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1.
Am J Transplant ; 18(4): 1007-1015, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29139621

RESUMO

Graft survival after small bowel transplantation remains impaired due to acute cellular rejection (ACR), the leading cause of graft loss. Although it was shown that the number of enteroendocrine progenitor cells in intestinal crypts was reduced during mild ACR, no results of Paneth and intestinal stem cells localized at the crypt bottom have been shown so far. Therefore, we wanted to elucidate integrity and functionality of the Paneth and stem cells during different degrees of ACR, and to assess whether these cells are the primary targets of the rejection process. We compared biopsies from ITx patients with no, mild, or moderate ACR by immunohistochemistry and quantitative PCR. Our results show that numbers of Paneth and stem cells remain constant in all study groups, whereas the transit-amplifying zone is the most impaired zone during ACR. We detected an unchanged level of antimicrobial peptides in Paneth cells and similar numbers of Ki-67+ IL-22R+ stem cells revealing cell functionality in moderate ACR samples. We conclude that Paneth and stem cells are not primary target cells during ACR. IL-22R+ Ki-67+ stem cells might be an interesting target cell population for protection and regeneration of the epithelial monolayer during/after a severe ACR in ITx patients.


Assuntos
Rejeição de Enxerto/fisiopatologia , Sobrevivência de Enxerto , Intestino Delgado/fisiopatologia , Intestino Delgado/transplante , Transplante de Órgãos/efeitos adversos , Celulas de Paneth/citologia , Células-Tronco/citologia , Adolescente , Adulto , Feminino , Seguimentos , Rejeição de Enxerto/etiologia , Humanos , Masculino , Celulas de Paneth/metabolismo , Prognóstico , Fatores de Risco , Células-Tronco/metabolismo , Adulto Jovem
2.
Am J Transplant ; 14(9): 2148-58, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25088845

RESUMO

Intestinal transplantation (ITx) faces the challenge of grafting a high immunogenic organ, which is certainly one of the major obstacles for intestinal allograft acceptance. The allograft has to guarantee the proper functioning of the mucosal immune machinery under immunosuppressive conditions. Recently, it has been elucidated that isolated lymphoid follicles (ILFs) are an indispensable part of mucosal immunity to maintain IgA synthesis and consequently to control commensal microflora. No data about these follicular structures in the setting of ITx are available so far. Therefore, we addressed the question whether constitution, integrity and function of allograft ILFs are disturbed by immunosuppressive regimen. We compared allograft ILFs from terminal ileum of transplant patients with ILFs from nontransplant patients via flow cytometry, quantitative real-time polymerase chain reaction and immunohistochemistry. We found that host leukocytes rapidly repopulate allograft ILFs and that maintenance immunosuppressive regimen, tacrolimus and corticosteroids, does not affect their cellular integrity and function. However, allograft ILFs revealed a higher maturation state than control samples and IgA positive plasma cells were increased in number in allograft mucosa. Our results open the path for a better understanding of allograft mucosal immunity.


Assuntos
Intestinos/transplante , Tecido Linfoide/imunologia , Adolescente , Adulto , Sequência de Bases , Criança , Pré-Escolar , Primers do DNA , Feminino , Humanos , Imunidade nas Mucosas , Imunoglobulina A/metabolismo , Lactente , Intestinos/imunologia , Masculino , Repetições de Microssatélites , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase em Tempo Real , Linfócitos T Auxiliares-Indutores/imunologia , Transplante , Adulto Jovem
3.
Clin Exp Immunol ; 162(1): 138-45, 2010 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-20831713

RESUMO

During intestinal transplant (ITx) operation, intestinal lymphatics are not reconstituted. Consequently, trafficking immune cells drain freely into the abdominal cavity. Our aim was to evaluate whether leucocytes migrating from a transplanted intestine could be recovered from the abdominal draining fluid collected by a peritoneal drainage system in the early post-ITx period, and to determine potential applications of the assessment of draining cellular populations. The cell composition of the abdominal draining fluid was analysed during the first 11 post-ITx days. Using flow cytometry, immune cells from blood and draining fluid samples obtained the same day showed an almost complete lymphopenia in peripheral blood, whereas CD3(+) CD4(+) CD8(-) , CD3(+) CD4(-) CD8(+) and human leucocyte antigen D-related (HLA-DR)(+) CD19(+) lymphocytes were the main populations in the draining fluid. Non-complicated recipients evolved from a mixed leucocyte pattern including granulocytes, monocytes and lymphocytes to an exclusively lymphocytic pattern along the first post-ITx week. At days 1-2 post-Itx, analysis by short tandem repeats fingerprinting of CD3(+) CD8(+) sorted T cells from draining fluid indicated that 50% of cells were from graft origin, whereas by day 11 post-ITx this proportion decreased to fewer than 1%. Our results show for the first time that the abdominal drainage fluid contains mainly immune cells trafficking from the implanted intestine, providing the opportunity to sample lymphocytes draining from the grafted organ along the post-ITx period. Therefore, this analysis may provide information useful for understanding ITx immunobiology and eventually could also be of interest for clinical management.


Assuntos
Intestinos/imunologia , Sistema Linfático/imunologia , Linfócitos/imunologia , Imunologia de Transplantes , Cavidade Abdominal/cirurgia , Antígenos CD19/metabolismo , Complexo CD3/metabolismo , Linfócitos T CD4-Positivos/citologia , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/metabolismo , Linfócitos T CD8-Positivos/citologia , Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/metabolismo , Movimento Celular/imunologia , Drenagem/métodos , Citometria de Fluxo , Antígenos HLA-DR/metabolismo , Humanos , Intestinos/transplante , Sistema Linfático/citologia , Sistema Linfático/metabolismo , Contagem de Linfócitos , Linfócitos/citologia , Linfócitos/metabolismo , Fatores de Tempo
4.
Transplant Proc ; 42(1): 57-61, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-20172281

RESUMO

INTRODUCTION: The diagnosis of rejection after intestinal transplantation is still performed by endoscopic biopsy monitoring. Less invasive diagnostic procedures are desirable, although they are not available so far. Calprotectin, a stable cytosolic granulocyte protein, which previously was used as a marker of inflammatory processes, has been proposed to be a biochemical marker for rejection. The aim of the present work was to analyze the concordance between calprotectin levels in intestinal content and the presence of graft rejection after small bowel transplantation. METHODS: Calprotectin level was measured using a commercial ELISA kit on 137 samples of intestinal content randomly collected during endoscopies performed on 11 intestinal transplantation patients during 2 years' follow-up. Calprotectin determinations were correlated with histological and clinical findings. The cut-off level was determined retrospectively by receiver-operator curve analysis. RESULTS: Based on histological findings and clinical records, samples were discerned as rejection positive (37 of 137), versus negative (35 of 137) samples or those with no clinical, endoscopic, or histological findings (65 of 137 samples). A cut-off value of 65 microg of calprotectin/g of intestinal content provided the best assay parameter according to the clinical findings: a 76% sensitivity and a 47% specificity. False positive results corresponded to patients with gastrointestinal infections (13%), systemic infections (13%), ulcers (10%), or nonspecific histological alterations of the mucosa (15%). The other false positive cases corresponded to postsurgical samples (4%), or patients with concomitant gastrointestinal symptoms (2%). Most false negative results (78%) were observed during recovery from severe acute rejection episodes, among successfully treated patients. In these cases, epithelial reconstitution and no mucosal infiltration was observed. If the latter group were discarded, sensitivity rose to 93%, and specificity, to 50% with a 96% negative predictive value. Furthermore, a weak correlation was observed between calprotectin levels and the severity of rejection. CONCLUSIONS: This study confirmed the results obtained by other groups: fecal calprotectin dosage showed a good sensitivity but low specificity for the diagnosis of intestinal rejection because high calprotectin levels can also be observed in other clinical conditions. Nevertheless, it might be used as a first line for continuous evaluation of intestinal transplantation status, like other biochemical parameters that are used in kidney or liver transplantation, before considering the need for a biopsy.


Assuntos
Rejeição de Enxerto/diagnóstico , Complexo Antígeno L1 Leucocitário/análise , Adolescente , Adulto , Biomarcadores/análise , Biomarcadores/metabolismo , Criança , Pré-Escolar , Ensaio de Imunoadsorção Enzimática/métodos , Fezes/química , Humanos , Lactente , Pessoa de Meia-Idade , Curva ROC , Estudos Retrospectivos , Transplante Homólogo
5.
Bioresour Technol ; 76(1): 71-3, 2001 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-11315814

RESUMO

Biomass production of fodder sorghum (Sorghum sp.) has been tested in a field trial over two harvesting periods under natural meteorological conditions using ammoxidized kraft lignin (AKL) as a slow-release fertilizer and urea as conventional reference. In the course of the first growth cycle, plants treated with urea gave higher biomass yields because of the better solubility of urea in the initial phase. However, during the second cycle AKL treated plants performed better than urea treated sorghum, indicating that nitrogen from AKL became readily available.


Assuntos
Lignina/metabolismo , Poaceae/metabolismo , Biomassa , Fertilização , Fertilizantes , Nitrogênio/química , Nitrogênio/metabolismo , Folhas de Planta/química , Poaceae/efeitos dos fármacos , Ureia/farmacologia
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