In vitro cytoskeleton changes of mouse neurons induced by purified HTLV-1, and PBMC from HAM/TSP patients and HTLV-1 carriers

Abstract

HTLV-1 is the causative agent of HAM/TSP. This neurological disease affects the CNS producing damage of the motor tracts at the spinal cord. The HAM/TSP pathogenesis remains undefined. It could include direct and indirect actions of HTLV-1. We studied the effect of purified HTLV-1 and the PBMC of 22 Chilean patients co-cultivated with fetal neurons of mouse (CNh cells): 8 HAM/TSP, 8 HTLV-1 carriers, and 6 non-infected controls. The viral antigens and provirus in CNh cells was evaluated with monoclonal and polyclonal antibodies reacting with HTLV-1 by immunofluorescence assay and PCR at 0, 7 and 15 days of co-cultures, respectively. Viral antigens were detected in 0.1-0.5%, and 0-0.3% of the neurons incubated with lymphocytes of HAM/TSP patients and HTLV-1 carriers, respectively. Neurons incubated with cells of 7 HAM/TSP patients, and 3 HTLV-1 carriers showed the presence of nucleotide sequences of tax gene. These results would be showing that CNh cells would express viral antigens and provirus. The HTLV-1 or their proteins were capable in vitro to produce structural and growth changes in the cytoskeleton of CNh neurons. In this series, the purified HTLV-1 was more effective in the neural changes than PBMC of HAM/TSP or HTLV-1 carriers.

Prevalence of human T-Cell lymphotropic virus type 1 and 2 among patients with malignant hematological diseases in South Chile

“…The region of the tax gene was amplified with the primers SK43: 5′‐CGG ATA CCC AGT CTA CGT GT‐3′ and SK44: 5′‐GAG CCG ATA ACG CGT CCA TCG‐3′. For the second round of nested PCR TAX1: 5′‐GTG TTT GGC GAT TGT GTA CA‐3′ and TAX2: 5′‐CCA TCG ATG GGG TCC CA‐3′ were used [Ramirez et al, 2004; Ramirez et al, 2007]. The primers were used at a final concentration of 0.3 µM in a reaction mixture of 25 µl containing 1–2 µg of DNA sample, 0.5 units of KOD HiFi DNA polymerase (KOD Hot Start, Novagen, TOYOBO), KOD Hot Start buffer 1×, 1.5 mM MgSO 4 and 0.2 mM of each dNTP.…”

Seroprevalence of human T-cell lymphotropic virus type 1 and 2 in blood donors from the regional hospital of Valdivia, Chile

“…Positive samples by ELISA were sent to the Public Health Institute of the Chilean Ministry of Health for confirmation. The now mandatory confirmation techniques in Chile are indirect immunofluorescence (IFI) performed in MT2 cells for HTLV-1 and PCR to confirm HTLV-1/2 infection in PBMC cells [22,23].…”

In vivo fluctuation of Tax, Foxp3, CTLA-4, and GITR mRNA expression in CD4+CD25+ T cells of patients with human T-lymphotropic virus type 1-associated myelopathy

“…All subjects resided in Santiago of Chile and had Spanish ethnic background. We confirmed HTLV-1 infection by immunofluorescence and PCR assays (18). The diagnosis of HAM/TSP was made according to the World Health Organization guidelines (19).…”