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91.
人乳头瘤病毒18型L1-E6、L1-E7嵌合基因表达载体的构建及表达 总被引:6,自引:1,他引:6
目的 构建HPV 18 L1-E6,L1-E7嵌合基因的表达载体,并在CHO细胞中表达。方法 克隆HPV18 L1-E6和L1-E7基因,插入中介载体pGEMT-Easy中并测序鉴定。采用PCR定点突变法,突变L1-E6,L1-E7基因序列中与转化作用相关的位点,分别与L1基因连接后插入真核表达载体pVAX1,构建真核表达质粒pVAX-1L1 E6Mxx,L1E7Mxx。用磷酸钙沉淀法,转染CHO细胞,以抗HPV-18L1,抗E6和抗E7特异性单克隆抗体(mAb)做ELISA和免疫细胞化学法检测。结果 ELISA检测显示,转染各种pVAX1-LIE6Mxx-L1E7Mxx融合蛋白表达质粒的细胞提取物的P-N值均>2.1;免疫细胞化学检测,在胞浆,胞核可见棕黄色颗粒。结论 我建的pVAX1-L1E6Mxx-E7Mxx融合蛋白质表达质粒,可在转当细胞内表达相应的L1-E6Mxx和L1-E7Mxx蛋白,为今后进行DNA疫苗的研究奠定了基础。 相似文献
92.
Komada H Ito M Nishio M Kawano M Ohta H Tsurudome M Kusagawa S O'Brien M Bando H Ito Y 《Medical microbiology and immunology》2000,189(1):1-6
cDNAs encoding human parainfluenza virus type 4B (hPIV-4B) hemagglutinin neuraminidase (HN) protein were cloned and the nucleotide
sequences were determined. A high degree of identity (81.4%) was observed between the nucleotide sequences of hPIV-4A and
-4B HN proteins, and an 87.3% identity was found between the deduced amino acid sequences. This degree of identity is considered
to be greater than immunological similarity between hPIV-4A and -4B HN proteins determined using monoclonal antibodies. To
elucidate the causes of the antigenic difference between HN proteins of hPIV-4A and -4B, we constructed three cDNAs of hPIV-4B
HN whose potential N-glycosylation sites were partially or completely the same as in hPIV-4A HN cDNA. We compared the antigenicity of the expressed
wild-type and mutant proteins, and found that the antigenicities of the mutant hPIV-4B HN proteins were more similar to the
hPIV-4A HN protein than to the non-mutant hPIV-4B HN protein. This study indicated that the antigenic diversity between hPIV-4A
and -4B was partly caused by deletion or creation of glycosylation sites, showing that the point mutations resulting in deletion
or creation of glycosylation sites is one of the initial steps leading to the division of virus into subtypes.
Received: 21 January 2000 相似文献
93.
Sugiura-Ogasawara M Ozaki Y Nakanishi T Kumamoto Y Suzumori K 《American journal of reproductive immunology (New York, N.Y. : 1989)》2005,53(1):50-53
PROBLEM: It is unclear whether chlamydia infection influences the miscarriage rate and immunological factors in patients with recurrent miscarriage. METHOD OF STUDY: Chlamydia DNA, IgA and IgG to Chlamydia trachomatis, natural killer cell activity, complement 3 (C3), C4, hemolytic complement, antinuclear antibodies, antiphospholipid antibodies, prolactin, activated partial thromboplastin time, prothrombin time and fibrinogen were examined in 504 patients with a history of two or more consecutive first-trimester miscarriages. Subsequent pregnancy outcomes were compared between cases with and without antibodies to C. trachomatis. RESULTS: Totals of 10 of 30 and 48 of 201 patients receiving no medication miscarried subsequently with and without chlamydia infection. Chlamydia IgA and/or IgG were associated with a high level of C3 but not other immunological and coagulatory parameters. CONCLUSION: Antibodies to C. trachomatis do not influence subsequent pregnancy outcome in patients with a history of recurrent miscarriage. 相似文献
94.
Telomere-specific fluorescence in situ hybridization analysis of couples with five or more recurrent miscarriages 总被引:1,自引:0,他引:1
Fluorescence in situ hybridization analysis using telomere specific probes has been used to detect cryptic translocations in the chromosomal telomeric regions. This study was performed in five clinically normal couples who have had five or more spontaneous abortions and whose karyotypes were found to be normal using conventional cytogenetic techniques. Using the telomere specific probes, in one couple we determined a cryptic translocation between chromosome 3 and 10, and, in another couple, the signal in chromosome 20 was detected in another chromosome, which was probably a D group chromosome. Additionally, in the latter and also in two other couples, we observed a polymorphism. The approach will be helpful for screening cryptic translocations using telomere specific multiple probe sets in couples with recurrent miscarriages. As prenatal diagnosis will be available for these couples for future pregnancies, it will be possible to help these families to have healthy fetuses. 相似文献
95.
Sputum cytology was performed in 179 cases of squamous-cell carcinoma of the lung; 134 cases were diagnosed as positive. There were no significant differences in diagnostic accuracy of sputum cytology between tumors sizes. In cases with tumors extending proximally into the main, lobar, or segmental bronchi, the diagnostic accuracy of sputum cytology was significantly higher than in cases where the proximal invasion of tumor was limited to the peripheral bronchi. In cases with tumors 3 cm or less in diameter, when tumors extended proximally into main, lobar, or segmental bronchi, the diagnostic accuracy of sputum cytology was significantly higher than in cases with tumors extending proximally into subsegmental or subsubsegmental bronchi. In peripherally located squamous-cell carcinoma, in cases in which the tumor arose in subsegmental or subsubsegmental bronchi, carcinoma could be detected by sputum cytology even when it was roentgenographically occult. 相似文献
96.
Takakuwa K Hataya I Arakawa M Kikuchi A Higashino M Yasuda M Kurabayashi T Tanaka K 《American journal of reproductive immunology (New York, N.Y. : 1989)》1999,42(4):233-239
PROBLEM: To clarify whether HLA-DP antigens are associated with patient population of unexplained recurrent abortion. METHOD OF STUDY: The frequency of HLA-DPB1 alleles in patients with unexplained recurrent abortion, and the compatibility of HLA-DPB1 alleles between patient couples, were studied using a polymerase chain reaction (PCR)-restricted fragment length polymorphism (RFLP) method. Thirty patients who had a history of unexplained primary recurrent abortion, and their husbands, were typed for HLA-DPB1 genotype. Two hundred and ninety-nine base pair fragments from the second exon of HLA-DPB1 genes were selectively amplified using the PCR-primers. After amplification, the DNAs were digested with restriction endonucleases, and subjected to electrophoresis in a 12% polyacrilamide gel to determine HLA-DPB1 genotype. RESULTS: The frequency of HLA-DPB1*0402 and DPB1*04 alleles in the patient group (n = 30) was significantly increased, as compared to that in the normal fertile women (n = 30). The frequency of HLA-DPB1*04 allele in the patient group was significantly increased, as compared to that in the general population (n = 112). No significant compatibility of HLA-DPB1 alleles could be observed between patient couples and normal fertile couples. CONCLUSION: These findings suggest a possible new class II association with patient population of unexplained recurrent abortion. 相似文献
97.
Uzi Gafter Benjamin Sredni Jacob Segal Yona Kalechman 《Journal of clinical immunology》1997,17(5):408-419
Spontaneous recurrent abortion (SRA) has been treated by means of immunization with paternal or third-party white blood cells, yet the immunological basis for SRA and for the role of immunization protocols in pregnancy outcome remains controversial. To elucidate this question, nine women with SRA were immunized with paternal mononuclear cells and studied before and 2 weeks after immunization. Seven women who became pregnant gave birth to live newborns. Secretion of the T helper 1 cytokines IL-2 and interferon- by patients' mononuclear cells decreased, while production of IL-10 increased. The levels of natural killer and lymphokine-activated killer cell-mediated cytotoxicity were markedly decreased. Monocyte functions such as secretion of IL-l, tumor necrosis factor a, IL-6, and cytotoxic activity decreased concurrently with elevations in IL-10 and transforming growth factor secretion. Production of IL-12, a pivotal regulatory cytokine, decreased. Furthermore, B7/1 expression on patients' mononuclear cells was downregulated. This resulted in a decrease in monocyte costimulatory activity of purified T cells with soluble anti-CD3, paralleled by a decline in allogeneic proliferative responses. These results suggest that the improved pregnancy success rate in women with SRA following immunization may be partly related to suppression of cell-mediated immunity and monocyte and natural killer cell activity. 相似文献
98.
99.
Hori S Nakano Y Furukawa TA Ogasawara M Katano K Aoki K Kitamura T 《American journal of reproductive immunology (New York, N.Y. : 1989)》2000,44(5):299-302
PROBLEM: The preconceptional natural-killer cell (NK) activity predicts subsequent miscarriage among women with unexplained recurrent spontaneous abortion (RSA). Psycho-neuro-immuno-endocrine network has recently been proposed as a mechanism for abortions. We therefore examined which psychosocial factors influenced the NK activity among women with RSA. METHOD OF STUDY: We measured the preconceptional NK activity of 61 women with a history two consecutive unexplained first-trimester miscarriages and no live births. We also administered semi-structured interviews and a battery of self-report questionnaires to assess their social support, personality, self-esteem and psychiatric symptoms. RESULTS: The preconceptional NK activity was negatively correlated with the women's neuroticism personality trait (r= -0.32, P = 0.01) and current depressive symptoms (r = -0.26, P= 0.05), and positively correlated with their self-esteem (r = 0.34, P = 0.01). CONCLUSIONS: In addition to several substances such as transforming-growth-factor beta and granulocyte-macrophase colony-stimulating factor, we found that low neuroticism, low depression scale score and high self-esteem contributed to high NK activity among women with RSA. 相似文献
100.
《American journal of reproductive immunology (New York, N.Y. : 1989)》1998,39(5):310-315
PROBLEM: β2 glycoprotein I (β2GPI) physiologically binds to negatively charged phospholipids (PLs) and is a natural regulator of the coagulation cascade. Thrombotic clinical complications and recurrent fetal loss associated with autoimmune antiphospholipid (aPL) antibodies are thought to be related to their binding to β2GPI-PL complex and interference with the physiological function of β2GPI. METHOD OF STUDY: To investigate the effect of aPL on β2GPI-PL interaction, we studied the binding of biotinylated β2GPI to cardiolipin (CL) by enzyme-linked immunosorbent assay (ELISA) in the presence and absence of purified aPL immunoglobulin G (IgG) antibodies. RESULTS: Adding five different aPL IgG antibodies with different levels of aPL activity isolated from the sera of five patients with aPL-associated recurrent fetal death greatly increased the binding of biotinylated β2GPI to CL-coated plates. The optical densities (ODs) were 0.635, 0.890, and 1.265 in the presence of three aPL IgG antibodies, compared to 0.425 in the absence of aPL IgG. In contrast, normal human IgG had no enhancing effect. The OD was 0.480 and 0.425, respectively. The enhancement of β2GPI binding to CL by aPL IgG correlated with the titers of aPL antibodies. The use of phosphate-buffered saline with increasing salt concentrations as a washing buffer for the ELISA resulted in more stable binding of β2GPI to PL in the presence of aPL IgG. CONCLUSIONS: These findings suggest that the binding of autoimmune aPL antibodies to β2GPI-PL complex results in abnormally tighter interaction between β2GPI and PLs, which may lead to physiological dysfunction of β2GPI as a regulator of coagulation. 相似文献