Objective: To characterize the immunogenicity and the induction of cross-reactive responses against Mycobacterium tuberculosis(M. tuberculosis) of a proteoliposome(PL)from Mycobacterium bovis Bacillus Calmette–Guerin...Objective: To characterize the immunogenicity and the induction of cross-reactive responses against Mycobacterium tuberculosis(M. tuberculosis) of a proteoliposome(PL)from Mycobacterium bovis Bacillus Calmette–Guerin(BCG) with and without alum hydroxide(AL) as adjuvant(PLBCG-AL and PLBCG, respectively) in BALB/c mice.Methods: BALB/c mice were inoculated with phosphate buffer solution, BCG, PLBCG and PLBCG-AL. The humoral immunogenicity was determined by ELISA [immunoglobulin G(Ig G), Ig G1 and Ig G2a] and the cellular immunogenicity was evaluated in vivo by delayed type hypersensitivity. The humoral cross-reactive response against M. tuberculosis was determined by Western blot.Results: Sera from animals immunized with PLBCG-AL and PLBCG showed significant increase in specific total Ig G and Ig G1 antibodies and the presence of cross-reactive antibodies against M. tuberculosis antigens, which were more intense with the use of alum as adjuvant. Mice immunized with PLBCG and PLBCG-AL also showed a specific cellular response in vivo.Conclusions: The cellular and humoral immunogenicity of PLBCG and the capacity to induce cross-reactive responses against M. tuberculosis is in agreement with the protective capacity previously demonstrated by this vaccine candidate and supports the continuation of its evaluation in further stages.展开更多
Heat shock protein 65 (HSP65) is one of the most important protective immunogens against the tuberculosis infection. The signal sequence of antigen 85B and the whole HSP65 DNA sequence of human Mycobacterium tuberculo...Heat shock protein 65 (HSP65) is one of the most important protective immunogens against the tuberculosis infection. The signal sequence of antigen 85B and the whole HSP65 DNA sequence of human Mycobacterium tuberculosis (M. tuberculosis) were amplified from BCG genome and plasmid pCMV-MTHSP65 respectively by polymerase chain reactions (PCR). These two sequences were cloned into the plasmid pBCG-2100 under the control of the promoter of heat shock protein 70 (HSP70) from human M. tuberculosis, yielding the prokaryotic shuttle expression plasmid pBCG-SP-HSP65. Results of restriction endonuclease analysis, PCR detection and DNA sequencing analysis showed that the two cloned DNA sequences were consistent with those previously reported, and the direction of their inserting into the recombinant was correct and the reading frame had been maintained. The recombinants were electroporated into BCG to construct the recombinant BCG vaccine and induced by heating. The induced expression detected by SDS-PAGE showed that the content of 65 kD protein expressed in recombinant BCG was 35.69 % in total bacterial protein and 74.09 % in the cell lysate supernatants, suggesting that the recombinant HSP65 gene could express in BCG with high efficiency and the expressed proteins were mainly soluble. Western-blot showed that the secretive recombinant proteins could specifically combine with antibody against M. tuberculosis HSP65, indicating that the recombinant proteins possess the biological activity of HSP65.展开更多
The cellular immune response elicited by Mycobacterium bovis Bacille Calmette-Guérin (BCG) has been carefully investigated, but the humoral immune response has been partially neglected. BALB/c mice were immunized...The cellular immune response elicited by Mycobacterium bovis Bacille Calmette-Guérin (BCG) has been carefully investigated, but the humoral immune response has been partially neglected. BALB/c mice were immunized with BCG strain used to immunize humans. Anti-BCG antibodies, as assayed by ELISA, began to appear in the sera after the third week of immunization and plateaued three weeks after the 8th immunization. The total immunoglobulins (Igs) were purified by caprylic acid method from pooled serum collected after the 8th immunization. Anti-BCG antigen antibodies were detected in the total Igs preparation as well as in IgG, IgM, IgA, IgG1, IgG2a, and IgG2b, but not in the IgG3. Distinct BCG proteins were recognized the IgGs in Western blot analysis. Opsonization of BCG bacilli by the purified Igs potentiated internalization of the bacteria by murine Raw 264.7 macrophages. The intracellular BCG elimination coincided with the induction of NO production, which was more pronounced in cells infected with opsonized BCG compared to those infected with the non-opsonized bacteria. Coincidently, the production of NO was also higher in macrophages infected with opsonized BCG (maximal NO production at 48 h of incubation). The obtained results demonstrate that repeated inoculations of BCG effectively activate the humoral immune response, justifying the use of BCG as a live recombinant vaccine vector to insert genes encoding virulence factors controlled by antibodies.展开更多
Recalcitrant warts can accurately be defined as warts that persist after six months of conventional therapy. Up to one-third of non-genital warts, especially periungual and plantar warts, become recalcitrant. Traditio...Recalcitrant warts can accurately be defined as warts that persist after six months of conventional therapy. Up to one-third of non-genital warts, especially periungual and plantar warts, become recalcitrant. Traditional treatment options for warts include topical salicylic acid, cryotherapy, and electrocautery;however, patients with recalcitrant warts remain a major therapeutic challenge. There is evidence that immunotherapy can clear recalcitrant warts if traditional treatment fails. Given this, clinical studies published in PubMed and Google Scholar that used Bacillus Calmette-Guerin (BCG), Mycobacterium Indicus Pranii (Mw vaccine), and purified protein derivative (PPD) as immunotherapy for wart, were reviewed in this study. Neither of these treatments has been subjected to a randomized controlled trial, thus to date, there are no standardized protocols to use them. Our review highlights the scientific facts in the clinical applications of the previous options to treat recalcitrant warts and investigate the differences among them, concerning efficacy, adverse effects, dosage, and route of administration.展开更多
We observed a 76-year-old man who presented “acute kidney-lung failure” 9 months after intravesical Bacillus Calmette-Guérin (BCG) adjuvant treatment for a T1 bladder cancer. He had inflammatory infiltration on...We observed a 76-year-old man who presented “acute kidney-lung failure” 9 months after intravesical Bacillus Calmette-Guérin (BCG) adjuvant treatment for a T1 bladder cancer. He had inflammatory infiltration on chest radiography and required dialysis for acute renal failure. A percutaneous renal biopsy was performed and revealed tubulointerstitial nephritis with a moderate eosinophilic infiltrate without granulomatous lesion. After a few days, an open lung biopsy was also done due to respiratory deterioration. The anatomopathologic specimen demonstrated moderate fibrosis with lympho-neutrophilic infiltration and few aspecific granulomatous lesions without caseous necrosis. Sarcoidosis was suspected and high dose oral methylprednisolone was started. Three weeks later, Mycobacterium bovis was identified by Polymerase Chain Reaction on open lung biopsy. He responded well to steroids and tuberculostatic tri-therapy. After one month of immunosuppressive treatment, renal function was resolved and hemodialysis could be discontinued. Despite the frequent use of adjuvant BCG immunotherapy, systemic complications such as hepatitis, pneumonitis, spondylodiscitis or multiorgan failure are rare (<1%). Hematogenous dissemination which occurs a few weeks after traumatic instillations is usually suspected but not demonstrated because of absence of mycobacterium in histological specimen. Our case differs from those previously reported by the simultaneous presence of acid-fast bacilli highlighted on lung samples. We discuss the pathophysiology of BCG complications, the use of prophylactic or therapeutic treatment and recommend guidelines to prevent such complications.展开更多
The BALB/c mice were immunized with rMS Sj26GST and rBCG Sj26GST vaccine in Schistosoma japonicum by subcutaneous injection After they were immunized for 8 weeks, the eyeballs were removed to get blood and macroph...The BALB/c mice were immunized with rMS Sj26GST and rBCG Sj26GST vaccine in Schistosoma japonicum by subcutaneous injection After they were immunized for 8 weeks, the eyeballs were removed to get blood and macrophages of abdominal cavity and spleen cells were harvested The lymphocytic stimulating index (SI) was used to measure the cellular proliferating ability and NO release was used to measure the phagocytic activity of the macrophages By using ELISA kit, the levels of interleukin 2 (IL 2) and interferon γ (IFN γ) in serum and the splenic lymphocytic cultured supernatant were detected The results showed that after the mice were immunized with 10 6 CFU of rMS Sj26GST and rBCG Sj26GST vaccine separately by subcutaneous injection, proliferating ability of splenic lymphocytes in the mice showed no difference ( P >0 05), but both were significantly increased as compared with that in the control group( P <0 05); The contents of NO in the intraperitoneal macrophages of rMS Sj26GST vaccine group were significantly lower than in the control group ( P <0 001) and rBCG Sj26GST vaccine group ( P <0 01); The levels of serum IL 2 in the rMS Sj26GST vaccine group were significantly increased as compared with that in the control group ( P <0 001), vector group ( P <0 01) and rBCG Sj26GST vaccine group ( P <0 05); The contents of serum IFN γ in the rMS Sj26GST vaccine group were significantly increased as compared with that in the control group ( P <0 01) and rBCG Sj26GST vaccine group ( P <0 05) The contents of IFN γ in the cultured supernatant were significantly lower than those of rBCG Sj26GST vaccine group ( P <0 001), but were significantly increased as compared with that in the control group ( P <0 01) It was indicated that both vaccines could enhance the immune response of the mice, but rMS Sj26GST vaccine had stronger immunogenicity than rBCG Sj26GST vaccine展开更多
基金Supported by the Long-Term Research Grant Scheme Grant,Department of Higher Education,Ministry of Education,Malaysia(Grant No.203.PPSK.67212002)as well as the Ministry of Science and Technology,Cuba
文摘Objective: To characterize the immunogenicity and the induction of cross-reactive responses against Mycobacterium tuberculosis(M. tuberculosis) of a proteoliposome(PL)from Mycobacterium bovis Bacillus Calmette–Guerin(BCG) with and without alum hydroxide(AL) as adjuvant(PLBCG-AL and PLBCG, respectively) in BALB/c mice.Methods: BALB/c mice were inoculated with phosphate buffer solution, BCG, PLBCG and PLBCG-AL. The humoral immunogenicity was determined by ELISA [immunoglobulin G(Ig G), Ig G1 and Ig G2a] and the cellular immunogenicity was evaluated in vivo by delayed type hypersensitivity. The humoral cross-reactive response against M. tuberculosis was determined by Western blot.Results: Sera from animals immunized with PLBCG-AL and PLBCG showed significant increase in specific total Ig G and Ig G1 antibodies and the presence of cross-reactive antibodies against M. tuberculosis antigens, which were more intense with the use of alum as adjuvant. Mice immunized with PLBCG and PLBCG-AL also showed a specific cellular response in vivo.Conclusions: The cellular and humoral immunogenicity of PLBCG and the capacity to induce cross-reactive responses against M. tuberculosis is in agreement with the protective capacity previously demonstrated by this vaccine candidate and supports the continuation of its evaluation in further stages.
文摘Heat shock protein 65 (HSP65) is one of the most important protective immunogens against the tuberculosis infection. The signal sequence of antigen 85B and the whole HSP65 DNA sequence of human Mycobacterium tuberculosis (M. tuberculosis) were amplified from BCG genome and plasmid pCMV-MTHSP65 respectively by polymerase chain reactions (PCR). These two sequences were cloned into the plasmid pBCG-2100 under the control of the promoter of heat shock protein 70 (HSP70) from human M. tuberculosis, yielding the prokaryotic shuttle expression plasmid pBCG-SP-HSP65. Results of restriction endonuclease analysis, PCR detection and DNA sequencing analysis showed that the two cloned DNA sequences were consistent with those previously reported, and the direction of their inserting into the recombinant was correct and the reading frame had been maintained. The recombinants were electroporated into BCG to construct the recombinant BCG vaccine and induced by heating. The induced expression detected by SDS-PAGE showed that the content of 65 kD protein expressed in recombinant BCG was 35.69 % in total bacterial protein and 74.09 % in the cell lysate supernatants, suggesting that the recombinant HSP65 gene could express in BCG with high efficiency and the expressed proteins were mainly soluble. Western-blot showed that the secretive recombinant proteins could specifically combine with antibody against M. tuberculosis HSP65, indicating that the recombinant proteins possess the biological activity of HSP65.
文摘The cellular immune response elicited by Mycobacterium bovis Bacille Calmette-Guérin (BCG) has been carefully investigated, but the humoral immune response has been partially neglected. BALB/c mice were immunized with BCG strain used to immunize humans. Anti-BCG antibodies, as assayed by ELISA, began to appear in the sera after the third week of immunization and plateaued three weeks after the 8th immunization. The total immunoglobulins (Igs) were purified by caprylic acid method from pooled serum collected after the 8th immunization. Anti-BCG antigen antibodies were detected in the total Igs preparation as well as in IgG, IgM, IgA, IgG1, IgG2a, and IgG2b, but not in the IgG3. Distinct BCG proteins were recognized the IgGs in Western blot analysis. Opsonization of BCG bacilli by the purified Igs potentiated internalization of the bacteria by murine Raw 264.7 macrophages. The intracellular BCG elimination coincided with the induction of NO production, which was more pronounced in cells infected with opsonized BCG compared to those infected with the non-opsonized bacteria. Coincidently, the production of NO was also higher in macrophages infected with opsonized BCG (maximal NO production at 48 h of incubation). The obtained results demonstrate that repeated inoculations of BCG effectively activate the humoral immune response, justifying the use of BCG as a live recombinant vaccine vector to insert genes encoding virulence factors controlled by antibodies.
文摘Recalcitrant warts can accurately be defined as warts that persist after six months of conventional therapy. Up to one-third of non-genital warts, especially periungual and plantar warts, become recalcitrant. Traditional treatment options for warts include topical salicylic acid, cryotherapy, and electrocautery;however, patients with recalcitrant warts remain a major therapeutic challenge. There is evidence that immunotherapy can clear recalcitrant warts if traditional treatment fails. Given this, clinical studies published in PubMed and Google Scholar that used Bacillus Calmette-Guerin (BCG), Mycobacterium Indicus Pranii (Mw vaccine), and purified protein derivative (PPD) as immunotherapy for wart, were reviewed in this study. Neither of these treatments has been subjected to a randomized controlled trial, thus to date, there are no standardized protocols to use them. Our review highlights the scientific facts in the clinical applications of the previous options to treat recalcitrant warts and investigate the differences among them, concerning efficacy, adverse effects, dosage, and route of administration.
文摘We observed a 76-year-old man who presented “acute kidney-lung failure” 9 months after intravesical Bacillus Calmette-Guérin (BCG) adjuvant treatment for a T1 bladder cancer. He had inflammatory infiltration on chest radiography and required dialysis for acute renal failure. A percutaneous renal biopsy was performed and revealed tubulointerstitial nephritis with a moderate eosinophilic infiltrate without granulomatous lesion. After a few days, an open lung biopsy was also done due to respiratory deterioration. The anatomopathologic specimen demonstrated moderate fibrosis with lympho-neutrophilic infiltration and few aspecific granulomatous lesions without caseous necrosis. Sarcoidosis was suspected and high dose oral methylprednisolone was started. Three weeks later, Mycobacterium bovis was identified by Polymerase Chain Reaction on open lung biopsy. He responded well to steroids and tuberculostatic tri-therapy. After one month of immunosuppressive treatment, renal function was resolved and hemodialysis could be discontinued. Despite the frequent use of adjuvant BCG immunotherapy, systemic complications such as hepatitis, pneumonitis, spondylodiscitis or multiorgan failure are rare (<1%). Hematogenous dissemination which occurs a few weeks after traumatic instillations is usually suspected but not demonstrated because of absence of mycobacterium in histological specimen. Our case differs from those previously reported by the simultaneous presence of acid-fast bacilli highlighted on lung samples. We discuss the pathophysiology of BCG complications, the use of prophylactic or therapeutic treatment and recommend guidelines to prevent such complications.
基金ThisprojectwassupportedbyagrantfromNationalNaturalSciencesFoundationofChina (No 39870 6 6 3)
文摘The BALB/c mice were immunized with rMS Sj26GST and rBCG Sj26GST vaccine in Schistosoma japonicum by subcutaneous injection After they were immunized for 8 weeks, the eyeballs were removed to get blood and macrophages of abdominal cavity and spleen cells were harvested The lymphocytic stimulating index (SI) was used to measure the cellular proliferating ability and NO release was used to measure the phagocytic activity of the macrophages By using ELISA kit, the levels of interleukin 2 (IL 2) and interferon γ (IFN γ) in serum and the splenic lymphocytic cultured supernatant were detected The results showed that after the mice were immunized with 10 6 CFU of rMS Sj26GST and rBCG Sj26GST vaccine separately by subcutaneous injection, proliferating ability of splenic lymphocytes in the mice showed no difference ( P >0 05), but both were significantly increased as compared with that in the control group( P <0 05); The contents of NO in the intraperitoneal macrophages of rMS Sj26GST vaccine group were significantly lower than in the control group ( P <0 001) and rBCG Sj26GST vaccine group ( P <0 01); The levels of serum IL 2 in the rMS Sj26GST vaccine group were significantly increased as compared with that in the control group ( P <0 001), vector group ( P <0 01) and rBCG Sj26GST vaccine group ( P <0 05); The contents of serum IFN γ in the rMS Sj26GST vaccine group were significantly increased as compared with that in the control group ( P <0 01) and rBCG Sj26GST vaccine group ( P <0 05) The contents of IFN γ in the cultured supernatant were significantly lower than those of rBCG Sj26GST vaccine group ( P <0 001), but were significantly increased as compared with that in the control group ( P <0 01) It was indicated that both vaccines could enhance the immune response of the mice, but rMS Sj26GST vaccine had stronger immunogenicity than rBCG Sj26GST vaccine
