The Cellient System for Cytohistology to Analyze p16 Positive Dyskeratocytes in Paraffin Sections of HPV-Positive Cervical Scrapes

Authors

  • Mathilde E. Boon Leiden Cytology and Pathology Laboratory (LCPL), Leiden, The Netherlands
  • Joke A.M. Dols Erasmus MC, University Medical Center Rotterdam, Rotterdam, The Netherlands
  • Nicodemus Butamanya Sekou-Toure Regional Hospital, Mwanza, Tanzania
  • Gregor Reid Lawson Health Research Institute, Canadian Research & Development Centre for Probiotics, Departments of Microbiology, Immunology and Surgery, the University of Western Ontario, London, Canada

DOI:

https://doi.org/10.6000/1927-7229.2012.01.01.13

Keywords:

Cellient, HPV, dyskeratosis, p16, HIV, BoonFix

Abstract

The Cellient™ Automated Cell Block System (Hologic) can be used to analyze cells of HPV-positive cervical scrapes staining positive with the biomarker p16. For this study fourteen cervical scrapes of Tanzanian women infected with HIV testing positive for HPV were selected. The paraffin Cellient sections were stained with the Papanicolaou method, with hematoxylin eosin (HE), and with the biomarker p16. This pilot study was limited to cases classified as atypical squamous lesion of unknown significance (ASCUS) and high-grade squamous lesion (HSIL) as diagnosed in the ThinPrep slide. The Cellient paraffin sections (cut from paraffin blocks prepared from the residual cervical sample) were classified into negative, atypical, CIN 1, CIN 2, and CIN 3. Multiple HPV genotypes were encountered in 79% of the scrapes. HPV16 was found in six scrapes and HPV52 in four. In the Papanicolaou sections, it was easy to detect dyskeratotic cells. Eleven of the 14 cases were p16 positive and five contained p16 positive dyskeratocytes. Of the 10 ASCUS scrapes, two contained p16 positive CIN 1 epithelial fragments. All four HSIL cases contained p16 positive CIN 3epithelial fragments. In HIV-positive HPV-positive women, the Cellient system resulted in high quality histology sections with perfect p16 images of dyskeratocytes.

References

Wagner DG, Russell DK, Benson JM, Schneider AE, Hoda RS, Bonfiglio TA. Cellient™ automated cell block versus traditional cell block preparation: A comparison of morphologic features and immunohistochemical staining. Diagn Cytopathol 2011; 39: 730-36. http://dx.doi.org/10.1002/dc.21457

Koss LG, Melamed MR, Eds. Koss’ Diagnostic Cytology and its Histopathologic Bases. 5th ed. Philadelphia–Tokyo: Lippincott Williams & Wilkins, 2006; p. 333.

Solomon D, Davey D, Kurman R, Moriarty A, O'Connor D, Prey M, Raab S, Sherman M, Wilbur D, Wright T Jr, Young N; Forum Group Members; Bethesda 2001 Workshop. The 2001 Bethesda System: terminology for reporting results of cervical cytology. JAMA 2002; 287: 2114-9. http://dx.doi.org/10.1001/jama.287.16.2114

Reid G, Anand S, Bingham MO, et al. Probiotics for the developing world. J Clin Gastroenterol 2005; 39: 485-88. http://dx.doi.org/10.1097/01.mcg.0000165648.32371.38

Dols JAM, Boon ME, Monachese M, et al. The impact of probiotic yogurt on HIV positive women in Tanzania. Int Dairy J 2011; (in press). http://dx.doi.org/10.1016/j.idairyj.2011.03.001

Kleter B, van Doorn LJ, Schrauwen L, et al. Development and clinical evaluation of a highly sensitive PCR-reverse hybridization line probe assay for detection and identification of anogenital human papillomavirus. J Clin Microbiol 1999; 37: 2508-2517.

Dols JAM, Reid G, Kort R, et al. PCR-based identification of eight Lactobacillus species and 18 hr-HPV genotypes in fixed cervical samples of South African women at risk of HIV and BV. Diagn Cytopathol 2011; (in press).

Boon ME, Suurmeijer AJH. The Pap Smear. 3rd ed. Amsterdam–Tokyo: Harwood Academic Publishers, 1996.

Risse EK, Ouwerkerk-Noordam E, Meijer-Marres EM, Boon ME. Exploiting the residual of cervical thin layer brush samples through cytohistology in cases with invasive carcinoma with application of antibodies. Acta Cytol 2010; 54: 175-82. http://dx.doi.org/10.1159/000325004

Lambert AP, Anschau F, Schmitt VM. p16INK4a expression in cervical premalignant and malignant lesions. Exp Mol Pathol 2006; 80: 192-96. http://dx.doi.org/10.1016/j.yexmp.2005.08.005

Missaoui N, Hmissa S, Frappart L, et al. p16INK4a overexpression and HPV infection in uterine cervix adenocarcinoma. Virchows Arch 2006; 448: 597-603. http://dx.doi.org/10.1007/s00428-005-0141-x

Yoshida T, Fukuda T, Sano T, Kanuma T, Owada N, Nakajima T. Usefulness of liquid-based cytology specimens for the immunocytochemical study of p16INK4a expression and human papillomavirus testing: a comparative study using simultaneously sampled histology materials. Cancer 2004; 102: 100-108. http://dx.doi.org/10.1002/cncr.20046

Galgano MT, Castle PE, Atkins KA, Brix WK, Nassau SR, Stoler MH. Using biomarkers as objective standards in the diagnosis of cervical biopsies. Am J Surg Pathol 2010; 34: 1077-87. http://dx.doi.org/10.1097/PAS.0b013e3181e8b2c4

Boon ME, Ouwerkerk-Noordam E, Suurmeijer AJH, Kok LP. Diagnostic parameters in liquid-based cervical cytology using a coagulant suspension fixative. Acta Cytol 2005; 49: 513-19. http://dx.doi.org/10.1159/000326197

Boon ME, Kok LP. Theory and practice of combining coagulant fixation and microwave histoprocessing. Biotech Histochem 2008; 83: 261-77. http://dx.doi.org/10.1080/10520290802553476

Boon ME, Kok LP. Microwave Cookbook of Pathology: The Art of Microscopic Visualization. Leiden: Coulomb Press Leyden, 1987.

Kok LP, Boon ME. Microwaves for the Art of Microscopy. 4th rev ed. Leiden: Coulomb Press Leyden, 2003.

Risse EKJ, Holierhoek JP, Meijer-Marres EM, Ouwerkerk-Noordam E, Boon ME. Increasing diagnostic accuracy of atypical glandular cells in cervical liquid-based cytology using cell blocks. Cytopathology 2011; 22: 253-60. http://dx.doi.org/10.1111/j.1365-2303.2010.00793.x

Downloads

Published

2012-01-28

How to Cite

Mathilde E. Boon, Joke A.M. Dols, Nicodemus Butamanya, & Gregor Reid. (2012). The Cellient System for Cytohistology to Analyze p16 Positive Dyskeratocytes in Paraffin Sections of HPV-Positive Cervical Scrapes . Journal of Analytical Oncology, 1(1), 88–94. https://doi.org/10.6000/1927-7229.2012.01.01.13

Issue

Section

Articles