title: Humoral factors in ALS patients during disease progression.
abstract: BACKGROUND
  Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease affecting upper and lower motor neurons in the CNS and leading to paralysis and death. There are currently no effective treatments for ALS due to the complexity and heterogeneity of factors involved in motor neuron degeneration. A complex of interrelated effectors have been identified in ALS, yet systemic factors indicating and/or reflecting pathological disease developments are uncertain. The purpose of the study was to identify humoral effectors as potential biomarkers during disease progression.
  
  
  METHODS
  Thirteen clinically definite ALS patients and seven non-neurological controls enrolled in the study. Peripheral blood samples were obtained from each ALS patient and control at two visits separated by 6 months. The Revised ALS Functional Rating Scale (ALSFRS-R) was used to evaluate overall ALS-patient functional status at each visit. Eleven humoral factors were analyzed in sera. Cytokine levels (GM-CSF, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, and TNF-α) were determined using the Bio-Rad Bio-Plex® Luminex 200 multiplex assay system. Nitrite, a breakdown product of NO, was quantified using a Griess Reagent System. Glutathione (GSH) concentrations were measured using a Glutathione Fluorometric Assay Kit.
  
  
  RESULTS
  ALS patients had ALSFRS-R scores of 30.5 ± 1.9 on their first visit and 27.3 ± 2.7 on the second visit, indicating slight disease progression. Serum multiplex cytokine panels revealed statistically significant changes in IL-2, IL-5, IL-6, and IL-8 levels in ALS patients depending on disease status at each visit. Nitrite serum levels trended upwards in ALS patients while serum GSH concentrations were drastically decreased in sera from ALS patients versus controls at both visits.
  
  
  CONCLUSIONS
  Our results demonstrated a systemic pro-inflammatory state and impaired antioxidant system in ALS patients during disease progression. Increased levels of pro-inflammatory IL-6, IL-8, and nitrite and significantly decreased endogenous antioxidant GSH levels could identify these humoral constituents as systemic biomarkers for ALS. However, systemic changes in IL-2, IL-5, and IL-6 levels determined between visits in ALS patients might indicate adaptive immune system responses dependent on current disease stage. These novel findings, showing dynamic changes in humoral effectors during disease progression, could be important for development of an effective treatment for ALS.
authors.affiliation: Saneron CCEL Therapeutics, Inc., Tampa, FL, USA. jehrhar1@health.usf.edu.
authors.forename: Jared
authors.initials: J
authors.lastname: Ehrhart
authors.affiliation: Center of Excellence for Aging & Brain Repair, University of South Florida, Morsani College of Medicine, Tampa, FL, USA. asmith1@health.usf.edu.
authors.forename: Adam J
authors.initials: AJ
authors.lastname: Smith
authors.affiliation: Saneron CCEL Therapeutics, Inc., Tampa, FL, USA. nkn@saneron-ccel.com.
authors.forename: Nicole
authors.initials: N
authors.lastname: Kuzmin-Nichols
authors.affiliation: Department of Neurology, University of South Florida, Morsani College of Medicine, Tampa, FL, USA. tzesiewi@health.usf.edu.
authors.forename: Theresa A
authors.initials: TA
authors.lastname: Zesiewicz
authors.affiliation: Department of Neurology, University of South Florida, Morsani College of Medicine, Tampa, FL, USA. ijahan@health.usf.edu.
authors.forename: Israt
authors.initials: I
authors.lastname: Jahan
authors.affiliation: Center of Excellence for Aging & Brain Repair, University of South Florida, Morsani College of Medicine, Tampa, FL, USA. dshytle@health.usf.edu.|Department of Neurosurgery and Brain Repair, University of South Florida, Morsani College of Medicine, 12901 Bruce B. Downs Blvd., Tampa, FL, 33612, USA. dshytle@health.usf.edu.
authors.forename: R Douglas
authors.initials: RD
authors.lastname: Shytle
authors.affiliation: Center of Excellence for Aging & Brain Repair, University of South Florida, Morsani College of Medicine, Tampa, FL, USA. skim13@health.usf.edu.
authors.forename: Seol-Hee
authors.initials: SH
authors.lastname: Kim
authors.affiliation: Saneron CCEL Therapeutics, Inc., Tampa, FL, USA. Cdh@saneron-ccel.com.
authors.forename: Cyndy D
authors.initials: CD
authors.lastname: Sanberg
authors.affiliation: Department of Neurology, University of South Florida, Morsani College of Medicine, Tampa, FL, USA. tvu6@health.usf.edu.
authors.forename: Tuan H
authors.initials: TH
authors.lastname: Vu
authors.affiliation: Department of Neurology, University of South Florida, Morsani College of Medicine, Tampa, FL, USA. cgooch@health.usf.edu.
authors.forename: Clifton L
authors.initials: CL
authors.lastname: Gooch
authors.affiliation: Center of Excellence for Aging & Brain Repair, University of South Florida, Morsani College of Medicine, Tampa, FL, USA. psanberg@usf.edu.|Department of Neurosurgery and Brain Repair, University of South Florida, Morsani College of Medicine, 12901 Bruce B. Downs Blvd., Tampa, FL, 33612, USA. psanberg@usf.edu.|Department of Pathology and Cell Biology, University of South Florida, Morsani College of Medicine, Tampa, FL, USA. psanberg@usf.edu.|Department of Psychiatry, University of South Florida, Morsani College of Medicine, Tampa, FL, USA. psanberg@usf.edu.
authors.forename: Paul R
authors.initials: PR
authors.lastname: Sanberg
authors.affiliation: Center of Excellence for Aging & Brain Repair, University of South Florida, Morsani College of Medicine, Tampa, FL, USA. sgarbuzo@health.usf.edu.|Department of Neurosurgery and Brain Repair, University of South Florida, Morsani College of Medicine, 12901 Bruce B. Downs Blvd., Tampa, FL, 33612, USA. sgarbuzo@health.usf.edu.|Department of Molecular Pharmacology and Physiology, University of South Florida, Morsani College of Medicine, Tampa, FL, USA. sgarbuzo@health.usf.edu.|Department of Pathology and Cell Biology, University of South Florida, Morsani College of Medicine, Tampa, FL, USA. sgarbuzo@health.usf.edu.
authors.forename: Svitlana
authors.initials: S
authors.lastname: Garbuzova-Davis
mesh_terms: D000690:Amyotrophic Lateral Sclerosis / Q000097:blood* / Q000175:diagnosis*; D015415:Biomarkers / Q000097:blood; D016022:Case-Control Studies; D018450:Disease Progression; D005260:Female; D005978:Glutathione / Q000097:blood; D006801:Humans; D007376:Interleukin-2 / Q000097:blood*; D015848:Interleukin-5 / Q000097:blood*; D015850:Interleukin-6 / Q000097:blood*; D016209:Interleukin-8 / Q000097:blood; D008297:Male; D008875:Middle Aged; D009573:Nitrites / Q000097:blood; D011379:Prognosis
chemical_list: D015415:Biomarkers; D007376:Interleukin-2; D015848:Interleukin-5; D015850:Interleukin-6; D016209:Interleukin-8; D009573:Nitrites; D005978:Glutathione