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Journal of Virology, October 2004, p. 10370-10377, Vol. 78, No. 19
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.19.10370-10377.2004

Analysis of Human Peripheral Blood Samples from Fatal and Nonfatal Cases of Ebola (Sudan) Hemorrhagic Fever: Cellular Responses, Virus Load, and Nitric Oxide Levels

Anthony Sanchez,1* Matthew Lukwiya,2 Daniel Bausch,1 Siddhartha Mahanty,1 Angela J. Sanchez,1 Kent D. Wagoner,1 and Pierre E. Rollin1

Special Pathogens Branch, Division of Viral and Rickettsial Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia,1 Saint Mary's Hospital, Gulu, Uganda2

Received 20 November 2003/ Accepted 28 May 2004

Peripheral blood samples obtained from patients during an outbreak of Ebola virus (Sudan species) disease in Uganda in 2000 were used to phenotype peripheral blood mononuclear cells (PBMC), quantitate gene expression, measure antigenemia, and determine nitric oxide levels. It was determined that as the severity of disease increased in infected patients, there was a corresponding increase in antigenemia and leukopenia. Blood smears revealed thrombocytopenia, a left shift in neutrophils (in some cases degenerating), and atypical lymphocytes. Infected patients who died had reduced numbers of T cells, CD8+ T cells, and activated (HLA-DR+) CD8+ T cells, while the opposite was noted for patients who survived the disease. Expression levels of cytokines, Fas antigen, and Fas ligand (TaqMan quantitation) in PBMC from infected patients were not significantly different from those in uninfected patients (treated in the same isolation wards), nor was there a significant increase in expression compared to healthy volunteers (United States). This unresponsive state of PBMC from infected patients despite high levels of circulating antigen and virus replication suggests that some form of immunosuppression had developed. Ebola virus RNA levels (virus load) in PBMC specimens were found to be much higher in infected patients who died than patients who survived the disease. Similarly, blood levels of nitric oxide were much higher in fatal cases (increasing with disease severity), and extremely elevated levels (≥150 µM) would have negatively affected vascular tone and contributed to virus-induced shock.

* Corresponding author. Mailing address: Centers for Disease Control and Prevention, 1600 Clifton Road N.E., Building 15, Room SB611, Mail Stop G-14, Atlanta, GA 30333. Phone: (404) 639-1119. Fax: (404) 639-1118. E-mail: ASanchez1{at}cdc.gov.

Journal of Virology, October 2004, p. 10370-10377, Vol. 78, No. 19
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.19.10370-10377.2004




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