Immunohistochemical and FISH Analysis of PD-L1/PD-L2 in Lung Cancer

Immunohistochemistry was performed for PD1 (clone NAT105, prediluted, CELL MARQUE, Rocklin, CA), PD-L1, PD-L2, phospho-STAT3 (PD-L1: clone E1L3N, 1:400 dilution, PD-L2: clone D7U8C, 1:20 dilution and phospho-STAT3: clone M9C6, 1:250 dilution; Cell Signaling Technology, Danvers, MA) and TTF1 (clone 8G7G3/1, prediluted, Ventana, Tucson, Arizona). Immunohistochemistry with the PD-L1 antibody was clinically validated internally against the PD-L1 22C3 clone from pharmDx and found to be comparable. Fluorescence in situ hybridization for JAK2/INSL6 and PD-L1/PD-L2 genes was performed as previously described (27 (link), 28 (link)). See supplementary methods for additional details.

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Gupta S., Cheville J.C., Jungbluth A.A., Zhang Y., Zhang L., Chen Y.B., Tickoo S.K., Fine S.W., Gopalan A., Al-Ahmadie H.A., Sirintrapun S.J., Blum K.A., Lohse C.M., Hakimi A.A., Thompson R.H., Leibovich B.C., Berger M.F., Arcila M.E., Ross D.S., Ladanyi M., Antonescu C.R, & Reuter V.E. (2019). JAK2/PD-L1/PD-L2 (9p24.1) Amplifications in Renal Cell Carcinomas with Sarcomatoid Transformation: Implications for Clinical Management. Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc, 32(9), 1344-1358.

Publication 2019

PD-L1: clone E1L3N PD-L2: clone D7U8C

Antibody Cell Clone Dilution Fluorescence in situ hybridization Genes Immunohistochemistry Jak2 Pd l1 Stat3 Ttf1

Corresponding Organization : Memorial Sloan Kettering Cancer Center

Other organizations : Mayo Clinic in Arizona, Mayo Clinic in Florida

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Variable analysis

independent variables

PD1 (clone NAT105, prediluted, CELL MARQUE, Rocklin, CA)
PD-L1 (clone E1L3N, 1:400 dilution, Cell Signaling Technology, Danvers, MA)
PD-L2 (clone D7U8C, 1:20 dilution, Cell Signaling Technology, Danvers, MA)
Phospho-STAT3 (clone M9C6, 1:250 dilution, Cell Signaling Technology, Danvers, MA)
TTF1 (clone 8G7G3/1, prediluted, Ventana, Tucson, Arizona)
JAK2/INSL6 (fluorescence in situ hybridization)
PD-L1/PD-L2 (fluorescence in situ hybridization)

dependent variables

Immunohistochemistry expression levels of PD1, PD-L1, PD-L2, phospho-STAT3, and TTF1
JAK2/INSL6 and PD-L1/PD-L2 gene expression (fluorescence in situ hybridization)

control variables

PD-L1 antibody (clone E1L3N) was clinically validated internally against the PD-L1 22C3 clone from pharmDx and found to be comparable

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