Bone marrow-derived dendritic cells pulsed with tumor lysates induce anti-tumor immunity against gastric cancer ex vivo

AIM： To investigate whether bone marrow-derived denritic cells pulsed with tumor lysates induce immunity against gastric cancer ex vivo. METHODS： c-kit＋ hematopoietic progenitor cells were magnetically isolated with a MiniMACS separator from BALB/c mice bone marrow cells. These cells were cultured with cytokines GM-CSF, IL-4, and TNFα to induce their maturation. They were analysed by morphological observation, phenotype analysis, and mixed lymphocyte reaction （MLR）. Bone marrowderived DCs （BM-DCs） were pulsed with tumor cell lysate obtained by rapid freezing and thawing at a 1：3 DC：tumor cell ratio. Finally, cytotoxic T lymphocyte （CTL） activity and interferon gamma （IFNγ） secretion was evaluated ex vivo. RESULTS： c-kit^＋ hematopoietic progenitor cells from mice bone marrow cells cultured with cytokines for 8 d showed the character of typical mature DCs.Morphologically, observed by light microscope, these cells were large with oval or irregularly shaped nuclei and with many small dendrites. Phenotypically, FACS analysis showed that they expressed.high levels of la, DEC-205, CD11b, CD80 and CD86 antigen, moderate levels of CD40, and negative for F4/80. Functionally, these cells gained the capacity to stimulate allogeneic T cells in MLR assay. However, immature DCs cultured with cytokines for 5 d did not have typical DCs phenotypic markers and could not stimulate allogeneic T cells. Ex vivo primed T cells with SGC-7901 tumor cell lysate-pulsed （TP） DCs were able to induce effective CTL activity against SGC-7901 tumor cells （E：T = 100：1, 69.55% ± 6.05% specific lysis）, but not B16 tumor cells, and produced higher levels of IFNγ, when stimulated with SGC-7901 tumor cells but not when stimulated with B16 tumor cells （1575.31 ± 60.25 pg/mL in SGC-7901 group vs 164.11± 18.52 pg/mL in B16 group, P 〈 0.01）. CONCLUSION： BM-derived DCs pulsed with tumor lysates Can induce anti-tumor immunity specific to gastric cancer ex vivo.

Gastric cancer cells induce human CD4^+ Foxp3^+ regulatory T cells through the production of TGF-β1

AIM: To elucidate the molecular and cellular features responsible for the increase of regulatory T cells (Tregs) in gastric cancer. METHODS: The frequencies of CD4 + Foxp3 + Tregs and the level of transforming growth factor-β1 (TGF-β1) were analyzed from 56 patients with gastric cancer byflow cytometry and enzyme-linked immunosorbent assay respectively. Foxp3 gene expression was analyzed by real-time polymerase chain reaction. The gastric cancer microenvironment was modeled by establishing the coculture of gastric cancer cell line, MGC-803, with sorting CD4 + T cells. The normal gastric mucosa cell line, GES-1, was used as the control. The production of TGF-β1 was detected in supernatant of MGC and GES-1. The carboxyfluorescein diacetatesuccinimidyl ester (CFSE) dilution assay was performed to evaluate the proliferation characteristics of induced Tregs. Neutralizing anti-TGF-β1 antibody was added to the co-culture system for neutralization experiments. RESULTS: The level of serum TGF-β1 in gastric cancer patients (15.1 ± 5.5 ng/mL) was significantly higher than that of the genderand age-matched healthy controls (10.3 ± 3.4 ng/mL) (P < 0.05). Furthermore, the higher TGF-β1 level correlated with the increased population of CD4 + Foxp3 + Tregs in advanced gastric cancer (r = 0.576, P < 0.05). A significant higher frequency of CD4 + Foxp3 + Tregs was observed in PBMCs cultured with the supernatant of MGC than GES-1 (10.6% ± 0.6% vs 8.7% ± 0.7%, P < 0.05). Moreover, using the purified CD4 + CD25 T cells, we confirmed that the increased Tregs were mainly induced from the conversation of CD4 + CD25 naive T cells, and induced Tregs were functional and able to suppress the proliferation of effector T cells. Finally, we demonstrated that gastric cancer cells induced the increased CD4 + Foxp3 + Tregs via producing TGF-β1. Gastric cancer cells upregulated the production of TGF-β1 and blockade of TGF-β1 partly abrogated Tregs phenotype. CONCLUSION: Gastric cancer cell can induce Tregs development via producing TGF-β1, by which the existence of cross-talk between the tumor and immune cells might regulate anti-tumor immune responses.

Gastric cancer-derived heat shock protein-gp96 peptide complex enhances dendritic cell activation

AIM To investigate the role of heat shock protein (HSP)glycoprotein (gp) 96 in dendritic cells (DCs) and lymphocytes induction in gastric cancer (GC). METHODS Human GC cell lines KATOIII, MKN-28 and SGC-7901 were infected with adenovirus gp96 at a multiplicity of infection of 100. gp96-GC antigen peptide complexes were purified. MTT (3-(4,5-dimethylthiazol-2-yl)2,5- diphenyltetrazolium bromide) assay, lactate dehydrogenase (LDH) release assay and enzyme-linked immunosorbent assay were used to determine allo-reactive T cell stimulation, natural killer (NK) cell activity and expression of cytokines (such as interleukin (IL)-10, IL-12, interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha), respectively. Effect of cytotoxic T lymphocyte (CTL) on DCs incubated with HSP-gp96 was also evaluated by LDH release. All assays were performed in triplicate and the average values were reported. Comparison between groups was conducted using Student's t test. RESULTS T cells incubated with HSP-gp96 exhibited a marked increase in proliferation in a dose-dependent manner (P < 0.05). NK cell activity after gp96-GC peptide complex treatment was significantly higher than that after antigen peptide treatment (P < 0.05). The activity of CTLs incubated with DCs from three GC cells lines was obviously higher than that stimulated by GC antigen at ratios of 50: 1, 25: 1, 10: 1, and 5: 1 (P < 0.05). Furthermore, the secretion of TNF-alpha, IL-10, IL-12 (P70) and IFN-alpha markedly increased after incubation with HSP-gp96 (P < 0.05). CONCLUSION HSP-gp96 promotes T cell response, enhances DC antigen presentation and induces cytokine secretion, as well. HSP-gp96 has potential as immunotherapy for elimination of residual GC cells.

Prognostic potential of an immune score based on the density of CD8＋ T cells, CD20＋ B cells, and CD33＋/p-STAT1＋ double-positive cells and HMGB1 expression within cancer nests in stage ⅢA gastric cancer patients

Objoctive： There is heterogeneity in the prognosis of gastric cancers staged according to the tumornodes-metastasis （TNM） system. This study evaluated the prognostic potential of an immune score system to supplement the TNM staging system. Mothodsg An immunohistochemical analysis was conducted to assess the density of T cells, B cells, and myeloid-derived suppressor cells （MDSCs） in cancer tissues from 100 stage IIIA gastric cancer patients; the expression of the high-mobility group protein B1 （HMGB1） was also evaluated in cancer cells. The relationship between the overall survival （OS）, disease-free survival （DFS）, and immunological parameters was analyzed.Results： An immune score system was compiled based on the prognostic role of the density ofT cells, B cells, MDSCs, and the expression of HMGB1 in cancer tissues. The median 5-year survival of this group of patient was 32%. However, the 5-year survival rates of 80.0%, 51.7%, 0%, 5.8%, and 0% varied among the patients with an immune score of 4 to those with an immune score of 0 based on the immune score system, respectively. Similarly, differences in DFS rates were observed among the immune score subgroups. Concluslons： An immune score system could effectively identify the prognostic heterogeneity within stage IliA gastric cancer patients, implying that this immune score system may potentially supplement the TNM staging system, and help in identifying a more homogeneous group of patients who on the basis of prognosis can undergo adjuvant therapy.

Relationship of serum neurotransmitters with anxiety depression, T lymphocyte subsets and NK cells in patients with lung cancer chemotherapy

Objective:To study the relationship of serum neurotransmitters with anxiety depression, T lymphocyte subsets and NK cells in patients with lung cancer chemotherapy.Methods: 56 cases of patients with advanced lung cancer who received chemotherapy in the First Affiliated Hospital of Chengdu Medical College between July 2013 and August 2016 were collected as observation group, and 50 healthy subjects who received physical examination in our hospital during the same period were selected as normal control group. Serum neurotransmitter, negative emotions and immune index levels were compared between the two groups of subjects. Pearson test was used to evaluate the relationship of serum neurotransmitter contents with negative emotions and immune index levels in patients with lung cancer chemotherapy. Results: Serum neurotransmitters DA, 5-HT and NE contents in observation group were lower than those in normal control group;SAS and SDS scores were higher than those of normal control group;peripheral blood CD4+T lymphocyte level, CD4+/CD8+ ratio and NK cell level were lower than those in normal control group while CD8+ T lymphocyte level was higher than that in normal control group. Pearson test showed that serum neurotransmitters DA, 5-HT and NE contents in patients with lung cancer chemotherapy were directly correlated with anxiety depression, T lymphocyte subset and NK cell levels.Conclusion: Serum neurotransmitter expression decrease in patients with lung cancer chemotherapy, and this is one of the important causes of anxiety depression and immune dysfunction in patients.

Prognostic value of programmed death.1, programmed death-ligand 1, programmed death-ligand 2 expression, and CD8(+) T cell density in primary tumors and metastatic lymph nodes from patients with stage T1.4N+M0 gastric adenocarcinoma

Background: Anti-programmed death-1/programmed death-ligand 1(PD-1/PD-L1) immunotherapy has been proved to be effective on gastric cancer in ongoing clinical trials. However, the value of PD-L1 in predicting responses of patients with gastric cancer to anti-PD-1/PD-L1 immunotherapy is controversial. Some studies suggested that intra-and inter-tumoral heterogeneity of PD-L1 expression might explain the controversy.This study aimed to analyze the expression of PD-L1, PD-L2, and PD-1 as well as CD8(+) T-cell density in primary tumors and lymph nodes from patients with stage T1-4 N+M0 gastric adenocarcinoma to explore the heterogeneity of PD-1 signaling pathway molecules.Methods: In primary tumors and metastatic as well as non-metastatic lymph nodes from patients with stage T1-4 N+M0 gastric adenocarcinoma, we detected PD-L1 and PD-L2 expression with immunohistochemistry. CD8(+)T-cell density in primary tumors and PD-1 expression on CD8(+)T cells were detected with immunofluorescence. Univariate analysis was used to determine the prognostic values of them. Cox proportional hazard regression model was used to identify independent risk factors that affect patients' overall survival and disease-free survival.Results: Among 119 eligible patients who had undergone surgical resection, the positive rate of PD-L1 was higher in metastatic lymph nodes than in primary tumors(45.4% vs. 38.7%, P = 0.005); the positive rate of PD-1 on CD8(+)T cells was significantly higher in primary tumors and metastatic lymph nodes than in tumor-free lymph nodes(both P < 0.001). The intensity of PD-1 expression on CD8(+) T cells in primary tumors and in metastatic lymph nodes were stronger than that in tumor-free lymph nodes from the same patient. Beside, the positive rate of PD-L2 did not show any differences between primary tumors and metastatic lymph nodes. In multivariate analysis, PD-L1 expression,PD-L2 expression, a low density of CD8(+) T cells in primary tumors, and PD-1 expression on CD8(+) T cells in primary tumors were associated with poor prognosis.Conclusion: The expression of PD-L1 is heterogeneous in primary tumors and in metastatic lymph nodes from patients with stageT1-4 N+M0 gastric adenocarcinoma, which might explain the inconsistent results in assessing the prognostic value of PD-L1 expression in previous studies.

Detection and analysis of T cell subsets, T cell cytokine and immunoglobulin in patients with oral cancer before and after operation

Objective:To investigate the changes of T cell subsets, T cell cytokine and immunoglobulin in patients with oral cancer before and after operation.Methods:A total of 40 cases of oral cancer treated in our hospital from January 2017 to December 2017 were selected as the observation group, and 40 healthy volunteers in our hospital were selected as the control group. The levels of T cell subsets, T cell cytokine and immunoglobulin in the observation group before and after operation were detected and compared with those in the control group.Results:The CD3+, CD4+ and CD4+/ CD8+ of the observation group before operation were lower than those of the control group, and CD8+ was higher than that of the control group, the differences were statistically significant. The CD3+, CD4+ and CD4+/CD8+ after operation in the observation group were (64.98 ± 3.88)%, (42.40 ± 3.53)% and (1.56 ± 0.41) respectively, which were all higher than those before the operation, but still lower than those in the control group. The CD8+ of the observation group was (27.25 ± 3.54)% lower than that before operation, but it was still higher than that of the control group. In the observation group, the levels of INF-γ+, CD4+/INF-γ+, IL-2+, and CD4+/IL-2+ before operation were lower in the observation group than those in the control group. In the observation group, INF-γ+, CD4+/INF-γ+, IL-2+, and CD4+/IL-2+ after surgery were (0.15±0.06)%, (0.07±0.03)%, (0.47±0.13)%, (0.16 ± 0.11)%, were higher than those before surgery, but still lower than those in the control group. The IgA and IgG before and after the surgery in the observation group were (2.01±0.13) g/L, (1.81±0.12) g/L, (10.53±3.12) g/L, and (8.41±2.01) g/L, respectively, compared with the control group, the difference was statistically significant. There was no significant change in IgA, IgG and IgM in the observation group before and after operation.Conclusion:Surgery can effectively improve the T lymphocyte subsets and T cell cytokine levels in oral cancer patients, and has a positive effect on improving immune function.

Gastric cancer and the epoch of immunotherapy approaches

The incidence of gastric cancer(GC) fell dramatically over the last 50 years, but according to IARC-Globocan 2008, it is the third most frequent cause of cancerrelated deaths with a case fatality GC ratio higher than other common malignancies. Surgical resection is the primary curative treatment for GC though the overall 5-year survival rate remains poor(approximately 20%-25%). To improve the outcome of resectable gastric cancer, different treatment strategies have been evaluated such as adjuvant or perioperative chemotherapy. In resected gastric cancer, the addition of radiotherapy to chemotherapy does not appear to provide any additional benefit. Moreover, in metastatic patients, chemotherapy is the mainstay of palliative therapy with a median overall survival of 8-10 mo and objective response rates of merely 20%-40%. Therefore, the potential for making key beneficial progress is to investigate the GC molecular biology to realize innovative therapeutic strategies, such as specific immunotherapy. In this review, we provide a panoramic view of the different immune-based strategies used for gastric cancer treatment and the results obtained in the most significant clinical trials. In detail, firstly we describe the therapeutic approaches that utilize the monoclonal antibodies while in the second part we analyze the cell-based immunotherapies.

Immunological milieu in the peritoneal cavity at laparotomy for gastric cancer

AIM： To investigate the immunological repertoire in the peritoneal cavity of gastric cancer patients. METHODS： The peritoneal cavity is a compartment in which immunological host-tumor interactions can occur. However, the role of lymphocytes in the peri- toneal cavity of gastric cancer patients is unclear. We observed 64 patients who underwent gastrectomy for gastric cancer and 11 patients who underwent lapa-roscopic cholecystectomy for gallstones and acted as controls. Lymphocytes isolated from both peripheral blood and peritoneal lavage were analyzed for surface markers of lymphocytes and their cytokine production by flow cytometry. CD4＋CD25h＋gh T cells isolated from the patient＇s peripheral blood were co-cultivated for 4 d with the intra-peritoneal lymphocytes, and a cytokine assay was performed. RESULTS： At gastrectomy, CCR7-CD45RA- CD8＋ effector memory T cells were observed in the peritoneal cavity. The frequency of CD4＋ CD25 h＋ghT ceils in both the peripheral blood and peritoneal cavity was elevated in patients at advanced stage [control vs stage IV in the peripheral blood： 6.89 （3.39-10.4） vs 15.34 （11.37-19.31）, P 〈 0.05, control vs stage IV in the peri- toneal cavity： 8.65 （5.28-12.0） vs 19.56 （14.81-24.32）, P 〈 0.05]. On the other hand, the suppression was restored with CD4＋ CD25h＋QhT cells from their own pe- ripheral blood. This study is the first to analyze lympho- cyte and cytokine production in the peritoneal cavity in patients with gastric cancer. Immune regulation at ad- vanced stage is reversible at the point of gastrectomy. CONCLUSION： The immunological milieu in the peri- toneal cavity of patients with advanced gastric cancer elicited a Th2 response even at gastrectomy, but this response was reversible.

T-regulatory lymphocytes in peripheral blood of gastric and colorectal cancer patients

AIM: To assess the absolute number of T-regulatory cells (Tregs; CD4+CD25+Foxp3+) in the peripheral blood of gastric and colorectal cancer patients. METHODS: We enrolled 70 cancer patients (33 gastric cancer, 37 colorectal cancer) and 17 healthy volunteers. The CD3+CD4+ lymphocytes and CD4+CD25+Foxp3+ Tregs in the peripheral blood were analyzed with flow cytometry. The absolute numbers of Tregs were calculated based on the CD4+CD25+Foxp3+ cells percent-age of CD3+CD4+ cells and the absolute numbers of CD3+CD4+ cells per microliter. RESULTS: The mean number of CD4+CD25+Foxp3+ cells per microliter in colorectal cancer patients was 15.7 (SD: 21.8), for gastric cancer patients 12.2 (SD: 14.3), and for controls 17.5 (SD: 11.4). The absolute number of Tregs was significantly lower in gastric cancer patients than in controls (P = 0.026). There was no statistically significant difference for gastric vs colorectal cancer or colorectal cancer vs controls. The absolute number of Tregs was also significantly depressed in N+ vs Ncancer patients [22.0 (27.7) vs 10.1 (9.0), P = 0.013], and in the subgroup of gastric cancer patients [30.3 (27.6) vs 9.6 (8.0), P = 0.003]. No statistical difference was observed in the proportion of Tregs in the CD4+ population between the groups. CONCLUSION: The absolute number of Tregs in peripheral blood of gastric cancer but not colorectal cancer patients was significantly decreased in comparison with that in healthy controls.

Role of RANTES and Its Receptor in Gastric Cancer Metastasis

This study examined the role of regulated upon activation normal T cell expressed and secreted（RANTES） and its receptor C-C chemokine receptor type 5（CCR5） in gastric cancer metastasis and the associated mechanism.The expression of RANTES and CCR5 was detected by using immunohistochemical staining and Western blotting in the gastric cancer tissues obtained from 60 gastric cancer patients with or without lymph node metastasis（n=30 in each）.The results showed that the expression levels of RANTES and CCR5 were higher in gastric cancer with lymph node metastasis than in that without metastasis（P0.05）.The expression levels of RANTES in 30 lymph nodes with cancerous invasion were higher than in 30 normal lymph nodes（P0.05）.Chemotactic test revealed that the number of migrating gastric cancer cells（n=295.0±54.6） induced by the protein of cancer-invading lymph nodes was greater than that by the protein mixture from cancer-invading lymph nodes and RANTES antibody（n=42.5±11.6）（P0.05）.RT-PCR showed that the expression levels of the main Th1 cytokines（IL-2,γ-IFN） were lower in gastric cancer with lymph node metastasis（2.22±0.90,3.26±1.15 respectively） than in that without metastasis（3.07±1.67,4.77±1.52 respectively）（P0.05）,but the expression level of the main Th 2 cytokine（IL-10） was higher in gastric cancer with lymph nodes metastasis（6.06±2.04） than in that without metastasis（4.88±1.87）（P0.05）.It was concluded that RANTES and its receptor CCR5 may contribute to gastric cancer metastasis through influencing the balance of Th1/Th2.RANTES and CCR5 may become a marker of gastric cancer metastasis.

基于T细胞和NK细胞协同作用的癌症免疫疗法

肿瘤异质性是包括免疫疗法在内癌症治疗方法面临的主要挑战之一。异质性可通过获得新突变而产生,从而产生抗治疗肿瘤亚克隆。此外,癌细胞可以通过改变其细胞状态来适应治疗压力。因此,我们面临的任务是如何积极设计免疫疗法,防止耐药肿瘤细胞的选择。T细胞和自然杀伤(NK)细胞在肿瘤免疫中具有互补作用,这些淋巴细胞群体使用不同的识别策略来识别癌细胞,NK细胞可以杀死逃脱CD8+T细胞识别的癌细胞。因此T细胞和NK细胞的双重攻击为加深免疫治疗的影响提供了机会。最近的研究还表明,NK细胞在向肿瘤招募树突状细胞中发挥重要作用,从而增强CD8+T细胞应答的诱导,而T细胞分泌的IL-2激活NK细胞。有趣的是,T细胞和NK细胞还共享几种重要的抑制和激活受体,这些受体可以作为增强T细胞和NK-细胞免疫作用的靶点。这些抑制性受体配体系统包括CD161-CLEC2D、TIGIT-CD155和NKG2A/CD94-HLA-E。此外,基于抑制和激活细胞因子的新的治疗策略也可以发挥T细胞和NK细胞的协同作用,这些基于T细胞和NK细胞协同作用的疗法将深刻影响目前的癌症免疫治疗。

Autologous Natural Killer Cell/Natural Killer T Cell Immunotherapy of Malignant Diseases

Adoptive immunotherapy, the therapeutic infusion of ex vivo activated cancer-fighting white blood cells that was pioneered by Dr. Steven Rosenberg over 30 years ago, has become more widespread due to outstanding published research documenting the clinical efficacy of this strategy. Based on the well-established in vivo functions of NK and NKT cells, their integral role in the efficacy of certain chemotherapeutic and immunomodulatory agents, and their direct therapeutic action as displayed in clinical trials, the use of autologous natural killer cell infusions is an appropriate and warranted therapeutic option for the treatment of malignant diseases, especially in patients whose disease is refractory to standard treatments such as chemotherapy and radiation.

基于生物信息学分析PTTG1和UBE2C与肝癌预后的相关性

目的利用GEO和TCGA数据库中的单细胞数据、基因表达数据和临床信息,挖掘与肝癌预后T/NK细胞相关的差异表达基因(T/NK cell-related differentially expressed genes,T/NK-DEGs),并构建肝癌预后模型。方法从GEO数据库中获得肝癌单细胞数据以及肝癌的基因表达矩阵。从TCGA数据库中获得肝癌基因组图谱(TCGA-LIHC)队列,其中包括mRNA表达数据、临床信息、生存信息、体细胞突变数据。基于以上数据集,通过生物信息学方法构建肝癌患者的预后模型,并预测该模型的性能。结果共鉴定出2个与肝癌预后相关的T/NK-DEGs,分别为PTTG1和UBE2C,并基于此构建了肝癌预后模型。根据风险评分,将患者分为高风险评分组和低风险评分组,其中高风险评分组患者预后明显较差。构建的肝癌预后模型在1年、3年和5年时间点的ROC曲线下面积(AUC^(ROC))分别为0.685、0.647和0.594,且较高的风险评分伴随较晚的临床分期(Ⅰ期、Ⅱ期、Ⅲ期)和T分期(T1、T2、T3),差异有统计学意义(P<0.05)。该预后模型能够预测肝癌患者的免疫细胞比例、免疫治疗和化疗药物的敏感性。结论构建的预后模型对肝癌患者的个体化生存预测和临床治疗反应具有重要的临床价值。

PD-1靶向纳米投递系统对胃癌细胞活性及T细胞免疫应答因子的影响

目的探讨PD-1靶向纳米投递系统对胃癌细胞活性及T细胞免疫应答因子的影响。方法取胃癌细胞,将其分为空白对照组、程序性细胞死亡蛋白-1(Programmed Cell Death-1,PD-1)抑制剂组和PD-1靶向组3组。用乳化溶剂挥发法制备PD-1靶向纳米投递系统,荧光倒置显微镜下观察转染情况,MTT法检测细胞增殖,流式细胞仪检测细胞凋亡,实时PCR法及免疫印迹法检测T细胞免疫应答因子表达。结果与空白对照组相比,PD-1抑制剂组、PD-1靶向组的转染率和凋亡率依次升高,且差异均有统计学意义(P<0.05)。与空白对照组相比,PD-1抑制剂组、PD-1靶向组细胞增殖率依次降低,且差异均有统计学意义(P<0.05)。与空白对照组相比,PD-1抑制剂组、PD-1靶向组细胞INF-γ、IL-2、CD80的mRNA表达及蛋白表达依次升高,TNF-α、TGF-β1、PD-1的mRNA表达及蛋白表达依次降低,且差异均有统计学意义(P<0.05)。结论PD-1靶向纳米投递系统可高效传送PD-1抑制剂到胃癌细胞,并有效改善细胞活性,调控T细胞免疫应答因子表达,可为胃癌治疗领域的进一步研究和临床应用提供新的思路和方向。

胃癌 癌周淋巴结 网膜及腹膜中T淋巴细胞亚群和NK细胞分布的研究

目的:通过检测胃癌、癌周正常胃粘膜、胃周淋巴结、大网膜、小网膜、腹膜及外周血中T淋巴细胞亚群和NK细胞数以探讨胃癌局部及全身的免疫状况。方法:利用流式细胞术测定40例胃癌患者的癌组织、癌周正常胃粘膜、胃周淋巴结、大网膜、小网膜、腹膜及外周血的CD4+、CD8+及NK细胞的含量。取22例良性疾病患者的胃粘膜和外周血做对照。结果:CD4+、CD8+、NK细胞计数在胃癌组织和胃周淋巴结中最高,均高于癌周正常胃粘膜、大网膜、小网膜、腹膜及外周血(P<0.05),但癌组织的CD4+/CD8+比值低于淋巴结和大网膜(P<0.05);良性疾病对照组的胃粘膜、外周血CD4+、CD8+、NK细胞计数和CD4+/CD8+比值均高于胃癌患者(P<0.01)。结论:胃癌患者局部及全身的免疫功能均低于对照组;胃癌组织内CD4+、CD8+、NK细胞较癌周正常胃粘膜、大网膜、小网膜、腹膜高聚集,但免疫功能呈现低下和抑制状态。

针灸预处理对摔跤运动员大负荷训练阶段NK细胞和T淋巴细胞亚群的影响

目的:探讨针灸预处理方法对男子摔跤运动员大负荷训练期间NK细胞、CD3+、CD4+、CD8+T淋巴细胞亚群的调节作用。方法:将20名摔跤运动员随机分为对照组和实验组,在8周大负荷训练期间进行实验,分别在大负荷运动训练前、大负荷运动训练后、调整训练后采取外周静脉血,用流式细胞术检测运动员NK细胞、T淋巴细胞亚群的相对含量。结果:运动员NK细胞数量变化,对照组W8和W9与W2相比明显降低,实验组W9与W8相比明显降低;CD3+T细胞数量变化,实验组W8明显高于同期对照组;CD4+T细胞数量变化,实验组W8、W9明显高于同期对照组;运动员CD4+/CD8+值变化,实验组表现为上升趋势,W4、W5与W2相比升高。结论:大负荷训练可引起摔跤运动员NK细胞数量下降,CD3+、CD4+、CD8+T细胞数量下降,针灸预处理可延缓或防止其下降,在一定程度上维持运动员免疫机能的稳态。

肺癌患者外周血T-淋巴细胞亚群NK细胞的基线数值及其与预后的关系

目的:对比肺癌患者与健康者之间淋巴细胞亚群差异,评估肺癌患者外周血T淋巴细胞亚群及自然杀伤细胞(NK)之基线值与预后的关系。方法:收集2006年2月至2013年3月就诊于天津医科大学肿瘤医院病例资料完整的肺癌患者105例,其中非小细胞肺癌(NSCLC)86例、小细胞肺癌(SCLC)19例,另选健康对照35例,对比接受治疗前肺癌患者和健康对照者外周血中的CD3+T细胞、CD4+T细胞、CD8+T细胞及NK细胞所占百分比,并回顾性分析86例NSCLC患者初治时外周血淋巴细胞亚群与预后的关系。结果:肺癌患者外周血CD3+T细胞、CD4+T细胞、NK细胞及CD4/CD8比值均明显低于健康对照组(P=0.011,P=0.007,P<0.001,P=0.025),而CD8+T细胞比例高于健康对照组(P=0.013)。当CD8+T≥31.8%及CD4/CD8<1.28时NSCLC患者可以获得一个更长的OS(中位OS分别为36.2 m vs.20.0 m,P=0.010;30.8 m vs.20.0 m,P=0.035)。而CD3+T细胞、CD4+T细胞及NK细胞百分比对NSCLC患者预后无显著影响。结论:外周血CD8+T细胞基线水平较高的NSCLC患者生存较长,此基线水平可能对NSCLC患者预后有指示作用。

吡酮莫特治疗对慢性鼻窦炎患者外周T淋巴细胞亚群、NK细胞活性以及炎症因子的影响

目的:探讨吡酮莫特治疗对慢性鼻窦炎者外周T淋巴细胞亚群、NK细胞活性以及炎症因子的影响。方法:选取武汉大学中南医院和潜江市中心医院耳鼻咽喉头颈外科住院患者确诊的慢性鼻窦炎患者176例作为研究对象,按照有无吡酮莫特治疗,分为实验组和对照组,检测两组外周T淋巴细胞亚群、NK细胞活性以及炎症因子并进行比较。结果:治疗后1个月、停药后6个月两组的CD3^+(%)、CD4^+(%)、CD4^+/CD8^+均明显增高,且实验组增高幅度均高于对照组(P=0.000)。治疗后1个月、停药后6个月两组的CD8^+(%)明显降低,且实验组降低幅度高于对照组(P=0.000)。治疗后1个月、停药后6个月两组的NK细胞活性明显增高,且实验组增高幅度高于对照组(P=0.000)。治疗后1个月、停药后6个月两组的CRP、IL-6、IL-8均明显降低,且实验组治降低幅度均高于对照组(P=0.000)。结论:吡酮莫特可以改善慢性鼻窦炎患者的免疫功能并发挥抗炎功能,从而发挥治疗作用。

原发性胃癌根治术后化疗对外周血T细胞亚群、NK细胞的影响及意义

目的：探讨原发性胃癌患者胃癌根治术后化疗对患者外周血T细胞亚群及NK细胞的影响及临床意义。方法：选择2010年11月~2013年11月入住我院35例行胃癌根治术的原发性胃癌患者作为观察组,采用EP化疗方案：100mg/m2足叶乙苷,d1-3,静脉滴注;25mg/m2顺铂,d1-3,静脉滴注。另外选择同期于我院进行体检的健康者35名作为对照组。比较化疗前后外周血白细胞总数、淋巴细胞百分比、淋巴细胞绝对值、外周血T细胞亚群相关指标（CD3＋、CD4＋、CD8＋、CD4＋/CD8＋及CD45RA＋/RO＋）及NK细胞。结果：（1）观察组化疗前后白细胞总数、淋巴细胞百分比及淋巴细胞绝对值差异均具有统计学意义（P〈0.05）,但观察组化疗后上述指标与对照组相比,差异无统计学意义（P〉0.05）;（2）观察组化疗后外周血T细胞亚群相关指标（CD3＋、CD4＋、CD8＋及CD4＋/CD8＋、CD45RA＋、CD45RA＋绝对值、CD45RO＋绝对值、CD45RA＋/RO＋）及NK细胞水平均显著高于化疗前（P〈0.05）。结论：原发性胃癌根治术后化疗可有效提高机体外周血T细胞亚群及NK细胞水平,对患者生活质量的改善及疾病的康复具有十分重要的临床意义。