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Objective To explore the anticancer mechanism of esophageal cancer vaccines and clinical effect.Methods Esophagus cancer cells (Eca109) were cultured and the soluble antigen were extracted from the cells.Esophagus cancer vaccine was constructed with the antigen and superantigen SEC.Lymphocytes were isolated from peripheral blood and then stimulated with the vaccine in vitro.Phenotypes of the cells were checked by FCM and killing activity was tested with cytotoxic assays.One hundred and six early esophageal cancer patients were selected after surgery,who were divided into the observation group of 53 cases with esophageal cancer vaccine therapy and 53 cases in the control group with conventional treatment.Then clinical effect was observated and 3 year follow-up survival rate was observed of the of two groups of patients.Results Proliferation of vaccine stimulated lymphocyte group was the strongest,and high peaked at 72 h (A =0.22),and raise CD8+ T cell populations of CTLs.The killing activity of lymphocyte group stimulated by the vaccine against target cells was significantly higher than that of lymphocyte group ((97.36±2.11) ％) vs.(79.27±5.57) ％,F =38.62,P＜0.01).Three years follow up shows that the survival rates of experiment group were 48.27％ (male) and 45.83％ (female) respectively,and control group were 21.43％ (male) and 24.00％ (female),and the difference was significant (x2 =5.06,6.28,P ＜ 0.05).Conclusion The tumor vaccine constructed with esophagus cancer antigen and superantigen SEC can induce PBMC to activate and proliferate into CD8+ CTL with specific cytotoxicity against the cells which the antigen comes from.The vaccine may raise the survival rate of the patients with Esophagus cancer.
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Objective:To analyze the effects of the clinical applications of vaccine for esophageal cancer to provide treatment ba-sis. Methods:Tumor-soluble antigen was extracted from clinical samples of esophageal cancer tissue. Vaccine for esophageal cancer was prepared using antigen and superantigen staphylococcal enterotoxin C (SEC). One group of patients with esophageal cancer was treated with vaccine after surgery (treatment group). Another group of patients with esophageal cancer was treated using routine meth-ods (control group). After five years, the patients were followed up to analyze survival. Results:After five years of follow up, the sur-vival rates of treatment and control groups were 173/328 (49.71%) and 67/326 (20.55%), respectively. Survival rates significantly dif-fered between the two groups (P<0.05). Furthermore, the survival rates of the female patients were higher than those of the male pa-tients (P<0.05). In the treatment group, the survival rates of patients with low-differentiation cancer were higher than those of patients with high-differentiation cancer (P<0.05). In the control group, the survival rates of patients with low-differentiation cancer were lower than those of patients with high-differentiation cancer (P<0.05). Conclusion:Tumor vaccine prepared with esophageal cancer antigen and superantigen SEC could increase the survival rate of patients with esophageal cancer, particularly female patients and those with low-differentiation cancer.
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INTRODUCTION: Different serum levels of the IgG/IgE for Paracoccidioides brasiliensis high mass molecular (hMM) fraction (~366kDa) in the acute and chronic forms of the disease have been reported. Considering the nonexistence of hMM fraction investigation involving clinical isolates of P. brasiliensis, the present study aimed to investigate the presence of the hMM fraction (~366kDa) in cell free antigens (CFA) from P. brasiliensis clinical isolates. METHODS: CFA from 10 clinical isolates and a reference strain (Pb18) were submitted to SDS-polyacrylamide gel electrophoresis (SDS-PAGE) followed by gel image capturing and densitometer analysis. Additionally, CFA from 20 isolates and Pb18 were analyzed by capture ELISA (cELISA) using polyclonal (polAb) or monoclonal (mAb) antibodies to the hMM fraction. RESULTS: The presence of the hMM component was observed in CFA of all samples analyzed by SDS-PAGE/densitometry and by cELISA. In addition, Pearson's correlation test demonstrated stronger coefficients between hMM fraction levels using pAb and mAb (R = 0.853) in cELISA. CONCLUSIONS: The soluble hMM fraction was present in all the P. brasiliensis clinical isolates analyzed and the reference strain Pb18, which could be used as a source of this antigen. The work also introduces for first time, the cELISA method for P. brasiliensis hMM fraction detection. Analysis also suggests that detection is viable using polAb or mAb and this methodology may be useful for future investigation of the soluble hMM fraction (~366kDa) in sera from PCM patients.
INTRODUÇÃO: Diferentes níveis sorológicos de IgG/IgE contra a fração de alta massa molecular (hMM) (~366kDa) de Paracoccidioides brasiliensis têm sido encontrados na PCM aguda e crônica. Considerando a inexistência de investigação sobre esta fração em isolados clínicos de P. brasiliensis, o objetivo deste estudo foi investigar a presença da fração hMM (~366kDa) no preparado livre de células (CFA) de P. brasiliensis obtidos de isolados clínicos. MÉTODOS: CFA de 10 isolados e de cepa de referência (Pb18) foram submetidas à eletroforese em gel de SDS-poliacrilamida (SDS-PAGE) seguida de captura de imagem e análise por densitometria. Adicionalmente, CFA de 20 isolados e de Pb18 foram analisados por ELISA captura (cELISA) utilizando anticorpos policlonal (polAb) ou monoclonal (mAb) para fração hMM. RESULTADOS: A presença do componente de hMM foi observada em todas as amostras analisadas por SDS-PAGE/densitometria e por cELISA. Adicionalmente, o teste de correlação de Pearson demonstrou forte relação entre os níveis de fração hMM usando pAb e mAb (R = 0.853) no cELISA. CONCLUSÕES: Conclui-se que a fração hMM está presente em todos os isolados clínicos de P. brasiliensis analisados e no isolado referencial, sugerindo a possibilidade dos mesmos serem utilizados como fonte desta fração antigênica. Este trabalho também introduz pela primeira vez o método de cELISA para detecção da fração hMM de P. brasiliensis, sugerindo que detecção utilizando anticorpos polAb ou mAb é viável e essa metodologia poderá ser útil para investigação futura desta fração solúvel (~366kDa) em soros de pacientes com PCM.
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Humans , Antibodies, Fungal/immunology , Antibodies, Monoclonal/immunology , Antigens, Fungal/immunology , Immunoglobulin G/immunology , Paracoccidioides/immunology , Paracoccidioidomycosis/parasitology , Chronic Disease , Electrophoresis, Polyacrylamide Gel , Enzyme-Linked Immunosorbent Assay , Molecular Weight , Paracoccidioides/isolation & purificationABSTRACT
Objective:To research the autoimmune mechanism of prolactin(PRL) in uveitis by observing prolaictin′s effects on peripheral blood mononuclear cells(PBMC) proliferation and activation at different concentration.Methods:1.The serum prolactin levels of uveitis and control were checked by chemical spectrometer. 2.To research the effects of different concentration PRL on uveitis PBMC′s proliferation and activation under the soluble antigen(S-Ag) stimulation. PBMC separated from uveitis and control,were cultured with S-Ag and different concentration PRL,and then were assayed the percentage of CD3~+CD25~+ positive cell through double-stain immunofluorescence flow cytometry after 108 h. After cultured 96 h, ~3H-TdR was mixed and counts per minute were assayed after 12 h.Results:1.There was no significance between the uveitis and control refer to the serum PRL level(t=1.936,P=0.051). 2.When uveitis′ PBMC was positive to S-Ag, there had promote proliferation and activation of PBMC with PRL concentration at 12.5-1 000 ?g/L,with the media concentration at 200 ?g/L,while it became weakened at 1 000 ?g/L(activation t=2.736,P=0.012;proliferation t=2.547,P=0.036).When uveitis′ PBMC was negative to S-Ag,there had no action as before.Conclusion:1.There is no significance between the uveitis and control refer to the serum PRL level. 2.Media concentration PRL(200 ?g/L) and S-Ag have costimulation to promote PBMC proliferation and activation in uveitis,though high concentration PRL loses the function.
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In the Philippines, passive haemagglutination and soluble antigen fluorescent antibody tests using Dirofilaria immitis antigens were carried out on the sera of 33 patients infected with Wuchereria bancrofti and on the sera of 24 uninfected persons. False positives were as high as 70.8% indicating that in areas where the incidence of D. immitis in dogs is high, W. bancrofti antigens must be used. D.A.Cz.
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Objective To induce islet grafting tolerance by intravenous injection of anti-CD4,anti-CD8 immunotoxins and donor soluble antigen.Methods 14 days or 7 days prior to transplantation,the immunotoxon 200 μg respectively,and donor soluble antigen 500 μg were injected intravenously into the recipients, then 500 donor islets were translanted under the left renal subcapsular space of diabetes reciPients (SD rats).Results The islet grafting survival time that pretreated with immunotoxon and dono soluble antigen was over 60 days(P<0.01).The immunotoxins or donor soluble antigen treatment alone only slightly prolonged the graft survival.Conclusion The anti-CD4,anti-CD8 immunotoxins combined with donor soluble antigen can induce donor specific immune tolerance.
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CTL responses by autologous colorectal cancer cells were observed in five cases. The specific CTL was induced from two cases, in which tumor associated antigen, CA-Hb_3, was strongly positive. The CTL could not induced from other three cases in which CA-Hb_3 antigen was weakly positive. Weakly positive tumor cells in the three cases were loaded CA-Hb_3 antigen into cytoplasmas by the cationic liposomes and tested by immunohistochemical assay. The results showed that CA-Hb_3 antigen of tumor cells in two cases changed to strongly positive, and could induce CTL responses successfully. With blocking agents BFA and anti-MHC-I monoclonal antibody, CTL responses could be blocked. Our results suggested that soluble CA-Hb_3 antigen could enhance the immunogenity of colorectal cancer cells and induce effective anti-tumor cellular immune responses.
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Objective To study the mucosal and systemic immune response after intranasal immunization with mucosal complex vaccine for Toxoplasma gondii,and to observe the protective effect on mice. Methods The mucosal complex vaccine was made of soluble tachyzoite antigen (STAg) and cholera toxin (CT),which were mixed and dissolved in PBS (1 ml PBS containing 1 mg STAg and 50 ?g CT). Fifty-two BALB/c mice were randomly divided into two groups: immunized group and control. Mice were intranasally immunized with 20 ?l mucosal complex vaccine (20 ?g STAg and 1?g CT) per mouse twice at an interval of two weeks,while the control mice were given PBS solution instead. Six mice of each group were killed by dislocation of cervical vertebra on day 14 after the last immunization. The specific IgG antibodies in serum and IgA in feces were detected by ELISA. Lymphocytes in spleen,Peyer's patches (PP) and intestinal intraepithelial lymphocyte(IEL) were isolated and counted. Percentage of CD4+ and CD8+ T cells was determined by immunocytochemistry. Other mice were challenged intragastrically each with 4?104 tachyzoites of RH strain Toxoplasma gondii on day 14 after the last immunization. Their health condition was observed and the number of tachyzoites in liver and brain was determined microscopically on the 30 th day after challenge. Results IgG antibodies in serum and IgA antibodies in feces of immunized mice were higher than the control (P
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Objective To develop and identify monoclonal antibodies (McAbs) against adult worm of Angiostrongylus cantonensis and observe its applicability. Methods BALB/c mice were immunized with soluble antigen of adult worms of A.cantonensis. The spleen cells of immunized mice were fused with myeloma cell, and the hybridoma secreting high titer of McAbs with high specificity was screened. By using the McAbs, serum of angiostrongyliasis patient and sera of the rats infected with A.cantonensis were detected by Western blotting and double antibody sandwich ELISA respectively. Results Three McAbs were established (2A2,3F1,4H2), which all showed no cross reaction with antigens of Schistosoma japonicum, Paragonimus westermani, Cysticercus cellulosae and Trichinella spiralis. Western blotting analysis demonstrated that the three McAbs recognized a Mr 15 000 soluble antigen of adult worm of A.cantonensis and recognized the Mr 24 000 and Mr 15 000 circulating antigens from the serum of angiostrongyliasis patient. The double antibody sandwich ELISA detection showed a positive rate of 76