Dissertation (Ph.D.)--University of South Florida, 2008. / Includes vita. Includes bibliographical references. Also available online.

Dissertation (Ph.D.)--University of South Florida, 2008. / Includes vita. Includes bibliographical references. Also available online.

A possible treatment for patients with lymphoma and myeloma is stem cell transplantation (SCT). SCT is preceded with cytostatic treatment. There are several side effects related to this treatment, for example fatigue, nausea, constipation/diarrhoea, pain, mucositis and loss of appetite. Aim: Investigate which side effects related to the treatment where most troubling after SCT and if nurse assessment and patient assessment differ. Methods: A quantitative empirical study with repeated measuring. The two groups of nurses and patients answered a form independently. Results: Loss of appetite and fatigue are the most troubling side effects according to both nurse and patient. Older patients tended to be more affected by fatigue. The nurses estimated the side effects such as loss of appetite, fatigue, diarrhoea and nausea lower than the patients did. Conclusion: No definitive conclusion could be made because of the small patient/nurse sample. However, there is a tendency showing difficulty for nurses to estimate correctly the side effects suffered by the patients. The nurses tend to estimate the side effects lower than the patients do.

Moklsiniame darbe tirtas chimerizmas vaikams po alogeninės kraujodaros kamieninių ląstelių transplantacijos. Tyrimas pradėtas Charité klinikose (Humboldto universitetas, Berlynas), kur chimerizmo tyrinėjimams pasirinkti recipientai, sergantys ūmia limfoblastine leukemija (ŪLL), Fanconi anemija (FA) ir adrenoleukodistrofija (ALD). Sergant šiomis ligomis nuoseklus chimerizmo stebėjimas ALP atliktas pirmą kartą. Darbas tęstas Vilniaus universitete, kuomet pirmą kartą buvo atlikti chimerizmo tyrinėjimai Lietuvoje transplantuotiems vaikams. Donoro ir recipiento ląstelėms atskirti naudoti polimorfiniai DNR žymenys, kurie tirti periferinio kraujo leukocituose (PKL) ir atskirose ląstelių populiacijose (ALP): CD3, CD19 ir CD34. Ląstelių frakcijoms atskirti naudotos imunomagnetinės dalelės. Atlikus chimerizmo analizę nutatyta, kad esant visiškam donoro chimerimui PKL, 74,4% ŪLL ir 66,7% ALD recipientų po mieloabliacinio kondicionavimo ląstelių populiacijose išlieka autologinė, aptinkama iki kelerių metų po transplantacijos. Po sumažinto intensyvumo kondicionavimo 45,5% FA recipientų autologinės kraujodaros pėdsakų nerandama nei PKL, nei ALP. FA ligoniams vystosi stabilus ilgalaikis mišrus chimerizmas, kuris susijęs su su ilgesniu išgyvenamumu po transplantacijos. Nustatėme, kad sergantiems FA galimybė išsivystyti mišriam chimerizmui yra 20 kartų didesnė perpylus kaulų čiulpus, nei periferinio kraujo kamienines ląsteles. / The doctoral thesis aimed to evaluate in a comparative way the benefit chimerism assessment in isolated cell populations (ICP) versus conventional monitoring in whole blood cells (WBC). The study was initiated in Pediatric Bone Marrow Transplant Service of Charité Children’s Hospital (Humboldt University, Berlin). Children suffered from: acute lymphoblastic leukemia (ALL), Fanconi anemia (FA) and adrenoleukodystrophy (ALD) were included into the study. Thereafter the doctoral thesis was accomplished at the Vilnius University where evaluation of the own experience of chimerism analysis in pediatric patients was performed. Chimerism was prospectively monitored in WBC and three cell subsets: CD3, CD19 and CD34. Cell populations were extracted from peripheral blood using immunomagnetic beads. Following polymorphisms of short tandem repeats between donor and recipient were compared. The analysis reveled that following myeloablative conditioning in patients with ALL and ALD analysis of ICP revealed persistent autologous hemopoiesis despite stable donor chimerism in WBC. In contrast after reduced intensity conditioning 45.5% of FA recipients had no evidence of autologous signals either in WBC, or in ICP. FA were found to develop stable long-lasting mixed chimerism associated with better overall survival. development of MC was related to the infusion of bone marrow but not to peripheral blood stem cells. Chimerism in ICP did no affected transplant outcome in any disease group.

The doctoral thesis aimed to evaluate in a comparative way the benefit chimerism assessment in isolated cell populations (ICP) versus conventional monitoring in whole blood cells (WBC). The study was initiated in Pediatric Bone Marrow Transplant Service of Charité Children’s Hospital (Humboldt University, Berlin). Children suffered from: acute lymphoblastic leukemia (ALL), Fanconi anemia (FA) and adrenoleukodystrophy (ALD) were included into the study. Thereafter the doctoral thesis was accomplished at the Vilnius University where evaluation of the own experience of chimerism analysis in pediatric patients was performed. Chimerism was prospectively monitored in WBC and three cell subsets: CD3, CD19 and CD34. Cell populations were extracted from peripheral blood using immunomagnetic beads. Following polymorphisms of short tandem repeats between donor and recipient were compared. The analysis reveled that following myeloablative conditioning in patients with ALL and ALD analysis of ICP revealed persistent autologous hemopoiesis despite stable donor chimerism in WBC. In contrast after reduced intensity conditioning 45.5% of FA recipients had no evidence of autologous signals either in WBC, or in ICP. FA were found to develop stable long-lasting mixed chimerism associated with better overall survival. development of MC was related to the infusion of bone marrow but not to peripheral blood stem cells. Chimerism in ICP did no affected transplant outcome in any disease group. / Moklsiniame darbe tirtas chimerizmas vaikams po alogeninės kraujodaros kamieninių ląstelių transplantacijos. Tyrimas pradėtas Charité klinikose (Humboldto universitetas, Berlynas), kur chimerizmo tyrinėjimams pasirinkti recipientai, sergantys ūmia limfoblastine leukemija (ŪLL), Fanconi anemija (FA) ir adrenoleukodistrofija (ALD). Sergant šiomis ligomis nuoseklus chimerizmo stebėjimas ALP atliktas pirmą kartą. Darbas tęstas Vilniaus universitete, kuomet pirmą kartą buvo atlikti chimerizmo tyrinėjimai Lietuvoje transplantuotiems vaikams. Donoro ir recipiento ląstelėms atskirti naudoti polimorfiniai DNR žymenys, kurie tirti periferinio kraujo leukocituose (PKL) ir atskirose ląstelių populiacijose (ALP): CD3, CD19 ir CD34. Ląstelių frakcijoms atskirti naudotos imunomagnetinės dalelės. Atlikus chimerizmo analizę nutatyta, kad esant visiškam donoro chimerimui PKL, 74,4% ŪLL ir 66,7% ALD recipientų po mieloabliacinio kondicionavimo ląstelių populiacijose išlieka autologinė, aptinkama iki kelerių metų po transplantacijos. Po sumažinto intensyvumo kondicionavimo 45,5% FA recipientų autologinės kraujodaros pėdsakų nerandama nei PKL, nei ALP. FA ligoniams vystosi stabilus ilgalaikis mišrus chimerizmas, kuris susijęs su su ilgesniu išgyvenamumu po transplantacijos. Nustatėme, kad sergantiems FA galimybė išsivystyti mišriam chimerizmui yra 20 kartų didesnė perpylus kaulų čiulpus, nei periferinio kraujo kamienines ląsteles.

The Royal College of Surgeons (RCS) rat was used as a model of human retinal degenerative disease, and for studying the efficacy of cell transplanation treatments. In order to characterize the spatial vision of the RCS strain, the visual acutiy and contrast sensitivity of adult non-dystrophic RCS rats was measured. The acuity and contrast sensitivity of these rats was normal. The acuity of dystrophic RCS rats was alos characterized to determine how photoreceptor degeneration affects vision. These rats progressively lost visual acuity from one month of age until elevn months of age when they were judged to be blind. The degeneration of vision in these animals was more protacted than would be predicted from previous anatomical and electrophysiological measures. Subretinal transplantation of human-derived Retinal Pigment Epithelial (RPE) cells and human Schwann cells into the dystrophic RCS rat significantly delayed the loss of visual acuity. These studies show that cell transplantation may be a viable method of limiting loss of vision in humans with retinal degenerative blinding diseases. / vii, 77 leaves ; 29 cm.

Chronic myelogenous leukemia (CML) is a clonal hematopoetic stem cell disorder characterized by proliferation of cells expressing BCR-ABL fusion protein. In the BCR-ABL+ leukemia murine model, 12B1, we explored the therapeutic applicability of chaperone-rich cell lysate (CRCL) in the context of syngeneic hematopoietic cell transplantation (HCT) to treat pre-existing leukemia. Our results demonstrate that tumor growth is significantly delayed in mice receiving syngeneic HCT from 12B1 tumor CRCL immunized donors compared to animals receiving HCT from non-immunized donors. CRCL immunization post-immune HCT further hindered tumor growth when compared to immune HCT without post-transplant vaccination. The magnitude of the immune response was consistent with the anti-tumor effects observed in vivo. We also demonstrated that cured mice had developed long-term tumor specific immunity against 12B1 tumor cells. In addition, we documented that both T cells and NK cells contributed to the anti-tumor effect of CRCL vaccination as depletion of either subset hampered tumor growth delay. Thus, our results suggest that CRCL represents a promising vaccine capable of generating specific immune responses. This anti-tumor immunity can be effectively transferred to a host via HCT and further enhanced post-HCT with additional tumor CRCL immunizations.CD4+CD25+ regulatory T cells (Treg) play a central role in the prevention of autoimmunity and in the control of immune responses by down-regulating the function of effector CD4+ or CD8+ T cells. The role of Treg in Mycobacterium tuberculosis infection and persistence is inadequately documented. Therefore, the current study was designed to determine whether CD4+CD25+ FoxP3+ regulatory T cells may modulate immunity against human tuberculosis (TB). Ourresults indicate that the number of CD4+CD25+FoxP3+ Treg increases in the blood or at the site of infection in active TB patients. The frequency of CD4+CD25+FoxP3+ Treg in pleural fluid inversely correlates with local MTB-specific immunity(p<0.002). These CD4+CD25+FoxP3+ T lymphocytes isolated from the blood and pleural fluid are capable of suppressing MTB-specific IFN-γ and IL-10 production in TB patients. Therefore, CD4+CD25+FoxP3+ Treg expanded in TB patients suppress Mycobacterium tuberculosis immunity and may therefore contribute to the pathogenesis of human TB.

BACKGROUND Adult bone marrow derived MSCs had been explored to treat myocardial infarction (MI) and heart failure, for which various beneficial paracrine effects had been suggested. Since MSCs in vitro express anti-inflammatory cytokines, we tested the hypothesis that changes in the pro-/anti-inflammatory cytokine ratio in the infarct microenvironment may provide such a paracrine mechanism to improve early cardiac function following acute coronary occlusion. / Methods Rats (n=88) underwent acute left coronary artery ligations and were randomized into groups M and C and then injected with culture media or MSCs, respectively. These rats underwent blinded echocardiography to evaluate left ventricular ejection fractions (LVEF). Real Time PCR was used to compare cytokine gene expression for IL-1beta, IL-6, IL-8 (pro-inflammatory) and IL-10 (anti-inflammatory) at various times. Extra-cellular matrix (ECM) deposition and inflammatory cell infiltration were also analyzed. / Results As early as 12 hours, the ratio of pro-/anti-inflammatory cytokine gene expression in group C was significantly lower than group M. Similar results were found at 24 hours, 1 and 2 weeks, respectively. LVEF improved significantly in group C (M=62% vs C=68% at 12 hours* , M=66% vs C=75% at 24 hours*, M=57% vs C=75% at 1 week *, and M=52% vs C=70% at 2 weeks*, *p&lt;0.01). The ratio of MMP-2/TIMP1 levels was lower in the Group C at all time frames, reaching significance at 12 and 24 hours and 2 weeks. In group C, histopathological analysis revealed significantly less ECM deposition (M=1.95% vs C=0.75% at 24 hours*, M=19.30% vs C=9.36% at 1 week*, M=24.46% vs C=7.57% at 2 weeks*, *p&lt;0.01). This was associated with significantly decreased inflammatory cell infiltration after 24 hours. / Conclusions The current data suggests that MSCs therapy decreases the pro-/anti-inflammatory cytokine ratio in the infarct microenvironment. This is associated with improved cardiac function, reduced ECM deposition, and decreased inflammatory cell infiltration. This paracrine mechanism of MSCs therapy may explain the early functional improvement after MI before cell transdifferentiation or other mechanisms takes place.

Hematopoietic stem cell transplantation (HSCT) is a potentially curative treatment for multiple sclerosis (MS) with now more than 600 documented cases in the medical literature. Long-term remission can be achieved with this therapy, but when is it justified to claim that a patient is cured from MS? In attempt to answer this question, the outcome of the Swedish patients is described, mechanisms behind the therapeutic effect are discussed and new tools for demonstration of absence of disease have been developed. In Swedish patients treated with HSCT for aggressive MS, disease free survival was 68 % at five years, and no patient progressed after three years of stable disease. Presence of gadolinium enhancing lesions prior to HSCT was associated with a favorable outcome (disease free survival 79 % vs 46 %, p=0.028). There was no mortality and no patient required intensive care. The immune system of twelve of these patients was investigated further. In most respects HSCT-treated patients were similar to healthy controls, demonstrating normalization. In the presence of a potential antigen, leukocytes from HSCT-treated patients ceased producing pro-inflammatory IL-17 and increased production of the inhibitory cytokine TGF-β1 suggesting restoration of tolerance. Cytokine levels and biomarkers of tissue damage were investigated in cerebrospinal fluid from a cohort of MS patients. The levels were related to clinical and imaging findings. A cytokine signature of patients with relapsing-remitting MS could be identified, characterized by increased levels of CCL22, CXCL10, sCD40L, CXCL1 and CCL5 as well as down-regulation of CCL2. Further, we could demonstrate that active inflammation in relapsing-remitting MS is a tissue damaging process, with increased levels of myelin basic protein and neurofilament light. Importantly, relapsing-remitting MS patients in remission displayed no tissue damage. In secondary progressive MS, moderate tissue damage was present without signs of active inflammation. From a clinical vantage point, it seems that we confidently can claim cure of relapsing-remitting MS patients after five years absence of disease activity. The new tools for evaluation of disease can strengthen this assertion and may enable earlier prediction of outcome.

Background: Epstein-Barr virus(EBV)-associated posttransplant lymphoproliferative disease (PTLD) is a serious complication after allogeneic hematopoietic stem cell transplantation (HSCT). Especially in cases with involvement of the central nervous system (CNS) treatment is difficult because the efficacy of most chemotherapeutic agents as well as EBV-specific cytotoxic donor T cells in liquor is uncertain. In the last years the anti-CD20 monoclonal antibody Rituximab was intensively investigated in the treatment of EBV-PTLD. However, only 8 patients with B-cell lymphoma and CNS involvement treated with Rituximab were reported. Case Report: A 24-year-old female patient with acute T-lymphoblastic leukemia in second complete remission had received allogeneic, unrelated, T-cell depleted HSCT. 10 months later an EBV-associated PTLD was diagnosed. Beside peripheral lymphomas and B symptoms the patient showed neurological symptoms. Examination of the cerebrospinal fluid (CSF) revealed a meningeosis lymphoblastica caused by the EBV lymphoma. Treatment with Rituximab and the antiviral drug Cidofovir led to complete remission with regression of the peripheral lymphomas and disappearance of the neurological symptoms. In addition, the PCR control on EBV DNA became negative in the plasma as well as in CSF. Conclusion: The combination of Rituximab and Cidofovir appears as an interesting alternative treatment in patients with EBV-associated PTLD and CNS involvement. / Hintergrund: Die Epstein-Barr-Virus(EBV)-assoziierte Posttransplantations-lymphoproliferative Disease (PTLD) ist eine gefürchtete Komplikation nach allogener hämatopoetischer Stammzelltransplantation (HSCT). Insbesondere bei Befall des zentralen Nervensystems (ZNS) ist die Behandlung auf Grund der unsicheren Liquorwirksamkeit der meisten Chemotherapeutika als auch von EBV-spezifischen zytotoxischen T-Spenderzellen schwierig. Der monoklonale Anti-CD20-Antikörper Rituximab wurde in den letzten Jahren bei Patienten mit EBV-PTLD intensiv untersucht. Allerdings wurde bislang lediglich von 8 Patienten mit ZNS-Befall eines B-Zell-Lymphoms berichtet, bei denen eine Therapie mit Rituximab erfolgte. Kasuistik: Eine 24-jährige Patientin hatte wegen einer akuten T-lymphoblastischen Leukämie in zweiter kompletter Remission eine allogen-unverwandte, T-Zelldepletierte HSCT erhalten. 10 Monate später wurde eine EBV-assoziierte PTLD diagnostiziert. Neben peripheren Lymphomen und B-Symptomen zeigte die Patientin neurologische Symptome. Die Liquoruntersuchung erbrachte den Befund einer Meningeosis lymphoblastica im Rahmen des EBV-Lymphoms. Die Behandlung mit Rituximab und dem Virustatikum Cidofovir führte zu einer kompletten Remission mit Rückbildung der peripheren Lymphome und Verschwinden der neurologischen Symptomatik. Außerdem wurde die PCR-Kontrolle auf EBV-DNA sowohl im Plasma als auch im Liquor negativ. Schlussfolgerung: Die Kombination von Rituximab und Cidofovir erscheint als eine interessante Therapiealternative für Patienten mit EBV-assoziierter PTLD und ZNS-Befall. / Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.