1. ¹Department of Pathology, The Ohio State University, Columbus, OH, USA
2. ²Department of Pathology, Santa Casa de Sáo Paulo Hospital, Sáo Paulo, Brazil
3. ³Department of Pathology, Instituto Nacional de Concerologia, Mexico City, Mexico

Correspondence: Dr G Nuovo, MD, Ohio State Medical Center, Dept Pathology –Div Anatomic Pathology, S305 Rhodes Hall; 450 W 10th Ave, Columbus, OH 43210-1228, USA. E-mail: nuovo-1@medctr.osu.edu

Received 24 May 2004; Revised 3 August 2004; Accepted 3 August 2004; Published online 24 September 2004.

Abstract

The purposes of this study were to elucidate the role of cytokine upregulation in the pathogenesis of rabies encephalitis and to compare the detection of Negri bodies with that of rabies protein by immunohistochemistry and rabies RNA by reverse transcriptase (RT) in situ PCR for its diagnosis. Negri bodies were evident in 4/7 of the documented rabies cases; viral protein and viral RNA were detected in each case. The average number of rabies-infected cells, determined by counting 150 neurons in serial sections in areas where viral protein was evident, with the three different detection methods was: Negri bodies (<1/150), immunohistochemistry (4/150), and RT in situ PCR (49/150). No rabies protein or RNA was detected in four control brain tissues that were read with the rabies cases in a blinded fashion. The ratio of cells expressing tumor necrosis factor (TNF) or inducible nitric oxide synthetase (iNOS) to 1 SSI-1/SOCS-1 (suppressors of cytokine signaling) expression, which is a novel class of negative feedback regulators of cytokine receptor signaling, was markedly increased only in the areas where many viral infected cells were present. Colabeling experiments showed that most of the cells expressing iNOS or TNF were not virally infected, but rather adjacent to rabies-infected neurons. We conclude that RT in situ PCR for rabies virus is the most accurate test for the determination of viral load in rabies encephalitis. Further, the disease is characterized by massive viral infection of neurons in a markedly focal distribution in conjunction with a concomitant upregulation of cytokine expression in adjacent, noninfected cells that may be due, in part, to SOCS downregulation.