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11.
Studies have demonstrated that lipid rafts ultimately regulate the endocytosis of anthrax toxin via clathrin dependent pathway. Interestingly, GPI-anchored protein rich rafts have also been shown to be transported down to the endocytic pathway to reducing late endosomes. Taking advantage of this parallelism, we tried translating the anthrax toxin natural intoxication mechanism by administering a DNA chimera that encoded protective antigen attached to a mammalian GPI-anchor sequence at its C-terminus (pGPI-PA63). We also designed a chimera that had an additional N-terminal TPA leader sequence (pTPA.GPI-PA63) with an aim to target GPI-PA63 to ER where new CD1 molecules are synthesized. Analysis of antibody titers demonstrated successful priming and potential IgG titers after the first boost. In vitro cell proliferation studies in the presence of GPI-attached PA63 peptides revealed that there was a clonal expansion of CD4+ NK1.1+ helper T cell population which rapidly produced IL-4 in response to T cell receptor ligation. These cells provided direct B cell help that aided IgG formation. Effector responses generated by NKT cells were found to be MHC II-independent and CD1d-restricted. In addition, the group pTPA.GPI-PA63 also displayed low magnitude MHC-II restricted (CD1d-independent) NKT cell and CD4+ T cell helper responses in response to non-GPI attached PA63 peptides which overall resulted in the heightened responses seen for this group. Importantly, DNA vaccination mediated the generation of high avidity neutralizing antibodies that mediated protection against lethal toxin challenge. 相似文献
12.
CD14+单核细胞人类白细胞抗原DR表达率与脓毒症关系的研究 总被引:15,自引:0,他引:15
目的了解烧伤延迟复苏时CDl4^+单核细胞人类白细胞抗原DR(HLA—DR)表达率的变化,分析其与脓毒症的关系。方法选择烧伤面积大于30%TBSA的25例烧伤延迟复苏患者,于伤后1、3、7、14、28d取外周血,其中7例患者住院期间并发脓毒症,于脓毒症发生后连续2d亦取其外周血。另取20例健康体检者外周血作为对照。流式细胞仪检测CD14^+单核细胞HLA.DR表达率,酶联免疫吸附测定法检测血浆中肿瘤坏死因子α(TNF—α)、白细胞介素10(IL-10)的浓度。结果非脓毒症患者伤后1、3、7、14、28dCD14^+单核细胞HLA—DR表达率分别为(15±6)%、(74±5)%、(264±17)%、(284-16)%、(474-16)%,明显低于健康体检者[(924±10)%,P〈0.01];脓毒症患者发生脓毒症后1、2d,该指标亦明显低于健康体检者及非脓毒症患者伤后1、7、14、28d(P〈0.01)。脓毒症患者TNF—d检出率及TNF—α、IL-10浓度,均高于非脓毒症患者和健康体检者(P〈0.05或P〈0.01)。伤后1、7、28d,外周血CD14^+单核细胞HLA—DR表达率与IL—10浓度呈显著负相关(r分别为-0.9963、-0.7459、-0.8474,P〈0.01)。结论烧伤延迟复苏患者免疫功能低下,促炎性介质释放量增加,并发脓毒症时则更为严重。外周血CD14^+单核细胞HLA—DR表达率可作为动态检测患者免疫功能状态的有效指标。 相似文献
13.
Permanent prostate brachytherapy for Japanese men: Results from initial 100 patients with prostate cancer 总被引:1,自引:1,他引:0
Toshikazu Okaneya Shuji Nishizawa Tsuyoshi Nakayama Takayuki Kamigaito Iwao Hashida Noriko Hosaka 《International journal of urology》2007,14(7):602-606
OBJECTIVE: To evaluate the initial results of brachytherapy for prostate cancer with permanent iodine-125 implant in Japan. METHODS: The results obtained with brachytherapy in the initial 100 Japanese patients treated at Nagano Municipal Hospital were reviewed. Patients with a prostate-specific antigen (PSA) level of less than 10 ng/mL and a Gleason's scores of 5, 6, 3 + 4 were classified as having a low risk of recurrence. Patients with a PSA level of 10-20 ng/mL and/or a Gleason's score of 4 + 3 were classified as having an intermediate risk for recurrence. Seventy-eight of the low-risk patients and 19 of the intermediate-risk patients were treated by seed implants alone, or seed implants combined with preceding external radiation, respectively. A total of 53 patients received neoadjuvant hormone therapy. The efficacy and morbidity of brachytherapy were investigated using the serum PSA, International Prostate Symptom Score, quality of life score and uroflowmetry data. RESULTS: The average V100 and D90 obtained by post-implant dosimetry was 94.3 and 113.7%, respectively. Serum PSA decreased gradually after treatment, although it had still not reached a nadir after 1 year. There was little difference of the PSA level between the patients with and without neoadjuvant hormone therapy even at 1 year after seed implantation. There were no PSA biochemical failure or clinical recurrence during the follow-up period. Voiding symptoms worsened until 3 months after treatment, and then gradually improved. Acute urinary retention occurred transiently in one patient (1%). Rectal bleeding and severe diarrhea did not occur. CONCLUSION: Brachytherapy is a feasible and effective option for the treatment of prostate cancer in Japanese men. Brachytherapy may have a different effect in Japanese patients with respect to voiding symptoms. Urinary retention was rare, but voiding symptoms were persistent in Japanese patients. Neoadjuvant hormone therapy deserves investigation to determine whether it can achieve better results, especially in patients with an intermediate risk. 相似文献
14.
H. Iwata T. Otoshi N. Takada T. Murai S. Tamano T. Watanabe Y. Katsura S. Fukushima 《Urological research》1995,23(1):27-32
A series of 8 rat and 16 mouse invasive bladder carcinomas were investigated for the presence of silverstained nucleolar organizer regions (AgNORs) to clarify whether this parameter is applicable to the estimation of their invasive character. With regard to number of AgNORs per cell, neither rat nor mouse carcinomas showed any difference between invasive and noninvasive sites within the same tumor. However, the frequency of cancer cells bearing bizarre dots, irregular in size and shape, was significantly higher at sites of actual invasion. Quantitative data generated using an image analyzer revealed significantly lower values for NOR roundness and significantly larger NOR size in invasive sites than in noninvasive sites in all groups. Double staining for the proliferation marker proliferating cell nuclear antigen (PCNA) and AgNORs was performed on eight rat carcinomas and a close correlation between the two was confirmed. Thus the number of AgNORs in PCNA-positive cells was significantly greater than in PCNA-negative cells. Furthermore, a particularly strong correlation was observed for incidences of PCNA-positive cells and bizarre dots (P<0.01). The quantitative data also demonstrated significant differences in size and shape of dots. It is concluded that AgNORs have diagnostic value for the invasive character of bladder carcinomas. 相似文献
15.
用快速双抗体夹心ELISA检测囊虫病人血清中的循环抗原,其阳性率为67.5%;正常人血有的假阳性率为5.33%;包虫、肝吸虫和弓形虫病人血清的交叉反应率分别为6.9%、7.5%和12.5%。结果表明.该法与常规双抗体夹心ELISA相比具有较为敏感、快速、稳定和节省血清、试剂用量的优点,但其特异性有待进一步提高。用快速双抗体夹心ELISA和间接型ELISA检测第1至10疗程囊虫病人血清中的循环抗原和抗体,表明检测循环抗原考核囊虫病人的治疗效果优于检测抗体 相似文献
16.
Purification of antigenically intact Ro ribonucleoproteins; biochemical and immunological evidence that the 52-kD protein is not a Ro protein. 总被引:3,自引:0,他引:3 下载免费PDF全文
G Boire M Gendron N Monast B Bastin H A Mnard 《Clinical and experimental immunology》1995,100(3):489-498
Anti-Ro sera immunoprecipitate Ro ribonucleoproteins (RNPs) from human cell extracts. Ro RNPs are biochemically heterogeneous particles whose functions are unknown and whose exact composition remains controversial. In addition to 60-kD Ro and to La proteins, a 52-kD polypeptide (p52) has been proposed to be a stable component of the Ro RNPs. To confirm the immunological studies supporting this hypothesis, we have biochemically purified Ro RNPs from HeLa cells using non-denaturing conditions. Ro RNPs segregated into three distinct populations, one of which only contained hY5 RNA (RohY5 RNPs). No p52 co-purified with Ro RNPs. Despite the absence of p52, purified Ro RNPs had biochemical and immunological properties identical to those of unfractionated Ro RNPs. Many anti-Ro sera only recognize p52 in immunoblots, and are said to be monospecific anti-p52. Preincubation with purified RohY5 RNPs (free of p52) of all human anti-Ro (including so-called monospecific anti-p52) sera abolished their capacity to immunoprecipitate Ro RNPs from unfractionated HeLa cell extracts. Conversely, preincubation of anti-Ro sera with purified p52 protein specifically inhibited recognition of p52 in immunoblots, but did not interfere with immunoprecipitation of Ro RNPs. Our data demonstrate that anti-p52 antibodies do not target intact Ro RNPs, nor do they target the native 60-kD Ro protein. Contrary to previous reports, p52 protein is not a stable component of antigenically intact Ro RNPs. 相似文献
17.
Macrophages, dendritic cells or B lymphocytes have been shownto play a major role in the presentation of soluble antigensto CD4+ T cells. In contrast, the capacity of these cells topresent particulate antigens such as bacterial or parasiticantigens to T cells remains controversial. To investigate thisquestion, well defined particulate antigens were prepared bycovalent linkage of proteins or peptides to 1 µm in diametersynthetic microspheres. The T cell immunogenicity of such particulateantigens was analyzed in vitro and in vivo. In vitro, a solubleprotein such as hen egg lysozyme (HEL) coupled to beads stimulateda strong proliferative T cell response of lymph node cells fromHEL-primed mice or of specific T cell hybridomas. HEL coupledto beads was presented to the specific T cell hybridomas bysplenocytes or by peritoneal macrophages, but not by lymphomaB cells. Immunization of mice with several different proteinantigens or with a synthetic peptide covalently linked to beadsinduced strong CD4+ T cell responses in the absence of adjuvant.The strong in vivo immunogenicity of proteins coupled to beadsdid not result from a non-specific adjuvant effect of beadssince covalent linkage of the antigen to beads was strictlyrequired to induce T cell responses in the absence of adjuvant.In vivo treatment by carrageenan showed that macrophages arerequired for the in vivo stimulation of T cell responses bythese particulate antigens. Thus, these results demonstratedthe role of phagocytic cells, especially macrophages, for invivo presentation of particulate antigens. These particulateantigens represent an interesting approach for the developmentof new vaccines, and for the in vivo analysis of the role ofvarious antigen presenting cells in T cell activation and differentiation. 相似文献
18.
PROBLEM: There is substantial data that support the efficacy of paternal leukocyte immunization (PLI) for the treatment of alloimmune mediated miscarriage; however, there is confusion regarding the laboratory test that should be performed to determine levels of maternal anti-paternal leukocyte antibodies (MAPLA). METHOD: Popular methodologies employed include: 1) microcytotoxicity (MCX), 2) mixed lymphocyte culture (MLC), and 3) cell flow cytometry crossmatch (FCXM). Cell flow cytometry crossmatch correlates well with the more difficult MLC assay although the former proves the more sensitive study. This work compares the MCX assays with FCXM. The study group consisted of ten women who had a history of three or more spontaneous abortions (SABs). All ten had very low levels (<10%) of MAPLA as measured by FCXM. Following PLI all subjects demonstrated elevated levels (>50%) of MAPLA by FCXM. At 12 weeks gestation, sera were simultaneously measured for MAPLA by MCX and FCXM. RESULTS: Although all ten patients had very high levels of MAPLA by FCXM during pregnancy, five of ten had antibodies to HLA Class I and two of ten had antibodies to HLA Class II paternal antigens by MCX. Furthermore, all patients who were positive by MCX to paternal Class I antigens were also positive to Class I antigens not seen in either parent. Both patients who were positive by MCX to paternal Class II antigens were also positive to maternal Class II antigens. Notable is that all ten women eventually delivered healthy infants. CONCLUSION: Based on this preliminary study, the MCX assay is neither sensitive or reliable enough to determine the need and/or to monitor the effectiveness of PLI. Flow cytometry should be the modality of choice when determining the need for alloimmunotherapy and to monitor the effectiveness of treatment. 相似文献
19.
含化复方新诺明引起过敏性休克一例患者,女,30岁,因咽峡炎于1992年8月7日晚10时自行含化复方新诺明(广州白云山制药厂)两片、约7分钟后,出现全身搔痒,当即肌注苯海拉明20mg,静推10%葡萄糖酸钙10ml,旋即全身泛起荨麻疹,奇痒难忍,眼睑轻度水肿,嘴唇发麻,咽部“发紧”,发音困难,烦躁,立即给予吸氧,静推地塞米松10mg,皮下注射肾上腺素0.5mg,与此同时,患者脸色苍白、鼻尖、未梢发凉,口唇紫绀,时吸暂停,意识丧失,脉搏140次/分,细数,血压测不到,再次给予皮下注射肾上腺素0.5mg,静点地塞米松15mg,并给予扩容、纠酸,约一分钟后,欲行气管切开时,患者呼吸渐恢复、继而意识清楚,紫绀减轻,血压10/16kPa,遂休克纠正。复方新诺明片致过敏性休克者尚属少见,该患者有青霉素过敏史,局部用药又易致过敏,考虑喉头痉挛与药物含化有关。(长治市人民医院王爱军,崔文华,郭天然)含化复方新诺明引起过敏性休克一例@王爱军,崔文华,郭天然$长治市人民医院 相似文献
20.
Eiji Takeuchi Yuji Nimura Shin-ichi Mizuno Hideaki Suzuki Shinsuke Iyomasa Masaki Terasaki Hiroshi Kuriki Keiko Tamiya-Koizumi Shonen Yoshida 《Journal of Hepato-Biliary-Pancreatic Surgery》1997,4(3):254-262
Hepatocyte regeneration has been widely investigated, with the mitotic index and the incorporation of [3H]thymidine being used as regeneration markers. We focused on the induction of DNA replication enzymes, particularly DNA polymerases
(pol) α, δ, and ε. Using rat models, we have shown that the activity of pol α in crude liver extract well represents the regenerating
capacity of hepatocytes. Using pol α as an indicator, we analyzed liver regeneration in rat models under various conditions:
obstructive jaundice, external or internal biliary drainage, and the obstruction of portal vein branches. It has been revealed
that the ligation of the common bile duct alone induces a certain amount of hepatocyte proliferation. It was striking that
external biliary drainage suppressed regeneration capacity in cholestatic rat liver after partial hepatectomy. The strong
regeneration in nonligated lobes induced by portal branch ligation was similar to the liver regeneration seen after partial
hepatectomy with respect to the induction of DNA polymerases. Taken together, the aspects of DNA replication, particularly
the induction of DNA polymerases, may contribute to shedding new light on the regeneration of human liver.
This work was supported in part by a Grant-in-Aid for General Scientific Research and for Cancer Research from the Ministry
of Education, Science and Culture, Japan, and by grants from the Uehara Memorial Foundation 相似文献