PT - JOURNAL ARTICLE AU - Kortekaas, Kim E. AU - Santegoets, Saskia J. AU - Abdulrahman, Ziena AU - van Ham, Vanessa J. AU - van der Tol, Marij AU - Ehsan, Ilina AU - van Doorn, Helena C. AU - Bosse, Tjalling AU - van Poelgeest, Mariëtte I. E. AU - van der Burg, Sjoerd H. TI - High numbers of activated helper T cells are associated with better clinical outcome in early stage vulvar cancer, irrespective of HPV or p53 status AID - 10.1186/s40425-019-0712-z DP - 2019 Dec 01 TA - Journal for ImmunoTherapy of Cancer PG - 236 VI - 7 IP - 1 4099 - http://jitc.bmj.com/content/7/1/236.short 4100 - http://jitc.bmj.com/content/7/1/236.full SO - J Immunother Cancer2019 Dec 01; 7 AB - Background Vulvar squamous cell carcinoma (VSCC) has been suggested to consist of three subtypes; HPV-positive, HPV-negative mutated TP53 or HPV-negative TP53 wildtype, with different clinical courses. To analyze the immune infiltrate in these molecular subtypes and its impact on clinical outcome, an in-depth study of the tumor immune microenvironment was performed.Methods Sixty-five patients with invasive VSCC matched for age, FIGO stage and treatment modality, were grouped according to the presence of HPV and p53 protein expression status. Archived tissues were analyzed for intraepithelial and stromal expression of CD3, CD8, Foxp3, PD-1, and pan-keratin in randomly selected areas using immunofluorescence. Additional phenotyping of T cells was performed ex-vivo on VSCC (n = 14) and blood samples by flow cytometry. Healthy vulvar samples and blood served as controls.Results Based on T-cell infiltration patterns about half of the VSCC were classified as inflamed or altered-excluded while one-third was immune-deserted. High intraepithelial helper T cell infiltration was observed in 78% of the HPV-induced VSCC, 60% of the HPVnegVSCC/p53wildtype and 40% of the HPVnegVSCC with abnormal p53 expression. A high intraepithelial infiltration with activated (CD3+PD-1+), specifically helper T cells (CD3+CD8−Foxp3−), was associated with a longer recurrence-free period and overall survival, irrespective of HPV and p53 status. Flow cytometry confirmed the tumor-specific presence of activated (CD4+PD-1++CD161−CD38+HLA-DR+ and CD8+CD103+CD161−NKG2A+/−PD1++CD38++HLA-DR+) effector memory T cells.Conclusion This is the first study demonstrating an association between intraepithelial T cells and clinical outcome in VSCC. Our data suggest that abnormal p53 expressing VSCCs mostly are cold tumors whereas HPV-driven VSCCs are strongly T-cell infiltrated.Mariëtte I. E. van Poelgeest and Sjoerd H. van der Burg contributed equally to this work.Abbreviations:Anti-CTLA-4Anti-cytotoxic T-lymphocyte-associated protein 4Anti-PD1Anti-programmed cell death protein 1BSABovine serum albuminCTLA-4Cytotoxic T-lymphocyte-associated protein 4FCSFetal calf serumFFPEFormalin-fixed paraffin embeddedHLAHuman leukocyte antigenHPVHuman papilloma virusHPVnegVSCCHuman papilloma virus negative vulvar squamous cell carcinomaHPVnegVSCC/p53abnHuman papilloma virus negative vulvar squamous cell carcinoma with p53 abnormal expressionHPVnegVSCC/p53wtHuman papilloma virus negative vulvar squamous cell carcinoma with p53 wildtype expressionHPVposVSCCHuman papilloma virus positive vulvar squamous cell carcinomaIDOIndoleamine 2,3-dioxygenaseIFNγInterferon-gammaIL-10Interleukin-10IL-4Interleukin-4IL-5Interleukin-5MAGEA1Melanoma-associated antigen 1MAGEA4Melanoma-associated antigen 4OSOverall survivalp53wtp53 wildtypep53abnp53 abnormalPBMCPeripheral blood mononuclear cellsPBSPhosphate buffered salinePD-1Programmed cell death protein 1PD-L1Programmed cell death ligand 1PHAPhytohemagglutininRFPRecurrence-free periodROCReceiver operating characteristicsSTINGStimulator of interferon genesTcmCentral memory T cellTemEffector memory T cellTemraEffector memory RA+ T cellsTh1T-helper 1Th2T-helper 2TILTumor infiltrating lymphocyteTNF-αTumor necrosis factor alphaTregRegulatory T cellVSCCVulvar squamous cell carcinoma