title: Automated human blood micronucleated reticulocyte measurements for rapid assessment of chromosomal damage.
abstract: This study evaluated the utility of human blood micronucleated reticulocyte (MNCD71+) frequency measurement as a cytogenetic damage biomarker. The analytical methodology was flow cytometry in conjunction with a previously described three color fluorescence labeling technique that includes anti-CD71 to focus analyses on the most immature fraction of reticulocytes [S.D. Dertinger, K. Camphausen, J.T. MacGregor, M.E. Bishop, D.K. Torous, S. Avlasevich, et al., Three-color labeling method for flow cytometric measurement of cytogenetic damage in rodent and human blood, Environ. Mol. Mutagen. 44 (2004) 427-435]. Blood specimens from 50 self-reported healthy adult volunteers were studied. In addition to MNCD71+ measurements, blood plasma folate and B12 levels were assessed, since these variables tend to influence other indices of cytogenetic damage. Time-course data are also provided for 10 cancer patients undergoing treatment. For these subjects, frequency of MNCD71+ was measured immediately before therapy, and daily during the first week of chemotherapy and/or fractionated radiotherapy. For the group of healthy volunteers, the variables of age, and folate and B12 levels demonstrated no significant effect on MNCD71+ frequency. In addition, no difference was observed between pre-treatment MNCD71+ values for cancer patients compared with healthy volunteers. Regarding chemotherapy and/or partial body radiotherapy, elevated frequencies were observed upon initiation of treatment for 9 of the 10 patients studied. Maximal effects were observed 3-5 days following initiation of therapy. The largest increases in frequency of MNCD71+ (up to 25.9-fold) were observed in those patients exposed to anti-neoplastic drugs, presumably due to the systemic red marrow exposure provided by these agents. Taken together, these data support the hypothesis that the MNCD71+ endpoint represents a valuable biomarker of cytogenetic damage that does not require cell culture or microscopy-based scoring.
authors.affiliation: Litron Laboratories, 200 Canal View Boulevard, Rochester, NY, United States. sdertinger@litronlabs.com
authors.forename: Stephen D
authors.initials: SD
authors.lastname: Dertinger
authors.forename: Richard K
authors.initials: RK
authors.lastname: Miller
authors.forename: Kelly
authors.initials: K
authors.lastname: Brewer
authors.forename: Therese
authors.initials: T
authors.lastname: Smudzin
authors.forename: Dorothea K
authors.initials: DK
authors.lastname: Torous
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Roberts
authors.forename: Svetlana L
authors.initials: SL
authors.lastname: Avlasevich
authors.forename: Steven M
authors.initials: SM
authors.lastname: Bryce
authors.forename: Siva
authors.initials: S
authors.lastname: Sugunan
authors.forename: Yuhchyau
authors.initials: Y
authors.lastname: Chen
mesh_terms: D000328:Adult; D000970:Antineoplastic Agents / Q000627:therapeutic use; D001331:Automation; D002869:Chromosome Aberrations; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D008297:Male; D008875:Middle Aged; D009369:Neoplasms / Q000097:blood* / Q000188:drug therapy; D012156:Reticulocytes / Q000166:cytology* / Q000648:ultrastructure
chemical_list: D000970:Antineoplastic Agents