Standardisation of Western blotting to detect HTLV-1 antibodies synthesised in the central nervous system of HAM/TSP patients

Mem Inst Oswaldo Cruz. 2013 Sep;108(6):730-4. doi: 10.1590/0074-0276108062013009.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Viral* / blood
  • Antibodies, Viral* / cerebrospinal fluid
  • Blotting, Western / standards*
  • Central Nervous System / immunology*
  • Central Nervous System / metabolism
  • Enzyme-Linked Immunosorbent Assay
  • Gene Products, env / immunology
  • Gene Products, gag / immunology
  • Human T-lymphotropic virus 1 / immunology*
  • Humans
  • Immunoglobulin G / blood
  • Immunoglobulin G / cerebrospinal fluid
  • Paraparesis, Tropical Spastic / blood
  • Paraparesis, Tropical Spastic / cerebrospinal fluid
  • Paraparesis, Tropical Spastic / immunology*
  • Sensitivity and Specificity

Substances

  • Antibodies, Viral
  • Gene Products, env
  • Gene Products, gag
  • Immunoglobulin G