Hemocyanin is an important high-molecular-weight allergen of Eriocheir sinensis / 中国免疫学杂志

Objective:To prove hemocyanin is an important high-molecular-weight allergen from Eriocheir sinensis.Methods:The proteins were extracted from Eriocheir sinensis tissue with lysate extraction method .The protein components were analyzed by SDS-PAGE,two-dimensional electrophoresis and Western blot.Serum IgE from allergic donors identified a candidate protein,which was char-acterized by MALDI-TOF/TOF.The immune property of the candidate protein was tested using Dot-blot.Results: By SDS-PAGE and two-dimensional electrophoresis analysis,we prove that the native protein components were completely.According to the Western blot result,at least 18 components could react with the positive serum.Among them, two 70-80 kD proteins had the same isoelectric point.So,may be they were the same protein.MALDI-TOF/TOF analysis results showed that the suspected proteins were hemocyanin.A sensitization frequency of 61%was observed in Eriocheir sinensis patients by Dot-blot.Conclusion:Hemocyanin was identified as an important high-molecular-weight allergen from Eriocheir sinensis.

Effect of different routes of keyhole limpet hemocyanin immunization on the T cell dependent antibody response in mice / 中国实验动物学报

Objective To study the influence of different routes of keyhole limpet hemocyanin ( KLH) immuniza-tion on the T-cell-dependent antibody response in mice.Methods SPF Kunming mice were divided into four groups: the intravenous injection group, subcutaneous injection group, intraperitoneal injection group and control group.Each mouse was injected 200 μg KLH intravenously, subcutaneously or intraperitoneally daily for consecutive 10 days, respectively. Mice in the control group were given solvent injection only.Serum concentration of IgG stimulated by KLH antigen was measured 7 days after the last dosing.Spleen was isolated to calculate the organ coefficient and examined by pathology u-sing hematoxylin and eosin staining.Results Intravenously, subcutaneously and intraperitoneally administered KLH stimu-lated the generation of secondary lymphoid follicles and germinal center to varying degrees, B cell apoptosis, increased a-mount of cells in the marginal zone and other pathological changes were observed in the spleen.Intravenous and intraperito-neal administration of KLH led to more pronounced pathological changes compared with that in the subcutaneous injection group.All of the three administration routes of KLH induced generation of IgG antibody, significantly higher than that in the control group (P<0.05).Intravenous injection of KLH generated the highest concentration of IgG and organ coefficient among the three administration routes ( P<0.05) .Conclusions Different immunization routes do affect the production of IgG antibody, organ coefficient and pathological changes in the spleen, and these differences should be taken into consider-ation when analyzing the T cell dependent antibody response in mice.

Hemocianinas, una herramienta inmunológica de la biomedicina actual / Hemocyanins as immunostimulants

Hemocyanins, the giant oxygen transporter glycoproteins of diverse mollusks, are xenogenic to the mammalian immune system and they display a remarkable immuno-genicity. Therefore they are ideal non-specific immunostimulants to treat some types of cancer. They are used as an alternative therapy for superficial urinary bladder cancer (SBC), that has been traditionally treated with Bacillus Calmette-Guerin (BCG). In contrast to BCG, hemocyanins do not cause side-effects, making them ideal for long-term repetitive treatments. Hemocyanins have also been exploited as carriers to develop antibodies against hapten molecules and peptides, as carrier-adjuvants for cutting-edge vaccines against cancer, drug addiction, and infectious diseases and in the diagnosis of parasitic diseases, such as Schistosomiasis. The hemocyanin from Megathura crenulata, also known as keyhole limpet hemocyanin (KLH), has been used for over thirty years for the purposes described above. More recently, hemoc yanin from the Chilean mollusk Concholepas concholepas (CCH) has proved to be a reliable alternative to KLH, either as carrier protein, and as a likely alternative for the immunotherapy of SBC. Despite KLH and CCH differ significantly in their origin and structure, we have demonstrated that both hemocyanins stimulate the immune system of mammals in a similar way by inducing a potent Thl-polarized cellular and humoral response.

Model of immune-mediated inner ear disease and efficacy of dexamethasone by systemic application or round window administration / 上海第二医科大学学报

Objective To establish a model of immune-mediated inner ear disease(IMIED) and to compare the efficacy of dexamethasone between systemic application and round window adminstration. Methods The guinea pigs were injected with keyhole limpet hemocyanin(KLH) mixed with complete Freund's adjuvant subcutaneouly in the back,and were boosted with KLH mixed with incomplete Freund's adjuvant two weeks later.Meanwhile,a small piece of gelatin sponge full of KLH was delivered on the round window through operation.Thus established a model of IMIED.Micro-osmotic pumps were used to deliver dexamethasone through the round window(PBS control group also set),and systemic application was implemented by intraperitoneal injection.The efficacy was compared by recording auditory brain-stem response(ABR) before and after the treatment. Results None of the 6 guinea pigs in the control group experienced significant hearing loss,while 22 of the 39 guinea pigs in the experiment group suffered from hearing loss beyond 10 dB.Seventeen guinea pigs with hearing loss no less than 15 dB were divided into three groups randomly and were given different treatment.Those(n=6) treated by local application through the round window enjoyed the efficacy and the mean ABR threshold decreased 13.3 dB.Four out of the 6 treated by systemic application enjoyed the efficacy and the mean ABR threshold decreased 13.7 dB.No efficacy was observed in the PBS group. Conclusion The model of IMIED can be successfully induce by KLH through the round window,and dexamethasone administered through the round window is as efficacious as systemic application.

Safety and Efficacy of Intravesical Keyhole-Limpet Hemocyanin Therapy for Superficial Transitional Cell Carcinoma: A Prospective, Multicenter Study / 대한비뇨기과학회지

PURPOSE: We wanted to determine the safety and efficacy 1 year after intravesical keyhole-limpet hemocyanin (KLH) therapy for the patients suffering with superficial transitional cell carcinoma. MATERIALS AND METHODS: A total of 60 patients with bladder cancer who had undergone transurethral resection were treated with a one-year protocol (once a week for 6 weeks and 12 further treatments every 4 weeks) of KLH 20mg. The local and systemic safety and efficacy were evaluated. RESULTS: In terms of the safety of KLH, 4 patients (6.7%) reported dizziness and slight fever, and 2 (3.3%) experienced urgency. The remaining 54 patients (90.0%) did not report any local side effects during the treatment period. In terms of efficacy, recurrence developed in 32 patients (53.3%) at a mean of 4.5 months after transurethral resection; 28 patients (46.7%) were disease free after 14 months. The univariate analysis showed a greater level of efficacy in the patients suffering with bacille Calmette-Gu rin (BCG)-refractory disease (p=0.0150). No significant differences were found between the low, the intermediate-risk and the high-risk patients in terms of efficacy. CONCLUSIONS: The results of this study have confirmed the good tolerability of intravesical KLH and better efficacy in the BCG-refractory patients. However, in terms of overall efficacy, intravesical KLH therapy had no superiority over that of other intravesical treatments in terms of the overall efficacy.

Effect of Inhibitor of TNF-alpha and Oxatomide on Experimental Immune Mediated Otitis Media with Effusion / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Local immune response, one of the pathogenesis of middle ear effusion (MEE) is associated with the development and persistence of effusion in the middle ear cavity and inflammatory mediators play a major role in the production of MEE. The purpose of this study was to determine the effects of TNF-alpha antagonist and oxatomide on the outcome of immune mediated otitis media with effusion (OME) in rats. MATERIALS AND METHOD: Otitis media was induced by injecting KLH (Keyhole lympet hemocyanin) transtympanically 7 days after systemic immunization. Phosphate-buffered saline solution was used as control. Other groups were pretreated with TNF-alpha antagonist (soluble TNF receptor type I, sTNF RI) or oxatomide respectively before transtympanic injection of KLH. Seventy-two hours after the transtympanic injection, temporal bones in each group were examined histopathologically and vascular permeability of the middle ear mucosa was measured by the Evans blue vital dye technique. RESULTS: In the KLH, sTNF RI and oxatomide groups, MEE was developed in 83%, 0%, 66% of the ears, respectively. The oxatomide group and sTNF RI group showed significant decrease in inflammation, mucosal thickening and vascular permeability as compared with KLH group and those parameters of sTNF RI group showed lower values than those of oxatomide group. CONCLUSION: Transtympanic administration of sTNF RI and oxatomide appears to suppress the development of immune mediated OME. In terms of inhibiting MEE, sTNF RI was more effective than oxatomide. This study suggests that TNF-alpha antagonist and oxatomide may have a adjunctive role in the treatment and prevention of otitis media with effusion.