Intraepithelial CD15 infiltration identifies high-grade anal dysplasia in people with HIV
Joaquín Burgos, Aleix Benítez-Martínez, Cristina Mancebo, Núria Massana, Antonio Astorga-Gamaza, Josep Castellvi, Stefania Landolfi, Adrià Curran, Jorge N. Garcia-Perez, Vicenç Falcó, María J. Buzón, Meritxell Genescà
Joaquín Burgos, Aleix Benítez-Martínez, Cristina Mancebo, Núria Massana, Antonio Astorga-Gamaza, Josep Castellvi, Stefania Landolfi, Adrià Curran, Jorge N. Garcia-Perez, Vicenç Falcó, María J. Buzón, Meritxell Genescà
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Research Article Immunology Infectious disease

Intraepithelial CD15 infiltration identifies high-grade anal dysplasia in people with HIV

Abstract

Men who have sex with men (MSM) with HIV are at high risk for squamous intraepithelial lesion (SIL) and anal cancer. Identifying local immunological mechanisms involved in the development of anal dysplasia could aid treatment and diagnostics. Here, we studied 111 anal biopsies obtained from 101 MSM with HIV, who participated in an anal screening program. We first assessed multiple immune subsets by flow cytometry, in addition to histological examination, in a discovery cohort. Selected molecules were further evaluated by immunohistochemistry in a validation cohort. Pathological samples were characterized by the presence of resident memory T cells with low expression of CD103 and by changes in natural killer cell subsets, affecting residency and activation. Furthermore, potentially immunosuppressive subsets, including CD15+CD16+ mature neutrophils, gradually increased as the anal lesion progressed. Immunohistochemistry verified the association between the presence of CD15 in the epithelium and SIL diagnosis for the correlation with high-grade SIL. A complex immunological environment with imbalanced proportions of resident effectors and immune-suppressive subsets characterized pathological samples. Neutrophil infiltration, determined by CD15 staining, may represent a valuable pathological marker associated with the grade of dysplasia.

Authors

Joaquín Burgos, Aleix Benítez-Martínez, Cristina Mancebo, Núria Massana, Antonio Astorga-Gamaza, Josep Castellvi, Stefania Landolfi, Adrià Curran, Jorge N. Garcia-Perez, Vicenç Falcó, María J. Buzón, Meritxell Genescà

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Figure 2

Frequency of NK lymphocyte populations associates with SIL.

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Frequency of NK lymphocyte populations associates with SIL. (A) Flow cyt...
(A) Flow cytometry gating strategy quantifying anal tissue CD3– NK subsets for a nonpathological sample (top) and for a high-grade squamous intraepithelial lesion sample (HSIL, bottom). Expression of CD16+CD56– (pink), CD16–CD56+ (green), and CD16+CD56+ (yellow) was used to identify NK subsets, in which the activation/resident memory marker (CD69) and the activation marker (HLA-DR) were assessed. (B) Frequency of CD3–CD16+CD56– NK cells out of all living CD45+ cells in normal (in green) versus pathological (H/L-SIL, in purple; HSIL are highlighted in brown) samples. (C) Frequency of HLA-DR expression in total NK cells in normal versus pathological (H/L-SIL). (D) Frequency of HLA-DR expression in CD56+CD16– NK cells in normal versus pathological (H/L-SIL). (E) Frequency of CD69 expression in total NK cells in normal versus pathological (H/L-SIL). (F) Frequency of CD56+CD16+/– NK cells expressing CD69 in normal versus pathological tissue samples. (G) Frequency of CD69+CD103+ cells in total NK cells in normal (green), LSIL (purple), and HSIL (brown) anal samples. Data are represented as a violin plot; horizontal lines are median and interquartile range. Statistical comparisons using nonparametric Kruskal-Wallis test with Dunn’s post hoc test for multiple comparisons and Mann-Whitney U test for 2-group analyses are shown: *P < 0.05.