Automated Multiplex IHC Analysis

Immunohistochemistry for TUNEL and Ki-67 was performed as previously described (19 (link)). For the PROSPECT samples 4μm- thick tissue sections were stained using an automated staining system (Leica Bond Max, Leica Microsystems, Vista, CA, USA), according to standard protocols. The38 Aperio Image Analysis Toolbox (Aperio, Leica Microsystems) was used for digital analysis of images obtained from scanned slides. PDL1 clone E1L3N from Cell Signaling Technologies, CD3 A0452 from Dako and CD8 C8/144B from Thermo Scientific were used.

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Koyama S., Akbay E.A., Li Y.Y., Aref A.R., Skoulidis F., Herter-Sprie G.S., Buczkowski K.A., Liu Y., Awad M.M., Denning W.L., Diao L., Wang J., Parra-Cuentas E.R., Wistuba I.I., Soucheray M., Thai T.C., Asahina H., Kitajima S., Altabef A., Cavanaugh J.D., Rhee K., Gao P., Zhang H., Fecci P.E., Shimamura T., Hellmann M.D., Heymach J.V., Hodi F.S., Freeman G.J., Barbie D.A., Dranoff G., Hammerman P.S, & Wong K.K. (2016). STK11/LKB1 deficiency promotes neutrophil recruitment and proinflammatory cytokine production to suppress T cell activity in the lung tumor microenvironment. Cancer research, 76(5), 999-1008.

Publication 2016

Leica Bond Max PDL1 clone E1L3N CD3 A0452 CD8 C8/144B

Clone Immunohistochemistry Pdl1 Tissue Tunel

Corresponding Organization : Brigham and Women's Hospital

Other organizations : The University of Texas MD Anderson Cancer Center, University of California, San Francisco, Duke Medical Center, Loyola University Chicago, Memorial Sloan Kettering Cancer Center

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Variable analysis

independent variables

TUNEL
Ki-67

dependent variables

Staining intensity and distribution of TUNEL and Ki-67
Expression of PDL1, CD3, and CD8

control variables

Tissue section thickness (4μm)
Automated staining system (Leica Bond Max)
Digital image analysis (Aperio Image Analysis Toolbox)
Antibodies used (PDL1 clone E1L3N, CD3 A0452, CD8 C8/144B)

controls

No positive or negative controls were explicitly mentioned in the input protocol.

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