CytoLyt fixation significantly inhibits MIB1 immunoreactivity whereas alternative Ki-67 clone 30-9 is not susceptible to the inhibition: Critical diagnostic implications Journal Article


Authors: Buonocore, D. J.; Konno, F.; Jungbluth, A. A.; Frosina, D.; Fayad, M.; Edelweiss, M.; Lin, O.; Rekhtman, N.
Article Title: CytoLyt fixation significantly inhibits MIB1 immunoreactivity whereas alternative Ki-67 clone 30-9 is not susceptible to the inhibition: Critical diagnostic implications
Abstract: Background: The Ki-67 proliferation marker has multiple diagnostic and prognostic applications. Although several clones to the Ki-67 antigen are commercially available, the MIB1 clone is widely recommended in the surgical pathology literature for neuroendocrine tumors. In our cytopathology practice, we have encountered unexpectedly low MIB1 immunoreactivity in CytoLyt-fixed cell blocks (CBs). The current study evaluated the impact of fixatives, CB processing, and immunocytochemical (ICC) procedures on Ki-67 immunoreactivity. Methods: Test CBs were prepared from freshly resected tumors, and multiple variables in the MIB1 ICC procedure were tested, including CytoLyt versus formalin collection media, MIB1 versus other Ki-67 clones including 30-9, and other variables. MIB1 versus Ki-67 30-9 clones were tested in parallel on CytoLyt-fixed CBs from clinical samples of small cell lung carcinoma (SCLC). Results: In the test CBs (n = 10), the mean MIB1 labeling index was 10% in CytoLyt versus 47% in formalin (P =.0116), with a mean loss of reactivity in matched CBs of 37% (up to 70%). None of the procedure modifications tested in 223 individual ICC reactions recovered MIB1 reactivity in CytoLyt except for switching to the Ki-67 30-9 antibody. In CytoLyt-fixed SCLC samples (n = 14), the Ki-67 30-9 antibody demonstrated expected ranges of reactivity (mean, 83%; range, 60%-100%), whereas MIB1 demonstrated markedly inhibited labeling (mean, 60%; range, 10%-95%) (P =.0058). Conclusions: CytoLyt fixation substantially inhibits MIB1 immunoreactivity, whereas the Ki-67 30-9 clone is not susceptible to inhibition. Markedly discrepant MIB1 reactivity may present a pitfall in the diagnosis of SCLC and may lead to the incorrect prognostic stratification of other tumor types. For laboratories using CytoLyt, we recommend using the Ki-67 30-9 antibody rather than the MIB1 antibody. © 2019 American Cancer Society
Keywords: immunohistochemistry; controlled study; human tissue; cancer surgery; unclassified drug; human cell; cancer diagnosis; ki 67 antigen; cytology; cancer susceptibility; immunoreactivity; molecular cloning; immunocytochemistry; quantitative analysis; ki-67; ex vivo study; cell adhesion; cytopathology; small cell carcinoma; small cell lung cancer; intracellular signaling; methanol; antibody labeling; antigen retrieval; human; priority journal; article; prognostic assessment; cytolyt; mib1
Journal Title: Cancer Cytopathology
Volume: 127
Issue: 10
ISSN: 1934-662X
Publisher: John Wiley & Sons  
Date Published: 2019-10-01
Start Page: 643
End Page: 649
Language: English
DOI: 10.1002/cncy.22170
PUBMED: 31398281
PROVIDER: scopus
PMCID: PMC8375359
DOI/URL:
Notes: Article -- Export Date: 1 November 2019 -- Source: Scopus
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MSK Authors
  1. Natasha Rekhtman
    424 Rekhtman
  2. Oscar Lin
    307 Lin
  3. Marcia Edelweiss
    104 Edelweiss
  4. Achim Jungbluth
    454 Jungbluth
  5. Denise Frosina
    123 Frosina
  6. Fumiko Konno
    11 Konno
  7. Miriam   Fayad
    14 Fayad