{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T11:55:14Z","timestamp":1785412514513,"version":"3.56.0"},"reference-count":42,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2026,5,31]],"date-time":"2026-05-31T00:00:00Z","timestamp":1780185600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004504","name":"Research Council of Lithuania","doi-asserted-by":"publisher","award":["S-MIP-23-97"],"award-info":[{"award-number":["S-MIP-23-97"]}],"id":[{"id":"10.13039\/501100004504","id-type":"DOI","asserted-by":"publisher"}]},{"award":["S-MIP-23-97"],"award-info":[{"award-number":["S-MIP-23-97"]}],"id":[{"id":"https:\/\/ror.org\/02gs16m83","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cancers"],"abstract":"<jats:p>Background: Bronchoscopic cryotherapy may enhance anti-tumor immunity and improve the effect of immune checkpoint blockade, but CD8+ T cell dynamics after cryotherapy combined with pembrolizumab-based therapy in metastatic non-small cell lung cancer (NSCLC) remain insufficiently characterized. Methods: In this prospective, exploratory, randomized, controlled, single-center study, metastatic NSCLC patients were assigned to bronchoscopic cryotherapy performed 7 \u00b1 1 days before first-line pembrolizumab-based therapy or to standard treatment alone. Peripheral blood mononuclear cells were analyzed by flow cytometry at baseline, week 3, and week 6. CD8+ T cell subsets defined by CD45RO, CD28, granzyme B (GzB), IFN\u03b3, Ki-67, and PD-1 were evaluated in relation to treatment group, radiologic response, progression-free survival (PFS), and overall survival (OS). Results: Flow cytometry was performed in 76 patients, including 34 in the cryotherapy group and 42 in the control group. Cryotherapy was associated with a treatment-specific increase in circulating GzB+ CD8+ T cells by week 6. In contrast, Ki-67+ CD8+ and GzB+Ki-67+ cells increased in both treatment groups, suggesting that early peripheral CD8+ proliferation was largely shared across pembrolizumab-based therapy. Radiologic responders demonstrated more sustained proliferative CD8+ dynamics, most consistently reflected by increased CD28+ Ki-67+ cells. In exploratory landmark survival analyses, a higher week-3-to-baseline GzB+ Ki-67+ CD8+ ratio showed a hypothesis-generating association with longer PFS and OS. Conclusions: In metastatic NSCLC patients receiving first-line pembrolizumab-based therapy, the addition of bronchoscopic cryotherapy was associated with peripheral CD8+ T cell remodeling toward enhanced cytotoxic activity. Proliferative CD8+ T cell changes were associated with radiologic response and better survival. These findings support bronchoscopic cryotherapy as a potential immune-modulating adjunct and warrant validation in larger studies.<\/jats:p>","DOI":"10.3390\/cancers18111793","type":"journal-article","created":{"date-parts":[[2026,6,1]],"date-time":"2026-06-01T12:19:42Z","timestamp":1780316382000},"page":"1793","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Peripheral CD8+ T Cell Dynamics and Clinical Outcomes in Metastatic Non-Small Cell Lung Cancer Following Bronchoscopic Cryotherapy and Pembrolizumab-Based Therapy"],"prefix":"10.3390","volume":"18","author":[{"given":"Gediminas","family":"Vasiliauskas","sequence":"first","affiliation":[{"name":"Department of Pulmonology, Lithuanian University of Health Sciences, 44307 Kaunas, Lithuania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Evelina","family":"\u017demait\u0117","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, Lithuanian University of Health Sciences, 44307 Kaunas, Lithuania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0413-6808","authenticated-orcid":false,"given":"Erika","family":"Skrodenien\u0117","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, Lithuanian University of Health Sciences, 44307 Kaunas, Lithuania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lina","family":"Po\u0161kien\u0117","sequence":"additional","affiliation":[{"name":"Department of Pathology, Lithuanian University of Health Sciences, 44307 Kaunas, Lithuania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-6130-0712","authenticated-orcid":false,"given":"Skaidrius","family":"Miliauskas","sequence":"additional","affiliation":[{"name":"Department of Pulmonology, Lithuanian University of Health Sciences, 44307 Kaunas, Lithuania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marius","family":"\u017demaitis","sequence":"additional","affiliation":[{"name":"Department of Pulmonology, Lithuanian University of Health Sciences, 44307 Kaunas, Lithuania"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,5,31]]},"reference":[{"key":"ref_1","first-page":"229","article-title":"Global cancer statistics 2022: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries","volume":"74","author":"Bray","year":"2024","journal-title":"CA Cancer J. 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