Quantitative in situ PCR assay of HTLV-1 infected cells in peripheral blood lymphocytes of patients with ATL, HAM/TSP and asymptomatic carriers.
We established an in situ PCR (IS-PCR) method that amplified the pX region of human T cell lymphotropic virus type 1 (HTLV-1) proviral DNA. The procedure was highly sensitive in accurately detecting the number of cells infected with HTLV-1. We estimated the number of HTLV-1 infected cells in peripheral blood lymphocytes (PBL) from patients with HAM/TSP, ATL and asymptomatic carriers. ATL patients (n = 5) had 8-93% IS-PCR positive cells for HTLV-1 and these percentages correlated with the clinical stages. Asymptomatic carriers (n = 3) had 0.8-3.8% (mean 1.1%, S.D. 1.7) positive cells. HAM/TSP patients (n = 10) had 3.1-8.5% (5.8% (5.8%, 2.7) positive cells. Patients with shorter duration of illness showed larger percentages compared with patients with longer duration. In one HAM/TSP patient, the number of IS-PCR positive cells decreased from 5.1% to 1.5% coincident with the times of lymphocytapheresis treatment. Our studies may suggest that an increased viral load initiates the pathogenic process of HAM/TSP and the estimation of HTLV-1 proviral load by IS-PCR method is useful to understand the clinical state of HAM/TSP.
Third Department of Internal Medicine, Kagoshima University, Japan.
K
K
Hashimoto
I
I
Higuchi
M
M
Osame
S
S
Izumo
D000328:Adult; D000368:Aged; D001483:Base Sequence; D002353:Carrier State / Q000175:diagnosis* / Q000276:immunology; D005260:Female; D015490:HTLV-I Infections / Q000175:diagnosis* / Q000276:immunology; D015368:Human T-lymphotropic virus 1 / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D015459:Leukemia-Lymphoma, Adult T-Cell / Q000175:diagnosis* / Q000276:immunology; D008214:Lymphocytes / Q000166:cytology / Q000821:virology*; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D015493:Paraparesis, Tropical Spastic / Q000175:diagnosis* / Q000276:immunology; D016133:Polymerase Chain Reaction / Q000379:methods; D012016:Reference Values; D012680:Sensitivity and Specificity
D015345:Oligonucleotide Probes