{"title":"BRAF V600E突变的分化型甲状腺乳头状癌中肿瘤浸润细胞毒性淋巴细胞的杀伤能力分析及其对淋巴结转移的影响","authors":"Xiaogang Liu, Honggang Liu, Lu Wang, Yubing Han, Linghong Kong, Xinpeng Zhang","doi":"10.1186/s13000-024-01454-9","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>Cytotoxic lymphocytes (CLs) express potent toxins, including perforin (P) and granzyme-B (G), which brings about target cell death. The purpose of this study was to evaluate the killing capacity of tumor-infiltrating CLs by means of P and G analysis, and explore the association with lymph node metastasis in papillary carcinoma of thyroid (PTC) without Hashimoto's thyroiditis (HT).</p><p><strong>Methods: </strong>Infiltration of lymphocytes in PTC was observed in frozen sections. Both fresh tumor tissues and paracancerous tissues with lymphocyte infiltration were collected and prepared into a single cell suspension. Flow cytometry was used to detect the percentages of CD3<sup>+</sup>P<sup>+</sup>, CD3<sup>+</sup>G<sup>+</sup>, CD8<sup>+</sup>P<sup>+</sup>, and CD8<sup>+</sup>G<sup>+</sup> T lymphocytes (TLs) and CD16-CD56<sup>+</sup>P<sup>+</sup> and CD16-CD56<sup>+</sup>G<sup>+</sup> natural killer (NK) cells. Finally, we investigated differential expression of P and G in NK cells and cytotoxic T lymphocytes (CTLs) in paired tumor tissues (group T, n = 44) and paracancerous tissues (group N, n = 44) from patients with PTC with the BRAF V600E mutation. Furthermore, patients were divided into two groups according to whether cervical central lymph node metastasis (CCLNM) existed: group A (with lymph node metastases, n = 27) and group B (with nonlymph node metastases, n = 17). Patients were also divided into three groups according to the total number of positive CCLNM: group B, group C (with low-level lymph node metastases, less than 5, n = 17) and group D (with high-level lymph node metastases, no less than 5, n = 10).</p><p><strong>Results: </strong>The percentage of CD3<sup>+</sup>P<sup>+</sup> CTLs was significantly higher in group N than in group T (P < 0.05). The percentage of CD8<sup>+</sup>G<sup>+</sup> CTLs was significantly higher in group T than in group N (P < 0.05). The percentages of CD3<sup>+</sup>G<sup>+</sup>, CD16-CD56<sup>+</sup>P<sup>+</sup>and CD16-CD56<sup>+</sup>G<sup>+</sup> NK cells showed no significant difference in either group T or group N (P > 0.05). The percentages of CD3<sup>+</sup>P<sup>+</sup> CTLs in group A and group C were significantly higher in the paracancerous tissue than in the tumor tissue (P < 0.05). The percentages of CD8<sup>+</sup>G<sup>+</sup> CTLs in group A and group C were significantly higher in the tumor tissues than in the paracancerous tissues (P < 0.05). The percentage of CD16-CD56<sup>+</sup>G<sup>+</sup> NK cells in group D was significantly higher in the tumor tissues than in the paracancerous tissues (P < 0.05).</p><p><strong>Conclusions: </strong>The killing capacity of infiltrating CLs in PTC differed between tumor tissues and paracancerous tissues. In cases with CCLNM, higher expression of CD16-CD56<sup>+</sup>G<sup>+</sup> NK cells in tumor tissues may be associated with a high risk of lymph node metastasis.</p>","PeriodicalId":11237,"journal":{"name":"Diagnostic Pathology","volume":null,"pages":null},"PeriodicalIF":2.4000,"publicationDate":"2024-02-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10858496/pdf/","citationCount":"0","resultStr":"{\"title\":\"Killing capacity analysis of tumor-infiltrating cytotoxic lymphocytes and impact on lymph node metastasis in differentiated papillary carcinoma of thyroid with the BRAF V600E mutation.\",\"authors\":\"Xiaogang Liu, Honggang Liu, Lu Wang, Yubing Han, Linghong Kong, Xinpeng Zhang\",\"doi\":\"10.1186/s13000-024-01454-9\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>Cytotoxic lymphocytes (CLs) express potent toxins, including perforin (P) and granzyme-B (G), which brings about target cell death. The purpose of this study was to evaluate the killing capacity of tumor-infiltrating CLs by means of P and G analysis, and explore the association with lymph node metastasis in papillary carcinoma of thyroid (PTC) without Hashimoto's thyroiditis (HT).</p><p><strong>Methods: </strong>Infiltration of lymphocytes in PTC was observed in frozen sections. Both fresh tumor tissues and paracancerous tissues with lymphocyte infiltration were collected and prepared into a single cell suspension. Flow cytometry was used to detect the percentages of CD3<sup>+</sup>P<sup>+</sup>, CD3<sup>+</sup>G<sup>+</sup>, CD8<sup>+</sup>P<sup>+</sup>, and CD8<sup>+</sup>G<sup>+</sup> T lymphocytes (TLs) and CD16-CD56<sup>+</sup>P<sup>+</sup> and CD16-CD56<sup>+</sup>G<sup>+</sup> natural killer (NK) cells. Finally, we investigated differential expression of P and G in NK cells and cytotoxic T lymphocytes (CTLs) in paired tumor tissues (group T, n = 44) and paracancerous tissues (group N, n = 44) from patients with PTC with the BRAF V600E mutation. Furthermore, patients were divided into two groups according to whether cervical central lymph node metastasis (CCLNM) existed: group A (with lymph node metastases, n = 27) and group B (with nonlymph node metastases, n = 17). Patients were also divided into three groups according to the total number of positive CCLNM: group B, group C (with low-level lymph node metastases, less than 5, n = 17) and group D (with high-level lymph node metastases, no less than 5, n = 10).</p><p><strong>Results: </strong>The percentage of CD3<sup>+</sup>P<sup>+</sup> CTLs was significantly higher in group N than in group T (P < 0.05). The percentage of CD8<sup>+</sup>G<sup>+</sup> CTLs was significantly higher in group T than in group N (P < 0.05). The percentages of CD3<sup>+</sup>G<sup>+</sup>, CD16-CD56<sup>+</sup>P<sup>+</sup>and CD16-CD56<sup>+</sup>G<sup>+</sup> NK cells showed no significant difference in either group T or group N (P > 0.05). The percentages of CD3<sup>+</sup>P<sup>+</sup> CTLs in group A and group C were significantly higher in the paracancerous tissue than in the tumor tissue (P < 0.05). The percentages of CD8<sup>+</sup>G<sup>+</sup> CTLs in group A and group C were significantly higher in the tumor tissues than in the paracancerous tissues (P < 0.05). The percentage of CD16-CD56<sup>+</sup>G<sup>+</sup> NK cells in group D was significantly higher in the tumor tissues than in the paracancerous tissues (P < 0.05).</p><p><strong>Conclusions: </strong>The killing capacity of infiltrating CLs in PTC differed between tumor tissues and paracancerous tissues. In cases with CCLNM, higher expression of CD16-CD56<sup>+</sup>G<sup>+</sup> NK cells in tumor tissues may be associated with a high risk of lymph node metastasis.</p>\",\"PeriodicalId\":11237,\"journal\":{\"name\":\"Diagnostic Pathology\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":2.4000,\"publicationDate\":\"2024-02-10\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10858496/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Diagnostic Pathology\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.1186/s13000-024-01454-9\",\"RegionNum\":3,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"PATHOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Diagnostic Pathology","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1186/s13000-024-01454-9","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"PATHOLOGY","Score":null,"Total":0}
引用次数: 0
摘要
背景:细胞毒性淋巴细胞(CLs)表达强效毒素,包括穿孔素(P)和颗粒酶-B(G),可导致靶细胞死亡。本研究的目的是通过P和G分析评估肿瘤浸润CL的杀伤能力,并探讨其与无桥本氏甲状腺炎(HT)的甲状腺乳头状癌(PTC)淋巴结转移的关系:方法:在冰冻切片中观察PTC中淋巴细胞的浸润情况。收集有淋巴细胞浸润的新鲜肿瘤组织和癌旁组织,制备成单细胞悬液。使用流式细胞术检测 CD3+P+、CD3+G+、CD8+P+ 和 CD8+G+ T 淋巴细胞(TL)以及 CD16-CD56+P+ 和 CD16-CD56+G+ 自然杀伤(NK)细胞的百分比。最后,我们研究了 BRAF V600E 突变的 PTC 患者配对肿瘤组织(T 组,n = 44)和癌旁组织(N 组,n = 44)中 NK 细胞和细胞毒性 T 淋巴细胞(CTLs)中 P 和 G 的不同表达。此外,根据是否存在宫颈中央淋巴结转移(CCLNM)将患者分为两组:A 组(有淋巴结转移,n = 27)和 B 组(无淋巴结转移,n = 17)。根据 CCLNM 阳性总数将患者分为三组:B 组、C 组(低位淋巴结转移,少于 5 个,n=17)和 D 组(高位淋巴结转移,不少于 5 个,n=10):N组CD3+P+ CTLs的百分比明显高于T组(P+G+ CTLs的百分比明显高于N组(P+G+、CD16-CD56+P+和CD16-CD56+G+ NK细胞在T组和N组均无明显差异(P>0.05))。A 组和 C 组 CD3+P+ CTLs 在癌旁组织中的比例明显高于肿瘤组织(P +G+ CTLs 在肿瘤组织中的比例明显高于癌旁组织(P +G+ NK 细胞在肿瘤组织中的比例明显高于癌旁组织(P 结论:CD3+P+ CTLs 在肿瘤组织中的比例明显高于癌旁组织(P +G+ NK 细胞在肿瘤组织中的比例明显高于癌旁组织(P 结论:CD3+P+ CTLs 在肿瘤组织中的比例明显高于癌旁组织(P +G+ CTLs 在肿瘤组织中的比例明显高于癌旁组织(PPTC中浸润CL的杀伤能力在肿瘤组织和癌旁组织之间存在差异。在CCLNM病例中,肿瘤组织中CD16-CD56+G+ NK细胞的高表达可能与淋巴结转移的高风险有关。
Killing capacity analysis of tumor-infiltrating cytotoxic lymphocytes and impact on lymph node metastasis in differentiated papillary carcinoma of thyroid with the BRAF V600E mutation.
Background: Cytotoxic lymphocytes (CLs) express potent toxins, including perforin (P) and granzyme-B (G), which brings about target cell death. The purpose of this study was to evaluate the killing capacity of tumor-infiltrating CLs by means of P and G analysis, and explore the association with lymph node metastasis in papillary carcinoma of thyroid (PTC) without Hashimoto's thyroiditis (HT).
Methods: Infiltration of lymphocytes in PTC was observed in frozen sections. Both fresh tumor tissues and paracancerous tissues with lymphocyte infiltration were collected and prepared into a single cell suspension. Flow cytometry was used to detect the percentages of CD3+P+, CD3+G+, CD8+P+, and CD8+G+ T lymphocytes (TLs) and CD16-CD56+P+ and CD16-CD56+G+ natural killer (NK) cells. Finally, we investigated differential expression of P and G in NK cells and cytotoxic T lymphocytes (CTLs) in paired tumor tissues (group T, n = 44) and paracancerous tissues (group N, n = 44) from patients with PTC with the BRAF V600E mutation. Furthermore, patients were divided into two groups according to whether cervical central lymph node metastasis (CCLNM) existed: group A (with lymph node metastases, n = 27) and group B (with nonlymph node metastases, n = 17). Patients were also divided into three groups according to the total number of positive CCLNM: group B, group C (with low-level lymph node metastases, less than 5, n = 17) and group D (with high-level lymph node metastases, no less than 5, n = 10).
Results: The percentage of CD3+P+ CTLs was significantly higher in group N than in group T (P < 0.05). The percentage of CD8+G+ CTLs was significantly higher in group T than in group N (P < 0.05). The percentages of CD3+G+, CD16-CD56+P+and CD16-CD56+G+ NK cells showed no significant difference in either group T or group N (P > 0.05). The percentages of CD3+P+ CTLs in group A and group C were significantly higher in the paracancerous tissue than in the tumor tissue (P < 0.05). The percentages of CD8+G+ CTLs in group A and group C were significantly higher in the tumor tissues than in the paracancerous tissues (P < 0.05). The percentage of CD16-CD56+G+ NK cells in group D was significantly higher in the tumor tissues than in the paracancerous tissues (P < 0.05).
Conclusions: The killing capacity of infiltrating CLs in PTC differed between tumor tissues and paracancerous tissues. In cases with CCLNM, higher expression of CD16-CD56+G+ NK cells in tumor tissues may be associated with a high risk of lymph node metastasis.
期刊介绍:
Diagnostic Pathology is an open access, peer-reviewed, online journal that considers research in surgical and clinical pathology, immunology, and biology, with a special focus on cutting-edge approaches in diagnostic pathology and tissue-based therapy. The journal covers all aspects of surgical pathology, including classic diagnostic pathology, prognosis-related diagnosis (tumor stages, prognosis markers, such as MIB-percentage, hormone receptors, etc.), and therapy-related findings. The journal also focuses on the technological aspects of pathology, including molecular biology techniques, morphometry aspects (stereology, DNA analysis, syntactic structure analysis), communication aspects (telecommunication, virtual microscopy, virtual pathology institutions, etc.), and electronic education and quality assurance (for example interactive publication, on-line references with automated updating, etc.).