3rd Annual BMRP Investigator Meeting - Abstract
A PCR Search for Evidence of Recurring or Persistent Viruses in Crohn’s Disease
Herbert J. Van Kruiningen1,a, Michelle Poulin2, Antonio E. Garmendia1, Pierre Desreumaux3, Jean-Frederic Colombel3, Gert De Hertogh4, Severine Vermeire4 and Gregory J. Tsongalis2
1Department of Pathobiology and Veterinary Science, University of Connecticut (Storrs, Connecticut, U.S.A); 2Hartford Hospital (Hartford, Connecticut, U.S.A.); 3Centre Hospitalier Régional, Universitaire de Lille (Lille, France); 4Department of Gastroenterology, University Hospital Gasthuisberg (Leuven, Belgium)
Background & Aim: New onset Crohn’s disease (CD) and acute flares are often associated with viral infections. Aphthous ulcers are a feature of a number of gastrointestinal viral infections and field studies link clustering of CD with contaminated ground water or the drinking of well water. The aim was to employ PCR and RT-PCR to search for evidence of persistent or recurrent viruses in the diseased tissues of patients.
Methods: Tissue blocks were obtained from surgical resection specimens from patients with Crohn’s disease and from a control population. They were quick frozen to -70ºC and later subdivided for DNA and RNA extraction. One hundred and seven samples were tested by PCR or RT-PCR, for EBV, CMV, HSV 1, HSV 2, HHV 8, pestivirus, and enteroviruses. From Belgium, there were specimens from 26 patients and 18 controls and from the north of France, there were specimens from 40 patients and 23 controls. Controls had had resection for diverticulosis, polyps, adenocarcinoma, or other intestinal neoplasia. Following some initial positive findings of EBV, serology for various EBV antigens was done on the sera of seven Belgian patients who had been followed after resection. Titers were measured at Clinical Laboratories, NYU Medical Center, New York, NY.
Results: Evidence of EBV was detected in 11 of 65 patients tested (17%) and in three of 41 controls (7.3%) (p=0.15). The proportions of positives were approximately equal among Belgian (15.4%) and French patients (17.9%). None of the 18 Belgian controls was positive, whereas three of 23 French controls were (13.0%). Only one patient was positive for pestivirus. One patient and one control were positive for CMV and HHS 1 and 2, HHV 8 and enteroviruses were not found. Five of the seven patients followed post-operatively for serologic evidence of EBV activity had antibodies to early viral antigen and to nuclear antigen, which were uninfluenced by surgery. None of the seven had IgM versus EBV. All seven had antibody to viral capsid antigen.
Conclusion: Our findings vary from those of Ruther et al. who demonstrated evidence of EBV in tissues from seven of 11 (64%) German CD patients. Antibodies to early EBV viral antigen and to nuclear antigen in five of seven other Belgian patients suggest persistent active viral infection.
aPrincipal Investigator