Human asymptomatic Ebola infection and strong inflammatory response.
BACKGROUND
Ebola virus is one of the most virulent pathogens, killing a very high proportion of patients within 5-7 days. Two outbreaks of fulminating haemorrhagic fever occurred in northern Gabon in 1996, with a 70% case-fatality rate. During both outbreaks we identified some individuals in direct contact with sick patients who never developed symptoms. We aimed to determine whether these individuals were indeed infected with Ebola virus, and how they maintained asymptomatic status.


METHODS
Blood was collected from 24 close contacts of symptomatic patients. These asymptomatic individuals were sampled 2, 3, or 4 times during a 1-month period after the first exposure to symptomatic patients. Serum samples were analysed for the presence of Ebola antigens, virus-specific IgM and IgG (by ELISA and western blot), and different cytokines and chemokines. RNA was extracted from peripheral blood mononuclear cells, and reverse transcriptase-PCR assays were done to amplify RNA of Ebola virus. PCR products were then sequenced.


RESULTS
11 of 24 asymptomatic individuals developed both IgM and IgG responses to Ebola antigens, indicating viral infection. Western-blot analysis showed that IgG responses were directed to nucleoprotein and viral protein of 40 kDa. The glycoprotein and viral protein of 24 kDa genes showed no nucleotide differences between symptomatic and asymptomatic individuals. Asymptomatic individuals had a strong inflammatory response characterised by high circulating concentrations of cytokines and chemokines.


CONCLUSIONS
This study showed that asymptomatic, replicative Ebola infection can and does occur in human beings. The lack of genetic differences between symptomatic and asymptomatic individuals suggest that asymptomatic Ebola infection did not result from viral mutations. Elucidation of the factors related to the genesis of the strong inflammatory response occurring early during the infectious process in these asymptomatic individuals could increase our understanding of the disease.
Leroy
E M
EM
Centre International de Recherches Médicales de Franceville, Gabon. leroy@cimf.sci.ga
Baize
S
S
Volchkov
V E
VE
Fisher-Hoch
S P
SP
Georges-Courbot
M C
MC
Lansoud-Soukate
J
J
Capron
M
M
Debré
P
P
McCormick
J B
JB
Georges
A J
AJ
D000914:Antibodies, Viral / Q000097:blood; D000956:Antigens, Viral / Q000097:blood; D015153:Blotting, Western; D018932:Chemokine CCL2 / Q000097:blood; D054407:Chemokine CCL4; D029043:Ebolavirus / Q000145:classification* / Q000235:genetics / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005500:Follow-Up Studies; D006023:Glycoproteins / Q000032:analysis; D019142:Hemorrhagic Fever, Ebola / Q000175:diagnosis* / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D007075:Immunoglobulin M / Q000097:blood; D016898:Interferon-alpha / Q000097:blood; D007375:Interleukin-1 / Q000097:blood; D018664:Interleukin-12 / Q000097:blood; D015850:Interleukin-6 / Q000097:blood; D019402:Macrophage Inflammatory Proteins / Q000097:blood; D009698:Nucleoproteins / Q000032:analysis; D009711:Nucleotides / Q000032:analysis; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000032:analysis; D013601:T-Lymphocytes / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis; D014764:Viral Proteins / Q000032:analysis; D014779:Virus Replication
D000914:Antibodies, Viral; D000956:Antigens, Viral; D018932:Chemokine CCL2; D054407:Chemokine CCL4; D006023:Glycoproteins; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D016898:Interferon-alpha; D007375:Interleukin-1; D015850:Interleukin-6; D019402:Macrophage Inflammatory Proteins; D009698:Nucleoproteins; D009711:Nucleotides; D012367:RNA, Viral; D014409:Tumor Necrosis Factor-alpha; D014764:Viral Proteins; D018664:Interleukin-12