Perivascular T cells are infected with HTLV-I in the spinal cord lesions with HTLV-I-associated myelopathy/tropical spastic paraparesis: double staining of immunohistochemistry and polymerase chain reaction in situ hybridization.
HTLV-I-infected cells play an important role in pathogenesis HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP). Our previous studies of quantitative polymerase chain reaction (PCR) and in situ PCR suggested that T cells infiltrating in the spinal cord lesion were infected with HTLV-I. To elucidate the localization of HTLV-I proviral DNA directly, we performed double staining using immunohistochemistry and PCR in situ hybridization (PCR-ISH). Fresh frozen sections of the spinal cord from four HAM patients taken at autopsy were first immunostained with antibodies to pan T cells (UCHL-1), macrophages (KP-1) and helper/inducer T cells (OPD4). Then PCR-ISH was carried out with specific primers and probe for the HTLV-I pX region. UCHL-1-positive cells were noted around perivascular areas and, to some extent, in the parenchyma. Of the UCHL-1-positive cells, 9.4% (case 1), 9.6% (case 2), 1.1% (case 3) and 6.7% (case 4) became positive in HTLV-I PCR-ISH. UCHL-1-negative cells were HTLV-I PCR-ISH negative and almost all KP-1-positive cells were HTLV-I negative. HTLV-I was localized to OPD4-positive cells in examined lesions of cases 2 and 4. These data are a direct demonstration of HTLV-I proviral DNA localizing to infiltrated T cells in HAM/TSP spinal cord lesions.
Matsuoka
E
E
The Division of Molecular Pathology and Genetic Epidemiology in Center for Chronic Viral Diseases, Kagoshima University School of Medicine, Sakuragaoka, Japan.
Takenouchi
N
N
Hashimoto
K
K
Kashio
N
N
Moritoyo
T
T
Higuchi
I
I
Isashiki
Y
Y
Sato
E
E
Osame
M
M
Izumo
S
S
D000368:Aged; D004279:DNA, Viral / Q000032:analysis; D006800:Deltaretrovirus Infections / Q000473:pathology / Q000821:virology*; D005260:Female; D015368:Human T-lymphotropic virus 1 / Q000302:isolation & purification*; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008297:Male; D008875:Middle Aged; D015493:Paraparesis, Tropical Spastic / Q000473:pathology / Q000821:virology*; D020286:Pericytes / Q000821:virology*; D016133:Polymerase Chain Reaction; D011533:Proviruses / Q000235:genetics; D012834:Silver; D013116:Spinal Cord / Q000473:pathology / Q000821:virology*; D013194:Staining and Labeling; D013601:T-Lymphocytes / Q000821:virology*
D004279:DNA, Viral; D012834:Silver