Different methods of cell quantification can lead to different results: a comparison of digital methods using a pilot study of dendritic cells in HIV-positive patients

• Diego Tetzner Fernandes ^[3] ; Willie F.P. van Heerden ^[1] ; Ana Carolina Prado Ribeiro ^[4] ; Thaís Bianca Brandão ^[5] ; Evandro Sobroza de Mello ^[6] ; Cesar Rivera ^[2] ; Marlene B. van Heerden ^[1] ; Rogerio Gondak ^[7] ; Alan Roger Santos-Silva ^[3] ; Pablo Agustin Vargas ^[8] ; Marcio Ajudarte Lopes ^[3]
  1. [1] University of Pretoria

     University of Pretoria

     City of Tshwane, Sudáfrica

     
  2. [2] Universidad de Talca

     Universidad de Talca

     Provincia de Talca, Chile

     
  3. [3] DDS, MSc, PhD. Oral Diagnosis Department, Piracicaba Dental School, University of Campinas (UNICAMP), Piracicaba, São Paulo, Brazil
  4. [4] DDS, MSc, PhD. Oral Diagnosis Department, Piracicaba Dental School, University of Campinas (UNICAMP), Piracicaba, São Paulo, Brazil; DDS, MSc, PhD. Dental Oncology Service, Instituto do Câncer do Estado de São Paulo, ICESP-FMUSP, São Paulo, Brazil
  5. [5] DDS, MSc, PhD. Dental Oncology Service, Instituto do Câncer do Estado de São Paulo, ICESP-FMUSP, São Paulo, Brazil
  6. [6] MD, MSc, PhD. Dental Oncology Service, Instituto do Câncer do Estado de São Paulo, ICESP-FMUSP, São Paulo, Brazil
  7. [7] DDS, MSc, PhD. Department of Pathology, Federal University of Santa Catarina, Florianopólis, Santa Catarina, Brazil
  8. [8] DDS, MSc, PhD. Oral Diagnosis Department, Piracicaba Dental School, University of Campinas (UNICAMP), Piracicaba, São Paulo, Brazil; DDS, MSc, PhD. Department of Oral Pathology and Oral Biology, School of Dentistry, University of Pretoria, Pretoria, South Africa

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• Localización: Medicina oral, patología oral y cirugía bucal. Ed. inglesa, ISSN-e 1698-6946, Vol. 25, Nº. 3 (May), 2020
• Idioma: inglés
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• Resumen

  • Although new digital pathology tools have improved the positive cell quantification, there is a heterogeneity of the quantification methods in the literature. The aim of this study was to evaluate and propose a novel dendritic cells quantification method in squamous cell carcinoma comparing it with a conventional quantification method.

    Twenty-six squamous cell carcinomas HIV-positive cases affecting the oropharynx, lips and oral cavity were selected. Immunohistochemistry for CD1a, CD83, and CD207 was performed. The immunohistochemical stains were evaluated by automated examination using a positive pixel count algorithm. A conventional quantification method (unspecific area method; UA) and a novel method (specific area method; SA) were performed obtaining the corresponding density of positive dendritic cells for the intratumoral and peritumoral regions. The Mann-Whitney U test was used to verify the influence of the quantification methods on the positive cell counting according to the evaluated regions. Data were subjected to the ANOVA and Student’s t-test to verify the influence of the tumour location, stage, histological grade, and amount of inflammation on the dendritic cells density counting.

    The cell quantification method affected the dendritic cells counting independently of the evaluated region (P-value <0.05). Significant differences between methods were also observed according to the tumour features evaluations.

    The positive cell quantification method influences the dendritic cells density results. Unlike the conventional method (UA method), the novel SA method avoids non-target areas included in the hotspots improving the reliability and reproducibility of the density cell quantification.