Levofloxacin combined with cellulase can eradicate bacille Calmette-Guerin biofilm infection / 南方医科大学学报

OBJECTIVE@#To investigate the inhibitory effects of levofloxacin (LEV) combined with cellulase against bacille CalmetteGuerin (BCG) biofilms in vitro.@*METHODS@#The mature growth cycle of BCG biofilms was determined using the XTT method and crystal violet staining. BCG planktonic bacteria and BCG biofilms were treated with different concentrations of LEV and cellulose alone or jointly, and the changes in biofilm biomass were quantified with crystal violet staining. The mature BCG biofilm was then treated with cellulase alone for 24 h, and after staining with SYTO 9 and Calcofluor White Stain, the number of viable bacteria and the change in cellulose content in the biofilm were observed with confocal laser scanning microscopy. The structural changes of the treated biofilm were observed under scanning electron microscopy.@*RESULTS@#The MIC, MBC and MBEC values of LEV determined by broth microdilution method were 4 μg/mL, 8 μg/mL and 1024 μg/mL, respectively. The combined treatment with 1/4×MIC LEV and 2.56, 5.12 or 10.24 U/mL cellulase resulted in a significant reduction in biofilm biomass (P < 0.001). Cellulase treatments at the concentrations of 10.24, 5.12 and 2.56 U/mL all produced significant dispersion effects on mature BCG biofilms (P < 0.001).@*CONCLUSION@#LEV combined with cellulose can effectively eradicate BCG biofilm infections, suggesting the potential of glycoside hydrolase therapy for improving the efficacy of antibiotics against biofilmassociated infections caused by Mycobacterium tuberculosis.

Bacille Calmette-Guérin vaccine induced lymphadenitis in infants and children masquerading with tubercular lymphadenitis at tertiary care hospital in North India

Background: Bacille calmette–guerin (BCG) vaccine containing live-attenuated Mycobacterium bovis was first used in humans to prevent tuberculosis in 1921. It is a safe vaccine. However, there can be minor adverse reactions such as pain, swelling, and redness at local injection site. The lesser known severe adverse reactions such as lymphadenitis, BCG osteitis, and disseminated BCG infection can be present after BCG vaccination. Aims and Objectives: The aim of the study was to report the cases of BCG lymphadenitis diagnosed by fine needle aspiration cytology (FNAC) after BCG vaccination in infants and children. Materials and Methods: All the infants and children (1 month–2 years) who presented with regional lymphadenopathy for FNAC after BCG vaccination were included in the study. Results: Total 21 patients with BCG lymphadenitis were recruited. In all the cases, the lymphadenitis involved ipsilateral left axillary lymph nodes. In three patient sites of involvement included the left supraclavicular lymph nodes and in other three patients left cervical lymph nodes along with left axillary lymph node. Sixteen patients had suppurative granulomatous lymphadenitis (SGLA) on FNAC and positive for Ziehl–Neelsen (ZN) staining for acid fast bacilli (AFB), while rest nine had non-suppurative granulomatous lymphadenitis (NSGLA) with negative for ZN staining for AFB. Conclusion: Early diagnosis of BCG lymphadenitis can help in proper management. Furthermore, it can prevent unnecessary anti tubercular treatment in children.

The Incidence Rate of Lymphadenitis after Bacille Calmette-Guérin (BCG) Vaccination

PURPOSE: Bacille Calmette-Guérin (BCG) lymphadenitis is a relatively frequent local adverse reactions after BCG vaccination. Its incidence rate is usually <1%. However, this rate may be different according to BCG strain, vaccination method or skill, etc. In the Republic of Korea, two BCG strains are used: intradermal Danish-1331 or percutaneous Tokyo-172. We surveyed the incidence rates of BCG lymphadenitis. METHODS: This survey was performed in total 25 centers (5 general hospitals, 20 private pediatric clinics). Immunized type of BCG strain in study subjects was verified by directly observing the scar. The occurrence of BCG lymphadenitis was asked to their parent. In cases of BCG lymphadenitis, location, diameter size, progression of suppuration, and treatment method were investigated, as well. RESULTS: The total number of study subjects was 3,342. Among these, the subjects suitable for enrollment criteria (total 3,222; Tokyo strain 2,501, Danish strain 721) were analyzed. BCG lymphadenitis regardless of its size developed in each five of subjects per strains, therefore, its incidence rate was 0.20% in Tokyo and 0.69% in Danish strain, respectively (P=0.086). However, when applying the WHO criteria - the development of lymph node swelling with diameter 1.5 cm or more, the incidence rate of BCG lymphadenitis was 0.16% (4 cases) in Tokyo and 0.42% (3 cases) in Danish strain, respectively. CONCLUSIONS: The incidence rate of lymphadenitis in two BCG types, percutaneous Tokyo and intradermal Danish strain BCG, is 0.20% and 0.69%, respectively. Both rates are acceptable.

BCG HSP70 gene transfection and its effect on immunogenicity of murine lymphocytic leukemia / 临床儿科杂志

Objective To assess the effect of heat shock protein 70 of BCG (BCG HSP70) gene transfection on tumorigenicity and immunogenicity of murine lymphocytic leukemia cells (L1210). Methods BCG HSP70 gene was transfected onto the surface of murine lymphocytic leukemia cells (L1210) by lipofectamine 2000. And then the positive clone (L1210-HSP70) highly expressing HSP70 was selected as the tumor vaccine to study the tumorigenicity experiments in nude mice and syngeneic mice, the therapeutic experiments, and the immunoprotective effects. Results The expression of BCG HSP70 on the L1210 cells surface was detected, and the L1210-HSP70 cells had the same tumorigenicity as the parental L1210 cells did. Tumorigenicity experiments in syngeneic mice:In L1210-HSP70 group, tumor growth was slow or without the formation of tumor. As compared with L1210 group and L1210-neo group the mice survival time was signiifcantly prolonged, showing a marked stimulating effect on L1210 specific Th1 cells,. Tumor-bearing mice showed complete coagulation necrosis and abundant CD8+T lymphocyte inifltration (P<0.05). The tumor vaccine of L1210-HSP70 cells had the antitumor therapeutic efifcacy and immune protection effect, demonstrating that the tumor growth was signiifcantly inhibited, tumor diameter was markedly reduced and the survival time of tumor-bearing DBA/2 mice was further prolonged. Conclusions BCG HSP70 gene transfection could effectively improve the immunogenicity of tumor cells, activate speciifc T cells and enhance the anti-tumor immunity in vivo. Meanwhile, the host anti-tumor immunity could be enhanced.

Preparation of leukemia cell vaccine expressing Bacille Calmette-Guérin heat shock protein 70 and its anti-leukemia therapeutic effect / 中华实用儿科临床杂志

Objective To culture the acute leukemia cells in vitro,and to prepare cancer vaccine expressing heat shock protein 70 (HSP70) of Bacille Calemette-Geérin(BCG) onto the cell surface,so as to study its anti-tumor effect and mechanism.Methods Acute myeloid leukemia (AML) cells were cultured in a serum-free Stemspan(H) culture supplemented with cytokines [stem cell factor(SCF),flt-3 ligand (FL),interleukin (IL)-3 and IL-6] in vitro.And B-lineage acute lymphoblastic leukemia (B-ALL) cells were cultured in a Iscove modified medium(IMDM) culture supplemented with cytokines (SCF,FL,IL-3 and IL-7) in vitro.Cellular morphology was observed by the microscopy and immunophenotype determination was used to verify the biological characteristics of acute leukemia cells after culture.Lipofectamine 2000 was used to transfect the pDisplay-HSP70 plasmid into acute leukemia cells.The expression of HSP70 on the cell surface was detected by fluorescene microscope.Then the immunogenicity of the leukemia cells expressing HSP70 were detected.The experimental groups were divided into 3 subgroups:the wide-type acute leukemia cells (wt-LC group),the pDisplay-leukemia cells (pDisplay-LC group),and the pDisplay-HSP70-leukemia cells (HSP70-LC group),respectively.The leukemia cells in different groups were cultured with autologous peripheral blood T cells for 72 hours.The proliferation indices of T cells were assayed by carboxyfluorescein diacetate succinimidyl ester (CFSE)-staining method,and the contents of interferon-γ(IFN-γ) were tested by enzyme-linked immunosorbent assay (ELISA).The leukemia cells in different groups were cultured with autologous peripheral blood T cells,and after 6 days,the fresh acute leukemia cells were added [in the different ratios of cytotoxicity T lymphocyte (CTL):leukemia cells were 10 ∶ 1,20 ∶ 1,40 ∶ 1 and 80 ∶ 1] and continued to be cultured for another 12 hours.Cytotoxicity assay was measured by lactate dehydrogenase (LDH) release.Results After short term culture in vitro,the leukemia cells were in colony-like suspension and maintain the proliferation characteristics were maintained.The cell proliferation was rapidly cultured for about 10 days and then was gradually slowed down.But there was no difference between the day 10 and day 0 in the expressions of CD13 and CD33 in fifteen cases of AML cells (P ＞ 0.05).Equally,there was no difference between the day 10 and day 0 in the expressions of CD19,CD10 and CD22 in fifteen cases of B-ALL cells (P ＞ 0.05).After BCG HSP70 gene transfection,the yellow-green fluorescence on the leukemia cells surface was observed under the confocal microscope.Detection of the immunogenicity:(1) Autologous T cell proliferation:the most significant T cell proliferation was observed in the group of HSP70-transfected leukemia cells (t =17.89,19.58,all P ＜0.05).There was no difference between the wt-LC group and pDisplay-LC group (P ＞ 0.05).(2) The contents of cytokines:the IFN-γ level in the group of HSP70-transfected leukemia cells was higher than those of wide-type acute leukemia cells and the pDisplay-transfected ones (t =24.72,24.81,all P ＜ 0.05).(3) Cytotoicity of CTL:the killing rate in HSP70-transfected leukemia cells was significantly higher than those of wide-type acute leukemia cells and pDisplay——transfected ones(F =13.66,P ＜ 0.05).And with the increase of the ratio from 10 ∶ 1 to 80 ∶ 1,the inhibiting activity of CTL in the HSP70-LC group was raising(F =19.69,P ＜ 0.05).Conclusions Fresh acute leukemia cells can be successfully cultured in vitro.Short-term culture can significantly increase the number of leukemia cells,but has little effect on surface antigen expression.So,the biological characteristics of the leukemia cells can be maintained.The leukemia cells vaccine expressing BCG HSP70 onto its surface was successfully prepared,and gene transfection of BCG HSP70 can significantly enhance the immunogenicity of leukemia cells.

Surveillance analysis on BCG vaccination and its safety for newborns in IPMCH / 上海预防医学

Objective ] To analyze the occurrence of adverse events following immunization (AEFI)of Bacille Calmette-Guerin vaccine(BCG)among neonates in IPMCH, and to evaluate the safety of BCG. [Methods] Data on BCG AEFI in 2010-2013 were collected through national AEFI information system.The descriptive methodology was used in this study . [ Results] During 2010 -2013, the coverage rate of the BCG vaccine among neonates in IPMCH was 95 .19%.And 21 BCG AEFI cases vaccinated were reported, with a reporting rate of 430.86 per 1 million doses, most of which were BCG lymphadenitis(20 cases with the incidence of 410.34 per 1 million doses). [Conclusion] The coverage rate of the BCG vaccination among neonates in IPMCH remained at high level .BCG vaccine proved safe . Most of rare vaccine reactions were BCG lymphadenitis , and the reporting rate of BCG lymphadenitis were lower than expected .

Adverse reactions to Mycobacterium bovis bacille Calmette-Guerin vaccination against tuberculosis in Iranian children

PURPOSE: There are considerable variations in the number of adverse reaction reports related to vaccine from different countries. The aim of this study was to review the development of adverse reactions to bacille Calmette-Guerin (BCG) vaccination among hospitalized patients in an Iranian referral hospital. MATERIALS AND METHODS: We identified hospitalized patients with BCG complications in Pediatric Infectious Disease Research Center, Tehran University of Medical Sciences, Tehran, Iran during January 2007-April 2009. Data on demographics, clinical features, laboratory findings, personal history (including vaccination history), family history, and outcomes were retrieved from medical records. RESULTS: There were 46 cases with BCG complication during the 2 years period. All of the children received vaccination at birth. Twenty-eight patients (61%) were male. The mean age of the patients was 13.5 +/-11.3 months (range, 1 to 52 months; median, 10 months). The majority of children (57%) with BCG complication were less than 1 year old. Among hospitalized patients due to BCG complications, suppurative lymphadenitis was occurred in 28 children (61%) and lymphadenopathy was seen in 9 children (20%). Disseminated BCG was detected in 8 patients (17%) and only 1 child (2%) was presented with abscess. In 7% (n = 3) of children, the family history of BCG complications were positive. CONCLUSION: The most common side effect of the BCG vaccine in our study was suppurative lymphadenitis. Disseminated BCG infection in complications leading to hospitalization in our study was 17%. With regard to the difficulty in implementing such a guideline in settings where BCG is given to all newborns, registration of Iranian primary immunodeficiency disorders (PID) patients would be helpful to increase the awareness of medical community of Iran to investigate underlying disease. In addition, BCG vaccination should postpone in each newborn with a family history of PID until the definite condition has been ruled out.

Enhancement of vitamin A combined vitamin D supplementation on immune response to Bacille Calmette-Guérin vaccine revaccinated in Chinese infants

OBJECTIVE@#To investigate whether there is an association between diameter of bacille Calmette-Guérin (BCG) scars and effect of purified protein derivative (PPD) reaction and to determine whether vitamin A (VA) combined vitamin D (VD) supplementation influences the immune response to BCG revaccinated in Chinese infants.@*METHODS@#A cross-section and 3-month community-randomised trial was conducted. A total of 5 629 infants at 3, 6 and 12 months of age in Junan County of China were examined for BCG scar formation. Then, 597 revaccinated infants were randomly assigned to supplementation (n=307) and control (n=290) groups. The supplementation group were daily assigned to 1 500 IU VA and 500 IU VD for 3 months. Then all infants were subjected to skin test with PPD.@*RESULTS@#The diameter of BCG scars was positively correlated with diameter of skin indurations of PPD (r=0.17, P<0.05) in the 5 629 infants. The rate of positive response to PPD was higher in the supplementation group than in the control group (96.1% versus 89.7%, P<0.05, prevalence ratio 1.07, 95% CI 1.02-1.12). The prevalence ratio of PPD response for the supplementation group compared with that for the control group was 1.07 (95% CI 1.01-1.13) for the males and 1.08 (95% CI 1.00-1.17) for the females. For the supplementation group, the males got larger tuberculin induration than the females [(0.73±0.21) cm versus (0.67±0.20) cm, P<0.05) after intervention.@*CONCLUSIONS@#The diameter of BCG scars was effectively correlated with PPD response, which indicates BCG scar formation may be an useful tool to evaluate the effect of tuberculosis prevention. VA combined VD supplementation may play an immuno-regulatory role in BCG revaccination. This may contribute to the prevention of childhood tuberculosis.

Tuberculous Prostatic Abscess Following Intravesical Bacillus Calmette-Guerin Instillation

Tuberculous granulomatous infection of the urinary tract is a rare complication of intravesical Bacille Calmette-Guerin (BCG) immunotherapy, which is commonly used for superficial urothelial cell carcinoma of the bladder. We present a case of tuberculous prostatic abscess in a patient who had undergone intravesical BCG immunotherapy for bladder carcinoma in situ.

Establishment of a Myelin Basic Protein Fragment-Induced Wistar Rat Model of Autoimmune Encephalomyelitis / 中国实验动物学报

Objective To establish an experimental autoimmune encephalomyelitis (EAE) model in Wistar rats with myelin basic protein fragment (MBP69-85) and observe its pathological changes. Methods MBP69-85 dissolved in normal saline was mixed with complete Freund's adjuvant (CFA) (including 6 mg bacille Calmette Guerin) to prepare the encephalitogenic emulsion. Ten out of 70 Wistar rats were chosen as a control group, the others were divided equally into group A,B,C according to the difference of the encephalitogenic emulsion. Rats in group A were immunized with 50 μg MBP69-85 +CFA (including 6 mg BCG). Rats in group B were immunized with 25 μg MBP69-85+CFA (including 6 mg BCG). Rats in group C were immunized with 25 μg MBP69-85+CFA (including 12 mg BCG). The pathological changes of brain and spinal cord tissues were examined by light microscopy after HE staining and immunohistochemistry of MBP and NF. Results Some of the Wistar rats immunized with 50 μg MBP69-85 showed disorder at 12～16 days after immunization. The clinical symptoms included tail acratia or paralysis of tail and limbs, head tilt, etc. and the mean score was 2.38±1.89. There were infiltration of inflammatory cells inside nervous tissue and perivascular cuffings in HE stained sections. The immunohistochemistry of MBP and NF showed demyelination in the white matter and axon injury. Conclusion To some extent, the establishment of EAE depends on the dose of the immunizing antigen. The BCG in CFA was not the major cause of morbility of the rats. The EAE model induced with MBP69-85 in Wistar rats, showing typical clinical symptoms and pathological changes of multiple sclerosis, is a reliable animal model for the study of pathogenesis and treatment of multiple sclerosis.

Establishment of and Effects of BPV on Experimental Allergic Encephalomyelitis in Lewis Rats / 医学研究杂志

Objective To establish the animal model of experimental allergic encephalomyelitis(EAE)in Lewis rats.Methods The EAE was induced by giving hypodermal injections in feet with spinal cords homogenate of guinea pigs(GPSCH)and complete Freund's adjuvant(CFA)containing 8mg.ml-1 bacille calmette guerin(BCG).The EAE model was evaluated by clinical manifestations and pathologic findings which was studied with the aid of light and electron microscope.Results The EAE in Lewis rats had an incidence of 9/10,a latency of 12.6?1.01d.Bordetella pertussis vaccine(BPV)could increase the morbidity,shorten the latency(P

The clinic observation of polysaccharide nucleicacid of bacille calmette-guerin injection on bronchus asthmatic patients / 中国基层医药

Objective To observe the influence on clinic symptom and pulmonary function in asthmatic patients treated with polysaccharide nucleicacid of calmette-guerin(BCG-PSN) injection in asthmatic patients.Methods 30 cases(treatment group) with additional BCG-PSN injection and 30 cases(control group) with only normal treatment were studied in contrast.They were observed in pulmonary function and clinic symptom before and after treatment.Results Symptom and pulmonary function improved in both groups.The validity rate in treatment group was 96.7%,and the control group was 80%.Conclusion BCG-PSN could alleviate the inflammation of bronchus and excellent effects of BCG-PSN injection in treatment on clinic symptom and pulmonary function of asthmatic patients were found.

Survey on the Association of the Number of BCG Scars and Atopic Diseases / 소아알레르기및호흡기학회지

PURPOSE: The prevalence of atopic diseases has been increasing remarkably. The less frequent opportunities for infection early in life, especially mycobacteria exposure, parallel this higher prevalence of atopic diseases. Bacille Calmette-Gu rin (BCG), a potent inducer of Th1 immune response, has been suggested to suppress Th2 response which is known to mediate IgE-mediated atopic disorders. This study was done to investigate whether there is any relation between the number of BCG scars and the prevalence of atopic disorders in early childhood. METHODS: We surveyed 393 parents with a children who were given percutaneous multi- puncture BCG vaccination within four weeks after birth. The main questions concerned the past history and present illness of physician-diagnosed atopic diseases (atopic dermatitis, bronchial asthma, and allergic rhinitis), the number of BCG scars (range; 0-18), and potential confounders such as gender, parental atopy, maternal smoking and environmental cofactors. The prevalence rate of each atopic disease was measured and analysed according to the number of BCG scars. RESULTS: Each prevalence rate was 18.1% for atopic dermatitis, 9.4% for bronchial asthma, 14.6% for allergic rhinitis, and 32.3% for any of them. All of them had received BCG vaccination during the first four weeks of life. The children with 15 or more BCG scars had a significantly lower prevalence of any atopic disease (22/99, 22.2%) as compared to those with four scars or less (51/125, 40.8%) by simple regression analysis. (P value=0.02) But this association was not significant after controlling for potential confounders. (P value= 0.26) CONCLUSION: This survey demonstrated a weak relation between a larger number of BCG scars and less atopy development at early childhood. But the relation was not so significant. Further studies are needed.

Effects of Bacille Calmette Guerin on immunological function of infants with bronchiolitis / 中国临床药理学与治疗学

AIM:To approach the effects of Bacille Calmette-Guerin(BCG)on immunological function of infants with bronchiolitis in vitro. METHODS: The monocytes were segregated from the peripheral blood of infants with acute bronchiolitis and healthy infants as control. The cytokines of IL-10 and IL-12 were measured by ELISA after interfered with different concentrations of BCG in vitro. IL-4, produced by lymphocytes treated with the cultural supernatant of BCG-stimulated monocytes were determined by ELISA. Acute bronchiolitis infants and health children treated with normal saline were as control. RESULTS: BCG stimulated IL-10 and IL-12 production in a dose response manner in a certain concentration range. The higher level of IL-10 was observed when BCG concentration reached 50 ?L/mL, while IL-12 had no significant increase. The level of IL-4 was markedly decreased compared with pre-treatment (P

Clinical study on bacille calmette-guerin alone or combined with interferon gamma and vitamin A in preventing asthma children after bronchiolitis / 实用儿科临床杂志

Objective To discuss the effects of bacille calmette guerin(BCG) alone of combined with interferon gamma(IFN-?) and vitamin A in preventing asthma after bronchiolitis.Methods Fourty-four bronchiolitis infants(RBI) were treated with BCG alone or combined with vitamin A/IFN-?.Protein purified derivative(PPD) skin test were observed before and 3 months after interference and all infants enrolled in the study were followed up for average one year to observe the wheezing episodes. Ninteen cases of RBI without BCG treatment were control group.Results The average diameter of PPD skin test in RBI after BCG inoculation was significantly larger than that of before inoculation and control group(P