PURPOSE: Forkhead box p3 (Foxp3) positive T regulatory cells (Tregs) have a functionally immunosuppressive property that prevents effector cells from acting against self in autoimmune diseases or a tumor. It is known that Tregs may be highly relevant in cancer progression. Dendritic cells (DCs) induce cutaneous immune response, however several studies have suggested that DCs are involved in immunosuppression. The aim of this study is to evaluate the prevalence of Tregs and DCs infiltration in cutaneous premalignant and malignant squamous lesions. MATERIALS AND METHODS: We evaluated Tregs and DCs in skin tissue samples obtained from 83 patients with actinic keratosis, Bowen's disease or squamous cell carcinoma by immunohistochemistry. RESULTS: The prevalence of Tregs and DCs was significantly higher in squamous cell carcinoma and Bowen's disease than in actinic keratosis. In addition, the number of DCs was closely correlated with the prevalence of Tregs, and DCs were also located in direct proximity to Tregs. CONCLUSION: Tregs is related to cutaneous squamous tumor progression.
Bowen's Disease/immunology/metabolism/pathology ; Carcinoma, Squamous Cell/immunology/metabolism/pathology ; Dendritic Cells/immunology/metabolism/pathology ; Forkhead Transcription Factors/immunology/*metabolism ; Humans ; Immune Tolerance ; Keratosis, Actinic/immunology/metabolism/pathology ; Skin Neoplasms/*immunology/metabolism/pathology ; T-Lymphocytes, Regulatory/*immunology/metabolism/pathology

Aged, 80 and over ; Antibodies, Monoclonal ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Diagnosis, Differential ; Granuloma ; metabolism ; pathology ; Humans ; Keratins ; immunology ; Male ; Mucin-1 ; metabolism ; Skin Neoplasms ; metabolism ; pathology

OBJECTIVETo analyze the expression of co-stimulatory molecules PD-1/PD-L1 in peripheral blood mononuclear cells in lung cancer patients, and to explore its biological significance.

METHODSOne hundred and thirty-three lung cancer patients, 25 lung infection patients and 23 healthy donors were enrolled in this study. 100 µl of whole blood from these subjects were collected. Multi-color immunofluorescence staining and flow cytometry were used to detect PD-1/PD-L1 expression. The results were statistically analyzed.

RESULTSThe expression level of CD3⁺CD8⁺ T cells in the lung cancer patients was (38.83 ± 1.74)%, significantly lower than that in the control group [(43.25 ± 3.35)%, P < 0.05]. CD8⁺CD28⁺ T cell subset in the peripheral blood of lung cancer patients was (17.73 ± 1.21)% significantly lower than that of the healthy donors [(27.96 ± 2.72)%, P < 0.01]. The CD8⁺CD28⁻ T cell subset was (21.19 ± 1.92)% in the lung cancer patients, significantly higher than that of the healthy control group [(15.18 ± 2.93)%, P < 0.05]. The expression level of PD-1 on the surface of CD8⁺CD28⁺ T cells was (10.67 ± 1.12)% in the group of lung cancer patients, significantly higher than that of the control group [(5.32 ± 1.58)%, P < 0.01]. It was also found that the expression of PD-1 on CD8⁺CD28⁻ T cells was up-regulated in the group of lung cancer patients (7.46 ± 1.25)%, significantly higher than that of the healthy control group [(2.68+1.07)%, P < 0.01]. The expression level of PD-L1 on CD68⁺ cells in the lung cancer patients was (16.03 ± 2.06)%, significantly higher than that of the healthy control group [(9.32 ± 2.00)%, P < 0.05].

CONCLUSIONUp-regulation of PD-1/PD-L1 on peripheral blood cells in lung cancer patients negatively regulates the lymphocytes, inhibits the immune response for killing tumor cells, and promotes tumor development and immune escape.

Adenocarcinoma ; blood ; pathology ; B7-H1 Antigen ; metabolism ; CD28 Antigens ; metabolism ; CD3 Complex ; metabolism ; CD8 Antigens ; metabolism ; Carcinoma, Large Cell ; blood ; pathology ; Carcinoma, Squamous Cell ; blood ; pathology ; Case-Control Studies ; Female ; Humans ; Lung Neoplasms ; blood ; pathology ; Male ; Middle Aged ; Programmed Cell Death 1 Receptor ; metabolism ; Small Cell Lung Carcinoma ; blood ; pathology ; T-Lymphocytes ; immunology ; metabolism ; Up-Regulation

OBJECTIVETo clone the variable region genes of the monoclonal antibody (McAb) against human heterogeneous nuclear ribonucleoprotein A2/B1 (HnRNPA2/B1), ligate them to assemble single chain Fv (ScFv) gene and express in Escherichia coli.

METHODSThe specificity of the anti-HnRNPA2/B1 McAb 3E8 to synthetic HnRNPA2/B1 peptide, HnRNPA2/B1 protein in lung cancer cells were examined by dot-immunobinding assay, Western blot and immunohistochemistry. The variable region genes of heavy chain (VH) and light chain (VL) were amplified from hybridoma cell by reverse transcription-polymerase chain reaction(RT-PCR), and then were linked by a linker peptide using SOE-PCR (splicing by overlap extension-PCR) to construct recombination ScFv gene. The latter was cloned into the expression vector pET28 (a+) and expressed in E coli BL21. The expressed product was identified by SDS-PAGE and competitive ELISA inhibition test.

RESULTSIt was shown that the McAb combined specifically with synthetic HnRNPA2/B1 peptide and HnRNPA2/B1 protein in three lung cancer cells. The cloned VH gene and VL gene were 345 bp and 309 bp respectively and were linked successfully to obtain ScFv gene. The ScFv protein was expressed in the form of inclusion body, with molecular weight of 28,000 and immunoreactivity to HnRNPA2/B1.

CONCLUSIONVH gene, VL gene and ScFv gene of anti-HnRNPA2/B1 antibody were cloned, constructed and functionally expressed in E coli. These results provide the experimental basis for elucidating the role of HnRNPA2/B1 in lung cancer.

Adenocarcinoma ; metabolism ; pathology ; Antibodies, Monoclonal ; genetics ; immunology ; metabolism ; Carcinoma, Small Cell ; metabolism ; pathology ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Line, Tumor ; Cloning, Molecular ; Escherichia coli ; metabolism ; Heterogeneous-Nuclear Ribonucleoprotein Group A-B ; immunology ; Humans ; Immunoglobulin Fragments ; genetics ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin Light Chains ; genetics ; Immunoglobulin Variable Region ; genetics ; Lung Neoplasms ; metabolism ; pathology

BACKGROUND AND OBJECTIVECytokine-induced killer (CIK) cells and autologous dendritic cells-CIK (DC-CIK) cells co-cultured with autologous dendritic cells (DCs) and CIK cells are commonly used for immunotherapy recently. We compared the anti-tumor immune response of CIK cells, autologous DC-CIK cells, and semi-allogeneic DC-CIK cells to explore a more effective anti-tumor adoptive immunotherapy approach.

METHODSPeripheral monocytes were isolated from patients with renal carcinoma, lung cancer, or maxillary squamous cell carcinoma and their healthy adult children. Isolated cells were cultured and induced as DCs and CIK cells in vitro. CIK cells from patients were co-cultured with autologous DCs and DCs from their children respectively, generating DC-CIK cells and semi-allogeneic DC-CIK cells. The anti-tumor activities of autologous CIK cells, autologous DC-CIK cells, and semi-allogeneic DC-CIK cells were measured by LDH assay. Intracellular staining was used to test the secretion of cytokines. Flow cytometry was applied for detecting the phonotype changes of these three types of cells. Cell proliferation and cell apoptosis were detected by 5,6-carboxyfluorescein diacetate succinimidyl ester (CFSE) and Annexin V/PI respectively.

RESULTSCompared with autologous CIK cells and DC-CIK cells, semi-allogeneic DC-CIK cells significantly enhanced the anti-tumor activity and IFN-gamma secretion, reduced IL-4 secretion, increased the ratio of CD3(+)CD56(+) cells and CD3(+)CD8(+) cells, decreased the number of CD4(+)CD25(+) cells, promoted cell proliferation, and lessened cell apoptosis.

CONCLUSIONSSemi-allogeneic DC-CIK cells had a stronger anti-tumor effect than did autologous CIK cells and DC-CIK cells. Our results provided experimental evidence for clinical application of DC-CIK cells.

Apoptosis ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Cells, Cultured ; Coculture Techniques ; Cytokine-Induced Killer Cells ; cytology ; immunology ; metabolism ; Cytokines ; metabolism ; Cytotoxicity, Immunologic ; Dendritic Cells ; cytology ; immunology ; metabolism ; Hep G2 Cells ; Humans ; Immunotherapy, Adoptive ; Interferon-gamma ; secretion ; Interleukin-4 ; secretion ; K562 Cells ; Kidney Neoplasms ; metabolism ; pathology ; L-Lactate Dehydrogenase ; metabolism ; Lung Neoplasms ; metabolism ; pathology ; Maxillary Neoplasms ; metabolism ; pathology

OBJECTIVE: Study on the role of mycoplasma infection and expression of CD44v6, CD44v9 protein in the pathogenesis, development and prognosis of laryngeal carcinoma (LC). METHOD: Immunohistochemistry was used to study 137 cases of laryngeal carcinoma tissues, 26 cases of precarcinoma tissues, 34 cases of vocal cord polypus and 15 cases of normal tissues adjacent to laryngeal carcinoma. RESULT: The positive rate of PD4 and CD44v6, CD44v9 was 45.3% (62/137), 72.3% (99/137) and 56.2% (77/137) in laryngeal carcinoma, which are much higher than those in non-carcinoma tissues. (2) The positive rate of PD4 and CD44v6. CD44v9 in the third and fourth stages of LC were higher than those in the first and second stages of LC. Laryngeal carcinoma with cervical metastasis had higher expression than those without cervical metastasis (P < 0.01). To 3 and 5 years survival, PD4 and CD44v6, CD44v9 positive cases had lower chance than those negative cases (P < 0.01 [Chinese character: see text] 0.05). (3) The overall positive rate of PD4 and CD44v6, CD44v9 was 27.7% (38/137) in laryngeal carcinoma tissues and 5.3% (4/75) in non-carcinoma tissues. CONCLUSION These results implicates that the pathogenesis, development and prognosis of laryngeal carcinoma maybe closely related to the high expression of PD4 and CD44v6, CD44v9 protein.
Adult ; Aged ; Antibodies, Monoclonal ; biosynthesis ; Apoptosis Regulatory Proteins ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; microbiology ; pathology ; Female ; Humans ; Hyaluronan Receptors ; metabolism ; Laryngeal Neoplasms ; metabolism ; microbiology ; pathology ; Male ; Middle Aged ; Mycoplasma ; immunology ; Neoplasm Staging ; Prognosis ; RNA-Binding Proteins ; metabolism

Adult ; Aged ; Aged, 80 and over ; Antibodies, Monoclonal ; metabolism ; Antigens, Neoplasm ; immunology ; metabolism ; Biomarkers, Tumor ; metabolism ; Carcinoma, Squamous Cell ; diagnosis ; metabolism ; pathology ; Cell Cycle Proteins ; immunology ; metabolism ; Cervical Intraepithelial Neoplasia ; diagnosis ; metabolism ; pathology ; Cyclin-Dependent Kinase Inhibitor p16 ; metabolism ; DNA Topoisomerases, Type II ; immunology ; metabolism ; DNA-Binding Proteins ; immunology ; metabolism ; Female ; Humans ; Immunohistochemistry ; Ki-67 Antigen ; metabolism ; Middle Aged ; Minichromosome Maintenance Complex Component 2 ; Nuclear Proteins ; immunology ; metabolism ; Uterine Cervical Neoplasms ; diagnosis ; metabolism ; pathology ; Young Adult

BACKGROUND/AIMS: Maximal duration of intravenous (IV) corticosteroid (CS) treatment and efficacy of cyclosporin A (CsA) have not been clarified for patients with severe ulcerative colitis. We aimed to evaluate and compare the effectiveness of CS and CsA combination therapy with prolonged CS therapy alone in patients with severe UC refractory to initial CS therapy. METHODS: We retrospectively reviewed the medical records of 84 episodes of severe UC in 59 patients between April 1999 and May 2005. RESULTS: Among 84 episodes with IV CS therapy, 45 (53.6%) experienced an early response, while 39 (46.4%) did not respond within 2 weeks. The remaining 36 episodes excluding 3 which underwent colectomy were assigned to either combination therapy of IV CS and CsA or prolonged IV CS treatment alone for additional 2 weeks. Twelve of 16 episodes (75.0%) responded to therapy with combinations of IV CsA and CS, and 16 of 20 episodes (80.0%) to prolonged IV CS treatment alone. There was no statistical difference in response and colectomy rate after 4 weeks between CsA-use group and CsA-non-use group (p=1.00). CONCLUSIONS: These results suggest that CS and CsA combination has no additional benefit over prolonged CS therapy alone in terms of short-term response and that CS can be safely prolonged even after the first 14 days of treatment for severe UC.
Aged ; Aged, 80 and over ; Antibodies, Monoclonal ; Carcinoma, Squamous Cell/*metabolism/pathology ; Cyclin D1/immunology/*metabolism ; Cyclin-Dependent Kinase Inhibitor p16/immunology/*metabolism ; Esophageal Neoplasms/*metabolism/pathology ; Esophagus/*abnormalities/metabolism/pathology ; Female ; G1 Phase ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Tumor Markers, Biological/metabolism ; Tumor Suppressor Protein p53/immunology/*metabolism

OBJECTIVETo elucidate the functional status of dendritic cells (DC) in the tissue of oral squamous cell carcinoma by analyzing characteristic phenotype of them.

METHODS34 specimens from oral squamous cell carcinoma cases primarily treated with surgery were selected as test group. In addition, 30 specimens of normal mucosa from oral mucocele cases were used as control. Distribution of DC expressing CD1a+, HLA-DR+ and CD83+ in tumor tissue and normal mucous membrane was observed by immunohistochemistry. The number of DC expressing the antigens, which represented the density of DC infiltrating into tissue, was counted by microscope. The density of DC and the rate of DC expressing HLA-DR in oral carcinoma group and control were statistically compared.

RESULTSThere was no CD83+ DC in all cases, but CD1a+ DC was found in all samples. The density of CD1a+ DC in tumor tissue was significantly lower than that in normal mucous membrane (P < 0.05). HLA-DR antigen expressed on the surface of DC in tumoral epithelium of 27-case carcinoma specimens and in normal mucous epithelium of 23 cases. The rate of HLA-DR positive expression of TIDC had no statistic significance between the two groups.

CONCLUSIONThe lower density of DC infiltrating in tumor tissue might reflect the microenviromental immunodeficiency of hosts with oral squamous cell carcinoma, and the functional mature of DC might be inhibited by the immunosuppressive action of oral squamous cell carcinoma.

Adult ; Aged ; Antigens, CD ; Antigens, CD1 ; analysis ; Carcinoma, Squamous Cell ; immunology ; pathology ; Cell Count ; Dendritic Cells ; immunology ; metabolism ; Female ; HLA-DR Antigens ; analysis ; Humans ; Immunoglobulins ; analysis ; Immunohistochemistry ; Male ; Membrane Glycoproteins ; analysis ; Middle Aged ; Mouth Mucosa ; immunology ; pathology ; Mouth Neoplasms ; immunology ; pathology ; Phenotype

OBJECTIVETo investigate the role of 67 000 laminin receptor (LN-R) in the processes of invasion and metastasis of laryngeal squamous cell carcinoma.

METHODSExpression of 67 000 LN-R mRNA in 20 cases with laryngeal squamous cell carcinomas and corresponding normal tissues was determined by RT-PCR, and the relationship between its expression level and patients' clinicopathological features was analyzed. Expression of 67 000 LN-R on the surface of AMC-HN-8 laryngeal carcinoma cells was examined by flow cytometry. The effect of 67 000 LN-R monoclonal antibody (MLuC5) on the adhesive and invasive abilities was observed by adhesion test and Boyden chamber invasiveness test in vitro.

RESULTSThe expression level of 67 000 LN-R mRNA in human laryngeal squamous cell carcinoma was significantly higher than that in normal tissues (P < 0.05). The expression level of 67 000 LN-R mRNA in laryngeal squamous cell carcinomas with cervical lymph node metastases was higher than in those without cervical lymph node metastases (P < 0.05). There was a significant negative correlation between the expression level of 67 000 LN-R mRNA and the degree of tumor differentiation, the level being higher in poorly differentiated tumors (P < 0.05). Flow cytometry showed that (80.9 +/- 0.9)% of AMC-HN-8 cells expressed 67 000 LN-R. MLuC5 inhibited the adhesion of AMC-HN-8 cells on LN, and after treated with MLuC5 for 60 and 120 minutes, the adhesion inhibition rate was 57.1% and 63.2%, respectively. The invasive ability to artificial basement membrane was reduced by MLuC5.

CONCLUSIONLaryngeal carcinoma overexpressing 67 000 LN-R has stronger invasiveness, and 67 000 LN-R monoclonal antibody may contribute to prevent metastasis of laryngeal carcinoma.

Aged ; Aged, 80 and over ; Antibodies, Monoclonal ; immunology ; Carcinoma, Squamous Cell ; metabolism ; secondary ; Cell Adhesion ; immunology ; Cell Differentiation ; Cell Line, Tumor ; Female ; Humans ; Laryngeal Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Invasiveness ; RNA, Messenger ; biosynthesis ; genetics ; Receptors, Laminin ; biosynthesis ; genetics ; immunology