{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,24]],"date-time":"2026-01-24T01:42:56Z","timestamp":1769218976988,"version":"3.49.0"},"reference-count":26,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2016,8,19]],"date-time":"2016-08-19T00:00:00Z","timestamp":1471564800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2016,8,19]],"date-time":"2016-08-19T00:00:00Z","timestamp":1471564800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>With the emergence of checkpoint blockade and other immunotherapeutic drugs, and the growing adoption of smaller, more flexible adaptive clinical trial designs, there is an unmet need to develop diagnostics that can rapidly immunophenotype patient tumors. The ability to longitudinally profile the tumor immune infiltrate in response to immunotherapy also presents a window of opportunity to illuminate mechanisms of resistance. We have developed a fine needle aspirate biopsy (FNA) platform to perform immune profiling on thoracic malignancies. Matching peripheral blood, bulk resected tumor, and FNA were analyzed from 13 mesothelioma patients. FNA samples yielded greater numbers of viable cells when compared to core needle biopsies. Cell numbers were adequate to perform flow cytometric analyses on T cell lineage, T cell activation and inhibitory receptor expression, and myeloid immunosuppressive checkpoint markers. FNA samples were representative of the tumor as a whole as assessed by head-to-head comparison to single cell suspensions of dissociated whole tumor. Parallel analysis of matched patient blood enabled us to establish quality assurance criteria to determine the accuracy of FNA procedures to sample tumor tissue. FNA biopsies provide a diagnostic to rapidly phenotype the tumor immune microenvironment that may be of great relevance to clinical trials.<\/jats:p>","DOI":"10.1038\/srep31745","type":"journal-article","created":{"date-parts":[[2016,8,19]],"date-time":"2016-08-19T11:26:53Z","timestamp":1471606013000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":25,"title":["Fine needle aspirate flow cytometric phenotyping characterizes immunosuppressive nature of the mesothelioma microenvironment"],"prefix":"10.1038","volume":"6","author":[{"given":"Patrick H.","family":"Lizotte","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Robert E.","family":"Jones","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lauren","family":"Keogh","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elena","family":"Ivanova","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hongye","family":"Liu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mark M.","family":"Awad","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Peter S.","family":"Hammerman","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ritu R.","family":"Gill","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"William G.","family":"Richards","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David A.","family":"Barbie","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Adam J.","family":"Bass","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Raphael","family":"Bueno","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jessie M.","family":"English","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mark","family":"Bittinger","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kwok-Kin","family":"Wong","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2016,8,19]]},"reference":[{"key":"BFsrep31745_CR1","doi-asserted-by":"publisher","first-page":"2455","DOI":"10.1056\/NEJMoa1200694","volume":"366","author":"JR Brahmer","year":"2012","unstructured":"Brahmer, J. R. et al. Safety and Activity of Anti\u2013PD-L1 Antibody in Patients with Advanced Cancer. N. Engl. J. Med. 366, 2455\u20132465 (2012).","journal-title":"N. Engl. J. Med."},{"key":"BFsrep31745_CR2","doi-asserted-by":"publisher","first-page":"4899","DOI":"10.4049\/jimmunol.1300271","volume":"190","author":"SR Goding","year":"2013","unstructured":"Goding, S. R. et al. Restoring Immune Function of Tumor-Specific CD4+ T Cells during Recurrence of Melanoma. J. Immunol. 190, 4899\u20134909 (2013).","journal-title":"J. Immunol."},{"key":"BFsrep31745_CR3","doi-asserted-by":"publisher","first-page":"711","DOI":"10.1056\/NEJMoa1003466","volume":"363","author":"FS Hodi","year":"2010","unstructured":"Hodi, F. S. et al. Improved Survival with Ipilimumab in Patients with Metastatic Melanoma. N. Engl. J. Med. 363, 711\u2013723 (2010).","journal-title":"N. Engl. J. Med."},{"key":"BFsrep31745_CR4","doi-asserted-by":"publisher","first-page":"6682","DOI":"10.1073\/pnas.1420370112","volume":"112","author":"J Liu","year":"2015","unstructured":"Liu, J. et al. Immune-checkpoint proteins VISTA and PD-1 nonredundantly regulate murine T-cell responses. Proc. Natl. Acad. Sci. 112, 6682\u20136687 (2015).","journal-title":"Proc. Natl. Acad. Sci"},{"key":"BFsrep31745_CR5","doi-asserted-by":"publisher","first-page":"2443","DOI":"10.1056\/NEJMoa1200690","volume":"366","author":"SL Topalian","year":"2012","unstructured":"Topalian, S. L. et al. Safety, Activity, and Immune Correlates of Anti\u2013PD-1 Antibody in Cancer. N. Engl. J. Med. 366, 2443\u20132454 (2012).","journal-title":"N. Engl. J. Med."},{"key":"BFsrep31745_CR6","doi-asserted-by":"publisher","first-page":"36","DOI":"10.1186\/1479-5876-12-36","volume":"12","author":"L Lu","year":"2014","unstructured":"Lu, L. et al. Combined PD-1 blockade and GITR triggering induce a potent antitumor immunity in murine cancer models and synergizes with chemotherapeutic drugs. J. Transl. Med. 12, 36 (2014).","journal-title":"J. Transl. Med."},{"key":"BFsrep31745_CR7","doi-asserted-by":"publisher","first-page":"640","DOI":"10.1053\/j.seminoncol.2015.05.014","volume":"42","author":"MF Sanmamed","year":"2015","unstructured":"Sanmamed, M. F. et al. Agonists of Co-stimulation in Cancer Immunotherapy Directed Against CD137, OX40, GITR, CD27, CD28, and ICOS. Semin. Oncol. 42, 640\u2013655 (2015).","journal-title":"Semin. Oncol."},{"key":"BFsrep31745_CR8","doi-asserted-by":"crossref","unstructured":"Boyd, J. D., Smith, G. D., Hong, H., Mageau, R. & Juskevicius, R. Fine-Needle Aspiration is Superior to Needle Core Biopsy as a Sample Aquisition Method for Flow Cytometric Analysis in Suspected Hematologic Neoplasms. Cytometry B Clin. Cytom (2014).","DOI":"10.1002\/cytob.21183"},{"key":"BFsrep31745_CR9","doi-asserted-by":"publisher","first-page":"311","DOI":"10.1097\/PAI.0b013e3181827da8","volume":"18","author":"A Demurtas","year":"2010","unstructured":"Demurtas, A. et al. Utility of flow cytometry immunophenotyping in fine-needle aspirate cytologic diagnosis of non-Hodgkin lymphoma: A series of 252 cases and review of the literature. Appl. Immunohistochem. Mol. Morphol. AIMM Off. Publ. Soc. Appl. Immunohistochem. 18, 311\u2013322 (2010).","journal-title":"Appl. Immunohistochem. Mol. Morphol. AIMM Off. Publ. Soc. Appl. Immunohistochem."},{"key":"BFsrep31745_CR10","doi-asserted-by":"publisher","first-page":"760","DOI":"10.1002\/dc.21618","volume":"40","author":"R Factor","year":"2012","unstructured":"Factor, R. & Layfield, L. J. Intraprocedural evaluation of fine-needle aspiration smears: how good are we? Diagn. Cytopathol. 40, 760\u2013763 (2012).","journal-title":"Diagn. Cytopathol."},{"key":"BFsrep31745_CR11","doi-asserted-by":"publisher","first-page":"381","DOI":"10.1002\/dc.20650","volume":"35","author":"R Gupta","year":"2007","unstructured":"Gupta, R., Naseem, S., Kashyap, R. & Paul, L. Role of fine-needle aspirate immunophenotyping by flow cytometry in rapid diagnosis of lymphoproliferative disorders. Diagn. Cytopathol. 35, 381\u2013385 (2007).","journal-title":"Diagn. Cytopathol."},{"key":"BFsrep31745_CR12","doi-asserted-by":"publisher","first-page":"3046","DOI":"10.1200\/JCO.2004.02.104","volume":"22","author":"ST Hehn","year":"2004","unstructured":"Hehn, S. T., Grogan, T. M. & Miller, T. P. Utility of fine-needle aspiration as a diagnostic technique in lymphoma. J. Clin. Oncol. Off. J. Am. Soc. Clin. Oncol. 22, 3046\u20133052 (2004).","journal-title":"J. Clin. Oncol. Off. J. Am. Soc. Clin. Oncol."},{"key":"BFsrep31745_CR13","doi-asserted-by":"publisher","first-page":"9029","DOI":"10.1200\/JCO.2005.01.5248","volume":"23","author":"RM Austin","year":"2005","unstructured":"Austin, R. M. et al. Fine Needle Aspiration Is a Feasible and Accurate Technique in the Diagnosis of Lymphoma. J. Clin. Oncol. 23, 9029\u20139030 (2005).","journal-title":"J. Clin. Oncol."},{"key":"BFsrep31745_CR14","first-page":"3016","volume":"31","author":"J Grosso","year":"2013","unstructured":"Grosso, J. et al. Association of tumor PD-L1 expression and immune biomarkers with clinical activity in patients (pts) with advanced solid tumors treated with nivolumab (anti-PD-1; BMS-936558; ONO-4538). ASCO Meet. Abstr. 31, 3016 (2013).","journal-title":"ASCO Meet. Abstr"},{"key":"BFsrep31745_CR15","doi-asserted-by":"publisher","first-page":"1019","DOI":"10.1007\/s00262-011-1172-6","volume":"61","author":"R-R Ji","year":"2012","unstructured":"Ji, R.-R. et al. An immune-active tumor microenvironment favors clinical response to ipilimumab. Cancer Immunol. Immunother. CII 61, 1019\u20131031 (2012).","journal-title":"Cancer Immunol. Immunother. CII"},{"key":"BFsrep31745_CR16","doi-asserted-by":"publisher","first-page":"1008","DOI":"10.1158\/2326-6066.CIR-14-0227","volume":"3","author":"SS Kwek","year":"2015","unstructured":"Kwek, S. S. et al. Preexisting Levels of CD4 T Cells Expressing PD-1 Are Related to Overall Survival in Prostate Cancer Patients Treated with Ipilimumab. Cancer Immunol. Res. 3, 1008\u20131016 (2015).","journal-title":"Cancer Immunol. Res."},{"key":"BFsrep31745_CR17","doi-asserted-by":"publisher","first-page":"31","DOI":"10.1186\/s40425-014-0031-3","volume":"2","author":"SJ Santegoets","year":"2014","unstructured":"Santegoets, S. J. et al. Myeloid derived suppressor and dendritic cell subsets are related to clinical outcome in prostate cancer patients treated with prostate GVAX and ipilimumab. J. Immunother. Cancer 2, 31 (2014).","journal-title":"J. Immunother. Cancer"},{"key":"BFsrep31745_CR18","doi-asserted-by":"publisher","first-page":"245","DOI":"10.1007\/s00262-012-1330-5","volume":"62","author":"SJAM Santegoets","year":"2013","unstructured":"Santegoets, S. J. A. M. et al. T cell profiling reveals high CD4+CTLA-4 + T cell frequency as dominant predictor for survival after prostate GVAX\/ipilimumab treatment. Cancer Immunol. Immunother. CII 62, 245\u2013256 (2013).","journal-title":"Cancer Immunol. Immunother. CII"},{"key":"BFsrep31745_CR19","doi-asserted-by":"publisher","first-page":"200ra116","DOI":"10.1126\/scitranslmed.3006504","volume":"5","author":"S Spranger","year":"2013","unstructured":"Spranger, S. et al. Up-Regulation of PD-L1, IDO, and Tregs in the Melanoma Tumor Microenvironment Is Driven by CD8+ T Cells. Sci. Transl. Med. 5, 200ra116 (2013).","journal-title":"Sci. Transl. Med."},{"key":"BFsrep31745_CR20","doi-asserted-by":"publisher","first-page":"226ra32","DOI":"10.1126\/scitranslmed.3008095","volume":"6","author":"D Zamarin","year":"2014","unstructured":"Zamarin, D. et al. Localized oncolytic virotherapy overcomes systemic tumor resistance to immune checkpoint blockade immunotherapy. Sci. Transl. Med. 6, 226ra32 (2014).","journal-title":"Sci. Transl. Med."},{"key":"BFsrep31745_CR21","doi-asserted-by":"publisher","first-page":"858","DOI":"10.1038\/nbt.2317","volume":"30","author":"B Bodenmiller","year":"2012","unstructured":"Bodenmiller, B. et al. Multiplexed mass cytometry profiling of cellular states perturbed by small-molecule regulators. Nat. Biotechnol. 30, 858\u2013867 (2012).","journal-title":"Nat. Biotechnol."},{"key":"BFsrep31745_CR22","doi-asserted-by":"publisher","first-page":"1627","DOI":"10.1056\/NEJMoa1507643","volume":"373","author":"H Borghaei","year":"2015","unstructured":"Borghaei, H. et al. Nivolumab versus Docetaxel in Advanced Nonsquamous Non-Small-Cell Lung Cancer. N. Engl. J. Med. 373, 1627\u20131639 (2015).","journal-title":"N. Engl. J. Med."},{"key":"BFsrep31745_CR23","doi-asserted-by":"publisher","first-page":"4286","DOI":"10.1158\/1078-0432.CCR-14-2607","volume":"21","author":"A Patnaik","year":"2015","unstructured":"Patnaik, A. et al. Phase I Study of Pembrolizumab (MK-3475; Anti-PD-1 Monoclonal Antibody) in Patients with Advanced Solid Tumors. Clin. Cancer Res. Off. J. Am. Assoc. Cancer Res. 21, 4286\u20134293 (2015).","journal-title":"Clin. Cancer Res. Off. J. Am. Assoc. Cancer Res."},{"key":"BFsrep31745_CR24","unstructured":"Murtagh, F. Multidimensional Clustering Algorithms. (Wuerzburg: Physica-Verlag, 1985)."},{"key":"BFsrep31745_CR25","doi-asserted-by":"publisher","first-page":"274","DOI":"10.1007\/s00357-014-9161-z","volume":"31","author":"F Murtagh","year":"2014","unstructured":"Murtagh, F. & Legendre, P. Ward\u2019s Hierarchical Agglomerative Clustering Method: Which Algorithms Implement Ward\u2019s Criterion? - Springer. J. Classif. 31, 274\u2013295 (2014).","journal-title":"J. Classif."},{"key":"BFsrep31745_CR26","first-page":"3221","volume":"15","author":"L Van Der Maaten","year":"2014","unstructured":"Van Der Maaten, L. Accelerating t-SNE Using Tree-based Algorithms. J Mach Learn Res 15, 3221\u20133245 (2014).","journal-title":"J Mach Learn Res"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/srep31745.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/srep31745","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/srep31745.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,1,5]],"date-time":"2023-01-05T05:37:44Z","timestamp":1672897064000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/srep31745"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2016,8,19]]},"references-count":26,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2016,8,30]]}},"alternative-id":["BFsrep31745"],"URL":"https:\/\/doi.org\/10.1038\/srep31745","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2016,8,19]]},"assertion":[{"value":"11 March 2016","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"26 July 2016","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"19 August 2016","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing financial interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"31745"}}