Airy-Gaussian vector beam and its application in generating flexible optical chains.

In recent years, the manipulation of structured optical beam has become an attractive and promising area. The Gaussian beam is the most common beam as the output beam of the laser, and the Airy beam is recently proposed with fascinating properties and applications. In this paper, for the first time to our knowledge, the polarization is used as a tool to design a new kind of Airy-Gaussian vector beam by connecting the Gaussian and Airy functions, which opens a new avenue in designing new beams based on the existed beams. We realize the Airy-Gaussian vector beam with space-variant polarization distribution in theory and experiment, and find that the vector beam can autofocus twice during propagation. The optical chains with flexible intensity peaks are achieved with the Airy-Gaussian vector beam, which can be applied in trapping and delivering particles including biological cells and Rydberg atoms. Such optical chains can significantly improve the trapping efficiency, reduce the heat accumulation, and sweep away the impurity particles.

PD-L1 blockade liberates intrinsic antitumourigenic properties of glycolytic macrophages in hepatocellular carcinoma.

OBJECTIVE: Patients with increased PD-L1+ host cells in tumours are more potent to benefit from antiprogrammed death-1/programmed death ligand-1 (PD-L1) treatment, but the underlying mechanism is still unclear. We aim to elucidate the nature, regulation and functional relevance of PD-L1+ host cells in hepatocellular carcinoma (HCC). DESIGN: A total of untreated 184 HCC patients was enrolled randomly. C57BL/6 mice are given injection of Hepa1-6 cells to form autologous hepatoma. ELISpot, flow cytometry and real-time PCR are applied to analyse the phenotypic characteristics of PD-L1+ cells isolated directly from HCC specimens paired with blood samples or generated from ex vivo and in vitro culture systems. Immunofluorescence and immunohistochemistry are performed to detect the presence of immune cells on paraffin-embedded and formalin-fixed samples. The underlying regulatory mechanisms of metabolic switching are assessed by both in vitro and in vivo studies. RESULTS: We demonstrate that PD-L1+ host macrophages, which constructively represent the major cellular source of PD-L1 in HCC tumours, display an HLA-DRhighCD86high glycolytic phenotype, significantly produce antitumourigenic IL-12p70 and are polarised by intrinsic glycolytic metabolism. Mechanistically, a key glycolytic enzyme PKM2 triggered by hepatoma cell derived fibronectin 1, via a HIF-1α-dependent manner, concurrently controls the antitumourigenic properties and inflammation-mediated PD-L1 expression in glycolytic macrophages. Importantly, although increased PKM2+ glycolytic macrophages predict poor prognosis of patients, blocking PD-L1 on these cells eliminates PD-L1-dominant immunosuppression and liberates intrinsic antitumourigenic properties. CONCLUSIONS: Selectively modulating the 'context' of glycolytic macrophages in HCC tumours might restore their antitumourigenic properties and provide a precise strategy for anticancer therapy.

B cells polarize pathogenic inflammatory T helper subsets through ICOSL-dependent glycolysis.

B cells constitute abundant cellular components in inflamed human tissues, but their role in pathogenesis of inflammatory T helper (TH) subsets is still unclear. Here, we demonstrate that B cells, particularly resting naïve B cells, have a previously unrecognized helper function that is involved in shaping the metabolic process and subsequent inflammatory differentiation of T-cell receptor-primed TH cells. ICOS/ICOSL axis-mediated glucose incorporation and utilization were crucial for inflammatory TH subset induction by B cells, and activation of mTOR was critical for T cell glycolysis in this process. Consistently, upon encountering ICOSL+ B cells, activated effector memory TH cells from patients with rheumatoid arthritis or systemic lupus erythematosus spontaneously differentiated into inflammatory TH subsets. Immunotherapy using rituximab that specifically depleted B cells in patients with rheumatoid arthritis efficiently abrogated the capabilities of memory TH cells to incorporate and use glucose, thereby impairing the pathogenic differentiation of inflammatory TH subsets.

[Effects of acute high altitude hypoxia on EEG power in different emotional states].

Objective: To investigate the effects of acute high altitude hypoxia on EEG power in different emotional states. Methods: This study was two-factor within-subject design (2 levels of oxygen environment ×4 levels of emotion type). Twelve male subjects aged between 20 and 25 years old were induced to produce four different types of emotions by emotional picture evoked paradigm: low valence and low arousal(LVLA), high valence and low arousal(HVLA), low valence and high arousal(LVHA), high valence and high arousal(HVHA). Brain Products 32 was used to collect EEG signals under different emotional states. The next day, a constant depressed oxygen chamber was used to simulate a 4 300 m plateau hypoxia environment, and the same group of subjects used the same experimental paradigm to collect EEG signals 10h after hypoxia. The collected EEG signals were analyzed by power spectrum (FFT), and the five frequency bands (Delta, Theta, Alpha, beta, gamma) of the frontal lobe (F3\Fz\F4) were analyzed by variance analysis of two-factor repeated measurements. Results: FFT analysis found that before and after acute hypoxia, the whole brain distribution of alpha wave in four emotional states was mainly concentrated in frontal and parietal leaves; the distribution of alpha wave in the whole brain was the least in relaxed emotional state. The results of the two-factor repeated measurement ANOVA showed that: ①the power of delta\ beta band was significantly affected by the oxygen environment(P＜0.05), and the power was enhanced under hypoxia. ②The power index of theta\ alpha band showed a significant interaction between the oxygen environment and emotional types(P＜0.05). Except for the HVLA emotional state, the power of theta alpha band was significantly enhanced under hypoxia. ③ The two factors had no significant influence on the gamma band(P＞0.05). Conclusion: Under the four kinds of emotional states, the difference of the influence of oxygen environment on brain activity was mainly in the frontal lobe, parietal lobe and part of temporal lobe. Of the four types of emotions, the oxygen environment had the least significant effect on brain activity in HVLA emotional states, while the rest showed significant differences.

[Intermittent hypoxic preconditioning relieves fear and anxiety behavior in post-traumatic stress model mice].

Intermittent hypoxia (IH) has preventive and therapeutic effects on hypertension, myocardial infarction, cerebral ischemia and depression, but its effect on post-traumatic stress disorder (PTSD) has not been known. In this study, we used inescapable electric foot shock combined with context recapture to build PTSD mouse model. The levels of fear and anxiety were valued by the open field, the elevated plus maze (EPM) and the fear conditioning tests; the level of spatial memory was valued by Y maze test; the number of Fos positive neurons in hippocampus, amygdala and medial prefrontal cortex was valued by immunohistochemical staining; and the protein expressions of hypoxia inducible factor-1α (HIF-1α), vascular endothelial growth factor (VEGF) and brain derived neurotrophic factor (BDNF) in these brain area were valued by Western blot. The results showed that IH and model (foot shock) had an interaction on percentage of entering open arms (OE%) in EPM and freezing time and the number of fecal pellets in fear conditioning test. IH increased OE% in EPM and reduced the freezing time and the number of fecal pellets in fear conditioning test in PTSD model mice. At the same time, IH reduced the number of Fos positive neurons in the hippocampus, amygdala and medial prefrontal cortex of PTSD model mice, and increased the protein expression levels of HIF-1α, VEGF and BDNF in these brain tissues. In conclusion, IH pretreatment can relieve fear and anxiety behavior in post-traumatic stress model mice, suggesting that IH may be an effective means of preventing PTSD.

B7-H6 protein expression has no prognostic significance in human gastric carcinoma.

B7-H6, a novel member of the B7 family which binds to NKp30 to trigger antitumor NK cell cytotoxicity and cytokine secretion. Recently, B7-H family has been reported to be a negative regulator of the immune response in patients with gastric carcinoma. However, no reports have investigated the clinical significance of B7-H6 expression in human gastric cancer. We present the first study to the clinicopathological and prognostic value of B7-H6 in primary gastric tumors and adjacent non-tumor tissues at the protein level. Here we show that B7-H6 immunoreactivity was expressed in 6/60 (10%) gastric tumors and 8/43 (18.60%) adjacent non-tumor tissues. No statistical difference was found between B7-H6 expression and various prognostic factors; however, B7-H6-positive carcinomas were significantly associated with a higher differentiation (p = 0.047). The survival analysis did not confirm the prognostic significance of B7-H6 expression in gastric cancer patients. Our data suggest that B7-H6, as detected by immunohistochemistry, is of limited value as a prognostic marker for gastric cancer.

Soluble B7-H3 elevations in hospitalized children with Mycoplasma pneumoniae pneumonia.

The purpose of this study is to explore the associations between soluble B7-H3 (sB7-H3) and cytokines, clinical characteristics and laboratory findings. Thirty-two children with Mycoplasma pneumoniae pneumonia diagnosed by both positive serology and PCR were admitted to Children's Hospital affiliated to Soochow University. These children were enrolled and evaluated from May 2012 through September 2012. Soluble B7-H3 level and cytokines (tumor necrosis factor-α (TNF-α), interferon-γ, interleukin (IL)-4, IL-10) were determined by enzyme-linked immunosorbent assay technique. Meanwhile, clinical parameters including laboratory findings were obtained. Soluble B7-H3 level was significantly increased in patients with M. pneumoniae pneumonia compared with the levels of sB7-H3 in control subjects (4.94 ± 2.69 vs. 3.42 ± 1.48, ng/mL; P = 0.032). Furthermore, level of sB7-H3 was correlated with TNF-α level in plasma in patients with M. pneumoniae pneumonia (rp = 0.667; P < 0.001) as well as level of sB7-H3 in M. pneumoniae pneumonia subjects was also correlated with duration of symptoms (rp = 0.607; P < 0.001), percentage of neutrophil cells (rp = 0.657; P < 0.001), and C-reactive protein level (rs = 0.445; P = 0.011). Level of sB7-H3 was decreased after treatment (6.08 ± 3.07 vs. 3.55 ± 1.58, ng/mL; P = 0.019). Soluble B7-H3 maybe plays an important role in immunopathogenesis of M. pneumoniae pneumonia, especially for increasing TNF-α concentration and activation neutrophils.

Therapeutic effects of anti-B7-H3 antibody in an ovalbumin-induced mouse asthma model.

BACKGROUND: B7 molecules play a key role in regulating allergen-induced T cell activation in asthma, which may occur through T cell recruitment and T helper cell differentiation on allergen provocation. Initial studies have shown that B7-H3 (CD276), a recently identified B7 family member, plays a critical role in the development of Th2 cells. OBJECTIVE: To investigate the effects of anti-B7-H3 monoclonal antibody (mAb) in a mouse model of allergic asthma. METHODS: The asthma model was established by ovalbumin (OVA) sensitization and challenging in female BALB/c mice. Total cell numbers in bronchoalveolar lavage fluid (BALF) were determined, and the expression levels of interferon gamma (IFN-γ), interleukin (IL)-4, and IL-17 in BALF were measured by enzyme-linked immunosorbent assay. Pulmonary eosinophil infiltration and mucus production were detected by hematoxylin and eosin (H&E) and periodic acid-Schiff (PAS), respectively. B7-H3 expression was detected by immunohistochemistry in frozen tissue sections. RESULTS: Anti-B7-H3 mAb treatment alleviated the asthmatic syndrome, decreased the levels of B7-H3-positive cells in the lung tissues, abrogated hypercellularity, eosinophil infiltration, and mucus production, and inhibited IL-4 and IL-17 production in BALF at the induction phase as compared with the immunoglobulin G (IgG) control group (P < .01). In addition, the treatment of anti-B7-H3 mAb at the induction phase could increase the expression levels of IFN-γ as compared with the IgG control group (P < .01). Anti-B7-H3 mAb treatment at the effector phase did not inhibit the asthma response. CONCLUSION: Blockade of B7-H3 signals may provide a novel therapeutic approach to the treatment of allergic asthma.

[The expression of peripheral Th1 and Th17 cells in inflammatory bowel disease and its potential clinical value].

OBJECTIVE: To explore the probable role of Th1 and Th17 cells in the pathogenesis of inflammatory bowel disease (IBD). METHODS: The peripheral blood mononuclear cells (PBMCs) from peripheral blood specimens were collected in the study, including 40 healthy controls, 42 ulcerative colitis (UC) and 39 Crohn's disease (CD). The proportion of Th1 and Th17 cells in the PBMCs was detected with flow cytometry after stimulated by PMA and ionomycin. The result and the clinical data were analyzed. RESULT: The Th1 cell expression was increased in CD (38.32 ± 16.18)% and UC group (34.23 ± 11.60)%, compared with the controls (24.58 ± 10.02)% (P < 0.01). During the convalescence, the Th1 expression in the CD and UC groups in vivo was significantly reduced without difference between the two groups (P > 0.05) . In the IBD group , significant difference in the frequency of Th17 cells could be found between the CD group (2.51 ± 1.59)% and the UC group (4.15 ± 2.75)%, while the Th17 cells were increased in both groups, compared with the controls (1.44 ± 0.73)% (P < 0.05) . Obvious difference in the frequency of Th17 cells could be found between patients at different activity stages and remission stages. The proportion of Th17 cells were higher in the UC patients than that in the CD patients (P < 0.01) . The Th17/Th1 ratio of CD patients, UC patients were 0.08 ± 0.06, 0.14 ± 0.11, which were both higher that in the controls (0.07 ± 0.06). Significant difference could be found between the UC group and the CD group (P < 0.01). CONCLUSIONS: The higher proportion of Th1 and Th17 cells are detected in the peripheral blood of IBD patients, which is correlated closely to the activity of the disease. Th1 and Th17 cells may play an important role in the pathogenesis of IBD.

Silencing of B7-H3 increases gemcitabine sensitivity by promoting apoptosis in pancreatic carcinoma.

In numerous types of cancer, the expression of a novel member of the B7 ligand family, the B7-H3 immunoregulatory protein, has been correlated with a poor prognosis. In the present study, we investigated the role of B7-H3 in chemoresistance in pancreatic carcinoma. Silencing of B7-H3, through lentivirus-mediated delivery of stable short hairpin RNA, was observed to increase the sensitivity of the human pancreatic carcinoma cell line Patu8988 to gemcitabine as a result of enhanced drug-induced apoptosis. Overexpression of B7-H3 caused the cancer cells to be more resistant to the drug. Subsequently, we investigated the underlying mechanisms of B7-H3-mediated gemcitabine resistance, and found that the levels of survivin decreased in cells in which B7-H3 had been knocked down. In vivo animal experiments demonstrated that tumors in which B7-H3 had been knocked down displayed a slower growth rate compared with the control xenografts. Notably, gemcitabine treatment led to a strong antitumor activity in mice with tumors in which B7-H3 had been knocked down; however, this effect was only marginal in the control group. Furthermore, survivin expression was weak in gemcitabine-treated tumors in which B7-H3 had been knocked down and apoptosis was increased, as revealed by terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick-end labeling (TUNEL) staining. In summary, the present study demonstrated that B7-H3 induces gemcitabine resistance in pancreatic carcinoma cells, at least partially by downregulating survivin expression. These results provide novel insights into the function of B7-H3 and encourage the design and investigation of approaches targeting this protein in treating pancreatic carcinoma.

B7-H3 overexpression in pancreatic cancer promotes tumor progression.

B7-H3, a member of the B7-family molecules, plays an important role in adaptive immune responses. In addition, B7-H3 is also expressed in several types of human cancers and is correlated with the poor outcome of cancer patients. However, its exact role in cancer is not known. In the present study, we compared B7-H3 expression in normal pancreas and pancreatic cancer tissue specimens, and determined the effects of low B7-H3 expression on the human pancreatic cancer cell line Patu8988 using lentivirus-mediated RNA interference. B7-H3 expression in pancreatic specimens was determined by enzyme-linked immunosorbent assay (ELISA). A Patu8988 cell line with low B7-H3 expression was established by lentivirus-mediated RNA interference to investigate the effect of B7-H3 on cell proliferation, migration and invasion in vitro. By establishing subcutaneous transplantation tumor and orthotopic transplantation pancreatic cancer mouse models, the effect of B7-H3 on cell proliferation, migration and invasion was studied in vivo. B7-H3 in tissue samples was significantly higher in the pancreatic cancer group than in the normal pancreas group (mean ± SD, 193.6±9.352 vs. 87.74±7.433 ng/g; P<0.0001). B7-H3 knockdown by RNA interference decreased cell migration and Transwell invasion up to 50% in vitro. No apparent impact was observed on cell proliferation in vitro. In the subcutaneous transplantation tumor mouse model, the tumor growth rate was reduced by the knockdown of B7-H3. In the orthotopic transplantation pancreatic cancer mouse model, the effect of inhibiting metastasis by knocking down B7-H3 was assessed in terms of the average postmortem abdominal visceral metastatic tumor weight. This demonstrated that inhibition of B7-H3 expression reduced pancreatic cancer metastasis in vivo. In conclusion, B7-H3 is aberrantly expressed in pancreatic cancer. In addition to modulating tumor immunity, B7-H3 may have a novel role in regulating pancreatic tumor progression.

Association between single nucleotide polymorphisms in the XRCC1 gene and susceptibility to prostate cancer in Chinese men.

BACKGROUND: Prostate cancer (Pca) is one of the most common complex and polygenic diseases in men. The X-ray repair complementing group 1 gene (XRCC1) is an important candidate in the pathogenesis of Pca. The purpose of this study was to evaluate the association between single nucleotide polymorphisms in the XRCC1 gene and susceptibility to Pca. MATERIALS AND METHODS: XRCC1 gene polymorphisms and associations with susceptibility to Pca were investigated in 193 prostate patients and 188 cancer-free Chinese men. RESULTS: The c.910A>G variant in the exon9 of XRCC1 gene could be detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing methods. Significantly increased susceptibility to prostate cancer was noted in the homozygote comparison (GG versus AA: OR=2.95, 95% CI 1.46-5.42, χ2=12.36, P=0.001), heterozygote comparison (AG versus AA: OR=1.76, 95% CI 1.12-2.51, χ2=4.04, P=0.045), dominant model (GG/AG versus AA: OR=1.93, 95% CI 1.19-2.97, χ2=9.12, P=0.003), recessive model (GG versus AG+AA: OR=2.17, 95% CI 1.33-4.06, χ2=8.86, P=0.003) and with allele contrast (G versus A: OR=1.89, 95% CI 1.56-2.42, χ2=14.67, P<0.000). CONCLUSIONS: These findings suggest that the c.910A>G polymorphism of the XRCC1 gene is associated with susceptibility to Pca in Chinese men, the G-allele conferring higher risk.

Changes of regulatory T cells and FoxP3 gene expression in the aging process and its relationship with lung tumors in humans and mice.

BACKGROUND: Immunosuppressive regulatory T cells (Tregs) participate in tumor immune evasion and the number and suppressive function of Tregs change with the aging process, but it is not clear whether such change leads to a higher incidence of tumors in the elderly. To this end, we designed experiments to explore the changes of Tregs and the functional gene Forkhead box P3 (FoxP3) in the aging process and its relationship with lung tumors in humans and mice. METHODS: The percentage of CD4(+)CD25(+)CD127(low) Tregs and expression of FoxP3 mRNA were analyzed using flow cytometry (FCM) and real-time fluorescence-based quantitative polymerase chain reaction (FQ-PCR). Markers were analyzed in the peripheral blood (PB) of 65 elderly patients (age ≥ 65 years) with primary non-small cell lung cancer (NSCLC), 20 younger patients (aged < 55 years) with NSCLC, 30 elderly healthy individuals and 30 young healthy individuals. Furthermore, we set up the Lewis lung cancer model with C57BL/6 female mice. Thirty-six mice were divided into a young healthy group, a middle-aged healthy group, an elderly healthy group, a young tumor group, a middle-aged tumor group, and an elderly tumor group. The percentage of CD4(+)CD25(+)FoxP3(+) Tregs and the expression level of FoxP3 mRNA in splenocytes were determined in the six groups. RESULTS: The percentage of peripheral CD4(+)CD25(+)CD127(low) Tregs and the expression of FoxP3 mRNA were significantly increased in elderly patients with NSCLC comparing with the other groups and in elderly healthy individuals compared with young healthy individuals. Further analysis showed that the percentage of CD4(+)CD25(+)CD127(low) Tregs and the expression of FoxP3 mRNA were closely associated with tumor node metastasis (TNM) staging in elderly patients with NSCLC. In the mouse model, the percentage of CD4(+)CD25(+)FoxP3(+) Tregs and the expression of FoxP3 mRNA in splenocytes of the tumor groups were significantly higher than in the healthy groups, with the highest expression in the elderly tumor group. In the healthy groups, the elderly healthy mice had the highest percentage of Tregs and expression of FoxP3 mRNA. The elderly mice had larger and heavier tumors than did the young and middle aged mice. CONCLUSIONS: The up-regulation of Tregs and the FoxP3 gene with aging may play an essential role in oncogenesis and development of lung tumors in an elderly population.

[Expression of B7-H3 costimulatory molecule in peripheral blood of myasthenia gravis patients].

AIM: To reveal the clinical significance of B7-H3 costimulatory molecule in myasthenia gravis (MG) patients by analyzing membranous B7-H3 (mB7-H3) and soluble B7-H3 (sB7-H3) expressions. METHODS: We collected peripheral blood samples of 35 MG patients and 44 health controls (HC) and detected the expression of mB7-H3 on peripheral blood mononuclear cells (PBMCs) using flow cytometry. ELISA was performed to analyze the levels of sB7-H3 in plasma samples from MG patients and HC. RESULTS: There was no significant difference in the expressions of mB7-H3 on T lymphocytes, monocytes or B cells between MG patients and HC. However, the level of sB7-H3 from MG patients was (2.166±0.958) ng/mL, significantly lower than that from HC (3.379±0.768) ng/mL. The level of sB7-H3 in general MG (GMG) patients (1.664±0.699) ng/mL was lower than that in ocular MG (OMG) patients (2.396±0.985) ng/mL. In MG patients complicated with abnormal thymus, the level of sB7-H3 was (1.593±0.441) ng/mL, also lower than that in MG patients with normal thymus (2.364±1.014) ng/mL. In addition, a significant negative correlation was found between the levels of sB7-H3 and QMGS in MG patients (r=-0.4189, P=0.012), but sB7-H3 was not associated with mB7-H3 in MG patients. CONCLUSION: In MG patients, down-regulation of sB7-H3 is finely correlated to the severity of the disease. Its different expression levels in various types of MG patients indicate that this costimulatory molecule may be involved in the immunopathogenesis of MG.

[Establishment of a sandwich ELISA for testing serum sB7-H3 and detection of sB7-H3 levels in liver disease].

AIM: To establish quantitative ELISA for soluble sB7-H3 and evaluate its clinical application. METHODS: Two mAbs of mouse anti-human sB7-H3(4H7 and 2E6)established by our lab were used. The mAb 4H7 was used as coating antibody and the 2E6 as a sandwich antibody, which recognized a different epitope and was labeled by biotin. The serum sB7-H3 levels of healthy volunteers and patients were measured by this method. RESULTS: Sandwich ELISA was established with the linear range covering from 8.192 ng/L to 2 000 ng/L. The outcomes of repeatability test, recovery test, stability test and specificity test were quite good. The increased levels of sB7-H3 were found in schistosomiasis cirrhosis, the reduced levels in severe hepatitis and the difference in both situations showed significance. CONCLUSION: A specific and sensitive sandwich ELISA is developed, the detection in liver disease indicates that the ELISA kit provides a potential tool in the clinical application.

[Preparation of anti-human 4-1BB monoclonal antibody and characterization of its biological activities].

AIM: To prepare an anti-human 4-1BB functional monoclonal antibody and to characterize its biological activities. METHODS: A stable human 4-1BB molecule transfected cell line 293T/4-1BB was used as an antigen to immunize BALB/c mice. By means of the cell fusion by hybridoma technique and multiple cell subcloning and repeated screening with 293T/4-1BB as the antibody screening positive cell while 293T/mock as the negative cell. The hybridoma cell lines specifically secreting anti-4-1BB monoclonal antibodies were selected. Then their characteristics and its biological activities were investigated by Western blot, fast-strip routine Ig subclass typing method, indirect immunofluorescence, competitive inhibition test, (3);H-TdR and cell apoptosis analysis. RESULTS: Three hybridoma cell lines 1G5, 4B11 and 9F11 with the property of secreting specific anti-4-1BB monoclonal antibody continuously and steadily were successfully obtained. These monoclonal antibodies could bind to human 4-1BB epitopes on activated T cells and monocytoes and DC. Additionally, mAb 4B11 could promote T proliferation and enhance the growth and maturation of Mo-DC. CONCLUSION: Three hybridoma cell lines which secrete anti-4-1BB monoclonal antibodies steadily have been established. These monoclonal antibodies could specifically recognize 4-1BB molecule and mAb 4B11 had a potent function to promote T proliferation cell as well as to enhance the growth and maturation of Mo-DC in vitro.

[Expression of mouse B7-H3-Fc fusion protein and characterization of its bioactivity].

AIM: To obtain mouse B7-H3-Fc fusion protein and to investigate its biological function and effects on T lymphocyte activation. METHODS: The genes coding extracellular domain of mouse B7-H3 and the Fc fragment of human IgG1 were amplified from pMD19-T/mouse B7-H3 and pMD19-T/human IgG1 vectors by PCR. The two genes were combined with mouse B7-H3-Fc fragment by overlap PCR. Then the resulting gene fragment was inserted into eukaryotic vector pIRES2-EGFP after digested with EcoR I and Bgl II to construct the recombinant vector pIRES2-EGFP/B7-H3-Fc. The recombinant vector was transfected into CHO cells with LipfectAMINE™ 2000, and the cells were further selected with G418. The collected supernatant of the transfected cell line cultured in serum-free media was ultrafiltrated and concentrated, then purified by Protein G column. The expression of mouse B7-H3-Fc fusion protein was confirmed by Western blot. Effects of fusion protein on T cells proliferation and cytokine production in vitro was studied by methods of CCK8 and ELISA. RESULTS: The results showed that the transfected CHO cell line secreting mouse B7-H3-Fc fusion protein was constructed successfully. In vitro, mouse B7-H3-Fc fusion protein obviously promoted the proliferation of T cells in a dose-dependent manner. CONCLUSION: A transfected CHO cell line stably expressing mouse B7-H3-Fc fusion protein has been obtained and the B7-H3-Fc fusion protein stimulates the proliferation of T cells and cytokines production in vitro.

B7-H3 and CD133 expression in non-small cell lung cancer and correlation with clinicopathologic factors and prognosis.

OBJECTIVE: To detect the expression of B7-H3 and CD133 in human non-small cell lung cancer (NSCLC) specimens and lung benign lesions, and to evaluate the correlation between the 2 biomarkers and clinicopathologic features. METHODS: This is a case-control study of 102 tissue specimens collected from NSCLC participants undergoing thoracic surgery in the Second Affiliated Hospital of Soochow University, Suzhou, China, between January 2006 and December 2008. From the 102 patients, 25 adjacent non-cancer samples were verified pathologically as normal tissue (positive group), and 24 benign inflammatory lesion tissues were used as control (negative group). Specimens from 126 participants were stained immunohistochemically using Image-Pro Plus software, and the cell number was measured in each section. RESULTS: Of the 102 specimens, 71 expressed B7-H3, and 51 expressed CD133, higher than that in benign lesions (p<0.001) or non-cancer tissues (p<0.001). B7-H3 expression in squamous cell carcinoma (SCC) was significantly higher than those in adenocarcinoma (p=0.048), while CD133 expression in large cell lung carcinoma was higher than that in SCC (p=0.023). The mean number of tumor-infiltrating lymphocytes (TILs) in the B7-H3-positive group was lower than that in the B7-H3-negative group (p=0.026). The mean TILs in the CD133-positive group was significantly lower than that in CD133-negative group (p=0.029). We found that CD133 was related to tumor cell differentiation degree and CD133 expression was negatively correlated with B7-H3 expression. The CD133 positive or B7-H3 negative was associated with poor prognosis of NSCLC patients by Cox regression analysis. CONCLUSION: Both CD133 and B7-H3 might induce apoptosis of TILs in NSCLC and tumor evading host immune surveillance. Either CD133 or B7-H3 might be an independent risk factor of NSCLC participants.

Clinical significance and regulation of the costimulatory molecule B7-H3 in human colorectal carcinoma.

B7-H3, a member of the B7-family molecules, plays an important role in adaptive immune responses, and was shown to either promote or inhibit T-cell responses in various experimental systems. B7-H3 was expressed in some human cancers and correlated with poor outcome of cancer patients. However, its exact role in cancer is not known. In the present study, we studied the expression of B7-H3 in the pathologic specimens of 102 patients treated for colorectal carcinoma (CRC) by immunohistochemistry. Strong B7-H3 expression was found in cancer tissues from 54.3% CRC patients, while minimal expression was found in adjacent normal colorectal tissues. Higher B7-H3 expression in tumor positively correlated with a more advanced tumor grade. In addition, consistent with a role of B7-H3 in suppressing tumor immune surveillance, the expression of B7-H3 in cancer cells negatively correlated with the intensity of tumor infiltrating T lymphocytes in both tumor nest and tumor stroma. Furthermore, we found that the level of soluble B7-H3 in sera from CRC patients was higher than healthy donors. TNF-alpha, an important cancer-promoting inflammatory molecule, was subsequently found to significantly increase the release of soluble B7-H3 in colon cancer cell lines. Therefore, our data suggest that both soluble and membranous B7-H3 proteins are involved in colon cancer progression and evasion of cancer immune surveillance.

Altered CD28 and CD95 mRNA expression in peripheral blood mononuclear cells from elderly patients with primary non-small cell lung cancer.

BACKGROUND: The expression of the co-stimulatory molecule CD28 and death receptor CD95 on T cells, which change with age, are considered as important immunological parameters of immunosenescence. It is well established that CD28 and CD95 are associated with tumorgenesis and tumor progression, but the relationship between the age-related changes of these two immunological markers and cancer in the elderly is largely unknown. METHODS: The levels of CD28 and CD95 mRNA in peripheral blood mononuclear cells (PBMCs) from sixty-three elderly patients (aged > or = 60 years) with primary non-small cell lung cancer (NSCLC) were analyzed by real-time fluorescence-based quantitative polymerase chain reaction (FQ-PCR). In addition, twenty young patients (aged < 60 years) with NSCLC, thirty elderly healthy donors and thirty young healthy donors were enrolled as controls. RESULTS: CD28 mRNA levels were significantly lower and CD95 mRNA levels were significantly higher in elderly patients with NSCLC than in the other groups. Similar results were found in elderly healthy donors comparing with young healthy donors. By Logistic regression analysis an increased risk of NSCLC was markedly associated with aging, down-regulation of CD28 mRNA and up-regulation of CD95 mRNA, and CD28 mRNA had an obvious negative correlation with the CD95 mRNA. In addition, the mRNA levels of CD28 and CD95 in the peripheral blood of the elderly patients was closely associated with the tumor node metastasis (TNM) stages, grade of cell differentiation and lymph node metastasis status, but not related to pathological types. CONCLUSIONS: The results suggest a close relationship between T cell senescence and NSCLC tumour progress in the elderly, and that up-regulation of CD28 mRNA or down-regulation of CD95 mRNA in peripheral blood T cells may play an important role in inhibiting oncogenesis and development of primary NSCLC in the elderly.