肿瘤相关三级淋巴结构在免疫治疗研究中的前沿进展

三级淋巴结构 （TLS） 是近年来在慢性炎症刺激的非淋巴组织中发现的异位淋巴结构,与淋巴结具有相似的结构和功能,主要由B淋巴细胞、T淋巴细胞和树突状细胞组成,也是抗肿瘤免疫应答启动的直接部位。肿瘤中TLS促进免疫细胞,特别是T细胞和B细胞等效应免疫细胞,向肿瘤微环境聚集,为抗肿瘤的细胞和体液免疫应答提供一个重要的局部场所,预示患者可能有良好的生存预后和积极的免疫治疗反应。本文概述了肿瘤相关TLS的评估鉴定、结构与组成、以及形成的驱动因素,并阐述了 TLS 作为潜在生物标志物在肿瘤免疫治疗中的作用及潜在临床运用价值。此外,本文还讨论了 TLS在肿瘤免疫治疗中的挑战和未来研究方向。     

MiR-520b inhibits proliferation,migration and invasion in gallbladder carcinoma by targeting RAB22A

IntroductionPrevious studies have reported that miR-520b exhibited inhibitory effects on various human tumors, whereas the effects of miR-520b on gallbladder carcinoma (GBC) have remained unclear. To investigate the effects of miR-520b on GBC progression and reveal the underlying mechanisms, this study was performed.Material and methodsMiR-520b and RAB22A mRNA levels were analyzed by quantitative real-time PCR (qPCR). RAB22A protein level was analyzed via Western blot and immunohistochemical (IHC) analysis. The proliferation, colony formation ability, migration and invasion of NOZ cells were measured via MTT, colony formation, wound healing and transwell invasion assay respectively.ResultsMiR-520b expression level was lower in human GBC tissues than that in neighboring normal tissues. MiR-520b mimic repressed NOZ cell proliferation, colony formation ability, migration and invasion, whereas miR-520b inhibitor exhibited opposite effects. Dual luciferase reporter assay confirmed that miR-520b could bind to the 3′-untranslated regions of RAB22A mRNA. Moreover, RAB22A overexpression significantly abolished the anti-tumor effects of miR-520b in a NOZ cell model. Western blot, qPCR and IHC analysis proved that human GBC tissues showed a higher RAB22A expression level than neighboring normal tissues. Additionally, there was a negative association between miR-520b and RAB22A expression.ConclusionsMiR-520b had suppressive effects on GBC via targeting RAB22A in vitro.     

Expression of normal and tumor-associated antigens in human breast carcinoma

The expression of six cytoplasmic/membrane antigens (beta 2-microglobulin, HLA, HLA-DR, carcinoembryonic antigen, and two breast tumor-associated antigens (TAAs), B6.2 and B72.3) was investigated in serial sections of 28 human breast carcinomas using monoclonal antibodies and the avidin-biotin complex immunoperoxidase technique. The frequency of expression and linkage between these antigens was determined, and antigenic expression was related to patient age, morphologic differentiation, cytologic grade, and estrogen receptor/progesterone receptor content of the tumor. The expression of beta 2-microglobulin and HLA correlated with morphologic differentiation, well-differentiated and moderately well-differentiated tumors expressing these antigens more often than poorly differentiated tumors. Expression of the TAAs, however, was not related to differentiation. There was no linkage between beta 2-microglobulin/HLA and the TAAs. Carcinoembryonic antigen was found to be linked to the TAA, B6.2. Expression of the TAA, B72.3, correlated with patient age. Eighty percent (23 of 28) of the tumors were positive for carcinoembryonic antigen or at least one of the TAAs. The estrogen receptor/progesterone receptor status of the tumor was not statistically related to the expression of any of the antigens studied. Analysis of tumor antigen profiles may provide important information relevant to prognosis, therapy, and early detection of cancer, as well as insights into the nature of the neoplastic process.     

Gallbladder neuroendocrine carcinoma: report of 10 cases and comparision of clinicopathologic features with gallbladder adenocarcinoma

Few cases of neuroendocrine carcinoma (NEC) of the gallbladder (GB-NEC) have been reported. Data obtained from the 10 patients with GB-NEC treated in our hospital between January 2008 and December 2012 were retrospectively analyzed and compared with those of 377 patients with gallbladder adenocarcinoma. GB-NEC accounted for 2.2% of all gallbladder cancers. The patients (8 females and 2 males) were 59.0 ± 10.0 years old. Four patients presented mixed adenocarcinoma, while six had pure NEC. Immunohistochemical examinations showed a positive rate of 100% for CgA, NSE, and CK; the positive rates for Syn, EMA, and CD56 were 88.9, 87.5, and 75%, respectively. TNM grades II, IVA, and IVB were found in 1, 2, and 7 patients, respectively. GB-NEC patients showed significantly higher N2 lymphatic metastasis rates than gallbladder adenocarcinoma patients (70.0 vs. 34.0%; P < 0.05). Two patients were treated with radical resection and the remaining 8 with palliative operation. The 1-, 2-, and 3-year survival rates were 20, 10, and 0%, respectively (median survival time, 3.0 m); the 1-, 2-, 3-, and 5-year survival rates for all gallbladder adenocarcinoma patients were 38.0, 31.0, 30.1, and 28.4%, respectively (median survival time, 6.0 m), the difference was statistically significant (P = 0.038). The results demonstrate that GB-NEC was mainly found in aged females and shows high malignancy. Its prognosis is poorer than that of gallbladder adenocarcinoma, and surgical resection combined with TACE, radiotherapy, and chemotherapy could increase patient survival.     

Induction of neovascularization in vivo and endothelial proliferation in vitro by tumor-associated macrophages

The role of macrophages in neovascularization of tumors was investigated by examining the ability of tumor-associated macrophages (TAM) and their conditioned culture media to induce neovascularization in the cornea of syngeneic rats and proliferation of bovine aortic endothelial cells in culture. TAM were isolated from a 3-methycholanthrene-induced fibrosarcoma propagated in F344 male rats by enzymatic dissociation and were purified by centrifugation through continuous Percoll density gradients, followed by adherence to fibronectin-coated dermal collagen gels. The angiogenic potential of (a) TAM and their 72-hour conditioned culture media, (b) whole tumor cell suspensions (WTCS), (c) tumor cell suspensions depleted of TAM (TCS), and (d) macrophage-depleted tumor cell suspensions reconstituted with TAM (TCS + TAM) were compared. Cells were injected directly; conditioned media were concentrated 10-fold, incorporated into slow-release Hydron pellets, and implanted intracorneally. Stimulation of bovine aortic endothelial cell growth by TAM was assayed in culture with TAM-conditioned media and compared with responses induced by conditioned media from peptone-elicited rat peritoneal exudate macrophages. TAM and their conditioned media induced neovascularization in 38 of 40 corneas (95%) and 15 of 17 corneas (88%), respectively. Maximal vessel ingrowth occurred by the 5th day of implantation. Neovascular responses induced by WTCS (24 of 26 corneas, 92%) and TCS (17 of 24 corneas, 71%) occurred on the 7th and 10th day, respectively. TCS + TAM induced neovascular responses comparable to those elicited by WTCS (19 of 20 corneas, 95%). Addition of TAM-conditioned media to bovine aortic endothelial cell cultures stimulated a 10-fold increase in cell number within 10 days. This growth stimulatory effect was comparable to or greater than responses induced by conditioned media from rat peritoneal macrophages. Our results demonstrate that TAM are potent stimulators of neovascularization and endothelial cell proliferation and that depletion of macrophages from tumor cell suspensions significantly decreased their angiogenic potential. This suggests that neovascularization of this tumor is mediated in part by macrophages.     

Crosstalk between tumor-associated macrophages and neighboring cells in hepatocellular carcinoma

The tumor microenvironment generally shows a substantial immunosuppressive activity in hepatocellular carcinoma (HCC), accounting for the suboptimal efficacy of immune-based treatments for this difficult-to-treat cancer. The crosstalk between tumor cells and various cell types in the tumor microenvironment is strongly related to HCC progression and treatment resistance. Monocytes are recruited to the HCC tumor microenvironment by various factors and become tumor-associated macrophages (TAMs) with distinct phenotypes. TAMs often contribute to weakened tumor-specific immune responses and a more aggressive phenotype of malignancy. Recent single-cell RNA-sequencing data have demonstrated the central roles of specific TAMs in tumorigenesis and treatment resistance by their interactions with various cell populations in the HCC tumor microenvironment. This review focuses on the roles of TAMs and the crosstalk between TAMs and neighboring cell types in the HCC tumor microenvironment.     

The role of tumor-associated macrophages in breast carcinoma invasion and metastasis

The main purpose of this study was to investigate the infiltration of tumor-associated macrophages (TAMs) in normal and malignant breast tissue and the draining lymph nodes, and to explore its effect on breast cancer invasion and metastasis. The infiltration densities of TAMs was observed using immunohistochemical staining of CD68 in 100 cases of breast cancer specimens and its paired adjacent non-cancer breast tissues and draining lymph modes, and then to evaluate the relation of TAMs to various clinicopathological features including patients prognosis in breast carcinoma. We observed the infiltration densities of TAMs were significantly higher in breast carcinoma tissue than in adjacent normal tissue and significantly higher in much larger size and higher stage cases. Furthermore, infiltration densities of TAMs have negative correlation with the 5-year survival rates of breast cancer patients. But in matched lymph-nodes, the infiltration densities of TAMs were significantly lower in cancerous metastatic lymph-node samples than in non-metastatic one. Therefore, our data suggests that TAMs infiltration in primary tumor promote invasion and lymphatic metastasis of breast cancer and have negative correlation with patients prognosis in breast cancer, but in lymph-node TAMs may play another role and need further study in the future.     

Hepatoid carcinoma of the gallbladder

Hepatoid carcinoma is a special type of extrahepatic tumor associated with hepatic differentiation, and has the morphological and functional features of hepatocellular carcinoma. Hepatoid carcinoma of the gallbladder is very rarely reported in the literature. We report a case of hepatoid carcinoma of the gallbladder in a 71-year-old female who presented with abdominal pain and was first diagnosed as cholelithiasis with cholecystitis. The microscopic findings of the gallbladder after cholecystectomy showed an area of tumor with polygonal cells, eosinophilic cytoplasm, distinct cell borders, round vesicular nuclei and prominent nucleoli, arranged in trabecular pattern resembling hepatocellular carcinoma intermingled with areas of adenocarcinoma or cholangiocarcinoma. The specimen from the pancreas also showed the same type of tumor cells. Histochemically, some of tumor cells were positive for Victoria Blue, Stein, and PAS. The immunohistochemistry for alpha-fetoprotein (AFP) showed strong intra cytoplasmic positivity, both in tumor cells with hepatic differentiation and tumor cells with bile duct epithelium differentiation. Based on these findings, this case was diagnosed as hepatoid carcinoma of the gallbladder with metastasis to the pancreas. This is the first case that has been reported in our department.     

Gastric carcinoma with lymphoid stroma

Summary A total of 626 surgically resected gastric carcinomas were reviewed, and 24 cases (3.8%) of gastric carcinoma with lymphoid stroma were identified. The tumour cells were consistently arranged in an anastomosing trabecular or alveolar pattern and were densely infiltrated by lymphoid cells. The specimens were studied using mucin histochemistry and the indirect immunoperoxidase method to determine the histochemical properties of this form of gastric carcinoma. The tumour cells were consistently positive for concanavalin A paradoxical staining, class III and almost devoid of acidic mucins, features demonstrating preferential differentiation toward pyloric glands or pseudopyloric glands. Immunohistochemically, positive reactions for Leu M1 and lysozyme, marker substances of (pseudo)pyloric gland cells, were often observed. Carcinoembryonic antigen was positive in focal areas without (pseudo)pyloric glandular patterns. Secretory component was focally positive. HLA-DR was strongly expressed in most cancer cells and 17 tumours (71%) showed positivity for interleukin 1 (IL-1). The lymphoid stroma contained a high percentage of UCHL1-reactive T cells both within and around the cancer cell nests, while SL26-reactive B cells clustered in lymphoid follicles. A considerable number of T-lymphoid cells were also reactive for IL-1. A number of plasma cells with a predominance of IgG-type were distributed around the cancer cell nests. S-100 protein-positive dendritic cells were not identified. We speculate that the prominent lymphoid stroma including intraepithelial lymphocyte-like T cells with IL-1 receptors is possibly induced by IL-1 related mediators released from the HLA-DR-positive gastric cancer cells of the (pseudo)pyloric gland-type.     

Proliferating cell nuclear antigen in gallbladder carcinoma

Proliferating cell nuclear antigen (PCNA) expression was studied in 103 gallbladder carcinomas and 23 metastatic lesions as well as in 25 control non-neoplastic gallbladder specimens. Positive nuclear staining was observed in 88% of controls, in 92% of carcinomas and in 70% of metastases. The mean number of positive cells was 21.2% in controls, 44.1% in primary carcinomas and 32% in metastatic cancer cells. Differences which were significant were control v . primary tumour, P <0.000001; control v . metastasis, P <0.01 and primary tumour v . metastasis, P <0.006. In 57 (60%) of the primary tumours there was positive staining in over 40% of tumour cells. We were not able to demonstrate any relationship between macroscopic or microscopic features and PCNA expression. However, tumours confined to the mucosa expressed PCNA more frequently than did more advanced tumours.     

Laminin和survivin的表达与胆囊癌临床病理特征的关系

目的:研究laminin和survivin蛋白在原发性胆囊癌组织中的表达情况,及其与癌组织类型、病理分级和转移状况的关系。 方法:应用免疫组织化学方法检测49例原发性胆囊癌、21例胆囊腺瘤和13例慢性胆囊炎组织中laminin和survivin蛋白的表达。 结果:胆囊黏膜内癌或原位癌细胞基底膜laminin线性染色完整；NevinⅡ期胆囊癌组织表现为基底膜不完整,变薄,断裂,或缺损；临床NevinⅢ、Ⅳ、Ⅴ期胆囊癌，则无基底膜形成，在肿瘤组织周围，laminin表达类型呈碎片状或断线状和连续线状，部分癌组织内laminin染色消失和癌细胞浆内有laminin弱染色。胆囊癌组织中survivin阳性表达率明显高于胆囊腺瘤和慢性胆囊炎组织，但survivin的阳性表达与胆囊癌细胞分化程度、病理分级和转移无关(P>0.05)。且胆囊癌组织中laminin的连续线断裂或缺失,与胆囊癌组织中survivin的表达情况无相关性。 结论:胆囊癌基质中laminin的表达类型与胆囊癌的侵袭和转移有关，而survivin在胆囊癌中表达增加，但两者之间似乎无关联。      

EGFR expression in gallbladder carcinoma in North America

BACKGROUND: Increased epidermal growth factor receptor (EGF receptor) expression has been noted in various cancers and has become a useful target for therapeutic interventions. Small studies from Asia and Australia have demonstrated EGFR over-expression in gallbladder cancer. We sought to evaluate the expression of EGFR in a series of 16 gallbladder cancer patients from North America.     

Mast cell regulatory effect on lymphoid cell proliferation

It has been shown rat mast cells (MC) can modulate lymphocyte proliferationin vitro. Depending on concentrations tested both serosal MC and their supernatants enhanced the spontaneous and T-mitogen-induced proliferation of spleen and lymph node cells. In addition T-mitogen-induced thymocyte proliferation was also increased. The enhancing effect of MC on lymphoid cell proliferation appeared after MC and lymphocytes were cocultured for 24, 48 or 72 h. The highest enhancing action of MC was observed when MC and lymphocytes were plated simultaneously. In contrast, when MC were added 24 or 48 h after the start of lymphocyte culture, the enhancing action of MC decreased or was abolished, respectively. No dependence was found between histamine concentration in MC supernatants and the enhancing activity of supernatants. After chromatographic separation of MC supernatants the fractions with molecular weights between 1–6 KDa augmented lymphoid cell proliferation.