1. Immune Cell Infiltration May Be a Key Determinant of Long-Term Survival in Small Cell Lung Cancer 
Prasuna Muppa, M.B.B.S., Simone Barreto Siqueira Parrilha Terra, MD, Anurag Sharma, MD, Aaron S. Mansfield, MD, Marie-Christine Aubry, MD, Kaustubh Bhinge, PhD, Michael K. Asiedu, PhD, Mariza de Andrade, PhD, Nafiseh Janaki, MD, Stephen J. Murphy, PhD, Aqsa Nasir, MD, Virginia Van Keulen, MS, George Vasmatzis, PhD, Dennis A. Wigle, MD, PhD, Ping Yang, PhD, Eunhee S. Yi, MD, Tobias Peikert, MD, Farhad Kosari, PhD 
Journal of Thoracic Oncology 
Volume 14, Issue 7, Pages (July 2019)
DOI: /j.jtho
Copyright © 2019 International Association for the Study of Lung Cancer Terms and Conditions

2. Figure 1
Immune-related transcripts were dominantly overexpressed in patients with long-term survival (LTS) compared with in patients with the expected survival time (EXS). Top left is a plot of the mean expression of transcripts measured by (log2) probeset signal intensities in patients with LTS versus in those with EXS. Red, orange, and green dots are Affymetrix probesets corresponding to immunoglobins (IGs), human leukocyte antigen (HLA), or other known (Kn) genes, respectively. Purple crosses and gray dots are probesets corresponding to unknown (UnK) genes and probesets with insignificant changes (fold change <2 or p > 0.05), respectively. Most of the differentially expressed genes was overexpressed in patients with LTS and had immune-related functions. C-X-C motif chemokine ligand 9 gene (CXCL9), CD3e molecule gene (CD3E), and beta-2-microglobulin gene (B2M) are shown as three representative overexpressed genes in LTS.
Journal of Thoracic Oncology  , DOI: ( /j.jtho )
Copyright © 2019 International Association for the Study of Lung Cancer Terms and Conditions

3. Figure 2
Immunohistochemistry scoring and data presentation scheme. (A) A representative case showing boxed areas for scoring immunohistochemistry data by using the Aperio software program. Green, yellow, and red boxes are selected areas for the intraepithelial (IE), stromal (ST), and interface (IF) regions, respectively. (B) Follow-up data for cases involving patients with the expected survival times (patients E1–E13) and cases involving the long-term survivors (patients L1–L17). (C) Median expression and confidence intervals for CD3 molecule (CD3) as a representative stain in patients with the expected survival time and long-term survivors in the three zones. Cell counts were generally much lower in the IE than in the ST and IF.
Journal of Thoracic Oncology  , DOI: ( /j.jtho )
Copyright © 2019 International Association for the Study of Lung Cancer Terms and Conditions

4. Figure 3
(A) Median counts of positive cell counts by each stain in the stroma in samples from long-term survivors (green) and samples from those with the expected survival time (brown). In each stain, counts in long-term survivors were compared with counts in those with the expected survival time by using a nonparametric test (*, **, and *** represent p < 0.05, p < 0.005, and p < 0.005, respectively). (B) A supervised clustering using CD3-positive and CD279-positive counts produced two clads, with 88% of those in one clad (13 of 17) being long-term survivors and 77% of those in the other clad (10 of 13) having the expected survival time. FoxP3, forkhead box p3; LYZ, lysozyme; MPO, myeloperoxidase.
Journal of Thoracic Oncology  , DOI: ( /j.jtho )
Copyright © 2019 International Association for the Study of Lung Cancer Terms and Conditions

5. Figure 4
Summary of the staining results in the intraepithelial (IE) (top panels), stroma (ST) (middle panels), and interface (IF) (lower panels) zones. (A) T-cell response estimated by CD4-positive (red) and CD8-positive (maroon) counts and major histocompatibility complex I (MHC I) expression (human leukocyte antigen 1) (top panel). In two cases (represented by na) MHC I stains were not available. (B) Antitumor humoral immune response estimated CD20-positive (blue) and CD138-positive (purple) cells. CD138-positive stains in the IE zone (gold lines) were predominantly in the tumor cells and not lymphocytes (see text). (C) Counts of lymphocytes associated with an immune-suppressive microenvironment, including CD14-positive myeloid lineage (olive), CD68-positive macrophages (yellow), and forkhead box p3 (FoxP3)-positive regulatory (salmon) cells. Antitumor immunity represented by CD4-positive and CD8-positive cells was generally much stronger in long-term survivors than in those with the expected survival time. However, in patient L6 (blue arrow) counts of CD4-positive and CD8-positive cells and MHC I expression were low, whereas CD138-positive counts in the stroma were relatively high, suggesting a predominant humoral antitumor immunity. Also in patients L10 and L13 (brown arrows), expression levels of MHC I were low, but counts of CD20-positive and CD138-positive cells were strong, suggesting significant contributions by the humoral antitumor immunity.
Journal of Thoracic Oncology  , DOI: ( /j.jtho )
Copyright © 2019 International Association for the Study of Lung Cancer Terms and Conditions

6. Figure 5
Ratios of CD68-positive macrophages to CD3-positive T lymphocytes (CD68/CD3) were lower in long-term survivors than in those with the expected survival time. Comparisons by nonparametric tests found differences to be significant in the stromal (ST) region and interface (IF) (p = and p = 0.047, respectively) and marginal in intraepithelial (IE) region (p = 0.06).
Journal of Thoracic Oncology  , DOI: ( /j.jtho )
Copyright © 2019 International Association for the Study of Lung Cancer Terms and Conditions