.
Abstract
With an increased need for blood donation and transfusion, blood banks have been of important use in ensuring that these processes take place. Blood banks are used to store and preserve blood for future use in blood transfusion. Blood storage methods have advanced with time. Blood is a fundamental component in the body. Currently, banks have involved in research and studies on blood cell therapies to develop stem cell treatment for varied types of diseases.s
Introduction
Cellular therapy is the varied use of mononuclear cells (MNCs), CD34+ lymphocytes or stem cells to prevent or cure different types of diseases. Cellar therapy is the logical extension of traditional blood banking, in which donation is done and transfusion done to the recipient with minimum modification. [1] The emerging forms of cellular therapy on the other hand rely on the ability to manipulate and select some specific types of cells and treat them (sometimes) in specific ways before infusing the cells into a recipient. The other difference between cellular therapy and the basic transfusion therapy is that cellular therapy involves infusion of a specific product intended tobe something potentially and substantially important once it has been infused for example destroy malignancy, repair a tissue or re-grow an organ while transfusion therapy focuses on supplying components important in supporting ongoing processes in the body for instance, platelet clot formation and carrying oxygen.
Modern forms of cellular therapy rely on apheresis technology in separation and harvesting of specific group of blood cells. Researchers have developed methods to specifically stimulate the production process of bone marrow stem cells more so granulocyte colony-stimulating factor, as well as the ability to detect the cells in the harvested product by means of CD34 detection on the cell surface, this lead to development of stemcellstherapy/progenitor cell transplantation. Different types of disorders can be treated by use of stem cell transplant with inclusion of many pre-malignancies and hematologic malignancies as well as hemoglobin, genetic metabolic and immunodeficiency diseases. The underlying principle involves patients own body marrow destruction together with destruction of the malignant cells followed by the stem cell transplant material infusion that is indented or designed to repopulate the bone marrow. Apheresis technology can be used to harvest the stem cell from the blood or from umbilical cord blood. All this is mostly carried out in blood banks.
The current development in cellular therapy can be used to collect cells to be used in designing and manufacturing patient specific vaccines for different types of tumor or cells that may be used to generate or regenerate missing organs. For example the advanced prostate cancer treatment which was recently approved. For treatment of advanced prostate cancer, the patient is supposed to undergo leukapheresis with respect to the extracting criteria in which the harvested antigen presenting cells stimulated in vitro by means of exposing them to prostatic acid phosphatase (PAP).Reinfusion of the cell is then done, aimed to attack malignant cells expressing Prostatic acid phosphatase. This is the first food and drug administration (FDA) approved autologous cellular immunotherapy. Other advancement in clinical trials are in process for vaccination for malignancies such as cervical, ovarian and breast cancer. The other cellular therapy methods under study include methods to repair damage to tissues and organs, which is known as regenerative medicine or tissue engineering.
In future, advancement in cellular therapy manipulation technology is expected may be to conveniently result in the ability to manufacture patient’s specific red blood cells or any other blood component. These changes are seen as potentially devastating to our industry by some blood banks but this should be looked in a way to acknowledge new technology. With the help of blood banks the field of cellular therapy is changing and growing very rapidly.
A phase III research by the Blood and Marrow Transplant Clinical Trials Network and the National Marrow Donor Program is currently under way to determine whether we have differences in patient outcomes or donor experiences between peripheral blood stem cells (PBSC)and marrow unrelated donor transplants. [2]
Methods to overcome the limited cell dose of cord blood to facilitate/ enable transplants for adult are recently being studied. These methods include combination of two or more closely matched cord blood units, ex-vivo expansion of the cells and the co-infusion of mesenchymal cells or peripheral blood stem cells. [3, 4, 5]
Laboratory Prove Laminaria’s Beneficial Effects
Dr. Hiroko Maruyama a Japanese scientist confirmed the efficiency and effectiveness of laminaria through clinical tests of Sprague Dawley rats. When given a cancer-causing chemical known as DMBA these rats get breast tumors. All the rats under study were given DMBA and half of them were given Laminaria in their food, by the end of seven months all the rats were autopsied with those which had laminaria intheir diets having fewer tumors.
Dr. Yamamoto reversed the study where he took the rats that had extensive tumors and added laminaria to their food. In duration of six months the tumor incidence that had occurred reduced by 35 %.
The study on the effectiveness and limitation of non-specific immunostimulation by Bacillus Calmette GuCrin (BCG) has been done. A high dose of BCG for a small tumor has been a successful therapy,its therapeutic effects, especially when used in conjunction with chemotherapy, were, prolongation of periods ofremission and survival for patients with postoperative breast cancer and with myelogenous and melanoma leukemia. Bacillus Calmette- GuCrin injection resulted in agranulocytomatous reaction at both the tumor site and at regional lymph nodes. The speculation on the mechanism of Bacillus Calmette GuCrin action is postulated that the “nonspecific” immunostimulants may lead to increase in “specific” immunity to tumors that share antigenic determinants with the bacterial adjuvant. Reports on previous remission of tumors in patients after streptococcal infection, namely erysipelas, and the successful in vitro treatment of Ehrlich ascites tumor prompted us to re-investigate the effect of living strepiococci on several transplantable murine tumors and on a spontaneously occurring lymphocytic leukemia that arise in AKR mice. The experiments discussed here include both, in vitro and in vivo streptococcal treatment of spontaneous and transplanted lymphocytic leukemias of AKR mice.
In vivo treatment
Mice with established spontaneous lymocytic leukemia (confirmed by their white blood cell and differential counts) were injected with 0.20 ml of a 20- hr streptococcal suspension, followed by three daily injections of 10, 000 U of penicillin to control the streptococcal infection. The procedure produced no prolongation of survival or cure compared to mice which did not under go treatment. However, injecting the mice with streptococci a day prior to or up to two days past a 1- 3 x106leukemic spleen cell injection, they showed some improvement in survival compared to recipient control mice that received no treatment. However, eventually all mice succumbed to leukemia. [6]
In vitro pretreatment
Following the protocol of Koshimura et al; spleen cell suspensions were incubated in vitro for 1.5- 2.5 hr at 370 C with a 20- hr culture of streptococcus pyogenes group A (of a strain designated “SA” by Koshimura). Subsequently, 1- 3 x 106 of these streptococcally pretreated spleen cells were injected into recipient mice groups of eight. The two control groups received either untreated spleen cells or streptococcal suspensions without spleen cells. The parameters followed in the recipient mice were leukemia incidence, survival time, peripheral lymphocyte and differential counts, and histopathologic findings of lymphoid tissue on autopsy.
In five similar experiments, it was found that 24of 40 mice that had received pretreated spleencells showed no signs of leukemia up to 4 %months later, whereas all 40control micesuccumbed to leukemia. Some of the former group died of infection several days after receivingthe streptococcal spleen cell mixture, whereas others proceeded to develop leukemia with some delay. There was an average increase in survival time from 14days in the controls to 30 days in recipients of pretreated cells and a delay in the increase of peripheral white blood cells and lymphoblasts. At autopsy, control mice typically showed greatly enlarged spleens (up to 1 g), livers, and lymph nodes. Histopathology of these enlarged organs showedextensive leukemic cell infiltration by lymphoblasts of lymph nodes, liver, spleen,thymus, and, sometimes, lungs.
Peripheral WBCsnumbered up to 299,000, ofwhich as many as 10% were immature blast cells. Leukemic infiltration ofbone marrow could not be ascertained. In contrast, animals that received streptococcally treated tumor cells had normal sized organs but showed an intensive inflammatory response in the lymphoid organs. This inflammatoryresponse may well have been a factor that contributed to the absence of leukemiain 24 of 40 mice and to prolongation of survival in others. (Autopsyfindings of these 24mice up to 4months after injection of leukemic cellsshowed no leukemic infiltration).
Another group of mice injected only with streptococci and penicillin alsoshowed inflammatory responses in spleen and lymphoid tissue. Pretreatment ofspleen cells of 49mice (in groups of eight) with either heat-killed streptococcior mixed bacterial toxins from Serratia and Streptococcus were totally ineffective.
The effectiveness of hemolytic streptococci against leukemia spleen cellsparalleled that found against other transplantable tumors.[7,8] Pretreatment ofsarcoma 37 or Krebs-2 carcinoma ascites tumor cells with several strainsof hemolytic streptococci resulted in oncolysis of the tumor cells within minutes,whereas treatment with Streptococcirs faecalis or Serrutia marcescens had noeffect. Purified streptococcal extracts and streptolysin Swere as effectiveas the living streptococci.
If streptococci are effective in vitro in lysing tumor or leukemic spleen cells,what are the possible reasons for the failure in our experiments to affect thefinal outcome in the in vivo treated leukemic host?The tumor load may have been too great for the number of streptococci injected.Rapp mentioned that BCG is only effective against a small number oftumor cells (105). [9]The injection of 1.5-3.5 x 105leukemic spleen cells per mouse may have provided a leukemic cell overload.A critical ratio of streptococci to tumor cells may be a prerequisite forsuccessful immunotherapy. This critical ratio may not have been reached in theunsuccessfully in vitro-pretreated cells and was not achieved in the in vivoexperiments. The ratio of streptococci to tumor cells measured in some experiments was approximately 190: 1and may have been close to borderline.
The treatment of the host with penicillin decreased the number of viablestreptococci, and this decrease further altered the ratio of bacterial to tumorcells, which reduced their effectiveness.
It is also known that penicillin suppresses the immune response. [10] Administrationof repeated injection of smaller dosages of streptococci might obviate thesubsequent penicillin treatment.
The lesson learned is: Obviously, preincubation of apatient’s leukemic cells with streptococci is not a practical measure, unless thistreatment would enhance their antigenicity for specific immunization. However,the demonstrated oncolytic ability, albeit in vitro, of certain strains of hemolyticstreptococci on several transplantable mouse tumors and on leukemic spleencells, probably due to the vast array of proteolytic enzymes, warrants furtherinvestigation of streptococci as possible oncolytic agents in the living host,despite the initial negative results inin vivo-treated mice. The intratumor injectionof streptococci, their use in combination with chemotherapy, shown to beeffective with BCG, also warrants further investigation.
Although we have achievedsome successes, we still neither understand the mode of action of these “nonspecific” immune- stimulants nor whether they are truly nonspecific. Agreateremphasis should be placed on studying the mechanism of action of the “nonspecific’’bacterial agents if we are to develop a rational approach to theimmunotherapy of cancer and leukemia.
Banking Umbilical Cord Tissue
Cord blood banking is storing umbilical cord blood for use in future. Cord blood is obtained after cutting the umbilical cord and then extracted from the fetal end of the cord giving up to 75 ml from the neonate.
Cryopreservation
After obtaining the blood, the blood unit is taken to the laboratory for processing the cryopreserved. Processing of the cord blood may be done to obtain the red blood cells while others may be indented to preserve them. A Cyropreservant is added to the cord blood to enable the cells undergo the cryogenic process. The cord blood unit is slowly cooled to – 900 C, it is added to liquid nitrogen in a tank to keep it frozen at around –196 0C. the slow freezing process is important in keeping the cells alive in the freezing process.
The food drug and administration organization govern the collection, storage, processing packaging, labeling and distribution of cord blood stem cells. The two standards which apply are:current good tissue practices and current good manufacturing practices. The current good tissue practices apply to processing, collection and storage of human tissues, cells and cellular or tissue based products which are regulated by the Center for Biologics Evaluation and Research. [10] The Current Good Manufacturing Practices standard is applied in the manufacturing of products considered as drugs.
During Cord blood transplant, less stringent matching between the tissue types of the recipient and the donor is required, this is known as Human Leukocyte Antigen (HLA). Six key antigens are required during bone marrow transplant, these are measures of graft versus host reaction, 6/6 match. If a 4/6 match occurs, cord blood transplant achieves the same medical success. Human Leukocyte Antigen is inheritable from both parents, therefore siblings are likely to have a match as well, and people from the same ethnic background are likely to match.People from minority ethnic groups find it difficult to find a perfectly matched donor for transplant, this has lead to the ability to transplant partially mismatched cord blood hence opening access to transplant therapy.
Researches have found that, allogeneic transplants give better outcomes for related recipients and donors; this is because in addition to matching the six key antigens, it gives a match of so many other antigens. [11]
Cord blood treatments
Currently we have three types of stem cell treatments done using cord blood, these are trials treatments, standard treatments and experimental treatments. Mostly, stem cells are used during normal or standard treatment. The problems which may occur during this process are problems in relation to blood cells. Standard treatment covers, leukemia treatment, lymphoma treatment, anemia, inherited red cell abnormalities, phagocyte disorders, bone marrow cancer and other types of cancers. Some diseases have been proven to be curable with the help of stem cells though the stem cell therapy has not yet been put in to practice to cure these diseases.That’s why they are called clinical trials but not the normal or standard therapies. In some cases, stem cell transplant lead to slowing down of the disease progression and a times these transplants may successfully lead to cure. However, the exact usage and dosage of the stem cell therapy has not yet been made clear.
Experimental treatments using stem cells have not yet been proven to cure or help human beings using stem cells treatments; many laboratory experiments are going on with cell animals. Some of the diseases being studied are Crohn’s disease and Juvenile Arthritis, central nervous system diseases (Parkinson’s and Alzheimer’s diseases).
Currently, stem cells treatments are limited; umbilical cord blood is only able to generally assist/help with bone marrow transplant.
Reference
- Snyder E. and Choate J. The emergence of cellular therapy: impact on transfusion medicine
- BMT CTN study 0201. A phase III randomized, multi-center trial comparing G-CSF mobilized peripheral blood stem cell with marrow transplantation from HLA compatible unrelated donors. 2004. https://web.emmes.com/study/bmt/protocp;/0201 protocal/ 0201 protocal.html
- Ooi J. Cord blood transplantation in adults. Bone Marrow Transplant. 2009; 44(10):661-666
- Brunstein CG, Gutman JA, Weisdorf DJ, et al. Allogeneic hematopoietic cell transplantation for hematological malignancy: related risks and benefits of umbilical cord blood. Blood. 2010; 116(22): 4693-4699
- Rocha V, Broxmeyer HE. New approaches for improving engraftment after cord blood transplantation. Biol Blood Marrow Transplant. 2010; 16(1, Suppl.): S126-S132
- HAVASH F.& A. J. DONNELLY 1963.Unpublished data
- HAVASH F. 1964.The Cytotoxic effects of Hemolytic Streptococci and Streptococcal Products on Ascites Tumor Cells. Third International Congress of Chemotherapy: 1096-1 103.George Thieme Verlag
- Havas F.A, Donnelly J & Porreca A.V (1963), The Cytotoxic Effects of Hemolytic Streptococci on Ascites, Tumor cells. Cancer Res 23: 700-706
- RAPP, H. S. J. KLEINSCHUSTERD, C. LUEKER & R. A. KAINER. (1976). Immunotherapy of experimental cancer as a guide to the treatment of human cancer
- Ann. N.Y. Acad. Sci. 276: 550-556.
- RAEBURJN. A. 1972. Antibiotics and immunodeficiency 19. Lancet
- Nietfeld JJ, Pasquini MC, Logan BR, Verter F, Horowitz MM. Lifetime probabilities of hematopoietic stem cell transplantation in the US. Biol Blood Marrow Transplant. 2008; 14:316-22
- Rocha V, Gluckman E, Boyer Chammard A, Locatelli F, Arcese W, Pasquini R, Ortega J, Souillet G et al. (1997). “Outcome of cord blood transplantation from related and unrelated donors”. New England Journal of Medicine337 (6): 373–381
