title: Severe anaemia is not associated with HIV-1 env gene characteristics in Malawian children.
abstract: BACKGROUND
  Anaemia is the most common haematological complication of HIV and associated with a high morbidity and a poor prognosis. The pathogenesis of HIV-associated anaemia is poorly understood and may include a direct effect of HIV on erythropoiesis. In vitro studies have suggested that specific HIV strains, like X4 that uses the CXCR4 co-receptor present on erythroid precursors, are associated with diminished erythropoiesis. This co-receptor affinity is determined by changes in the hypervariable loop of the HIV-1 envelope genome. In a previous case-control study we observed an association between HIV and severe anaemia in Malawian children that could not be fully explained by secondary infections and micronutrient deficiencies alone. We therefore explored the possibility that alterations in the V1-V2-V3 fragment of HIV-1 were associated with severe anaemia.
  
  
  METHODS
  Using peripheral blood nucleic acid isolates of HIV-infected children identified in the previous studied we assessed if variability of the V1-V2-V3 region of HIV and the occurrence of X4 strains were more common in HIV-infected children with (cases, n = 29) and without severe anaemia (controls, n = 30). For 15 cases bone marrow isolates were available to compare against peripheral blood. All children were followed for 18 months after recruitment.
  
  
  RESULTS
  Phylogenetic analysis showed that HIV-1 subtype C was present in all but one child. All V1-V2-V3 characteristics tested: V3 charge, V1-V2 length and potential glycosylation sites, were not found to be different between cases and controls. Using a computer model (C-PSSM) four children (7.8%) were identified to have an X4 strain. This prevalence was not different between study groups (p = 1.00). The V3 loop characteristics for bone marrow and peripheral blood isolates in the case group were identical. None of the children identified as having an X4 strain developed a (new) episode of severe anaemia during follow up.
  
  
  CONCLUSIONS
  The prevalence of X4 strains in these young HIV-1-subtype-C-infected children that were most likely vertically infected and naïve to anti-retroviral therapy can be considered high compared to previous results from Malawi. It is unlikely that V1-V2-V3 fragment characteristics and HIV co-receptor affinity is an important feature in the development of severe anaemia in Malawian children.
authors.affiliation: Emma Children's Hospital, Academic Medical Centre, Amsterdam, The Netherlands. Job.Calis@gmail.com
authors.forename: Job C J
authors.initials: JC
authors.lastname: Calis
authors.forename: Hellen P
authors.initials: HP
authors.lastname: Rotteveel
authors.forename: Antoinette C
authors.initials: AC
authors.lastname: van der Kuyl
authors.forename: Fokla
authors.initials: F
authors.lastname: Zorgdrager
authors.forename: David
authors.initials: D
authors.lastname: Kachala
authors.forename: Michaël Boele
authors.initials: MB
authors.lastname: van Hensbroek
authors.forename: Marion
authors.initials: M
authors.lastname: Cornelissen
mesh_terms: D000740:Anemia / Q000150:complications* / Q000401:mortality / Q000821:virology; D001853:Bone Marrow / Q000821:virology; D016022:Case-Control Studies; D002675:Child, Preschool; D003430:Cross-Sectional Studies; D017931:DNA Primers; D005260:Female; D005500:Follow-Up Studies; D015752:Genes, env / Q000235:genetics*; D015658:HIV Infections / Q000097:blood / Q000150:complications* / Q000821:virology; D015497:HIV-1 / Q000145:classification / Q000235:genetics*; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008295:Malawi; D008297:Male; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D015995:Prevalence; D019718:Receptors, CXCR4 / Q000235:genetics / Q000378:metabolism*; D011795:Surveys and Questionnaires
chemical_list: D017931:DNA Primers; D019718:Receptors, CXCR4