Specific Colon Cancer Cell Cytotoxicity Induced by Bacteriophage E Gene Expression under Transcriptional Control of Carcinoembryonic Antigen Promoter

Colorectal cancer is one of the most prevalent cancers in the world. Patients in advanced stages often develop metastases that require chemotherapy and usually show a poor response, have a low survival rate and develop considerable toxicity with adverse symptoms. Gene therapy may act as an adjuvant therapy in attempts to destroy the tumor without affecting normal host tissue. The bacteriophage E gene has demonstrated significant antitumor activity in several cancers, but without any tumor-specific activity. The use of tumor-specific promoters may help to direct the expression of therapeutic genes so they act against specific cancer cells. We used the carcinoembryonic antigen promoter (CEA) to direct E gene expression (pCEA-E) towards colon cancer cells. pCEA-E induced a high cell growth inhibition of human HTC-116 colon adenocarcinoma and mouse MC-38 colon cancer cells in comparison to normal human CCD18co colon cells, which have practically undetectable levels of CEA. In addition, in vivo analyses of mice bearing tumors induced using MC-38 cells showed a significant decrease in tumor volume after pCEA-E treatment and a low level of Ki-67 in relation to untreated tumors. These results suggest that the CEA promoter is an excellent candidate for directing E gene expression specifically toward colon cancer cells.     

Impact of Soluble HLA-G Levels and Endometrial NK Cells in Uterine Flushing Samples from Primary and Secondary Unexplained Infertile Women

The aim of this research was to determine the levels of human leukocyte antigen G (HLA-G) and endometrial Natural Killer ((e)NK) cell percentages in uterine flushing samples from primary and secondary infertile women. sHLA-G levels were lower in the uterine flushing samples from primary infertile women in comparison with women with secondary infertility. Lower CD56⁺KIR2DL4⁺ (e)NK cell percentages were detected in primary infertile women compared with secondary infertile women. This is the first study demonstrating that primary and secondary unexplained infertilities are characterized by different basal sHLA-G levels and CD56⁺KIR2DL4⁺ (e)NK cell percentages.     

Tn Antigen Expression Defines an Immune Cold Subset of Mismatch-Repair Deficient Colorectal Cancer

Colorectal cancer (CRC) cells often express Tn antigen, a tumor-associated truncated immature O-glycan (GalNAcα-O-Ser/Thr) that can promote tumor progression. Immunotherapies against Tn antigen have been developed and are being evaluated in clinical trials. Tn antigen can also be considered a novel immune checkpoint that induces immunosuppressive signaling through glycan-biding lectins to lead effector T cell apoptosis. We evaluated the correlation of Tn antigen expression by immunohistochemistry with mismatch-repair (MMR) status, tumor-infiltrating lymphocytes, tumor cell PD-L1 expression, and clinicopathological characteristics in 507 CRC patients. Although 91.9% of CRCs showed negative or weak Tn antigen staining (Tn-negative/weak), we identified a small subset of CRCs (8.1%) that displayed particularly intense and diffuse distribution of Tn antigen immunoreactivity (Tn-strong) that closely related to deficient MMR (dMMR). Moreover, 40 dMMR CRCs were stratified into 24 Tn-negative/weak dMMR tumors (60.0%) exhibiting dense CD8+ lymphocyte infiltrate concomitant with a high rate of PD-L1 positivity, and 16 Tn-strong dMMR tumors (40.0%) that demonstrated CD8+ T cell exclusion and a lack of PD-L1 expression, which was comparable to those of proficient MMR. Our finding suggests that the immune cold subset of patients with Tn-strong dMMR CRC may be effectively treated with immune checkpoint blockade therapy or cellular immunotherapy targeting Tn antigen.     

Parameters optimization of recombinant hepatitis B antigen adsorption–desorption in purification process on three different kinds of diatomaceous earth matrix

The purpose of this study was to compare adsorptive capacity of three kinds of celite for r-HBsAg. The experimental design methodology was used to optimize the parameters which affect adsorption efficiency. The examined parameters were the adsorption/desorption temperature, pH, time, agitation speed and antigen concentration.     

应用荧光微球技术进行免疫检测研究

以聚苯乙烯羧基荧光微球为固相载体,将人免疫球蛋白G(IgG)和猪血红蛋白(Hb)分别与微球偶联,应用Luminex-100TM荧光微球检测系统检测抗体IgG和抗原Hb与荧光微球的偶联效率.结果表明,采用100μg·mL-1抗体或抗原浓度可以获得较高的偶联效率,且检测快速灵敏.应用荧光微球进行抗原抗体免疫检测可行.     

Study on the Expression and Clinical Significances of Lewis y Antigen and Integrin αv, β3 in Epithelial Ovarian Tumors

Objective

To detect the expression and clinical significances of Lewis y antigen and integrin αv, β3 in epithelial ovarian tumors, and to explore the expression correlation between Lewis y antigen and integrin αv, β3.

Methods

Immunohistochemical staining was performed in 95 cases of epithelial ovarian cancer, 37 cases of borderline tumors, 20 cases of benign tumors, and 20 cases of normal ovarian tissue, for the detection of Lewis y antigen and integrin αv, β3 expressions, and to analyze the relationship between Lewis y antigen and integrin, and the relationship between clinical and pathological parameters of ovarian cancer. In addition, immunofluorescence double labeling was utilized to detect the expression correlation between Lewis y antigen and integrin αv, β3 in ovarian cancer.

Results

In epithelial ovarian tumors, the expression rate of Lewis y antigen was 81.05%, significantly higher than that of borderline (51.53%) (P < 0.05) and benign (25%) (P < 0.01) tumors, and normal ovarian tissues (0) (P < 0.01). The expression rate of integrin αv, β3 in malignant epithelial ovarian tumors was 78.95% and 82.11%, respectively, significantly higher than that of the borderline (45.94%, 40.54%) (both P < 0.05), benign group (10.00%, 15.00%) (both P < 0.01) and normal ovary group (5%, 15%) (both P < 0.01).

Conclusions

Lewis y and integrins αv, β3 are relevant to pelvic and abdominal diffusion and metastasis of ovarian cancer cells, suggesting that these two molecules mediate a boosting function for tumor metastasis.     

Rapid screening and identification of dominant B cell epitopes of HBV surface antigen by quantum dot-based fluorescence polarization assay

A method for quickly screening and identifying dominant B cell epitopes was developed using hepatitis B virus (HBV) surface antigen as a target. Eleven amino acid fragments from HBV surface antigen were synthesized by 9-fluorenylmethoxy carbonyl solid-phase peptide synthesis strategy, and then CdTe quantum dots were used to label the N-terminals of all peptides. After optimizing the factors for fluorescence polarization (FP) immunoassay, the antigenicities of synthetic peptides were determined by analyzing the recognition and combination of peptides and standard antibody samples. The results of FP assays confirmed that 10 of 11 synthetic peptides have distinct antigenicities. In order to screen dominant antigenic peptides, the FP assays were carried out to investigate the antibodies against the 10 synthetic peptides of HBV surface antigen respectively in 159 samples of anti-HBV surface antigen-positive antiserum. The results showed that 3 of the 10 antigenic peptides may be immunodominant because the antibodies against them existed more widely among the samples and their antibody titers were higher than those of other peptides. Using three dominant antigenic peptides, 293 serum samples were detected for HBV infection by FP assays; the results showed that the antibody-positive ratio was 51.9% and the sensitivity and specificity were 84.3% and 98.2%, respectively. In conclusion, a quantum dot-based FP assay is a very simple, rapid, and convenient method for determining immunodominant antigenic peptides and has great potential in applications such as epitope mapping, vaccine designing, or clinical disease diagnosis in the future.     

Hesperidin inhibits development of atopic dermatitis-like skin lesions in NC/Nga mice by suppressing Th17 activity

Hesperidin (previously called vitamin P) is a predominant flavanone present in citrus fruits, and is presumed to have a role in their beneficial effect for human health because it possesses various physiological activities. In this study, we investigated the anti-allergic and anti-inflammatory effects of hesperidin and α-glucopyranosyl (αG)-hesperidin, its derivative with enhanced water-solubility, in NC/Nga mice, a human-like mouse model of atopic dermatitis. NC/Nga mice were fed a 0.1% αG-hesperidin or hesperidin diet for 8 weeks. αG-hesperidin and hesperidin feeding effectively inhibited skin lesions and immunoglobulin E (IgE) elevation. At the end of the 8-week-experimental period, the production of inflammatory cytokine interleukin (IL)-17 and interferon-gamma (IFN-γ) from splenocytes was lower in the αG-hesperidin/hesperidin-fed group than in the control group. Changes in mRNA expression in splenocytes are also examined using DNA microarray and real-time RT-PCR. It was revealed that cytotoxic T-lymphocyte antigen 4 (CTLA4), a regulatory T-cell (Treg) marker, was markedly upregulated in splenocytes, particularly by αG-hesperidin feeding. These results suggest that αG-hesperidin attenuated exacerbation of AD-like symptoms, decreased systemic immune hyper-responsiveness in part through the reduction of IgE, IL-17 and IFN-γ, and also modulated Th17/Treg balance in NC/Nga mice. Therefore, αG-hesperidin may be useful in the management of Th17-mediated allergic disorders.     

抗人红细胞表面抗原glycophorin A杂交瘤细胞株的建立

应用杂交瘤技术,用纯化人红细胞表面抗原glycophorin A“N”型免疫纯系BALB/c小鼠,用glycophorin A“M”型作为固相抗原,用EIA间接筛选出识别“M”、“N”血凝抗原以外的抗glycophorin A的抗体,进而利用Coombs原理,加入抗免疫球蛋白,筛选出非凝集型抗glycophorin A抗体,得到61株非凝集型抗glycophorin A杂交瘤,其中12株有限稀释法克隆4次后得到5株稳定分泌非凝集型抗体的杂交瘤并进行初步鉴定。单抗上清血凝效价可达1:20～50,腹水血凝效价为1:800～1:1 600。     

糖类抗原19-9在膀胱癌患者尿液中异常表达

通过检测膀胱癌患者尿液中糖类抗原 1 9- 9(CA1 9- 9)的含量 ,分析并评估其对诊断膀胱癌的临床价值。用全自动化学荧光系统检测膀胱癌患者 (3 0例 ) ,泌尿系统常见良性疾病 (5 3例 )、恶性疾病 (2 2例 )和其它系统恶性肿瘤 (3 5例 )患者以及 3 0例为既往无各系统恶性肿瘤和泌尿系统病史的志愿者 (对照组 )尿液中CA1 9- 9含量。结果显示 ,膀胱癌组CA1 9- 9含量水平为 1 5 9.0± 1 2 8.0U/mL。对照组的尿CA1 9- 9含量水平为 1 2 .4± 8.4U/mL。以对照组均值± 1 .96SD为临界点 ,即大于 2 8.9U/mL为阳性。膀胱移行细胞癌诊断灵敏度 86.7%、特异性 68.2 %。膀胱癌组与对照组的尿CA1 9- 9含量差异有显著统计意义 (P <0 .0 0 1 ) ,与其它各组差异也有显著统计意义 (P <0 .0 0 1 )。良性泌尿系统疾病组尿CA1 9- 9含量水平为 5 3 .9± 77.9U/mL ,明显高于对照组 (P =0 .0 0 1 ) ,但与泌尿系统其它恶性肿瘤组和其它系统恶性肿瘤组之间差别无显著意义