RESUMO
INTRODUCTION: Different strategies have been developed to identify those refractory celiac disease (RCD) patients who are at risk to develop an enteropathy associated T-cell lymphoma (EATL). Flow cytometric analysis of intra-epithelial lymphocytes (IEL) with an aberrant phenotype is considered the golden standard but is not widely available. Immunohistochemistry (IHC) and T-cell receptor (TCR) rearrangement studies are commonly available but may lack sensitivity and specificity. Here, we compared the three different methods in the workup of patients suspected for RCD. METHODS: Duodenal biopsies from control patient (n = 5), RCD patients with moderately increased aberrant IEL populations (20-50 %: n = 14), and RCD patients with high numbers of aberrant IEL (>50 %: n = 5) as determined by flow cytometry were analysed by IHC and TCR-γ chain rearrangement analysis. Three pathologists scored the slides independently. RESULTS: Sensitivity of IHC and TCR-γ rearrangement analysis in RCD patients with high numbers of aberrant IELs was 100 and 71 %, respectively. RCD patients with aberrant cells between 25 and 50 % however, were missed by IHC and TCR in 50 and 57 % of cases, respectively. In addition, inter-rater reliability analysis of the IHC scoring revealed coder-pair Kappa coefficients between 0.28 and 0.85. CONCLUSION: Immunohistochemistry and to a lesser extent TCR-γ clonality analysis are sensitive in identifying patients with high numbers of aberrant IEL populations, yet miss half of RCD patients with moderately increased numbers. In addition, IHC has a high inter-observer variability. Therefore, patients suspected for RCD should undergo flow cytometric analysis of the duodenum.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Doença Celíaca/diagnóstico , Mucosa Intestinal/imunologia , Linfoma de Células T/diagnóstico , Adulto , Idoso , Doença Celíaca/complicações , Doença Celíaca/imunologia , Separação Celular/métodos , Resistência a Medicamentos , Feminino , Citometria de Fluxo , Rearranjo Gênico da Cadeia gama dos Receptores de Antígenos dos Linfócitos T/genética , Humanos , Imuno-Histoquímica , Mucosa Intestinal/patologia , Linfoma de Células T/etiologia , Linfoma de Células T/imunologia , Masculino , Pessoa de Meia-Idade , Variações Dependentes do Observador , Receptores de Antígenos de Linfócitos T/genética , Recidiva , Sensibilidade e Especificidade , Adulto JovemRESUMO
UNLABELLED: Brain death donors are frequently used for transplantation. Previous studies showed that brain death (BD) negatively affects the immunological and inflammatory status of both liver and kidney. OBJECTIVE: Therefore we studied the inflammatory and morphological changes in donor small intestine after brain death induction. METHODS: BD was induced in rats by slow inflation of an epidural balloon catheter. Three groups (n = 6) were compared, 1 hour, 4 hours BD and sham operated controls. The liver was used as a reference to confirm our previous findings. Intestinal injury was determined using the Park score. Polymorphonuclear cells (PMNs) were counted in intestine and liver as a marker for inflammatory response. Real time PCR was used to demonstrate the effects of BD on ICAM-1 expression in the jejunum. RESULTS: The morphology of the intestine was compromised after 1 and 4 hours BD. In brain dead rats, apical lifting of epithelial cells was clearly present, which resulted in higher Park scores compared to controls (P < .05). Liver morphology remained intact. In small intestine and liver an increased PMN influx in the 1 hour BD group was observed in comparison to controls. Hepatic PMN influx increased further in the 4 hours BD group (P < .05). ICAM-1 was upregulated in jejunum in both the 1 hour BD and 4 hours BD groups compared to controls (P < .05). In conclusion, the early occurrence of intestinal damage after BD induction may negatively influence transplant outcome.