Programmed death ligand 1 expression and tumor infiltrating lymphocytes in neurofibromatosis type 1 and 2 associated tumors

Abstract

Immune checkpoint inhibitors targeting programmed cell death 1 (PD-1) or its ligand (PD-L1) have been shown to be effective in treating patients with a variety of cancers. Biomarker studies have found positive associations between clinical response rates and PD-L1 expression on tumor cells, as well as the presence of tumor infiltrating lymphocytes (TILs). It is currently unknown whether tumors associated with neurofibromatosis types 1 and 2 (NF1 and NF2) express PD-L1. We performed immunohistochemistry for PD-L1 (clones SP142 and E1L3N), CD3, CD20, CD8, and CD68 in NF1-related tumors (ten dermal and six plexiform neurofibromas) and NF2-related tumors (ten meningiomas and ten schwannomas) using archival formalin-fixed paraffin-embedded tissues. Expression of PD-L1 was considered positive in cases with > 5% membranous staining of tumor cells, in accordance with previously published biomarker studies. PD-L1 expression in tumor cells (using the SP142 and E1L3N clones, respectively) was assessed as positive in plexiform neurofibromas (6/6 and 5/6) dermal neurofibromas (8/10 and 6/10), schwannomas (7/10 and 10/10), and meningiomas (4/10 and 2/10). Sparse to moderate presence of CD68, CD3, or CD8 positive TILs was found in 36 (100%) of tumor specimens. Our findings indicate that adaptive resistance to cell-mediated immunity may play a major role in the tumor immune microenvironment of NF1 and NF2-associated tumors. Expression of PD-L1 on tumor cells and the presence of TILs suggest that these tumors might be responsive to immunotherapy with immune checkpoint inhibitors, which should be explored in clinical trials for NF patients.

Keywords: Meningioma; Neurofibroma; Neurofibromatosis type 1; Neurofibromatosis type 2; Programmed death-ligand 1; Schwannoma; Tumor infiltrating lymphocytes.

Figure 1. Immunohistochemistry staining of PD-L1 expression in NF associated tumors

This figure illustrates the PD-L1 expression at low magnification (100×) and high magnification (400×) with monoclonal antibodies, clone SP142 (E to H) vs. E1L3N (I to L) in NF2 meningioma (E vs. I), NF2 schwannoma (F vs. J), NF1 cutaneous neurofibroma (G vs. K), and NF1 plexiform neurofibroma (H vs. L). H&E stains (A to D) indicate the morphology of NF related tumors: NF2 meningioma (A), NF2 schwannoma (B), NF1 cutaneous neurofibroma (C), and NF1 plexiform neurofibroma (D) at low magnification (100×) and high magnification (400×).

Figure 2. Tumor-infiltrating lymphocytes and macrophages in NF-related tumors

Immunohistochemistry stain of CD3, CD8, CD68, and CD20 positive lymphocytes and macrophages in NF2 meningioma (A–D), NF2 schwannoma (E-H), NF1cutanous neurofibroma (I-L), and NF1 plexiform neurofibroma (M-P). This figure reveals the infiltration of CD3+ (A, E, I, and M) and CD8+ (B, F, J, and N) T lymphocytes and CD68+ (D, H, L, and P) macrophages in tumor tissues, not CD20+ B cells (C, G, K, and O). “Leaking” of CD20+ B cells in meningioma and schwannoma is shown in (C and G), with arrows indicating the vascular structures.