title: Molecular characterization of HTLV-1 gp46 glycoprotein from health carriers and HAM/TSP infected individuals.
abstract: BACKGROUND
  Human T-cell Leukemia Virus type 1 (HTLV-1) is the etiological agent of tropical spastic paraparesis/HTLV-associated myelopathy (HAM/TSP) that can be identified in around 0.25%-3.8% of the infected population. Disease progression can be monitored by the proviral load and may depend on genetic factors, however, it is not well understood why some HTLV-1 infected people develop the disease while others do not. The present study attempts to assess the molecular diversity of gp46 glycoprotein in HAM/TSP patients and Health Carrier (HC) individuals.
  
  
  METHODS
  Blood samples were collected from 10 individuals, and DNA was extracted from PBMCs to measure the HTLV-1 proviral load. The gp46 coding sequences were amplified PCR, cloned and sequenced. The molecular characterization was performed using bioinformatics tools.
  
  
  RESULTS
  The median HTLV-1 proviral load of HC (n = 5) and HAM/TSP (n = 5) patients was similar (average 316,227 copies/106 PBMCs). The gp46 molecular characterization of 146 clones (70 HC and 76 HAM/TSP) revealed an overall diversity, within HC and HAM/TSP clones, of 0.4% and 0.6%, respectively. Five frequent mutations were detected among groups (HAM/TSP and HC clone sequences). A single amino acid (aa) substitution (S35L) was exclusive for the HC group, and three gp46 substitutions (F14S, N42H, G72S) were exclusive for the HAM/TSP group. The remaining frequent mutation (V247I) was present in both groups (p = 0.0014). The in silico protein analysis revealed that the mutated alleles F14S and N42H represent more hydrophilic and flexible protein domains that are likely to be less antigenic. The Receptor Binding Domain is quite variable in the HAM/TSP group. Two other domains (aa 53-75 and 175-209) that contain multiple linear T-cell epitopes showed genetic diversity in both HAM/TSP and HC groups. Further analysis revealed 27 and 13 T-cell epitopes for class I HLA alleles and class II HLA alleles, when analyzing the entire gp46.
  
  
  CONCLUSIONS
  The most common gp46 mutations were not associated clinical status because they were found in only one individual, except for the V247I mutation, that was found at viral clones from HAM/TSP ad HC individuals. Because of this, we cannot associate any of the gp46 found mutations with the clinical profile.
authors.affiliation: Centro de Pesquisa Gonçalo Moniz, Fundação Oswaldo Cruz, CPqGM/FIOCRUZ, Rua Waldemar Falcão 121, Brotas, Salvador, Bahia, 40295-001, Brazil.
authors.forename: Aline C A
authors.initials: AC
authors.lastname: Mota-Miranda
authors.forename: Fernanda K
authors.initials: FK
authors.lastname: Barreto
authors.forename: Maria F C
authors.initials: MF
authors.lastname: Amarante
authors.forename: Everton
authors.initials: E
authors.lastname: Batista
authors.forename: Joana P
authors.initials: JP
authors.lastname: Monteiro-Cunha
authors.forename: Lourdes
authors.initials: L
authors.lastname: Farre
authors.forename: Bernardo
authors.initials: B
authors.lastname: Galvão-Castro
authors.forename: Luiz C J
authors.initials: LC
authors.lastname: Alcantara
mesh_terms: D000328:Adult; D000368:Aged; D000595:Amino Acid Sequence; D002353:Carrier State / Q000276:immunology / Q000821:virology*; D018984:Epitopes, T-Lymphocyte / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D005260:Female; D015686:Gene Products, env / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D015368:Human T-lymphotropic virus 1 / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D006801:Humans; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D009154:Mutation; D015493:Paraparesis, Tropical Spastic / Q000276:immunology / Q000821:virology*; D017434:Protein Structure, Tertiary; D015682:Retroviridae Proteins, Oncogenic / Q000737:chemistry / Q000235:genetics* / Q000276:immunology
chemical_list: D018984:Epitopes, T-Lymphocyte; D015686:Gene Products, env; D015682:Retroviridae Proteins, Oncogenic; C055266:gp46 protein, Human T-cell leukemia virus type I