Prevalencia de anticuerpos post-vacunales anti-antígeno de superficie del Virus de Hepatitis B / Prevalence of post-vaccine antibodies against surface antigen of Hepatitis B Virus

INTRODUCCIÓN. Los riesgos biológicos deben ser tratados de una manera especial; como tomar en cuenta la cantidad de Anticuerpos Anti Hbs de Hepatitis B de acuerdo al número de vacunas administradas y al puesto de trabajo. OBJETIVO. Determinar la prevalencia de anticuerpos post-vacunales anti-antígeno de superficie del virus de la Hepatitis B, en los trabajadores de salud, de acuerdo al número de dosis recibidas MATERIALES Y MÉTODOS. Estudio observacional retrospectivo de la prevalencia de anticuerpos post-vacunales anti-antígeno de superficie del virus de la Hepatitis B, en el personal de salud de acuerdo al número de vacunas, en el Hospital de Especialidades Carlos Andrade Marín, en el período comprendido entre marzo y octubre de 2017, analizado en el Sistema Informático AS 400. RESULTADOS. Predominó el sexo femenino. La edad fue 31,88% , (90:414) (30 y 40 años de edad), como área mayoritaria fue el Servicio de Cirugía con el 42,51% del total que generaron más anticuerpos a 100UI/ml y se consideraron seroprotegidos. En los trabajadores a los que no se les suministro la dosificación estándar se generaron niveles menores a los que si culminaron esta dosificación, CONCLUSION. Se determinó la prevalencia de los niveles séricos de anticuerpos anti antígeno de superficie, se estableció de acuerdo al número de dosis, la vacunación fue la principal fuente de generación de anticuerpos anti antígeno de superficie o anti ­ Hbs.

INTRODUCTION. Biological risks must be treated in a special way; how to take into account the amount of Antibodies Anti Hbs Hepatitis B according to the number of vaccines administered and the workplace. OBJECTIVE. To determine the prevalence of postvaccine anti-antigen antibodies to Hepatitis B virus surface, in health workers, according to the number of doses received. MATERIALS AND METHODS. Retrospective study of the prevalence of post-vacunal antibodies against antigen. of surface of the Hepatitis B virus, in the health personnel according to the number of vaccines, in the Hospital of Specialties Carlos Andrade Marín, in the period between March and October of 2017, analyzed in the Computer System AS 400. RESULTS. Female sex predominated. The age was 31,88% (32; 414), (30 and 40 years of age), as the majority area was the Surgery Service with 42,51% (43; 414) of the total that generated more antibodies at 100UI / ml and were considered seroprotected. In workers who were not given the standard dosage levels were generated lower than those that culminated this dosage, CONCLUSION. The prevalence of serum levels of antibodies against surface antigen was determined, was established according to the number of doses, vaccination was the main source of generation of antibodies against surface antigen or anti - Hbs

Purification and characterization of monoclonal IgG antibodies recognizing Ebola virus glycoprotein

PURPOSE: The goal of this study was to purify and characterize Ebola virus glycoprotein (GP)-specific IgG antibodies from hybridoma clones. MATERIALS AND METHODS: For hybridoma production, mice were injected by intramuscular-electroporation with GP DNA vaccines, and boosted with GP vaccines. The spleen cells were used for producing GP-specific hybridoma. Enzyme-linked immunosorbent assay, Western blot assay, flow cytometry, and virus-neutralizing assay were used to test the ability of monoclonal IgG antibodies to recognize GP and neutralize Ebola virus. RESULTS: Twelve hybridomas, the cell supernatants of which displayed GP-binding activity by enzyme-linked immunosorbent assay and the presence of both IgG heavy and light chains by Western blot assay, were chosen as a possible IgG producer. Among these, five clones (C36-1, D11-3, D12-1, D34-2, and E140-2) were identified to secrete monoclonal IgG antibodies. When the monoclonal IgG antibodies from the 5 clones were tested for their antigen specificity, they recognized GP in an antigen-specific and IgG dose-dependent manner. They remained reactive to GP at the lowest tested concentrations (1.953–7.8 ng/mL). In particular, IgG antibodies from clones D11-3, D12-1, and E140-2 recognized the native forms of GP expressed on the cell surface. These antibodies were identified as IgG1, IgG2a, or IgG2b kappa types and appeared to recognize the native forms of GP, but not the denatured forms of GP, as determined by Western blot assay. Despite their GP-binding activity, none of the IgG antibodies neutralized Ebola virus infection in vitro, suggesting that these antibodies are unable to neutralize Ebola virus infection. CONCLUSION: This study shows that the purified IgG antibodies from 5 clones (C36-1, D11-3, D12-1, D34-2, and E140-2) possess GP-binding activity but not Ebola virus-neutralizing activity.

Genetic identification and serological evaluation of commercial inactivated foot-and-mouth disease virus vaccine in pigs

Vaccination is considered a frequently used tool to prevent and control foot-and-mouth disease (FMD). However, the effectiveness of conventional FMD virus (FMDV) vaccines in pigs has been controversial because the massive prophylactic vaccination could not elicit proper immune response nor prevent the broad spread of FMD outbreak, mainly in pig farms, in South Korea during outbreaks of 2014. In addition, there has been little information on the efficacy of inactivated, high potency, multivalent, oil-based FMDV vaccine in pigs, because an evaluation of FMDV vaccines had been mainly carried out using cattle. In this study, we evaluated the genetic identification of commercial inactivated FMDV vaccine and monitored the immune responses in pigs under the field condition. Results implied that it contained three different serotypes with a high level of antigen payload. However, serological results showed low mean percentage of inhibition, and positive rate reached its peak at 6-week post-vaccination, indicating current FMDV vaccine need to improve for a prophylactic vaccination policy in pigs. Therefore, there is an imperative need to develop FMDV vaccine that can provide rapid and long-lasting protective immunity in pigs.

Prevalência de aloimunização eritrocitária em pacientes portadores de anemia falciforme / Prevalence of red blood cell alloimmunization in patients with sickle cell anemia

Embora as transfusões de concentrado de hemácias sejam importantes para o tratamento de pacientes com anemia falciforme, elas acarretam riscos imunológicos tais como a aloimunização a antígenos eritrocitários. Aproximadamente 50% dos pacientes de anemia falciforme recebem transfusões no decorrer da vida, e entre 5% a 10% destes pacientes são submetidos a um programa de transfusão crônica. A aloimunização eritrocitária é uma complicação séria da transfusão, mas relativamente comum. Esta condição pode inclusive levar a reações transfusionais hemolíticas tardias e contribuir para aumentar as comorbidades da doença. Importantes medidas para prevenção destas complicações nestes pacientes são o uso de hemácias previamente fenotipadas, além da fenotipagem do próprio receptor de concentrado de hemácias, determinando seu correto perfil fenotípico e possibilitando a escolha de concentrado de hemácias com antígenos correspondentes ao do paciente a ser transfundido. Extensa genotipagem eritrocitária profilática para selecionar doadores para pacientes que receberão repetidas transfusões durante um longo período é uma aplicação atraente de tipagem de sangue baseados em DNA. Isto é, particularmente relevante para pacientes com doença falciforme onde a taxa de aloimunização é elevada.

Although packed red blood cells transfusions are important for treating patients with sickle cell anemia, this intervention may lead to immunological disturbs, such as alloimmunization by erythrocyte antigens. Approximately 50% of patients with sickle cell anemia receive blood transfusions during their life span, and about 5 to 10% of them require a chronic transfusion scheme. The red blood cell alloimmunization is a serious but common transfusion reaction. This condition could lead to delayed hemolytic transfusion reactions, contributing to increase comorbidities of the disease. Important measures to prevent these complications in patients are the use of previously phenotyped red blood cells, in addition to the phenotyping of red blood cells from the acceptor patient, determining the correct phenotypic profile and enabling the choice of red blood cells with corresponding antigens to the patient to be transfused. Extensive prophylactic red blood cell genotyping to select donors for patients receiving repeated transfusions over a long period of time is a compelling application of DNA-based blood typing. This is particularly relevant for patients with sickle cell disease where the rate of alloimmunization is high.

Reappraisal of the Immunogenicity and Safety of Three Hepatitis A Vaccines in Adolescents

Although the overall incidence of hepatitis A in Korea has been decreasing, adolescents remain highly vulnerable to its outbreaks. This study was conducted to compare the immunogenicity and safety of three hepatitis A vaccines in Korean adolescents. Healthy anti-hepatitis A virus seronegative subjects aged 13 to 19 yr were randomized in three equal groups to receive two doses of Avaxim(TM), Epaxal(R), or Havrix(R), 6 to 12 months apart. Seroconversion rates one month after the first dose were 98%, 95%, and 93% for Avaxim(TM), Epaxal(R), and Havrix(R), respectively. Seroconversion rates reached 100% for all vaccine groups one month after the second dose. Anti-HAV geometric mean concentrations (GMCs) were 7,207.7 mIU/mL (95% CI, 6023.1-8684.7), 1,750.5 mIU/mL (95% CI, 1362.9-2248.3), and 1,953.5 mIU/mL (95% CI, 1459.4-2614.7) after two doses of Avaxim(TM), Epaxal(R), and Havrix(R) respectively. Avaxim(TM) was significantly more immunogenic than Epaxal(R) and Havrix(R), whereas there were no significant differences in antibody responses between Epaxal(R) and Havrix(R). Local and systemic solicited adverse events (AEs) were mostly of mild-to-moderate intensity and resolved within 5 days. No serious AEs were reported. In conclusion, all three vaccines are highly immunogenic and well-tolerated in Korean adolescents. (Clinical Trial Registry NCT00483470)

Circulating Anti-Elastin Antibody Levels and Arterial Disease Characteristics: Associations with Arterial Stiffness and Atherosclerosis

PURPOSE: Elastin is a major arterial structural protein, and elastin-derived peptides are related to arterial change. We previously reported on a novel assay developed using aortic elastin peptides; however, its clinical implications remain unclear. In this study, we assessed whether anti-elastin antibody titers reflect the risk of coronary artery disease (CAD) or its characteristics. MATERIALS AND METHODS: We included 174 CAD patients and 171 age- and sex-matched controls. Anti-elastin antibody titers were quantified by enzyme-linked immunosorbent assay. Parameters of arterial stiffness, including the augmentation index (AI) and heart-to-femoral pulse wave velocity (hfPWV), were measured non-invasively. The clinical and angiographic characteristics of CAD patients were also evaluated. Associations between anti-elastin levels and vascular characteristics were examined by linear regression analysis. RESULTS: The median blood level of anti-elastin was significantly lower in the CAD group than in the controls [197 arbitrary unit (a.u.) vs. 63 a.u., p<0.001]. Levels of anti-elastin were significantly lower in men and in subjects with hypertension, diabetes mellitus, hyperlipidemia, or high hfPWV. Nevertheless, anti-elastin levels were not dependent on atherothrombotic events or the angiographic severity of CAD. In a multivariate analysis, male sex (beta=-0.38, p<0.001), diabetes mellitus (beta=-0.62, p<0.001), hyperlipidemia (beta=-0.29, p<0.001), and AI (beta=-0.006, p=0.02) were ultimately identified as determinants of anti-elastin levels. CONCLUSION: Lower levels of anti-elastin are related to CAD. The association between antibody titers and CAD is linked to arterial stiffness rather than the advancement of atherosclerosis.

Effect of α-glycan isolated from Isatis indigotica on humoral and cellular immunities in mice immunized with H1N1 influenza vaccine / 中国药理学与毒理学杂志

OBJECTlVE To investigate the effect of α-glycan isolated from Isatis indigotica on humoral immunity and cellular immunity functions in mice immunized with H1N1 influenza vaccine. METHODS BALB/c mice were immunized intramuscularly once with H1N1 influenza vaccine ( 3 μg) plusα-glycan ( 100μg) each mouse. The serum total antibody titer and its isotype antibody titer of immu-nized mice were analyzed by ELlSA at 5, 8, 10, 12 and 14 d after injection at vaccine. The proliferation activities of spleen T and B lymphocytes were determined with MTT method. The levels of cytokines interferon-γ( lFN-γ) , tumor necrosis factorα( TNF-α) , interleukin-4( lL-4) and lL-12 were measured by ELlSA kits. The populations of CD4+, CD8+, CD3+ and CD19+ lymphocytes were determined by flow cytometry. Furthermore, the proliferation rate of macrophages was studied with MTT method in vitro. RESULTS The α-glycan from I.indigotica could gradually induce high specific-antibody production 5-14 d after immunization with H1N1 influenza antigen plus theα-glycan in mice compared to immunization with antigen alone ( P<0.01) . After injection of antigen withα-glycan for 5 d, the main lgG isotype was lgM, and the titer levels of total lgG, lgG1 , lgG2a and lgG2b were also significantly raised following 5-14 d after immunization. The α-glycan significantly promoted the spleen T and B lymphocytes proliferation ( growth rate 44.2%and 37.8%) , stimulated the secretion of lFN-γand lL-12 of splenocytes ( P<0.01, P<0.05) , and also promoted lL-4 secretion of thymocytes (P<0.01). The polysaccharide significantly raised the percent age of CD3+T cells ( P<0.01) , CD3+/CD19+ T lymphocytes ( P<0.01) , and CD8+ T cells ( P<0.01) but decreased the percentage of CD4+/CD8+ T lymphocytes compared with antigen alone group ( P<0.01) . Furthermore, the α-glycan exhibited significant effects on the proliferation and TNF-α secre-tion of MH-S macrophages. CONCLUSlON Theα-glycan isolated from I.indigotica can improve humoral and cellular immunity response in mice immunized with H1N1 influenza vaccine.

Sensitization of baicalin in guinea pigs and its possible mechanism / 中国药理学与毒理学杂志

OBJECTlVE To elucidate the sensitization mechanism of baicalin in guinea pigs. METHODS Baicalin antigens were prepared by activated ester method. A guinea pig model of allergic shock induced by the coupling was established. Passive cutaneous anaphylaxis ( PCA) was used to ana-lyze the regularity of the specific antibody. The lgG antibodies of baicalin in serum among 19 sensitized guinea pigs were determined by ELlSA. RESULTS UV scanning spectrum couplings occurred between baicalin and protein ( bovine serum albumin and human serum albumin) . The conjugation ratio was 9∶1 and 5∶1, respectively. The guinea pig model of allergic shock was successfully established and suggested that the antigen induced a typical immediate hypersensitivity response. The antibody could be determined on the 19th day and reached the maximum on the 31st day and then degraded gradually. Both specific lgE and lgG existed in the sensitized serum, but there was no correlation between lgG and lgE. The pos-itive rate of specific lgG antibody to baicalin was 84%. The degranulation rate of peritoneal mast cells was (72.6±11.4)%and higher than that incubated with plasma from control (26.8±6.9)%(P<0.05). lsolated ileum from sensitized guinea pigs contracted conspicuously after being challenged by baicalin-human serum albumin. CONCLUSlON Baicalin can induce allergic reaction in guinea pigs and the reaction is correlated with lgE and lgG production.

Influence of immunosuppressants on C4d deposition in renal allografts of rats with chronic allograft nephropathy / 第二军医大学学报

Objective To observe C4d deposition in renal allografts of rats undergoing chronic allograft nephropathy (CAN), and to analyze the effects of immunosuppressants on deposition of C4d in peritubular capillaries. Methods The renal grafts of Fisher 344 rats were ortho topically transplanted into Lewis rats to create CAN models, and all the recipients were given cyclosporine A (CsA) 10 mg/(kg �� d) X10d after operation. The models were then divided into 5 groups (each n=9): Group A was normal saline control group, only receiving vehicle orally; Group B, C, D, and E received CsA 6 mg/(kg �� d), RAPA0. 8 mg/(kg �� d), FK506 0. 15 mg/(kg �� d), and MMF 20 mg/(kg �� d), respectively. The renal allografts were harvested after three rats were sacrificed at the 4th, 8th and 12th weeks post-transplantation. The histological changes were assessed according to Banff 97 standard. The deposition of C4d was detected by immunofluorescence method. Results C4d deposition in peritubular capillary (PTC) was found in all the allografts at the 4th week after transplantation, while there were no obvious clinical pathological changes of CAN in all groups, and the Banff scores were not significantly different among different groups (P > 0. 05). CAN manifestations of different degrees were observed 8 weeks after operation, with increased C4d deposition in the PTC. Severest CAN was observed at the 12th week after operation, accompanied by the most C4d deposition in the PTC. C4d deposition was positively correlated with the severity of CAN (r=0. 894, P = 0. 000). Compared with the control group, CsA and FK506 showed no significant effect on C4d deposition (P>0. 05); however, MMF and RAPA significantly decreasedC4d deposition (P<0. 05). Conclusion Deposition of C4d in PTC may appear in allografts earlier than the pathological changes of CAN, and the deposition is associated with the progression of CAN. MMF and RAPA can inhibit the progression of CAN, while CsA and FK506 can not.

Early results of serum metal ions change and its impact on host immunity after metal-on-metal total hip arthroplasty / 中华骨科杂志

Objective To compare the serum metal ion levels and host immunologic response of patients received non-modular,large-head metal-on-metal total hip arthroplasty (THA,Durom) with those received modular titanium acetabular component (Trilogy) using conventional head sizes and metal-on-highly cross-linked polyethylene articulations at mean 2-year follow-up interval.Methods A continuous series of 32 consecutive patients (32 hips) accepted THA with a Durom large head metal-on-metal articulation between January 2008 and December 2010 was as study group (Durom Group).From all the patients who received THA with Trilogy prosthesis at the same period,32 patients matched the Durom group in age,gender and body mass index (BMI) were chosen to form the Trilogy group.Another 32 healthy volunteers also matched the Durom group were chosen to form the control group.At the final follow-up,the serum metal ion levels,host immunologic response were evaluated.Results The mean follow-up time for Durom group was 24.9 months (range,15-34 months) compared to 25.53 months (range,15-48 months) for Trilogy group.Co and Cr levels in Durom group were 4.33 and 1.95 times as high as those in Trilogy group.There was no statistically significant difference in serum C3,C4,IgA,IgG,IgM value in three groups.CD3+,CD4+ and CD8+cells in Durom group decreased significantly compared with Trilogy and control groups.The interferon-γ(IFN-γ) level in Durom group was significantly higher than that in Trilogy and control groups.Conclusion The serum cobalt and chromium levels in patients received Durom metal-on-metal THA elevated significantly in early period.Meanwhile,some amount of impact on host cell mediated immunity occurred.

Analysis of the poor answer situation in chronic hepatitis B with lamivudine treatment / 中国基层医药

Objective To explore the use of lamivudine alone induced the poor response,especially the related factors of the occurrence of YMDD.Methods Retrospective analysis of 160 cases of the poor answering cases after lamivudine treatment,the baseline parameter values,the time appearing to the poor answering and the HBVDNA load in treatment were analyzed,to study their relationship with poor response.Results The lamivudine prolonged,the poor response to the probability gradually increase.The mutation rate in the 12nd month was 19.67％ and in the 36th month was 49.18％ ( P ＜ 0.05 ).Response to the many reasons,the YMDD mutation is the more common one.The relationship among HBVDNA load,ALT level on baseline and the rate of poor response emergence,the YMDD mutation rate was clear.By comparison in groups,the difference was significant (P＜0.05).Conclusion The lower baselins of ALT level,the higher of HBVDNA load,the greater probability of the poor response and the YMDD mutation.

Effect of tertamethylpyrazine on perioperative humoral immune function in patients required for autologous blood transfusion / 中华麻醉学杂志

ObjectiveTo investigate the effect of tertamethylpyrazine on perioperative humoral immune function in patients required for autologous blood transfusion.MethodsSixty ASA Ⅰ or Ⅱ patients aged 20-60 yr weighing 50-75 kg undergoing elective spinal surgery were randomly divided into 2 groups ( n =30 each):test group and control group.In test group,tertamethylpyrazine 2 mg/kg was infused intravenously over 5 min at 30 min before the autologous blood was collected.Tertamethylpyrazine was added to the heparinized saline and washing saline at the same time (25 mg per 500 ml) until the final concentration reached 0.005 ％.Tertamethylpyrazine was not given in control group.Venous blood samples were taken before anesthesia induction (T0) and at 1 h after operation (T1),and on day 1 and 5 after operation (T2.3) for measurement of serum IgG and IgM concentrations by ELISA.The operation time,intraoperative blood loss and amount of salvaged blood reinfused were recorded.ResultsThere was no significant difference in the operation time,intraoperative blood loss and amount of salvaged blood reinfused between the two groups (P ＞ 0.05).Compared with the baseline value at T0,serum IgG and IgM concentrations were significantly decreased at T1-3 in control group and the serum IgG concentration was significantly decreased at T1.2 in test group ( P ＜ 0.05 or 0.01 ).The serum IgG concentration at T2.3 and serum IgM concentration at T1-3 were significantly higher in test group than in control group ( P ＜ 0.05 or 0.01 ).ConclusionTertamethylpyrazine can reduce humoral immunosuppression to some extent and improve the balance of humoral immunity in patients required for autologous blood transfusion.

Cutaneous cytomegalovirus infection in a child with hyper IgE and specific defects in antibody response to protein vaccines

Cytomegalovirus (CMV) infection is a common opportunistic systemic infection in immunocompromised patients, but skin involvement is rare. Herein, we report a 10 year-old girl from consanguineous parents who was referred to our center because of disseminated maculopapular rash. She had history of upper and lower respiratory tract infections. In immunological studies, increased serum IgE level and decreased responses to tetanus and diphtheria were detected. Polymerase chain reaction (PCR) examination of bronchoalveolar lavage and serum sample revealed the presence of CMV. Early diagnosis of cutaneous CMV and appropriate treatment are the key actions in management of patients with underlying immunodeficiencies to avoid further complications.

Study on the relationship between the single nucleotide polymorphism of programmed cell death-1 gene and early virologic response of interferon-alpha in patients with chronic hepatitis B / 中华传染病杂志

Objective To investigate the relationship between the single nucleotide polymorphisms (SNP) of programmed cell death-1 (PD-1) gene and early virologic response of interferon-α (IFN-α) in patients with chronic hepatitis B (CHB). Methods A total of 135 CHB patients were prospectively enrolled in this study. SNP of PD-1.1 and PD-1.2 genes were examined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in these patients.Then its relationship with early virologic response to IFN-α treatment was analyzed. The data were analyzed by x2 test. Results Among the 135 patients, 33 (24.4%) achieved early virologic response to IFN-α. There were 35, 77, and 23 patients with AA, AG, GG genotgpe of PD-1.1. The early virologic response was achived in 5(14.3%), 25(32.5%) and 3(13.0%) among patients with AA,AG, GG genotypes of PD-1.1, respectively. There were statistically different (x2 = 6. 258, P =0. 044). The subjects with AG genotype showed higher response rate than those with AA or GG genotypes (x2 = 6. 246, P= 0. 012). However, the early virologic response rates were not significant different among subjects with AA, AG or GG genotype of PD-1. 2 ( x2= 3.957, P= 0. 138).Conclusion SNP of PD-1.1 gene may be used as a marker to predict the early virologic response to IFN-α treatment in Chinese CHB patients.

Immunoprotection analysis of hepatitis C virus recombinant protein / 中华传染病杂志

Objective To investigate immune responses and protective effect induced by two recombinant proteins of hepatitis C virus(HCV)in BALB/c mice.Methods BALB/c mice were immunized with recombinant proteins HCV-T and(or)HCV-F4HVR1 three times.Specific antibodies in sera were tested by enzyme-linked immunosorbent assay(ELISA).Five mice were sacrificed after 14 days of the last immunization.Splenic cells were isolated and levels of interferon(IFN)-γ,interleukin(IL)-4 and cytotoxic T lymphocyte(CTL)cytotoxicity assay were measured in vitro.The remaining mice were subcutaneously injected with 1.0×106 SP2/0-NS3 cells on the back to investigate the protective effects.The differences of means between groups were compared by LSD-t test.Results Compared with phosphate bufter saline(PBS)group,combined immunization with HCV-T and HCV-F4HVR1 induced higher levels of specific IgG against HCV-F4HVR1(t=3.815,3.762,P＜0.05),HCV-NS3-specific CTL response(t=3.971,P＜0.05)and IL-4(t=3.512,3.417,P＜0.05)and IFN-γ(t=3.813,3.426,3.671,P＜0.05)secretions.Conclusion High levels of specific humoral immunity and cellular immunity are induced in vivo after combined immunization with HCV-T and HCV-F4 HVR1,which could effectively prevent from the attack of SP2/0-NS3 cells.

Detection of serum antibodies to polymorphic membrane proteins (Pmp) in patients with urogenital Chlamydia trachomatis infection / 中华皮肤科杂志

Objecfive To detect serum antibodies to Pmp in patients with urogenital Chlamydia trachomatis infection and to assess the relationship between Pmp and urogenital C.traehomatis infection.Methods Twenty healthy adults and 77 patients with urogenital C. trachomatis infection were recruited into this study.A 3-month foilow-up was carried out in 43 patients,who were classified into persistent infection group(n=19)and negative-conversion group(n=24).Western-blot was performed to detect serum antibodies to Pmp in all subjects.Results The positivity rate of anti-Pmp antibodies was 90.20% (71/77) in patients,significantly higher than that in the normal controls[20% (4/20),P<0.05].All the 9 types of anti-Pmp antibodies were detected in patients with a varying positivity rates,which were 61.04% (47/77),88.31% (68/77),63.63% (49/77),28.57% (22,77),63.63% (49/77),75.32% (58/77),62.34% (48/77),77.92% (60/77)and 70.13% (54/77) for antibodies against PmpA,PmpB,PmpC,PmpD,PmpE,PmpF,PmpG,PmpH and PmpI respectivelyThe prevalence was highest for anti-Pmp B antibodies and lowest for anti-Pmp D antibodies.There was no significant difference in the positivity rate of anti-Pmp antibodies between persistent infection group and negativeconversion group.Conclusions Anti-Pmp antibodies could be generated in patients infected with C. trachomatis.The immunogenicity of different Pmps is different,and the immunoprotective activity of Pmps is rather weak.Individual differences exist in serum anti-Pmp antibodies among patients.Nine types of Pmps are expressed in patients with urogenital C. trachomatis infection.

Resposta humoral ao vírus dengue em mães/filhos: relação com a sequência de Mõbius / Humoral immune response to dengue virus in mothers/children: relation with Mõbius sequence

OBJETIVO: Investigar a resposta humoral para o vírus dengue em pacientes com sequência de Mõbius e suas mães, analisando a relação entre a infecção na gestação e o nascimento de portadores da sequência de Mõbius. MÉTODOS: Foram revisados os prontuários dos pacientes atendidos em centro de referência. Aplicou-se um questionário estruturado a cada genitora. Sorologia por Elisa para IgG de dengue foi efetuada em 35 pacientes e suas mães. O teste de neutralização por redução em placas foi feito nas que referiram infecção viral na gestação e em seus filhos, para determinar o sorotipo viral causador da infecção. RESULTADOS: Febre, cefaléia e/ou dor retro-orbitária foi referida por 18 (51,4 por cento) mães. Três (8,6 por cento) referiram dengue no primeiro ou início do segundo trimestre de gestação. Cruzando-se as informações das sorologias das mães e crianças observou-se que em 57,1 por cento dos casos há positividade sorológica para o vírus do dengue na mãe e no filho. Das três mães com infecção na gestação, uma e seu filho, apresentou teste de neutralização por redução em placa positivo para o vírus dengue tipo três (DENV-3), entretanto o sorotipo apenas foi introduzido em Pernambuco em 2002, o que exclui a possibilidade de ter ocorrido transmissão vertical da doença. CONCLUSÃO: As sorologias dos casos que tiveram diagnóstico clínico de dengue na gravidez revelaram-se incompatíveis com a hipótese de ter ocorrido transmissão vertical da doença. Portanto, na amostra estudada, a infecção pelo vírus do dengue não pode ser considerada como fator implicado na gênese da sequência de Mõbius.

PURPOSE: To investigate the humoral immune response to dengue virus in patients with Mõbius sequence and their mothers, assessing the relation between this infection during pregnancy and Mõbius sequence. METHODS: The medical records were reviewed, and a questionnaire was answered by each mother. IgG ELISA was performed in 35 patients and their mothers. A plaque reduction neutralization test was further done in the mothers who reported a viral infection during pregnancy and in their children for determining which dengue serotype virus had caused the infection. RESULTS: Fever, headache and/or retrobulbar pain during pregnancy was referred by eighteen (51.4 percent) mothers. Three (8.6 percent) reported dengue during the first or second quarteers of pregnancy. When cross analyzing the mothers and their respective children serological results, 57.1 percent of them matched. Of the three mothers with clinical diagnosis of viral infection during pregnancy, one and its respective children had a positive plaque reduction neutralization test for dengue serotype three (DENV-3), this serotype reached Pernambuco in 2002, which excludes the possibility of a vertical transmission to the children. CONCLUSION: The serology of the cases that had clinical diagnosis of dengue during pregnancy proved to be incompatible with the hypothesis of the disease vertical transmission. Therefore, in the present serie, the dengue virus infection can not be considered as a factor involved in the genesis of Mõbius sequence.

Expression of t antigen fusion protein of JC virus and preparation of its polyclonal antibody / 中华传染病杂志

Objective To construct prokaryotic expression vector carrying jc virus(JCV)t-antigen gene,express and purify this fusion protein.Methods The JCV t-antigen gene from a cerebrospinal fluid sample was amplified using polymerase chain reaction(PCR)method.After sequencing.the gene was cloned into plasmid pET32a(+)to construct recombinant prokaryotic expression vector pET32a(+)-t.The t-antigen fusion protein was expressed by isopropy-~D-thiogalactoside(IPTG)induction and prepared in large scale,then purified by Ni+affinity column chromatography.The polyclonal antibody was obtained from the BAI.B/C mouse immunity by the purified protein.Results The relative molecular nlass of recombinant protein expressed by pET32a(+)-t was about 41 000.Sodium dodeeylsulfate-polyaerylamide gel electrophoresis(SDS-PAGE)showed that the fusion protein W&S highly expressed after 3.5～20.Oh of IPTG induction.The antigenicity of the purified protein Was well confirmed by Western blot.The anti-mousepolyclonal antibody was obtained successfully from immunized BALB/c mice.Conclusions The prokaryotic expression vector pET32a(+)-t is successful constructed and the fusion protein is expressed and purified.Furthermore,the antibody of JCV small envelop protein t is successfully prepared.This work provides vMuable information for further study on epidemiology and biological function of t antigen.

Preparation of the polyclonal antibody against KIAA0649 and its cellular localization / 北京大学学报(医学版)

Objective: To prepare and characterize the polyclonal antibody against KIAA0649 and identify the localization and the functional motif of KIAA0649. Methods: Three polypeptides were synthesized based on the bioinformatics analysis of KIAA0649 protein. New Zealand rabbits were immunized with the mixture of the three KIAA0649 peptides coupled with keyhole limpet hemocyanin ( KLH). The titer of the antisera was detected with ELISA. The antisera were purified with immuno-affinity chromatography when the titer reached 1:10~5. Western blot was performed with the purified antisera on the cell lysates of U2OS cells transfected with either Flag-KIAA0649 or KIAA0649-targeting siRNA. Immunofluorescence was performed with the purified antisera and anti-Flag antibody on the cells transfected with FlagKIAA0649. A series of Flag-K1AA0649 deletion mutants was constructed by PCR cloning. The cellular compartmentation of full-length Flag-KIAA0649 and its deletion mutants were analyzed with immunofluorescence. Results: The results of Western blot and immunofluorescence demonstrated that the antisera from the KIAA0649 polypeptides-immunized rabbits specifically recognized endogenous and exogenous KIAA0649. The full-length Flag-K1AA0649 displayed specific nuclear foci. The Flag-KIAA0649 deletion mutant containing PENF motif showed the same nuclear foci as the full length of Flag-KJAA0649, suggesting that the PENF motif could be the minimum functional motif of KIAA0649. Conclusion: We have obtained anti-KIAA0649 polyclonal antibody which will be useful for further investigation. The PENF motifcould be the minimum functional domain of KIAA0649.