Barriers and supportive factors in engaging cancer patients in physical activity programmes - a literature review.

BACKGROUND: Regardless of cancer type or stage of treatment, physical activity (PA) has been shown to reduce the risk of cancer recurrence and death. It is associated with a range of positive effects on patients' physical and psychological well-being, particularly in the areas of aerobic fitness, fatigue, mental health and perceived overall quality of life. However, in current oncology practice, the combination of its indication with treatment is still relatively rare. At the same time, cancer patients' participation in regular physical activity is usually very low. However, as PA is an effective method to support cancer treatment and plays an important role in prevention, it is necessary to find effective strategies to involve patients more widely in physical activities. To this end, physical activity programmes organised directly by facilities providing comprehensive cancer care appear to be very suitable. PURPOSE: This literature review maps the main barriers and facilitators to cancer patients' participation in physical activity programmes. In particular, economic factors related to health policy, reflected in the availability of this type of supportive care for patients, the level of health literacy, the organization of PA programs, health care providers - both physicians and health care workers, social support and intrapsychic influences on the part of patients play a major role. Since the implementation of physical activity programmes into the existing cancer care system is a rather challenging process, the paper also deals with the possibilities of using the Health Belief Model. In the given context, this model allows the prediction and identification of barriers and supportive factors to patients' involvement in PA programs in order to maximize their effectiveness and adapt them to the needs of patients and, at the same time, to the capabilities of a specific medical facility.

Selective depletion of alloreactive donor T cells leads to elimination of graft-versus-host reactivity and stimulates graft-versus-leukaemia/myeloma effect.

Graft-versus-host disease is a severe complication of allogeneic stem cell transplantation. The major role is played by alloreactive donor T-cell clones leading to host tissue damage. Selective depletion is a strategy to eliminate host-reactive donor T cells from haematopoietic stem cell allografts to prevent graft-versus-host disease while conserving useful donor immune functions. We have used irradiated peripheral blood mononuclear cells from cancer patients and healthy donor cells as responder cells in primary mixed leukocyte reaction. To prepare graft-versus leukaemia/myeloma-specific T cells, alloreactive T cells in primary mixed leukocyte reaction were depleted with anti-CD25 immunotoxin. The remaining T cells had insignificant alloreactivity in secondary mixed leukocyte reaction. Then, allodepleted donor T cells were repeatedly stimulated using purified leukaemia/tumour cells from the same cancer patient. Leukaemia/tumour-reactive donor T cells were purified using cell sorter on the basis of CD4 and CD8 activation. Their specificity was tested in nonradioactive cytotoxicity test. We performed 22 reactions (15 samples with leukemic and 7 samples with multiple myeloma cells). Selective depletion of alloreactive donor T cells with anti-CD25 immunotoxin led to significant depletion (99.2-100 %, median 99.7%). The effect of donor T cells was well preserved, while the graft-versus-host reactivation of donor cells was negligible, even after repeated stimulation with patient's non-tumour cells. Thus, it is possible to selectively deplete donor alloreactive T cells with anti-CD25 immunotoxin. In the cases of leukaemia patients, a strong graft-versus-leukaemia reactivity was noticed in allodepleted donor T cells; in myeloma patients, graft-versus-myeloma reactivity was less significant.

Sample processing and methodological pitfalls in multiple myeloma research.

In this paper, initial processing of biological material, cell separation algorithms and other procedures are discussed. For samples with initial infiltration of plasma cells > 5%, CD138 MicroBeads and Auto-Magnetic-Activated Cell Sorting program are used. Fluorescence-Activated Cell Sorting is used exclusively for cell populations with low-abundance; these samples are detected using fluorescently labeled antibodies only. Isolated plasma cells are further processed for molecular biological studies, for cytogenetics and protein analyses. Furthermore, this work examines the pitfalls of research related to multiple myeloma; some of them we have overcome, while the others are still problematic.

[Plasma cell separation algorithm from bone marrow samples]. / Algoritmus separace plazmatických bunek ze vzorku kostní drene.

BACKGROUNDS: The aim of this paper is to present an algorithm for plasma cell separation from bone marrow samples of multiple myeloma patients. The main prerequisite for applying modern research methods in this disease is gaining pure cell populations. MATERIAL AND METHODS: Bone marrow samples were collected from outpatients or inpatients of the Internal Haematology and Oncology Clinic of the Faculty Hospital Brno, after they had signed an Informed Consent Form. The bone marrow was first depleted of red cells (by density gradient centrifugation or erythrolysis), plasma cells were labelled by monoclonal antibody against syndecan-1 (CD138) and separated either magnetically or by cell sorter. The purity of separated population was evaluated by flow cytometry or, alternatively, morfologically. RESULTS: We processed 28 bone marrow samples, in parallel, by magnetic or fluorescence-based separation, and we evaluated the purity of the separated fractions. Based on a statistical evaluation of resulting purities in the entire sample set as well as the individual groups divided according to the initial plasma cell content, a separation algorithm was proposed with a cut-off value of 5% of plasma cells in mononuclear fraction of bone marrow: samples with less than 5% of plasma cells are henceforth separated on cell sorter, samples with more than 5% are separated magnetically.The effectiveness of this algorithm was evaluated after the first year of its application on a dataset of 210 bone marrow samples: median purity of the separated plasma cells increased from 62.4% (0.4-99.6%) to 94.0% (23.9-100%). CONCLUSION: The introduction of a fluorescence-based separation markedly increased the effectiveness of plasma cell separation from bone marrow samples, mainly in samples with low plasma cell content where magnetic separation used thus far is not sufficient. This finding also opened a door for plasma cell separation of bone marrow samples from patients with monoclonal gammopathy of undetermined significance, where plasma cell count is typically below or just over one percent.

Optimization of immunomagnetic selection of myeloma cells from bone marrow using magnetic activated cell sorting.

Plasma cells (PCs) enrichment from bone marrow samples of multiple myeloma (MM) patients is frequently performed by immunomagnetic separation (magnetic activated cell sorting, MACS) using anti-CD138 MicroBeads. The aim of our work was to find optimal strategy for immunomagnetic separation of PCs and determine optimal algorithm of separation techniques for samples with various percentage of neoplastic cells. From 2007 to 2008, selection of PCs using separation programs Possels and Posseld(2) was carried out on 234 bone marrow samples obtained from 208 MM patients. In 2008, an optimal algorithm for separation programs was introduced based on the analysis of the previous experiments. The Possels program is applicable for samples with >10% PCs in the mononuclear fraction, while the Posseld(2) program is used for samples with 5-10% PCs in the mononuclear fraction. Median purity of 92.6% for the positive fraction of cells (range 14.5-99.6%) and median recovery of 60.4% (range 25.7-99.5%) were obtained when the Possels program was applied (n = 45). A total of 80% (36/45) of processed samples had purity of >70%. Median purity for the positive fraction of 83.7% (range 14.3-99.7%) and median recovery of 14.3% (range 3.6-50.0%) were achieved using the Posseld(2) program (n = 99). A total of 68% (67/99) of processed samples reached >70% purity. This separation strategy enabled us to obtain sufficient amounts of highly purified PCs required for subsequent research purposes. The MACS method has been unsuccessful if the percentage of PCs in the initial sample was <5%. These samples were processed by fluorescence activated cell sorting (FACS).

Phenotype of plasma cells in multiple myeloma and monoclonal gammopathy of undetermined significance.

Flow cytometry is a useful tool for the analysis of plasma cells in monoclonal gammopathies. The aim of this study was to find possibilities and limits of multicolour flow cytometry in diagnostics of monoclonal gammopathy of undetermined significance (MGUS) and multiple myeloma (MM) and to identify parameters that could be used to differentiate between these two disorders. Surface markers CD38 and CD138 were used for identification of plasma cells, CD19 and CD56 further distinguished normal and abnormal plasma cells, respectively. The percentage of circulating plasma cells in peripheral blood was lower in MGUS patients then in MM (p<0,001) In bone marrow, the percentage of residual polyclonal CD19 plasma cell was higher (p<0,001) and the percentage of malignant monoclonal CD56 plasma cell was lower (p<0,001) in MGUS than in MM. In conclusion, flow cytometry is relatively quick and effective method for analysis of plasma cells thus immunophenotyping can significantly contribute to the differential diagnosis of plasma cell proliferations.