title: ELISA quantification of CCI-103F binding in canine tumors prior to and during irradiation.
abstract: OBJECTIVE
  The purpose of this work was to evaluate multiple injections of CCI-103F, a marker of hypoxia, as a method to quantify alterations in tumor hypoxia during irradiation.
  
  
  METHODS
  Twelve dogs with spontaneous solid tumors were given intravenous CCI-103F, and tumor biopsies were taken at various times after injection. Two tumor samples were taken at each biopsy procedure. CCI-103F antigen concentration was quantified by ELISA. Four of the dogs were given one injection of CCI-103F, and the other eight received two injections. In dogs receiving two injections, CCI-103F was administered before irradiation and 7 days later, following a total dose of 15.0 Gy. Plasma CCI-103F pharmacokinetics were assessed in dogs receiving two injections.
  
  
  RESULTS
  CCI-103F antigen was detectable in the initial biopsy in each of the four dogs receiving one injection, and the amount of detectable antigen decreased in subsequent biopsies with an initial half life of approximately 19 h. This suggests that multiple injections of CCI-103F could be used in the same subject to monitor tumor hypoxia as a function of time or during a course of treatment. In the eight dogs receiving two injections of CCI-103F, the CCI-103F antigen concentration in the 24 h samples ranged from 4.66-151.9 mumol CCI-103F antigen/kg tumor, a difference of a factor of approximately 33. The ratio of maximum to minimum concentration of CCI-103F antigen in 51 paired biopsy samples ranged from 1.01-4.07, with a mean (+/- s.d.) of 1.67 +/- 0.67. Seventy-five percent of the ratios were < or = 2.02. There was no apparent relationship between the magnitude of the ratio, i.e., intratumoral variation, and tumor volume or the absolute tumor concentration of CCI-103F antigen. Absolute radiobiologic hypoxic fraction was not known but the pattern of change in amount of intratumoral CCI-103F antigen in dogs given two injections of CCI-103F was consistent with little change in pretreatment oxygen status in six dogs, and an increase in tumor oxygenation in two dogs.
  
  
  CONCLUSIONS
  It appears possible to obtain an estimate of the change in tumor hypoxia in an individual tumor over time by assaying biopsy samples for CCI-103F antigen concentration.
authors.affiliation: Department of Anatomy, Physiological Sciences and Radiology, College of Veterinary Medicine, North Carolina State University, Raleigh 27606.
authors.forename: D E
authors.initials: DE
authors.lastname: Thrall
authors.forename: M C
authors.initials: MC
authors.lastname: McEntee
authors.forename: J M
authors.initials: JM
authors.lastname: Cline
authors.forename: J A
authors.initials: JA
authors.lastname: Raleigh
mesh_terms: D000818:Animals; D015687:Cell Hypoxia / Q000502:physiology*; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay; D009374:Neoplasms, Experimental / Q000378:metabolism* / Q000532:radiotherapy*; D009593:Nitroimidazoles / Q000378:metabolism; D005466:Fluorocarbons
chemical_list: D009593:Nitroimidazoles; C049948:CCI 103F; D005466:Fluorocarbons