RESUMO
BACKGROUND: Mantle cell lymphomas (MCL) represent a rare subclass of Non-Hodgkin Lymphoma affecting the lacrimal gland (GL). AIM: To extensively describe the immunohistochemical profile of GL-MCL. MATERIAL UND METHODS: Single center, retrospective electronic records review of 3 patients with biopsy proven LG-MCL. RESULTS: The herein presented case series of three patients comprises a focal case involving solely the lacrimal gland, a symptomatic LG-MCL manifesting as the first sign of a systemic disease as well as a case of LG-MCL presenting as a relapsed systemic lymphoma. The three patients presented positive CD19 and CD20, negative CD10 and CD23. One patient showed an uncommon negativity for CD5. The increased expression of cyclin D1 caused by the classical translocation t(11;14) (q13;q32) in the fluorescence-in-situ-hybridisation were observed in all cases. B-cell-lymphoma-2 protein (BCL-2) and transcription factor SOX-11 (SOX-11) were also overexpressed. DISCUSSION: LG-MCL show an immunohistochemical profile corresponding to the classical profile of MCL. Overexpression of molecules for target therapies was found in all cases (CD20 for rituximab, BCL2 for Bruton-kinase-inhibitors and CD19 for CAR-T cell therapy). The removal of the GL can potentially drive to severe complications, even if aimed to confirm diagnosis. Therefore, the choice between GL-biopsy and exstirpation should be carefully evaluated, especially in cases of suspected lymphoma.
Assuntos
Aparelho Lacrimal , Linfoma de Células B , Linfoma de Célula do Manto , Adulto , Humanos , Linfoma de Célula do Manto/diagnóstico , Linfoma de Célula do Manto/genética , Linfoma de Célula do Manto/metabolismo , Estudos Retrospectivos , Linfoma de Células B/patologia , Biologia MolecularRESUMO
T-lymphocytic enteral leiomyositis (T-lel) is a rare disorder causing chronic intestinal pseudo-obstruction (CIPO), with cases predominantly being reported in the field of veterinary and pediatric medicine. Here, we present a case of T-lel-associated CIPO in an adult female, who initially presented with a paralytic ileus 2 weeks after a common gastroenteritis. The histological diagnosis was established through full-thickness bowel biopsy, exhibiting a dense lymphocytic infiltrate in the lamina muscularis of the intestinal wall. This case shows that T-lel can be a cause of chronic intestinal pseudo-obstruction not only in children but also in adults. A subsequent induction of an immunosuppressive therapy with steroids, azathioprine, and ultimately TNF-alpha-inhibiting antibodies led to a slow recovery and stable disease.
Assuntos
Gastroenterite/diagnóstico , Pseudo-Obstrução Intestinal/diagnóstico , Intestino Delgado/patologia , Adulto , Biópsia , Doença Crônica , Feminino , Humanos , Pseudo-Obstrução Intestinal/etiologiaRESUMO
Pediatric histiocytic sarcoma (HS) clonally related to anteceding leukemia is a rare malignancy with poor outcome. We performed a molecular characterization of HS and the corresponding leukemia by methylation arrays and whole-exome sequencing and found a variety of aberrations in both entities with deletions of CDKN2A/B as a recurrent finding. Furthermore, data from genome-wide mutation analysis from one patient allowed the reconstruction of a sequence of tumorigenesis of leukemia and HS lesions including the acquisition of a putatively activating KRAS frameshift deletion (p.A66fs). Our results provide an insight into the genetic landscape of pediatric HS clonally related to anteceding leukemia.
Assuntos
Análise Mutacional de DNA/métodos , Genoma Humano , Sarcoma Histiocítico/genética , Leucemia Mieloide Aguda/genética , Criança , Inibidor de Quinase Dependente de Ciclina p15/genética , Inibidor p16 de Quinase Dependente de Ciclina/genética , Mutação da Fase de Leitura , Sarcoma Histiocítico/patologia , Humanos , Leucemia Mieloide Aguda/patologia , Prognóstico , Proteínas Proto-Oncogênicas p21(ras)/genética , Deleção de Sequência , Sequenciamento Completo do GenomaAssuntos
Acetaminofen/efeitos adversos , Analgésicos não Narcóticos/efeitos adversos , Doença Hepática Induzida por Substâncias e Drogas/etiologia , Atrofia Muscular Espinal/tratamento farmacológico , Acetaminofen/administração & dosagem , Administração Intravenosa , Adulto , Analgésicos não Narcóticos/administração & dosagem , Biópsia , Doença Hepática Induzida por Substâncias e Drogas/diagnóstico , Doença Hepática Induzida por Substâncias e Drogas/patologia , Humanos , Fígado/diagnóstico por imagem , Fígado/efeitos dos fármacos , Fígado/patologia , Masculino , Tomografia Computadorizada por Raios XRESUMO
Persistently elevated eosinophil granulocytes in the peripheral blood in children is challenging because of a complex diagnosis especially after solid organ transplantation and can lead to difficulties in finding an underlying causative factor.We report a 12-year-old boy who developed severe hypereosinophilia 11 years after liver transplantation due to biliary atresia. Accompanying symptoms were recurrent fever, fatigue, elevated liver enzymes, abdominal pain, and significant weight loss. After exclusion of secondary causes of eosinophilia, an idiopathic hypereosinophilic syndrome (I-HES) was diagnosed. Treatment with prednisolone resulted in an immediate response with rapid reduction of eosinophils, normalization of liver enzymes, and amelioration of any clinical symptoms. A hypereosinophilic syndrome in patients after liver transplantation is rare, and a broad differential diagnosis has to be considered. Prednisolone may lead to a prompt amelioration of eosinophilia and associated symptoms.
Assuntos
Síndrome Hipereosinofílica/etiologia , Transplante de Fígado , Complicações Pós-Operatórias , Criança , Humanos , Síndrome Hipereosinofílica/diagnóstico , Masculino , Complicações Pós-Operatórias/diagnósticoRESUMO
BACKGROUND: The pathogenesis of intestinal dysmotility in gastroschisis is not completely understood. Peel formation and disorganization of interstitial Cajal cells (ICC) have been proposed in humans. The aim of this study was to evaluate the impact of prenatal coverage of gastroschisis on gut inflammation and expression of ICC in a fetal lamb model. METHODS: Twenty-one German blackhead sheep with an abdominal wall defect that was created fetoscopically on day 77 of 145 days gestation were used in this study. Intrauterine surgery with the aim to cover the defect was performed 3 weeks later; two fetuses were covered completely, 5 partially and 11 remained uncovered. Three fetuses without gastroschisis were used as controls. All fetuses were retrieved by cesarean section at day 135. Samples of the small intestine were stained with hematoxylin and eosin for histologic analysis of peel formation and serosal and muscular thickness. For ICC detection, immunohistochemistry using anti-CD117 (c-Kit) antibody was used. RESULTS: In all samples with exposure to amniotic fluid, peel formation and significantly decreased ICC were found. Complete coverage reduced peel formation and disorganization of ICC compared to uncovered animals almost to the level of controls. CONCLUSIONS: Peel formation and ICC derangement were significantly reduced by prenatal coverage of gastroschisis. Moreover, this animal model mimics the histopathological bowel changes as seen in human gastroschisis and may, therefore, be used for further research on the pathophysiology and fetal therapy of this malformation.
Assuntos
Fetoscopia , Gastrosquise/cirurgia , Inflamação/patologia , Células Intersticiais de Cajal/metabolismo , Animais , Contagem de Células , Modelos Animais de Doenças , Feminino , Gastrosquise/patologia , Imuno-Histoquímica , Inflamação/metabolismo , Mucosa Intestinal/patologia , Gravidez , OvinosAssuntos
Anemia Falciforme/virologia , Hepatite Autoimune/virologia , Infecções por Parvoviridae/complicações , Anemia Falciforme/sangue , Anemia Falciforme/fisiopatologia , Pré-Escolar , DNA Viral/sangue , Feminino , Hepatite Autoimune/sangue , Hepatite Autoimune/fisiopatologia , Humanos , Infecções por Parvoviridae/sangue , Infecções por Parvoviridae/fisiopatologia , Infecções por Parvoviridae/virologia , Parvovirus B19 Humano , Reação em Cadeia da Polimerase Via Transcriptase Reversa , ViremiaAssuntos
Antineoplásicos/efeitos adversos , Benzenossulfonatos/efeitos adversos , Carcinoma Hepatocelular/cirurgia , Doença Hepática Induzida por Substâncias e Drogas/diagnóstico , Hepatite C/cirurgia , Neoplasias Hepáticas/cirurgia , Transplante de Fígado/imunologia , Piridinas/efeitos adversos , Doença Aguda , Antineoplásicos/uso terapêutico , Benzenossulfonatos/uso terapêutico , Bilirrubina/sangue , Carcinoma Hepatocelular/tratamento farmacológico , Humanos , Neoplasias Hepáticas/tratamento farmacológico , Masculino , Pessoa de Meia-Idade , Niacinamida/análogos & derivados , Compostos de Fenilureia , Piridinas/uso terapêutico , Sorafenibe , Resultado do TratamentoRESUMO
BACKGROUND: We aimed to compare two techniques of heterotopic heart transplantation in rats. Non-volume-loaded (NL) and volume-loaded (VL) models were tested for their physiologic and immunologic properties to assess their suitability for transplant studies. METHODS: Syngeneic heterotopic heart transplants were performed according to the techniques previously described by Ono (NL) and Yokoyama (VL). Grafts were followed over 90 days with sequential echocardiography. Ex-vivo Langendorff perfusion was used to gain functional data. Allogeneic heart transplants were done to determine whether chronic allograft vasculopathy (CAV) develops at a different pace in both transplant models. RESULTS: The ischemic time during surgery was significantly longer using the VL model (p<0.001). The LV diameter of NL hearts decreased over time while that of the VL model significantly increased (p=0.004 on POD 90). Mean LV developed pressure and (dP/dt)max were significantly higher with the NL model (61.1+/-8.5 mmHg and 4261.7+/-419.6 mmHg/s) than with VL hearts (19.9+/-16.5 mmHg; p=0.011 and 924.8+/-605.6 mmHg/s; p<0.001). The mean weight of NL hearts (0.45+/-0.03 g) was significantly less than that of VL hearts (1.21+/-0.16 g, p<0.001). Histology of syngeneic NL grafts showed healthy, but partly atrophic myocardium, whereas the LV myocardium of VL hearts showed dilation and scarring typical for chronic ischemic injury. Heart allografts similarly developed CAV with luminal narrowing of 37.2+/-16.6% (NL) and 34.4+/-21.4% (VL), respectively by POD 90 (p=0.807). CONCLUSIONS: Since the coronary arteries in the VL model get perfused with partly deoxygenated blood, the myocardium suffers from chronic ischemic injury. We recommend using the NL model in preclinical transplant studies.
Assuntos
Volume Cardíaco/fisiologia , Transplante de Coração/métodos , Transplante Heterotópico , Animais , Aorta Abdominal/transplante , Aorta Torácica/diagnóstico por imagem , Diástole , Ecocardiografia , Coração/fisiopatologia , Masculino , Modelos Animais , Miocárdio/patologia , Ratos , Veia Cava Inferior/patologiaRESUMO
We report a Caucasian neonate with chronic non-spherocytic hemolytic anemia due to a class I G6PD deficiency. A novel mutation missense mutation in exon eight of the G6PD gene was detected (c.827C>T p.Pro276Leu). Bilirubin peaked on day 5 at 24 mg/dl with a conjugated bilirubin of 17 mg/dl. Jaundice resolved within 4 weeks. A detailed work-up failed to reveal other specific factors contributing to cholestasis. Severe hemolytic disease of the newborn may cause cholestasis even in the absence of associated primary hepato-biliary disease.
Assuntos
Anemia Hemolítica Congênita não Esferocítica/complicações , Colestase/etiologia , Deficiência de Glucosefosfato Desidrogenase/complicações , Recém-Nascido Prematuro , Icterícia Neonatal/etiologia , Kernicterus/complicações , Mutação de Sentido Incorreto , Anemia Hemolítica Congênita não Esferocítica/genética , Anemia Hemolítica Congênita não Esferocítica/patologia , Colestase/patologia , Deficiência de Glucosefosfato Desidrogenase/genética , Deficiência de Glucosefosfato Desidrogenase/patologia , Humanos , Recém-Nascido , Icterícia Neonatal/patologia , Kernicterus/genética , Kernicterus/patologia , MasculinoAssuntos
Proctoscopia/efeitos adversos , Reto/microbiologia , Sepse/microbiologia , Choque Séptico/microbiologia , Infecções Estreptocócicas/microbiologia , Streptococcus pyogenes/patogenicidade , Adulto , Antibacterianos/uso terapêutico , Biópsia , Humanos , Masculino , Mucosa/microbiologia , Mucosa/patologia , Pelve , Reto/patologia , Sepse/tratamento farmacológico , Choque Séptico/tratamento farmacológico , Infecções Estreptocócicas/tratamento farmacológico , Infecções Estreptocócicas/transmissão , Streptococcus pyogenes/isolamento & purificaçãoRESUMO
BACKGROUND: Interleukin-12 (IL-12) is well characterized to induce cellular antitumoral immunity by activation of NK-cells and T-lymphocytes. However, systemic administration of recombinant human IL-12 resulted in severe toxicity without perceptible therapeutic benefit. Even though intratumoral expression of IL-12 leads to tumor regression and long-term survival in a variety of animal models, clinical trials have not yet shown a significant therapeutic benefit. One major obstacle in the treatment with IL-12 is to overcome the relatively low expression of the therapeutic gene without compromising the safety of such an approach. Our objective was to generate an adenoviral vector system enabling the regulated expression of very high levels of bioactive, human IL-12. RESULTS: High gene expression was obtained utilizing the VP16 herpes simplex transactivator. Strong regulation of gene expression was realized by fusion of the VP16 to a tetracycline repressor with binding of the fusion protein to a flanking tetracycline operator and further enhanced by auto-regulated expression of its fusion gene within a bicistronic promoter construct. Infection of human colon cancer cells (HT29) at a multiplicity of infection (m.o.i.) of 10 resulted in the production of up to 8000 ng/106 cells in 48 h, thus exceeding any published vector system so far. Doxycycline concentrations as low as 30 ng/ml resulted in up to 5000-fold suppression, enabling significant reduction of gene expression in a possible clinical setting. Bioactivity of the human single-chain IL-12 was similar to purified human heterodimeric IL-12. Frozen sections of human colon cancer showed high expression of the coxsackie adenovirus receptor with significant production of human single chain IL-12 in colon cancer biopsies after infection with 3*107 p.f.u. Ad.3r-scIL12. Doxycycline mediated suppression of gene expression was up to 9000-fold in the infected colon cancer tissue. CONCLUSION: VP16 transactivator-mediated and doxycycline-regulated expression of the human interleukin-12 gene enables highly efficient and tightly controlled cytokine expression in human cancer. These data illustrate the potential of the described adenoviral vector system for the safe and superior expression of therapeutic genes in the treatment of colorectal cancer and other malignancies.
Assuntos
Adenoviridae/genética , Clonagem Molecular/métodos , Doxiciclina/administração & dosagem , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Interleucina-12/genética , Interleucina-12/metabolismo , Transdução Genética/métodos , Relação Dose-Resposta a Droga , Células HT29 , HumanosRESUMO
Hepatitis B virus (HBV) DNA is detectable in a number of liver transplant candidates who are negative for hepatitis B surface antigen (HBsAg). After liver transplantation (LT), such patients may have molecular and/or serologic evidence of HBV replication. However, clinical disease from reactivation of occult HBV infection after LT has not been described. We report a patient who underwent LT for cryptogenic cirrhosis and had to be retransplanted twice for hepatic artery thrombosis. The patient was negative for HBsAg and positive for anti-hepatitis B core (HBc) and anti-HBs before all LT procedures and developed acute hepatitis B shortly after receiving the third graft. The HBV strain isolated at that time exhibited an unusual in frame insertion of a CAG motif within the HBV polymerase (HBV(INS+)). HBV(INS+) was detected retrospectively as a minor species in pretransplantation sera and the explanted native liver by insertion-specific polymerase chain reaction. This case in an occult HBV carrier shows that clinically apparent, endogenous reinfection of the graft may occur with minor HBV variants that are not detectable in pretransplantation samples by standard diagnostic procedures. This has implications for the analysis of sources of acute hepatitis B in patients after LT and possibly for consideration of antiviral prophylaxis in anti-HBc/anti-HBs/HBV DNA-positive patients.
Assuntos
Portador Sadio , Vírus da Hepatite B/metabolismo , Transplante de Fígado , Ativação Viral , Portador Sadio/microbiologia , Hepatite B/fisiopatologia , Hepatite B/transmissão , Antígenos de Superfície da Hepatite B/análise , Vírus da Hepatite B/genética , Humanos , Cirrose Hepática/cirurgia , Masculino , Pessoa de Meia-Idade , Filogenia , ReoperaçãoRESUMO
OBJECTIVE: Current hormone therapy in postmenopausal women is associated with uterotrophic activity and cancer-promoting effects. In this experimental study, we compared the effects of the selective estrogen-receptor (ER) beta agonist biochanin A, and the selective ERalpha agonist ethinylestradiol, on the development of intimal hyperplasia after balloon injury and on uterus morphology. DESIGN: Female F344 rats with or without prior ovariectomy were used for aortic denudations. Animals remained untreated or received oral biochanin A (100 mg/kg) or ethinylestradiol (100 microg/kg). After 14 days, aortas and uteri were harvested for histologic and immunohistochemical analyses. Computerized assessments of aortic adhesion molecule expression, and isometric relaxation experiments, and uteri were analyzed. In vitro studies with smooth muscle cells and endothelial cells were performed to further investigate the effects of hormone treatment on cell proliferation, migration and adhesion molecule expression. RESULTS: Among untreated rats, ovariectomized animals tended to show greater neointimal hyperplasia and increased expression of the adhesion molecules 1 (ICAM-1) and vascular cell adhesion molecule 1 (VCAM-1). Biochanin A treatment reduced neointima formation, inhibited VCAM-1 up-regulation, and improved the vascular relaxation response. No effect was observed on uterus growth or histology. Ethinylestradiol also reduced aortic neointima formation and inhibited VCAM-1 up-regulation, but failed to improve endothelial function and significantly induced uterus growth. Both agents showed antiproliferative and weak antimigratory effects on smooth muscle cells, and reduced VCAM-1 expression on stimulated endothelial cells in vitro. CONCLUSIONS: The ERbeta agonist biochanin A shows vasculoprotective effects without uterotrophic activity. Because hormone therapy may have cancer-promoting side effects, administration of ERbeta-selective agents might be alternatively used to reduce the risk of cardiovascular disease in postmenopausal women.
Assuntos
Receptor beta de Estrogênio/agonistas , Etinilestradiol/farmacologia , Genisteína/farmacologia , Útero/efeitos dos fármacos , Administração Oral , Animais , Aorta/efeitos dos fármacos , Cateterismo , Endotélio Vascular/efeitos dos fármacos , Endotélio Vascular/lesões , Etinilestradiol/administração & dosagem , Feminino , Genisteína/administração & dosagem , Imuno-Histoquímica , Molécula 1 de Adesão Intercelular/metabolismo , Ratos , Ratos Endogâmicos F344 , Útero/patologia , Molécula 1 de Adesão de Célula Vascular/metabolismoRESUMO
BACKGROUND: Since the two estrogen receptor isoforms ERalpha and ERbeta have been discovered it is unclear by which receptor immunomodulating or feminizing effects are mediated. In this study, the effects of the two selective ERalpha- and ERbeta-agonists ethinylestradiol and biochaninA, respectively, on acute cardiac allograft rejection, uterus growth, vascular adhesion molecule and MHC-II expression were investigated and verified using in vitro cell culture. METHODS: Heterotopic Lewis to ovarectomized F344 cardiac transplantations were performed. The study groups received supplemental biochaninA or ethinylestradiol, the control group received no treatment. Grafts and uteri were harvested on the fifth postoperative day and blood was taken for hormone plasma level quantifications. Purified Lewis aortic endothelial cell cultures were pre-treated with biochaninA or ethinylestradiol and stimulated with TNF-alpha or IFN-gamma for quantification of ICAM-1/VCAM-1 and MHC-II expression. Endothelium-lymphocyte adhesion assays were performed using purified F344 lymphocytes. RESULTS: Both biochaninA and ethinylestradiol treatment significantly reduced graft mononuclear infiltration of CD8(+) and ED1(+) cells and markedly reduced ISHLT grading compared to untreated controls. Either agent significantly inhibited lymphocyte adhesion, endothelial VCAM-1 upregulation during graft rejection and during TNF-alpha-stimulation in vitro, whereas no effect was observed for ICAM-1 upregulation. BiochaninA but not ethinylestradiol significantly reduced endothelial MHC-II upregulation in vivo and in vitro. Only ethinylestradiol treatment strongly affected uterus growth in ovarectomized recipients. CONCLUSIONS: Only the treatment with the phytoestrogen biochaninA reduced endothelial MHC-II expression in vivo and in vitro and weakened allograft rejection without affecting the reproductive system. Supplemental phytoestrogens may therefore provide further benefits in the clinical setting.
Assuntos
Genisteína/uso terapêutico , Rejeição de Enxerto/prevenção & controle , Transplante de Coração/imunologia , Fitoestrógenos/uso terapêutico , Doença Aguda , Animais , Adesão Celular , Moléculas de Adesão Celular/metabolismo , Células Endoteliais/imunologia , Células Endoteliais/metabolismo , Feminino , Genisteína/sangue , Genisteína/farmacologia , Antígenos de Histocompatibilidade Classe II/imunologia , Linfócitos/imunologia , Tamanho do Órgão/efeitos dos fármacos , Fitoestrógenos/sangue , Fitoestrógenos/farmacologia , Ratos , Ratos Endogâmicos Lew , Receptores de Estrogênio/agonistas , Receptores de Estrogênio/análise , Útero/anatomia & histologia , Útero/efeitos dos fármacos , Útero/imunologiaRESUMO
BACKGROUND: The malononitrilamide FK778 is a novel derivate of leflunomide and interacts with T- and B-cell function by inhibiting de novo pyrimidine synthesis. We investigated the effects of FK778 upon acute cardiac allograft rejection and upon adhesion molecule upregulation in experimental transplantation and by using in vitro cell culture. METHODS: Heterotopic, abdominal cardiac transplantations were performed in the Brown Norway (BN) to Lewis (Lew) rat model. The study groups received daily low- or high-dose FK778 immunosuppression. FK778 plasma levels were quantified by HPLC. Grafts were harvested on the fifth postoperative day for histologic and immunohistologic examinations using computerized morphometry. Purified BN aortic endothelial cell cultures were pretreated with low- or high-dose FK778 according to FK778 plasma levels and were stimulated with tumor necrosis factor (TNF)-alpha. Adhesion molecule expression was quantified by immunofluorescence, FACS analysis, and Western blotting. Lymphocyte-endothelium adhesion assays were performed using purified Lew lymphocytes and radiolabeled TNF-alpha was used for receptor binding assays. RESULTS: FK778 treatment dose-dependently reduced graft mononuclear infiltration of CD4(+), CD8(+), and ED1(+) cells, but only high-dose FK778 treatment significantly reduced early upregulation of ICAM-1 and VCAM-1 in vivo. FK778 also dose-dependently reduced TNF-alpha-stimulated endothelial adhesion molecule upregulation in vitro, whereas the effect on VCAM-1 was more dominant. We did not find evidence that FK778 interferes with surface receptor binding of TNF-alpha. Lymphocyte adhesion to endothelial cell monolayers was significantly attenuated by FK778. CONCLUSION: Besides its inhibitory effect on pyrimidine synthesis, FK778 directly reduces endothelial adhesion molecule upregulation and attenuates lymphocyte-endothelium interaction, which is a critical step in graft rejection.
Assuntos
Adesão Celular/efeitos dos fármacos , Células Endoteliais/citologia , Rejeição de Enxerto/tratamento farmacológico , Transplante de Coração , Isoxazóis/farmacologia , Linfócitos/citologia , Doença Aguda , Alcinos , Animais , Antineoplásicos/farmacocinética , Adesão Celular/imunologia , Células Endoteliais/metabolismo , Rejeição de Enxerto/imunologia , Rejeição de Enxerto/metabolismo , Técnicas In Vitro , Molécula 1 de Adesão Intercelular/metabolismo , Radioisótopos do Iodo , Isoxazóis/sangue , Linfócitos/metabolismo , Nitrilas , Ratos , Ratos Endogâmicos BN , Ratos Endogâmicos Lew , Receptores do Fator de Necrose Tumoral/metabolismo , Fator de Necrose Tumoral alfa/farmacocinética , Molécula 1 de Adesão de Célula Vascular/metabolismoRESUMO
Transplantation of retinal pigment epithelial (RPE) cells is discussed as a possible therapeutic approach for retinal degeneration. Xenogeneic transplantation of human RPE cells in animal models has been studied extensively. Various methods have been used to identify the graft cells, but these methods interfere with cell behaviour so that the monitored physiological post-transplantation course may be influenced. In the present study, we applied a method for an unequivocal identification of the graft cells without interfering cell metabolism or behaviour using in situ hybridisation (ISH) of human specific Alu sequences. Visualisation of the strong extended nuclear signal of Alu sequences was much easier than that of the small nuclear signals of donor-specific sex chromosome probes. With Alu probe, even single graft cells can be identified and their development can be observed in short-term and long-term studies. With this procedure, we could prove that donor cells were injected correctly into the subretinal space by a special injection technique that we developed previously. In combination with immunohistochemistry, donor cells could be clearly discriminated from macrophages, which contained phagocytosed donor cell fragments. Application of these ISH methods for species-specific identification was valuable for follow-up-studies of RPE transplantation.
Assuntos
Elementos Alu , Hibridização In Situ , Modelos Animais , Epitélio Pigmentado Ocular/transplante , Transplante Heterólogo , Animais , Anticorpos Monoclonais , Núcleo Celular/química , DNA/análise , Humanos , Imuno-Histoquímica , Macrófagos/imunologia , Fagocitose , Epitélio Pigmentado Ocular/ultraestrutura , Ratos , Doadores de TecidosRESUMO
Allogeneic stem cell transplantation (allo-SCT) after reduced-intensity conditioning was evaluated in 22 patients (median age 53, range 36-66 years) with multiple myeloma with progression after an autologous SCT. Seven patients received a transplant from a human leucocyte antigen (HLA)-identical sibling and 15 patients (68%) from an unrelated donor [including 3/22 (14%) from a HLA-mismatched unrelated donor]. Graft-versus-host disease (GVHD) prophylaxis consisted of serotherapy with antithymocyte globulin (ATG) and cyclosporine (CSA) (n = 12) or CSA plus mycophenolate mofetil (n = 10). Despite of heavy pretreatment, the transplant-related mortality (TRM) for all grafted patients was acceptable at 5/22 patients (23%). Seven of 21 patients (33%) that were evaluated developed grade II GVHD and one (5%) patient developed grade III/IV acute GVHD. Seven patients developed chronic GVHD (cGVHD), but only one was extensive. Eleven patients died of progressive disease within a median of 7 months (2-19 months) post transplant. Thirteen of all 22 patients (59%) achieved a partial or complete remission with six of these 13 patients (46%) remaining event free at a median of 24 months (range 8-36 months) post allografting. Estimated 2 year overall and event-free survival was, respectively, 25.5% and 22.0% for the whole patient group, and 62.5% and 57.1% for patients with chemosensitive disease. Chemorefractory disease prior to allogeneic stem cell transplantation (P = 0.0182) and absence of cGVHD (P = 0.069) were associated with shorter event-free survival. Thus long-term disease control can be achieved, but is restricted to patients responding to prior salvage chemotherapy.