Abstract
Intrathecal synthesis of human T-lymphotropic virus type 1 (HTLV-1) antibodies (Abs) represents conclusive evidence of a specific immune response in the central nervous system of HTLV-1 associated myelopathy/tropical spastic paraparesis (HAM/TSP) patients. Western blotting (WB) for HTLV Abs in serum is a confirmatory test for HTLV-1 infection. The aim of this study was to standardise the Western blot to demonstrate the intrathecal pattern of Abs against HTLV-1 proteins in HAM/TSP patients. Paired cerebrospinal fluid (CSF) and serum samples were selected from 20 patients with definite HAM/TSP, 19 HTLV-1 seronegative patients and two HTLV-1 patients without definite HAM/TSP. The presence of reactive bands of greater intensity in the CSF compared to serum (or bands in only the CSF) indicated the intrathecal synthesis of anti-HTLV-1 Abs. All definite HAM/TSP patients presented with an intrathecal synthesis of anti-HTLV-1 Abs; these Abs were not detected in the control patients. The most frequent intrathecal targets of anti-HTLV-1 Abs were GD21, rgp46-I and p24 and, to a lesser extent, p19, p26, p28, p32, p36, p53 gp21 and gp46. The intrathecal immune response against env (GD21 and rgp46-I) and gag (p24) proteins represents the most important humoral pattern in HAM/TSP. This response may be used as a diagnostic marker, considering the frequent association of intrathecal anti-HTLV-1 Ab synthesis with HAM/TSP and the pathogenesis of this neurological disease.
Highlights
HTLV proteins in the central nervous system (CNS) Luiz Claudio Pereira Ribeiro et al 731 toxoplasmosis) and group 3 contained samples from two patients who were seropositive for human T-lymphotropic virus type 1 (HTLV-1) [Parkinson’s disease and probably HTLV-1 associated myelopathy/tropical spastic paraparesis (HAM/TSP) characterised by Babinski sign and HTLV-1 Abs in the serum and cerebrospinal fluid (CSF), according to only the criteria by de Castro-Costa et al (2006)]
The results indicated the intrathecal synthesis of anti-HTLV-1 Abs against the same viral proteins (GD21, p19/gp21, p24, p26, p28, p32, p36, gp46, p53 and rgp46-I) as those found in the trial conducted at HUGG (Figure)
The Western blotting (WB) test has previously been used to confirm the presence of anti-HTLV-1 Abs in serum and CSF (Osame et al 1987, Ceroni et al 1988, Gessain et al 1988, Link et al 1989)
Summary
HTLV proteins in the CNS Luiz Claudio Pereira Ribeiro et al 731 toxoplasmosis) and group 3 contained samples from two patients who were seropositive for HTLV-1 [Parkinson’s disease and probably HAM/TSP characterised by Babinski sign and HTLV-1 Abs in the serum and CSF, according to only the criteria by de Castro-Costa et al (2006)]. Screening for anti-HTLV-1 Abs - All patients underwent serological screening for HTLV-1 using the ELISA method (Vironostika HTLV-1, Organon, Teknika, Durham, NC) with subsequent confirmation of the reactive cases by WB (HTLV-BLOT 2.4, MP Diagnostics, MP Biomedicals Asia Pacific Pte Ltd, Singapore). CSF samples were screened for anti-HTLV-1 Abs using the ELISA method (Vironostika HTLV-1, Organon, Teknika, Durham, NC). To interpret the WB test, samples that reacted to at least one protein from the gag gene and one protein from the env gene, including rgp46-I, were considered positive for HTLV-1. WB test to evaluate the intrathecal synthesis of antiHTLV-1 Abs - To detect the intrathecal synthesis of antiHTLV-1 Abs we applied the WB test (HTLV-BLOT 2.4, MP Diagnostics, MP Biomedicals Asia Pacific Pte Ltd, Singapore) according to the manufacturer’s instructions, except in the dilution of the samples. The intrathecal synthesis of anti-HTLV-1 Abs against gag genes (p19, p24, p26, p28, p32, p36 and p53) and env genes (GD21, gp, gp46 and rgp46-I) was analysed
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