title: A novel data mining system points out hidden relationships between immunological markers in multiple sclerosis.
abstract: BACKGROUND
  Multiple Sclerosis (MS) is a multi-factorial disease, where a single biomarker unlikely can provide comprehensive information. Moreover, due to the non-linearity of biomarkers, traditional statistic is both unsuitable and underpowered to dissect their relationship. Patients affected with primary (PP=14), secondary (SP=33), benign (BB=26), relapsing-remitting (RR=30) MS, and 42 sex and age matched healthy controls were studied. We performed a depth immune-phenotypic and functional analysis of peripheral blood mononuclear cell (PBMCs) by flow-cytometry. Semantic connectivity maps (AutoCM) were applied to find the natural associations among immunological markers. AutoCM is a special kind of Artificial Neural Network able to find consistent trends and associations among variables. The matrix of connections, visualized through minimum spanning tree, keeps non linear associations among variables and captures connection schemes among clusters.
  
  
  RESULTS
  Complex immunological relationships were shown to be related to different disease courses. Low CD4IL25+ cells level was strongly related (link strength, ls=0.81) to SP MS. This phenotype was also associated to high CD4ROR+ cells levels (ls=0.56). BB MS was related to high CD4+IL13 cell levels (ls=0.90), as well as to high CD14+IL6 cells percentage (ls=0.80). RR MS was strongly (ls=0.87) related to CD4+IL25 high cell levels, as well indirectly to high percentages of CD4+IL13 cells. In this latter strong (ls=0.92) association could be confirmed the induction activity of the former cells (CD4+IL25) on the latter (CD4+IL13). Another interesting topographic data was the isolation of Th9 cells (CD4IL9) from the main part of the immunological network related to MS, suggesting a possible secondary role of this new described cell phenotype in MS disease.
  
  
  CONCLUSIONS
  This novel application of non-linear mathematical techniques suggests peculiar immunological signatures for different MS phenotypes. Notably, the immune-network displayed by this new method, rather than a single marker, might be viewed as the right target of immunotherapy. Furthermore, this new statistical technique could be also employed to increase the knowledge of other age-related multifactorial disease in which complex immunological networks play a substantial role.
authors.affiliation: Semeion Research Center, Via Sersale 117, Rome, 00128, Italy. enzo.grossi@bracco.com.
authors.forename: Maira
authors.initials: M
authors.lastname: Gironi
authors.forename: Marina
authors.initials: M
authors.lastname: Saresella
authors.forename: Marco
authors.initials: M
authors.lastname: Rovaris
authors.forename: Matilde
authors.initials: M
authors.lastname: Vaghi
authors.forename: Raffaello
authors.initials: R
authors.lastname: Nemni
authors.forename: Mario
authors.initials: M
authors.lastname: Clerici
authors.forename: Enzo
authors.initials: E
authors.lastname: Grossi