title: Surface antigen profiles of leukocytes and melanoma cells in lymph node metastases are associated with survival in AJCC stage III melanoma patients.
abstract: There is an urgent need to identify more accurate prognostic biomarkers in melanoma patients, particularly in those with metastatic disease. This study aimed to identify melanoma and leukocyte surface antigens predictive of survival in a prospective series of AJCC stage IIIb/c melanoma patients (n = 29). Live cell suspensions were prepared from melanoma metastases within lymph nodes (LN). The suspensions were immuno-magnetically separated into CD45(+) (leukocyte) and CD45(-) (non-hematopoietic, enriched melanoma cell) fractions. Surface antigens on CD45(-) and CD45(+) cell populations were profiled using DotScan™ microarrays (Medsaic Pty. Ltd.) and showed differential abundance levels for 52 and 78 antigens respectively. Associations of the surface profiles with clinicopathologic and outcome data (median follow-up 35.4 months post LN resection) were sought using univariate (log-rank test) and multivariate (Wald's test; modelled with patient's age, gender and AJCC staging at LN recurrence) survival models. CD9 (p = 0.036), CD39 (p = 0.004) and CD55 (p = 0.005) on CD45(+) leukocytes were independently associated with distant metastasis-free survival using multivariate analysis. Leukocytes with high CD39 levels were also significantly associated with increased overall survival (OS) in multivariate analysis (p = 0.016). LNs containing leukocytes expressing CD11b (p = 0.025), CD49d (p = 0.043) and CD79b (p = 0.044) were associated with reduced OS on univariate analysis. For enriched melanoma cells (CD45(-) cell populations), 11 surface antigens were significantly correlated with the disease-free interval (DFI) between diagnosis of culprit primary melanoma and LN metastasis resection. Nine antigens on CD45(+) leukocytes also correlated with DFI. Following validation in independent datasets, surface markers identified here should enable more accurate determination of prognosis in stage III melanoma patients and provide better risk stratification of patients entering clinical trials.
authors.affiliation: School of Molecular Bioscience, University of Sydney, Sydney, NSW, 2006, Australia. kim.kaufman@sydney.edu.au.
authors.forename: Kimberley L
authors.initials: KL
authors.lastname: Kaufman
authors.affiliation: School of Molecular Bioscience, University of Sydney, Sydney, NSW, 2006, Australia.
authors.forename: Swetlana
authors.initials: S
authors.lastname: Mactier
authors.affiliation: Garvan Institute of Medical Research, Darlinghurst, NSW, 2010, Australia.|School of Mathematics and Statistics and Prince of Wales Clinical School, University of New South Wales, Kensington, NSW, 2052, Australia.
authors.forename: Nicola J
authors.initials: NJ
authors.lastname: Armstrong
authors.affiliation: School of Molecular Bioscience, University of Sydney, Sydney, NSW, 2006, Australia.
authors.forename: Duthika
authors.initials: D
authors.lastname: Mallawaaratchy
authors.affiliation: Discipline of Infectious Diseases and Immunology Sydney Medical School, University of Sydney, Sydney, NSW, 2006, Australia.|Discipline of Dermatology, Bosch Institute, Faculty of Medicine, Sydney Medical School, Sydney, NSW, 2006, Australia.
authors.forename: Scott N
authors.initials: SN
authors.lastname: Byrne
authors.affiliation: Melanoma Institute Australia, North Sydney, NSW, 2060, Australia.|Discipline of Surgery, Sydney Medical School, The University of Sydney, Sydney, NSW, 2006, Australia.
authors.forename: Lauren E
authors.initials: LE
authors.lastname: Haydu
authors.affiliation: Melanoma Institute Australia, North Sydney, NSW, 2060, Australia.
authors.forename: Valerie
authors.initials: V
authors.lastname: Jakrot
authors.affiliation: Melanoma Institute Australia, North Sydney, NSW, 2060, Australia.
authors.forename: John F
authors.initials: JF
authors.lastname: Thompson
authors.affiliation: Melanoma Institute Australia, North Sydney, NSW, 2060, Australia.|Westmead Institute of Cancer Research, The University of Sydney at Westmead Millennium Institute, Westmead, NSW, 2145, Australia.
authors.forename: Graham J
authors.initials: GJ
authors.lastname: Mann
authors.affiliation: Melanoma Institute Australia, North Sydney, NSW, 2060, Australia.|Department of Tissue Pathology and Diagnostic Oncology, Royal Prince Alfred Hospital, Camperdown, NSW, 2050, Australia.|Discipline of Pathology, Sydney Medical School, The University of Sydney, Sydney, NSW, 2006, Australia.
authors.forename: Richard A
authors.initials: RA
authors.lastname: Scolyer
authors.affiliation: School of Molecular Bioscience, University of Sydney, Sydney, NSW, 2006, Australia.
authors.forename: Richard I
authors.initials: RI
authors.lastname: Christopherson
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000954:Antigens, Surface / Q000378:metabolism*; D015153:Blotting, Western; D005260:Female; D006801:Humans; D007962:Leukocytes / Q000378:metabolism* / Q000473:pathology; D008207:Lymphatic Metastasis; D008297:Male; D008545:Melanoma / Q000378:metabolism / Q000401:mortality* / Q000473:pathology; D008875:Middle Aged; D009367:Neoplasm Staging; D011379:Prognosis; D011446:Prospective Studies; D012878:Skin Neoplasms / Q000378:metabolism / Q000401:mortality* / Q000556:secondary; D015996:Survival Rate; D014407:Tumor Cells, Cultured
chemical_list: D000954:Antigens, Surface
keywords: Antibody microarray; CD antigen; Metastatic melanoma; Prognosis; Survival