scholarly journals Prevalence of tumor-infiltrating lymphocytes and PD-L1 expression in the soft tissue sarcoma microenvironment

2015 ◽  
Vol 46 (3) ◽  
pp. 357-365 ◽  
Author(s):  
Sandra P. D’Angelo ◽  
Alexander N. Shoushtari ◽  
Narasimhan P. Agaram ◽  
Deborah Kuk ◽  
Li-Xuan Qin ◽  
...  
Keyword(s):  
Soft Tissue ◽  
Soft Tissue Sarcoma ◽  
Tumor Infiltrating Lymphocytes ◽  
Infiltrating Lymphocytes

scholarly journals Expanded Tumor-infiltrating Lymphocytes From Soft Tissue Sarcoma Have Tumor-specific Function

2021 ◽  
Vol Publish Ahead of Print ◽  
Author(s):  
John E. Mullinax ◽  
MacLean Hall ◽  
Matthew Beatty ◽  
Amy M. Weber ◽  
Zachary Sannasardo ◽  
...  
Keyword(s):  
Soft Tissue ◽  
Soft Tissue Sarcoma ◽  
Tumor Infiltrating Lymphocytes ◽  
Specific Function ◽  
Infiltrating Lymphocytes

scholarly journals 179 CD8+CD69+ Expanded Tumor Infiltrating Lymphocytes from Soft Tissue Sarcoma Have Increased Tumor-Specific Functional Capacity

2021 ◽  
Vol 9 (Suppl 3) ◽  
pp. A191-A191
Author(s):  
Jonathan Hensel ◽  
Alejandro Alfaro ◽  
Mary Rau ◽  
Patricio Perez-Villarroel ◽  
Zachary Sannasardo ◽  
...  
Keyword(s):  
Soft Tissue ◽  
Soft Tissue Sarcoma ◽  
Cell Lines ◽  
Functional Capacity ◽  
Tumor Infiltrating Lymphocytes ◽  
Cellular Immunotherapy ◽  
Autologous Tumor ◽  
Specific Reactivity ◽  
Immortalized Cell ◽  
Infiltrating Lymphocytes

BackgroundAdoptive cell therapy (ACT) utilizing tumor infiltrating lymphocytes (TIL) has demonstrated durable responses in patients with metastatic melanoma and offers potential for other solid tumors. Preclinical experience with expanded TIL from soft tissue sarcoma (STS) demonstrates less frequent tumor-specific reactivity compared to melanoma samples, limiting the potential for efficacy.1 We hypothesized that CD69+ TIL have increased tumor-specific reactivity, which can be manipulated in culture, thereby offering an opportunity to enhance the antitumor effect of this cellular immunotherapy product.MethodsPatients were enrolled on an IRB-approved protocol and TIL were expanded from fresh surgical specimens. After enzymatic digestion, tumor single cell suspensions were cultured in media containing 10% human serum and IL-2 (6000IU/mL). Expanded TIL were then enriched for CD8+ using magnetic bead isolation and CD69+ by flow cytometry cell sorting (FACS). After co-culture with autologous tumor digest, functional capacity was compared between bulk TIL and enriched TIL by evaluation of IFN-gamma (IFNg) and Granzyme B (GzB) secretion. Capacity for direct tumor cytotoxicity was assessed by Cr51 assay after co-culture of autologous immortalized cell lines with expanded TIL subpopulations after enrichment.ResultsFollowing co-culture with autologous tumor digest, CD69+ TIL demonstrated increased IFNg secretion compared to CD69- TIL in 6 samples (1.4–4.2x, p<0.05). CD8+ enriched TIL (75% compared to bulk) had higher relative IFNg secretion in both CD69+ and CD69- subsets (4.2 and 5.8x, respectively, p<0.001). Maximal IFNg secretion was seen from TIL that were both CD69+ sorted and CD8+ enriched, demonstrating an synergistic effect (16.3x vs Bulk CD69-, 4.2x vs Bulk CD69+, 2.8x vs CD8 enriched CD69- ; p<0.001). Functional capacity was also assessed by GzB secretion with similar results. CD69+ TIL had increased relative secretion (1.8–2.2x) compared to CD69- TIL (p< 0.01). CD8+ enriched TIL had increased relative GzB secretion in both CD69- and CD69+ sorted fractions (1.4x, 1.2x, respectively, p<.05). CD69+ sorted and CD8+ enriched TIL demonstrated an additive effect (2.6x vs Bulk CD69-, p<0.01; 1.2x vs Bulk CD69+, p<0.05; 1.8x vs CD8 enriched CD69-, p<0.01). CD8+ enriched CD69+ sorted TIL had greater relative cytotoxicity (3x, p<0.05) at 40:1 E:T ratio against autologous tumor cell lines compared to bulk expanded TIL (figure 1).Abstract 179 Figure 1Functional capacity of CD69+ TIL is demonstrated by increased secretion of GzB (A) and IFNg (B) after co-culture with autologous tumor digest. CD69+ TIL have greater cytotoxicity against autologous immortalized cell lines compared to bulk TIL at 40:1 E:T ratio (C).ConclusionsTIL expanded from STS demonstrate greater tumor-specific functional capacity and cytotoxicity after CD8 enrichment and CD69+ FACS compared to bulk expanded TIL. These data validate the strategy to enhance CD8+CD69+ TIL during culture to yield a more efficacious cellular immunotherapy product.AcknowledgementsThis work was funded by NIH K08CA252642Trial Registration n/aReference1. Mullinax JE, Hall M, Beatty M, Weber AM, Sannasardo Z, Svrdlin T, Hensel J, Bui M, Richards A, Gonzalez RJ, Cox CA, Kelley L, Mulé JJ, Sarnaik AA, Pilon-Thomas S. Expanded Tumor-infiltrating Lymphocytes From Soft Tissue Sarcoma Have Tumor-specific Function. J Immunother 2021 Feb-Mar 01;44(2):63–70.Ethics ApprovalAbstract cites IRB-approved protocol in methods section.Consent n/a

503 Tumor infiltrating lymphocytes in soft tissue sarcomas upregulate the exhaustion marker TIGIT and are reinvigorated by IL-15 stimulation and TIGIT blockade

2020 ◽  
Vol 8 (Suppl 3) ◽  
pp. A538-A538
Author(s):  
Sean Judge ◽  
Morgan Darrow ◽  
Steven Thorpe ◽  
Alicia Gingrich ◽  
Edmond O’Donnell ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Soft Tissue ◽  
Ex Vivo ◽  
Soft Tissue Sarcomas ◽  
Tumor Infiltrating Lymphocytes ◽  
Cd8 T Cell ◽  
Cell Phenotype ◽  
And Function ◽  
Infiltrating Lymphocytes

BackgroundAlthough the presence and activity of tumor infiltrating lymphocytes (TILs) have been shown to be important factors for survival and response to immunotherapy for multiple cancer types, the benefits of immunotherapy in soft tissue sarcomas (STS) have been limited, and novel approaches are needed. In this study, we sought to characterize the phenotype and function of tumor infiltrating natural killer (NK) and T cells in STS patients and to evaluate clinically relevant strategies to augment TIL function.MethodsUsing both prospectively collected blood and tumor tissue from STS patients undergoing surgical resection (n = 21) and archived specimens (n = 45), we performed flow cytometry and immunohistochemistry to evaluate the extent of peripheral and intratumoral CD3-CD56+ NK and CD8+ T cell phenotype and function as predictors of outcome. We also analyzed TCGA data and the peripheral blood of dogs with spontaneous osteosarcoma receiving inhaled IL-15 on a clinical trial to evaluate the association of CD3-NKp46+ NK and CD8+ T cell activation as well as TIGIT upregulation with outcome. Finally, we stimulated patient PBMCs and TILs ex vivo with IL-15 and a novel human anti-TIGIT antibody to assess the impact of combination therapy on NK and T cell phenotype and function. Parametric and non-parametric statistical tests were used where appropriate. Univariate and multivariate survival analyses were performed by Cox proportional hazards models.ResultsCompared to peripheral expression, intratumoral NK and T cells showed an activated and exhausted phenotype by CD69 and TIGIT, respectively. Ex vivo TIL stimulation with IL-15 further increased markers of activation and function including CD69, Ki67, IFNg, and granzyme B, while increasing expression of exhaustion marker TIGIT. Analysis of a retrospective STS cohort and TCGA STS gene expression confirmed the association of TILs with improved prognosis. Dogs with metastatic osteosarcoma receiving inhaled IL-15 exhibited upregulation of activation markers and TIGIT. In vitro, IL-15 and TIGIT blockade of both peripheral and intratumoral NK cells increased cytotoxicity against sarcoma cell lines and increased expression of degranulation marker CD107a compared to IL-15 alone.ConclusionsTILs are associated with improved survival in STS, and tumor infiltrating NK and T cells show features of both increased activation and increased exhaustion. Tumor-infiltrating NK and T cells respond to IL-15 stimulation, but simultaneously further upregulate TIGIT with the combination of IL-15 and TIGIT blockade showing greatest cytotoxic effects. Overall, our data suggest that the combination of IL-15 and TIGIT blockade is a promising clinical strategy in STS.Ethics ApprovalAll experiments involving human and canine patients were approved by the respective Institutional Review Boards at the University of California, Davis, Schools of Medicine (Protocol #218204-9) and Veterinary Medicine (IACUC #20179).

scholarly journals PD-L1 and PD-1 and characterization of tumor-infiltrating lymphocytes in high grade sarcomas of soft tissue – prognostic implications and rationale for immunotherapy

2017 ◽  
Vol 7 (3) ◽  
pp. e1389366 ◽  
Author(s):  
Melanie Boxberg ◽  
Katja Steiger ◽  
Ulrich Lenze ◽  
Hans Rechl ◽  
Rüdiger von Eisenhart-Rothe ◽  
...  
Keyword(s):  
Soft Tissue ◽  
Tumor Infiltrating Lymphocytes ◽  
High Grade ◽  
Prognostic Implications ◽  
Infiltrating Lymphocytes

Morphological Examination of a Herpes-type Virus Indigenous for Tree Shrews

1970 ◽  
Vol 28 ◽  
pp. 160-161
Author(s):  
J. P. Brunschwig ◽  
R. M. McCombs ◽  
R. Mirkovic ◽  
M. Benyesh-Melnick
Keyword(s):  
Electron Microscopy ◽  
Soft Tissue ◽  
Chick Embryo ◽  
Soft Tissue Sarcoma ◽  
Cell Cultures ◽  
Tree Shrew ◽  
Type Virus ◽  
Morphological Examination ◽  
Tree Shrews ◽  
Embryo Cells

A new virus, established as a member of the herpesvirus group by electron microscopy, was isolated from spontaneously degenerating cell cultures derived from the kidneys and lungs of two normal tree shrews. The virus was found to replicate best in cells derived from the homologous species. The cells used were a tree shrew cell line, T-23, which was derived from a spontaneous soft tissue sarcoma. The virus did not multiply or did so poorly for a limited number of passages in human, monkey, rodent, rabbit or chick embryo cells. In the T-23 cells, the virus behaved as members of the subgroup B of herpesvirus, in that the virus remained primarily cell associated.

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