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Improved Swiss-rolling Technique for Intestinal Tissue Preparation for Immunohistochemical and Immunofluorescent Analyses
Published on: July 13, 2016
Whipple's disease: immunospecific and quantitative immunohistochemical study of intestinal biopsy specimens
Hubert Lepidi1, Florence Fenollar, Rene Gerolami
1Unité des Rickettsies, Université de la Méditerranée, and Hôpital de la Conception, Marseille, France.
Insights
Immunohistochemistry effectively detects Tropheryma whipplei in intestinal biopsies for Whipple's disease diagnosis. This method offers high specificity and sensitivity, aiding in both active infection detection and retrospective studies.
Area of Science:
- Gastroenterology
- Infectious Diseases
- Pathology
Background:
- Whipple's disease diagnosis traditionally relies on methods like PAS staining and electron microscopy.
- Accurate detection of Tropheryma whipplei is crucial for diagnosing and managing Whipple's disease.
Purpose of the Study:
- To evaluate the diagnostic utility of immunohistochemistry (IHC) for detecting Tropheryma whipplei in intestinal biopsies.
- To quantify infected cells and assess the diagnostic value of IHC compared to PAS staining.
Main Methods:
- Analysis of 29 duodenal biopsy specimens from 15 Whipple's disease patients (untreated and treated) and controls.
- Quantitative image analysis of PAS staining and IHC using CD68 and T. whipplei antibodies.
- Comparison of staining surface areas before and after antibiotic therapy.
Main Results:
- Immunohistochemistry detected T. whipplei in all 15 Whipple's disease patients.
- Massive intestinal macrophagic infiltration (CD68+) was observed in patients (20.3% pre-therapy, 13.4% post-therapy) vs. controls (2.1%).
- While IHC and PAS staining were similar pre-therapy, IHC showed significantly lower surface area post-therapy (2.8% vs. 7.9%).
Conclusions:
- Immunohistochemistry is a highly specific and sensitive method for detecting T. whipplei in intestinal tissues.
- IHC is a valuable diagnostic tool for active Whipple's disease and retrospective analyses.
- Quantitative image analysis aids in assessing treatment response and disease burden.
Abstract:
Whipple's disease may be diagnosed by periodic acid-Schiff (PAS) staining, electron microscopy, or polymerase chain reaction of intestinal biopsy specimens. The aim of this study was to evaluate the diagnostic value of immunohistochemistry and the quantification of infected cells in intestinal Whipple's disease. A total of 29 duodenal biopsy specimens from 15 patients with untreated and treated Whipple's disease were examined and compared with biopsy specimens from control patients with normal intestinal mucosa or various pathologic processes. Percentages of staining surfaces with PAS stain and antibodies directed against CD68, a macrophage marker, or the Whipple bacillus, Tropheryma whipplei, were studied quantitatively using a computerized system of image analysis. Positive detection of T. whipplei was obtained using immunohistochemistry in all 15 patients with Whipple's disease. No bacteria were detected in any of the negative controls. The use of quantitative image analysis showed a massive intestinal macrophagic infiltration before (20.3%) and after (13.4%) antibiotic therapy completion as compared with controls (2.1%). The 2 detection methods for T. whipplei, PAS stain and immunohistochemistry, were quantitatively similar before therapy (19.9% versus 17.5%), but the immunodetection-based surface area was significantly lower than the PAS staining surface area after therapy (2.8% versus 7.9%). Our findings indicate that immunohistochemistry is highly specific and sensitive and is applicable as a diagnostic method on intestinal tissue specimens to detect T. whipplei during active infection or in retrospective studies.
