肺癌患者外周血CD+8 自然杀伤T细胞表面受体NKG2D和NKG2A表达的临床意义

 【摘要】　目的　检测肺癌患者外周血CD+8 自然杀伤（NK）T细胞表面受体NKG2D和NKG2A的表达,探讨二者表达失衡与肺癌免疫逃逸的关系。方法　选择95例原发未治疗的肺癌患者,采用流式细胞术检测CD+8 NKT细胞表面受体NKG2D和NKG2A的表达,以50名健康人为对照。结果　肺癌组CD+8 NKT细胞NKG2D+表达率[（77.07±5.77）%]明显低于对照组[（84.13±4.49）%],差异有统计学意义（t＝8.14,P＜0.05）;在TNM分期中,Ⅰ～ⅢA、ⅢB、Ⅳ期患者CD+8 NKT细胞NKG2D+表达率依次为（81.07±5.02）%、（76.95±4.70）%、（72.80±5.16）%,差异有统计学意义（F＝18.74,P＜0.05）。肺癌组CD+8 NKT细胞NKG2A+表达率[（33.58±8.82）%]明显高于对照组[（25.31±8.38）%],差异有统计学意义（t＝-5.46,P＜0.05）;在TNM分期中,Ⅰ～ⅢA、ⅢB、Ⅳ期患者CD+8 NKT细胞NKG2A+的表达率依次为（25.10±6.93）%、（33.24±3.76）%、（43.64±6.10）%,差异有统计学意义（F＝75.73,P＜0.05）。结论　肺癌患者CD+8 NKT细胞表面NKG2A和NKG2D表达失衡可能抑制该细胞的杀伤功能,而这可能是肿瘤免疫逃逸的机制之一。     

Reversed CD4/CD8 ratios of tumor-infiltrating lymphocytes are correlated with the progression of human cervical carcinoma.

BACKGROUND: To investigate the clinical significance of tumor-infiltrating lymphocytes (TILs) within the tumor milieu of human cervical carcinoma, the authors quantitatively measured and compared the subpopulations of lymphocytes infiltrating the neoplastic cervix. METHODS: A total of 30 patients with Stage Ia-IIa cervical carcinoma were enrolled. TILs were isolated from tissue specimens by means of a mechanical dispersal technique, and the immunocyte subsets were quantified with dual-color flow cytometry. Bulky tumor was defined as tumor size >4 cm in greatest dimension according to the 1995 staging of the International Federation of Gynecology and Obstetrics. RESULTS: The CD4/CD8 ratios of TILs were reversed in both cervical squamous cell carcinoma (n = 20) and cervical adenocarcinoma (n = 10). The proportion of CD4(+) T cells was significantly lower in tumors from patients with lymph node metastasis (n = 8) than in those from patients without lymph node metastasis (n = 22) (24.5 vs. 32.7, P = 0.001), as was the reversed CD4/CD8 ratio (0.50 vs. 0.81, P = 0.001). The proportion of CD4(+) T cells was much lower in bulky tumors (n = 5) than in nonbulky tumors (n = 25) (21.4 vs. 32.5, P < 0.001), reflecting in a more strongly reversed CD4/CD8 ratio (0.41 vs. 0.81, P = 0.001). CONCLUSIONS: Decreased proportions of tumor-infiltrating CD4+ T cells with reversed CD4/CD8 ratios are highly correlated with rapid tumor growth and lymph node metastasis in cervical carcinoma. The regional immune escape is of prognostic importance with regard to cancer progression.     

Altered CD94/NKG2A and perforin expression reduce the cytotoxic activity in malignant pleural effusions

CD94/NKG2A is an inhibitory receptor expressed by NK cells and cytotoxic lymphocytes and, upon activation by HLA-E, downregulates the cytolytic activities of these cells thus representing a tumour immune escape mechanism.This study was aimed at assessing whether cytotoxic lymphocytes (CD8+) and NK cells from malignant pleural effusions have a deregulated expression of CD94/NKG2A.The expression of membrane CD94/NKG2A and perforin was evaluated by flow-cytometry in CD8+ and NK cells from pleural effusions and autologous peripheral blood of cancer (n = 19) and congestive heart failure (CHF) (n = 11) patients. Intracellular CD94/NKG2A expression was evaluated by flow-cytometry in pleural effusion CD8+ and NK cells from cancer patients (n = 10). Cytotoxic activity against cancer cells exerted by pleural and autologous peripheral blood T lymphocytes from cancer patients was assessed by flow-cytometry assay.Pleural CD8+ from cancer patients showed a reduced expression of membrane CD94/NKG2A and perforin when compared to autologous peripheral blood and CHF pleural effusions. Reduced numbers of NK cells were present in pleural effusions from both cancer and CHF patients. Pleural NK from cancer patients showed a reduced expression of membrane CD94/NKG2A and perforin when compared to autologous peripheral blood. Pleural T lymphocytes from cancer patients exhibited a reduced cytotoxic activity against cancer cells when compared to autologous peripheral blood T lymphocytes. The intracellular expression of CD94/NKG2A in CD8+ and NK cells from cancer patients was higher than membrane expression.In conclusion, this study provides compelling evidences of new mechanisms underlying the reduced host defence against cancer within the pleural space.     

抑制性杀伤细胞受体在NK细胞和T细胞上的表达及其与急性移植物抗宿主病的关系

 目的 研究异基因造血干细胞移植（allo-HSCT）患者移植前后外周血NK及T细胞上4种抑制性杀伤细胞受体（CD158a、CD158b、NKB1和CD94／NKG2A）的表达及其与急性移植物抗宿主病（aGVHD）的关系。方法 采用流式细胞术检测NK及T细胞上抑制性杀伤细胞受体的表达。结果 NK细胞上CD158a和CD158b的表达于移植后3～4个月、NKB1和CD94／NKG2A的表达于移植后2个月恢复移植前水平。移植前后CD158a和NKB1在CD+3 T细胞上持续低水平表达;移植前CD158b和CD94／NKG2A在CD+3 T细胞上的表达水平较高,且主要表达于CD+8 T细胞上,移植后其在CD+8 T细胞上的表达增加。CD+8 T细胞上CD158b的表达在发生Ⅰ度aGVHD时显著增高,与无aGVHD组及Ⅱ~Ⅳ度aGVHD组相比,差异均有统计学意义（P均＜0.05）;在发生Ⅱ~Ⅳ度aGVHD时增高不明显,与无aGVHD组相比,差异无统计学意义（P＞0.05）。结论 CD158b在CD+8 T细胞上高表达可能有助于降低T细胞同种反应性,减轻aGVHD的程度。     

Characterization of the cytolytic activity of CD4+ and CD8+ tumor-infiltrating lymphocytes in human renal cell carcinoma

Previously we showed that IL2 expanded tumor-infiltrating lymphocytes (TILs) from renal cell carcinoma mediated non-major histocompatibility complex-restricted cytotoxicity. Phenotypic analysis showed that cultured TILs were composed mostly of T-lymphocytes with varying numbers of CD4+, CD8+, and CD56+ (Leu19+) populations. Here we compared the cytolytic activity of the two predominant TIL subsets, CD3+CD4+ and CD3+CD8+, to that of the CD56+ populations. Using magnetic beads coated with antibodies to either CD4 or CD8, CD3+CD4+, and CD3+CD8+ TILs were isolated in a highly enriched form (greater than 92%) and could be expanded for over 40 days in vitro with 1000 units/ml IL2. In a 4-h 51Cr release assay the CD4+ and CD8+ TILs showed minimal lytic activity, whereas unseparated cells exhibited significant levels of non-major histocompatibility complex-restricted cytotoxicity. The lytic activity seen in the 4-h assay with unseparated TILs appeared to be related to the presence of CD56+ populations. With one exception none of the purified CD4+ or CD8+ TILs expressed any significant levels of CD56, while the unseparated TILs contained varying numbers of CD3+CD56+ and CD3-CD56+ populations. Cell-sorting experiments verified that the CD56+ populations were responsible for most of the lytic activity in 4 h even though CD3+CD56- cells represented the predominant cell type. Although CD3+CD56- TILs were minimally lytic in 4 h, we show here that both CD3+CD4+ and CD3+CD8+ subsets displayed substantial cytotoxicity in long-term assays. In the 18-h 51Cr release assay 5 of 6 CD4+ and 2 of 3 CD8+ TILs were lytic for the autologous tumor. In two cases, restimulation with the autologous tumor induced augmented cytolytic activity of TIL subsets and in one case induced lytic activity in 4 h. The cytotoxic activity of TIL subsets was further examined using a 72-h assay in which TILs were cocultured with a confluent layer of tumor cells. The degree of cytotoxicity was quantitated by measuring the amount of crystal violet dye that was incorporated by tumor cells which remained after the incubation period. CD4+ and CD8+ TILs typically caused greater than a 50% reduction of tumor cells in 3 days and the level of reduction was increased when IL2 was added to the cultures. All the CD4+ and CD8+ subset preparations were cytotoxic in the 3-day assay even though some were not lytic for certain targets in the 18-h 51Cr release assay.(ABSTRACT TRUNCATED AT 400 WORDS)     

CD8~+NKT细胞表面活化性受体NKG2D在肺癌患者外周血中的表达及临床意义

背景与目的 NKT细胞活化性受体NKG2D及sMICA是近来肿瘤免疫研究领域的热点之一。本研究旨在观察肺癌患者外周血中CD8+NKT细胞受体NKG2D表达水平的变化,并对NKG2D及sMICA进行相关性分析,探讨它们在肺癌免疫监视中的作用及临床意义。方法选择82例初发未治疗的肺癌患者,采用流式细胞术检测外周血CD8+NKT细胞活化性受体NKG2D的表达,并以45例健康人作对照,采用酶联免疫吸附法检测肺癌患者血清中sMICA的表达,分析NKG2D与肺癌临床生物学特征的关系。结果肺癌患者外周血中CD8+NKT细胞表面活化性受体NKG2D水平均低于对照组,差异有统计学意义(P<0.001)。随TNM分期的增加,NKG2D的表达率逐渐降低。其中IV期肺癌患者NKG2D的表达明显低于I期-II期及III期患者该受体的表达,差异有统计学意义(P<0.001)。肺Ca患者中吸烟人群外周血中CD8+NKT细胞受体NKG2D的表达较非吸烟者低,差异有统计学意义(P<0.05)。CD8+NKT细胞受体NKG2D与sMICA呈负相关(r=-0.598,P<0.001)。结论肺癌患者CD8+NKT细胞表面受体NKG2D在外周血中低表...     

Enhancement by monocytes of perforin production and its gene expression by human CD8+ T cells stimulated with interleukin-2.

Pore-forming protein (PFP) is an important effector molecule for cytotoxicity mediated by cytotoxic T cells and NK cells. In the present study, the effect of monocytes on PFP production by interleukin-2 (IL-2)-stimulated T lymphocytes was examined. Highly purified lymphocytes (> 99%) and monocytes (> 90%) were isolated by centrifugal elutriation from peripheral blood of healthy donors, and, CD4+ and CD8+ cells were isolated from the purified lymphocytes by using antibody-bound magnetic beads. PFP production was quantitated with a universal microspectrophotometer in combination with immunostaining using anti-PFP antibody. Monocytes did not produce any PFP. High levels of PFP production were observed in CD8+ cells, but not CD4+ cells after incubation for 4 days with IL-2. Addition of monocytes to cultures of CD8+ cells resulted in significant augmentation of PFP production after 3 days' stimulation with IL-2. Monokines (TNF alpha and IL-6) caused a significant increase in PFP production by IL-2-stimulated CD8+ cells. Northern blot analysis revealed that the PFP mRNA levels was enhanced by stimulation with IL-2, and that addition of monocytes to cultures of CD8+ cells plus IL-2 augmented their PFP mRNA expression. These observations suggest that monocytes are important in in situ regulation of the CD8+ T cell-mediated cytotoxic response through production of PFP.     

Identification of an enriched CD4+ CD8α++ CD8β+ T-cell subset among tumor-infiltrating lymphocytes in human renal cell carcinoma

Three-color immunofluorescence flow-cytometric analysis of freshly isolated tumor-infiltrating lymphocytes (TIL) from patients with primary renal cell carcinoma (RCC) revealed a unique, not previously described TIL subset with a CD3₊ CD4₊ CD8α₊₊ CD8β₊ phenotype. This subset represented at least 5% of CD3₊ TIL in 15 of 21 patients with clear cell RCC, whereas it was not or only marginally represented in patients with papillary RCC or sarcomatoid RCC. In one-third of the patients with clear cell RCC, more than 20% of CD3₊ TIL and in one patient more than half of the CD3₊ TIL displayed this phenotype. The occurrence of this subset was not associated with pathological stage, tumor diameter, nuclear grade, cytogenetic abnormalities or vascular invasion in this patient cohort. When present, the CD3₊ CD8α₊₊ CD8β₊ subset was detected in similar proportions in tumor tissue and tumor capsula, and it was also detected in adjacent non-tumoral renal tissue, albeit in much lower proportions. Despite strong cell surface expression of various activation markers (CD69, CD54 and HLA-DR), CD3₊ CD4⁺ CD8α₊₊ anti-CD3-redirected cytotoxicity assay. In contrast with CD3₊ CD4₊ CD8₋ cells from the same tumor sample, they were markedly deficient in IL-2Rα up-regulation following anti-CD3 triggering. The possibility that these cells represent either anergic cells or a highly specialized effector population with a discrete, as yet undescribed function is discussed. Int. J. Cancer 71:178–182, 1997. © 1997 Wiley-Liss, Inc.     

An increase in CD4+CD25+FOXP3+ regulatory T cells in tumor-infiltrating lymphocytes of human glioblastoma multiforme

The subpopulation of CD4+CD25+ immunoregulatory T (Tr) cells constitutes 5%-10% of CD4+ cells in humans. These cells play a crucial role in the control of tumor immune response. In this study, we evaluated the distribution of Tr cells in tumor-infiltrating lymphocytes of human glioblastoma multiforme and examined the difference between the brain and autologous blood with respect to Tr cells. Glioma samples from 10 patients were classified as WHO grade IV astrocytoma. Control samples were obtained from patients undergoing resection of a seizure focus. The samples were analyzed by flow cytometry to determine the frequency of Tr cells and by real-time PCR for forkhead box P3 (FOXP3) expression. We then examined the expression of CD62L, CD45RO, and cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) and assessed the functionality of Tr cells in vitro. There was a significant difference in the number of FOXP3-expressing CD4+CD25+ T cells within glioma-infiltrating lymphocytes as compared to controls (P < 0.01). This difference was further observed in studies of autologous patient blood and control blood. The expression level of FOXP3 mRNA was high in Tr cells and weak in CD4+CD25-T cells. Moreover, the expression of CD62L and CTLA-4 was elevated in glioma Tr cells as compared to that in the controls. These cells were also CD45RO positive. Functional assays confirmed the suppressive activity of Tr cells in patients with glioma. The expression of CD4+CD25+FOXP3+ T cells was significantly higher in patients with glioblastoma multiforme than in controls. This increase in the frequency of Tr cells that display suppressive activity might play a role in modulation of the immune response against glioma. In light of these findings, Tr cells may represent a potential target for immunotherapy of malignant brain tumors.     

Ex vivo expansion of CD8+CD56+ and CD8+CD56- natural killer T cells specific for MUC1 mucin

Prostate cancers express MUC1, but nearly all metastatic cells lack HLA class I molecules. Thus, a lymphocyte population that can sense its antigenic environment, while also able to react to stimuli of natural killer (NK) cells, may be a more versatile effector cell population for antitumor immune responses. Herein, we report that tumor-specific MUC1 peptide, interleukin 2, and interleukin 12 act synergistically to stimulate the ex vivo expansion of CD8(+)CD56(-) T cells and CD8(+)CD56(+) natural killer T (NKT) cells from the peripheral blood mononuclear cells of prostate cancer patients, as well as healthy male and female donors. Both the CD56(+) NKT cells and CD56(-) T cells lysed allogeneic mucin-bearing target cells, as well as NK target cells, but not lymphokine-activated killer target cells. However, the CD56(+) NKT cells displayed a 2-fold greater cytolytic activity than the CD56(-) T cells. The mucin-specific cytolytic activity and NK cytolytic activities for both lymphocyte populations were independent of HLA class I and CD1 molecules. The CD56(-) T cells up-regulated CD56 with continued antigenic stimulation in the presence of interleukin 12, suggesting that CD8(+)CD56(-) T cells are NKT cells. However, CD56(+) NKT cells expand poorly to continued stimulation. All mucin-stimulated NKT cells exhibited the activated/memory CD45RO phenotype. The NKT cell lines express the alpha/beta T-cell receptor (TCR). The TCR repertoire was limited and varied with cell line, but was not the V alpha 24V beta 11 TCR typically associated with NKT cells. Whereas CD161 is generally considered a marker of NKT cells, the mucin-stimulated NKT cells did not express this marker. Thus, we have described two phenotypically distinct NKT types that do not display a biased TCR repertoire, but do display specificity for a tumor-specific peptide antigen (CTL-like activity), as well as HLA class I-deficient target cells (NK-like activity).     

A correlation between the expression of CD 8 antigen and specific cytotoxicity of tumor-infiltrating lymphocytes

Tumor-infiltrating lymphocytes (TIL) from six gynecologic malignant tumors (two uterine cervical cancers, two ovarian serous cystadenocarcinomas, and two uterine corpus cancers), cultured in the presence of recombinant interleukin 2, were assayed for their cytotoxic activities against various fresh tumor cells including autologous tumors. A clear correlation between phenotype and cytotoxic activity of TIL was observed. Four of six TIL preparations exhibited strong cytotoxic activity against autologous fresh tumor target cells, and were all CD8+. In contrast, cytotoxic activity was not detected in any of the CD4+ TIL preparations. The cytotoxic activities of the CD8+ TIL preparations were highly specific; only autologous fresh tumor cells were lysed. This result is consistent with the notion that TIL are of a different cell lineage from lymphokine-activated killer cells which are antigen-nonspecific and CD8-. Instead, TIL appear to be of cytotoxic T cell lineage that is highly antigen-specific and CD8+. To explore the potential for clinical use, we have attempted to augment the cytotoxic activities of these CD8+ TIL by treatment of the target tumor cells with gamma interferon (IFN) in vitro, hoping that elevated expression of MHC class I gene products on the cell surface would enhance their recognition. It was observed that brief treatment of freshly prepared tumor cells in vitro with gamma-IFN resulted in augmentation of the expression of MHC class I gene products, and the treated tumor cells were more susceptible to lysis by TIL than untreated cells.     

CD3+ CD4+ and CD3+ CD8+ Lymphocyte Subgroups and their Surface Receptors NKG2D and NKG2A in Patients with Nonsmall Cell Lung Cancer

Background: To explore the prevalence of lymphocyte subgroups CD3+ CD4+ and CD3+ CD8+ and their surfacereceptors NKG2D and NKG2A in patients with non-small cell lung cancer (NSCLC). Materials and Methods: Atotal of 40 patients with NSCLC were divided into different groups according to different clinical factors (TNMstaging, pathological patterns and genders) for assessment of relations with CD3+ CD4+ and CD3+ CD8+ and thesurface receptors NKG2D and NKG2A of T lymphocytes in peripheral blood by flow cytometry. Results: Patientsin the advanced group had evidently lower levels of CD3+ CD4+ but markedly higher levels of CD3+ CD8+ inperipheral blood than those with early lesions (p<0.05). In addition, NSCLC patients in the advanced group hadobviously higher CD3+ CD4+ NKG2D and CD3+ CD8+ NKG2A expression rates but lower CD3+ CD4+ NKG2A andCD3+ CD8+ NKG2D expression rates (p<0.05). However, there were no significant differences between NSCLCpatients with different genders and pathological patterns in expression levels of lymphocyte subgroups CD3+CD4+ and CD3+ CD8+ and their surface receptors NKG2D and NKG2A. Conclusions: Unbalanced expressionof surface receptors NKG2D and NKG2A in CD3+ CD4+ and CD3+ CD8+ lymphocytes may be associated witha poor prognosis, greater malignancy and immunological evasion by advanced cancers, related to progressionof lung cancer.     

CD4+/CD8+ T淋巴细胞在肝细胞癌组织中的浸润及与预后的相关性研究

目的 检测CD4⁺/CD8⁺ T淋巴细胞在肝细胞癌（hepatocellular carcinoma,HCC）组织中的浸润程度,并分析其与预后的相关性。方法 收集行肝切除术的HCC患者215例,采用免疫组化技术检测CD4⁺/CD8⁺ T淋巴细胞在HCC癌组织中的浸润程度,根据浸润情况比较患者肝切除术后无瘤生存率和总生存率。结果 CD4⁺ T淋巴细胞高浸润和低浸润比例分别为60.9%和39.1%。CD4⁺ T淋巴细胞高浸润组患者总生存率和无瘤生存率均显著高于低浸润组（P=0.015,P=0.038）。CD8⁺ T淋巴细胞高浸润和低浸润比例分别为34.9%和65.1%。CD8⁺ T淋巴细胞高浸润组患者的总生存率和无瘤生存率亦显著高于低浸润组患者（P=0.033,P=0.047）。结论 CD4⁺或CD8⁺ T淋巴细胞低浸润可能与HCC患者术后不良预后相关。     

High number of intraepithelial CD8+ tumor-infiltrating lymphocytes is associated with the absence of lymph node metastases in patients with large early-stage cervical cancer

In a prospective study, we have examined the tumor-specific immune response in a group of 59 patients with human papillomavirus (HPV) 16-positive (HPV16(+))-induced or HPV18(+)-induced cervical cancer. Local antitumor immunity was analyzed by the enumeration of tumor-infiltrating dendritic cells and CD4+, CD8+, and regulatory T cells as well as by calculation of the ratio of CD8+/CD4+ T cells and CD8+/regulatory T cells. Systemic tumor-specific immunity was assessed by determination of the HPV E6- and/or E7-specific T-cell response in the blood of these patients. Finally, these variables were evaluated with respect to known histopathologic prognostic variables, including the absence (LN-) or presence (LN+) of lymph node metastases. Stratification according to the lymph node status of patients revealed a significantly stronger CD8+ T-cell tumor infiltration, a higher CD8+/CD4+ T-cell ratio, and higher CD8+/regulatory T-cell ratio in the group of patients in which the tumor failed to metastasize to the tumor-draining lymph node. Subdivision according to the presence (IR+) or absence (IR-) of circulating HPV-specific T cells disclosed that the highest number of tumor-infiltrating CD8+ T cells was found in the group of LN- patients displaying a concomitant systemic tumor-specific immune response (LN-IR+). CD8+ T-cell infiltration in LN-IR- patients was comparable with that of LN+ patients. In cervical cancer, the absence of lymph node metastases is strongly associated with a better prognosis. Our data indicate that, especially in a subgroup of LN- patients, a strong and effective interaction between immune system and tumor exists. This subgroup of cervical cancer patients may have the best prognosis.     

光动力对小鼠鼻咽癌移植瘤survivin表达及CD+8细胞毒性T细胞的影响

目的 观察氨基酮戊酸(5-Aminolevulinic acid,ALA)介导的光动力疗法(photodynamic therapy,PDT)对小鼠鼻咽癌移植瘤survivin蛋白表达及CD+8细胞毒性T细胞(CD+8 CTLs)的影响.探讨ALA-PDT治疗鼻咽癌的机制.方法 32只 SPF级BALB/c 小鼠皮下接种鼻咽癌CNE 2细胞建立荷瘤鼠模型,随机分为2组:PDT组、对照组.PDT治疗后定期采用免疫组化法检测肿瘤组织survivin蛋白表达及CD+8细胞毒性T细胞.结果 治疗后24h,PDT组survivin蛋白表达明显低于对照组(P＜0.05),肿瘤局部CD+8细胞数量较对照组无明显变化(P＞0.05).治疗后72h,PDT组肿瘤局部CD+8细胞数量均明显高于对照组(P＜0.05).结论 抑制survivin蛋白的表达以及产生局部免疫效应均可能是ALA-PDT治疗鼻咽癌的机制之一.     

Analysis of radiotherapy-induced alteration of CD8+ T cells and PD-L1 expression in patients with uterine cervical squamous cell carcinoma

Radiotherapy induces an immune response in the cancer microenvironment that may influence clinical outcome. The present study aimed to analyse the alteration of CD8⁺ T-cell infiltration and programmed death-ligand 1 (PD-L1) expression following radiotherapy in clinical samples from patients with uterine cervical squamous cell carcinoma. Additionally, the current study sought to analyse the association between these immune responses and clinical outcomes. A total of 75 patients who received either definitive chemoradiotherapy or radiotherapy were retrospectively analyzed. CD8⁺ T-cell infiltration and PD-L1 expression were determined by immunohistochemistry using biopsy specimens before radiotherapy (pre-RT) and after 10 Gy radiotherapy (post-10 Gy). The PD-L1⁺ rate was significantly increased from 5% (4/75) pre-RT to 52% (39/75) post-10 Gy (P<0.01). Despite this increase in the PD-L1⁺ rate post-10 Gy, there was no significant association between both pre-RT and post-10 Gy and overall survival (OS), locoregional control (LC) and progression-free survival (PFS). On the other hand, the CD8⁺ T-cell infiltration density was significantly decreased for all patients (median, 23.1% pre-RT vs. 16.9% post-10 Gy; P=0.038); however, this tended to increase in patients treated with radiotherapy alone (median, 17.7% pre-RT vs. 24.0% post-10 Gy; P=0.400). Notably, patients with high CD8⁺ T-cell infiltration either pre-RT or post-10 Gy exhibited positive associations with OS, LC and PFS. Thus, the present analysis suggested that CD8⁺ T-cell infiltration may be a prognostic biomarker for patients with cervical cancer receiving radiotherapy. Furthermore, immune checkpoint inhibitors may be effective in patients who have received radiotherapy, since radiotherapy upregulated PD-L1 expression in cervical cancer specimens.