s proprietary and validated manufacturing protocols will assist ViaCord in offering the very best in stem cell banking.”

New Stem Cells Found in Ovary

Parte et al. Stem Cells Dev.

Very small embryonic like cells (VSEL) are a type of stem cell that appears to be found in bone marrow and other tissues of the body, presumably as a remnant of embryonic or embryonic-like cells left over from development. In a recent paper it was demonstrated that these cells may be found in the ovary surface epithelium in adult rabbit, sheep, monkey and menopausal human.

Indian scientists found two distinct populations of putative stem cells of variable size were detected in the ovary surface epithelium: one being smaller in size around the range of 1-3 micrometers and the other being of a size approximate to the surrounding erythrocytes.

The smaller cells resembled VSELs and were pluripotent in nature with nuclear Oct-4 and cell surface SSEA-4. The larger cells were 4-7micrometers and possessed cytoplasmic localization of Oct-4 and minimal expression of SSEA-4. The scientists believed that the larger cells were possibly the progenitor germ cells.

The VSEL cells were capable of spontaneously differentiating into oocyte-like structures, parthenote-like structures, embryoid body-like structures, cells with neuronal-like phenotype and embryonic stem (ES) cell-like colonies. They expressed Oct-4, Oct-4A, Nanog, Sox-2, TERT, and Stat-3 as detected by RT-PCR.

Germ cell markers like c-Kit, DAZL, GDF-9, VASA and ZP4 were immuno-localized in oocyte-like structures formed from the VSEL.

These studies are interesting because prior to this there were reports of bone marrow derived cells being implicated in production of oocytes. Specifically, Jonathan Tilley from Harvard reported that bone marrow transplantation can give rise to new oocytes that are donor derived http://www.ncbi.nlm.nih.gov/pubmed/17664466.

If these studies are reproducible it may be that adult stem cells could be useful in the treatment of infertility. Conversely it may be possible to repair oocytes of women who have undergone chemo/radiation therapy. Interestingly, Tilly’s group also published that ovarian tissue contains VSEL-like cells http://www.ncbi.nlm.nih.gov/pubmed/20188358

Scientists identify and isolate adult mammary stem cells in mice

(Times of India) It is well-known that stem cells exist in adult tissues. The most commonly known stem cell, the bone marrow stem cell, plays the physiological role of generating billions of blood cells per hour while being capable of making copies of itself. Subsequent to the discovery of the bone marrow stem cell in the 1960s by Till and McCulloch, other types of stem cells were subsequently identified in other tissues. For example, the brain contains a stem cell compartment term the “dentate gyrus” which is capable of creating new neurons at a basal rate, with acceleration of new neuron formation during pregnancy or after stroke. Other tissue specific stem cells include those found the in liver, the heart, and the spleen. One common characteristic amongst stem cells is their ability to efflux various drugs through expression of the multi-drug resistance (MDR) protein, as well as preferential state of quiescence in absence of growth factor activation.
One important reason to seek tissue-specific stem cells is that if they could be expanded in large numbers they may theoretically be superior to other stem cell types for therapeutic uses. For example, culture-expanded cardiac specific stem cells are superior to bone marrow stem cells at accelerating healing of the heart muscle after a myocardial infarction. These types of stem cells are actually in clinical trials at present.
The other reason for identifying tissue-specific stem cells is that they may be useful in identifying molecular events that occur in the process of normal tissue changing to cancer. This is of interest because cancer stem cells are believed to originate from tissue-specific stem cells acquiring numerous mutations.
Currently researchers from the Fred Hutchinson Cancer Research Center have identified a tissue-specific stem cell in the breast. The scientists developed genetically engineered mice in which the green marker protein GFP was used to identify only breast cells that express the stem cell phenotype. The findings appeared in peer-reviewed journal Genes and Development.
“Until now, we have not been able to identify stem cells in mammary tissue. They have never been detected before with such specificity. It is extraordinary. You can see these green stem cells under the microscope in their pure, natural state,” said Larry Rohrschneider of the Hutchinson Center.
It was demonstrated that the activity of the mammary stem cells is modulated during times associated with breast growth such as puberty and pregnancy.
“We have found that those transplanted green stem cells can generate new mammary tissue and this tissue can produce milk, just like normal mammary epithelial cells,” said co-author Lixia Bai.
“Identification of the exact stem cell and its location is the first critical and fundamental step toward understanding the regulatory mechanisms of these important cells,” she said.
The technology described in the publication may be useful in isolating and expanding human breast specific stem cells. If these studies are reproducible, it will be of great interest to see whether they still possess ability to home to injured tissue, which to date has been clearly demonstrated in bone marrow stem cells but not with too much clarity with cardiac –specific stem cells or other types of tissue-specific stem cells.