T-Cell response profiling to biological threat agents including the SARS coronavirus.
The emergence of pathogens such as SARS and the increased threat of bioterrorism has stimulated the development of novel diagnostic assays for differential diagnosis. Rather than focusing on the detection of an individual pathogen component, we have developed a T cell profiling system to monitor responses to the pathogens in an array format. Using a matrix of antigens specific for different pathogens, a specific T cell profile was generated for each individual by monitoring the intracellular production of interferon-gamma by flow cytometry. This assay allows for the testing of multiple proteins or peptides at a single time and provides a quantitative and phenotypic assessment of CD4(+) and CD8(+) responding cells. We present profiling examples for several positive individuals, including those vaccinated with the smallpox and anthrax vaccines. We also show antigen optimization for the SARS-hCoV, as studies revealed that these proteins contain peptides which cross-react with more common coronaviruses, a cause of the common cold. The T cell array is an early and sensitive multiplex measure of active infection, exposure to a pathogen, or effective, recent vaccination.
Gioia
C
C
Unit of Cellular Immunology, National Institute for Infectious Diseases (I.N.M.I.) Lazzaro Spallanzani I.R.C.C.S., Rome, Italy.
Horejsh
D
D
Agrati
C
C
Martini
F
F
Capobianchi
M R
MR
Ippolito
G
G
Poccia
F
F
D000956:Antigens, Viral / Q000276:immunology; D023482:Bioterrorism; D002478:Cells, Cultured; D018352:Coronavirus Infections / Q000276:immunology* / Q000821:virology; D005434:Flow Cytometry; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis; D015759:Ionomycin / Q000494:pharmacology; D007476:Ionophores / Q000494:pharmacology; D007963:Leukocytes, Mononuclear; D015240:Phorbol 12,13-Dibutyrate / Q000494:pharmacology; D045169:Severe Acute Respiratory Syndrome / Q000276:immunology* / Q000821:virology; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology*
D000956:Antigens, Viral; D007476:Ionophores; D015240:Phorbol 12,13-Dibutyrate; D015759:Ionomycin; D007371:Interferon-gamma