75 results about "Bacille Calmette Guerin" patented technology

A vaccine against Mycobacteria tuberculosis (Mtb) is provided. The vaccine comprises a recombinant Bacille Calmette-Guerin (BCG) subunit-based vaccine in which one or more Mtb antigens and one or more Mtb resuscitation or reactivation antigens are overexpressed, and in which at least a portion of the DosR regulon is up-regulated. The vaccine is protective against active Mtb infection both pre- and post-exposure to Mtb, and thus prevents disease symptoms due to the recurrence of a latent Mtb infection.

A vaccine against Mycobacteria tuberculosis (Mtb) is provided. The vaccine comprises a recombinant Bacille Calmette-Guerin (BCG) subunit-based vaccine in which one or more Mtb antigens and one or more Mtb resuscitation or reactivation antigens are overexpressed, and in which at least a portion of the DosR regulon is up-regulated. The vaccine is protective against active Mtb infection both pre- and post-exposure to Mtb, and thus prevents disease symptoms due to the recurrence of a latent Mtb infection.

Methods and compositions for use of human dendritic cells to activate T cells for immunotherapeutic responses against primary and metastatic cancer are disclosed. In one embodiment, human dendritic cells exposed to a tumor associated antigen, or an antigenic fragment thereof in combination with bacillus Calmette-Guerin (BCG), are administered to a cancer patient to activate a predominantly CD8+T cell response in vivo. In an alternate embodiment, human dendritic cells are exposed to a tumor associated antigen or a specific antigenic peptide in combination with BCG in vitro and incubated or cultured with primed or unprimed T cells to activate a predominantly CD8+T cell response in vitro. The activated T cells are then administered to a cancer patient. Antigen in combination with BCG is processed by dendritic cells through the MHC-CLASS I compartment which provides for a predominantly CD8+T cell response. The addition of LPS provides for a greater number of mature dendritic cells enhancing the T cell response to antigen. Methods and compositions for human dendritic cells with extended life span and cryopreserved dendritic cells are disclosed.

The invention discloses a construction method of mycobacterium tuberculosis fusion protein Mtb 10.4-Hsp16.3, an expression method thereof, a purification method thereof and application thereof. The construction method comprises the following steps: performing the PCR (polymerase chain reaction) amplification to Mtb 10.4 gene and Hsp16.3 gene, and sequentially inserting the amplified genes into a multi-clone site of a cloning vector to construct a recombinant vector; then expressing in the colibacillus; and finally purifying to obtain the fusion protein Mtb 10.4-Hsp16.3, wherein the fusion protein can be applied in a tuberculosis subunit vaccine. The invention has the advantages that the fusion protein vaccine can induce the higher cell of the animal to react with body fluid immunoreaction; the animal challenge test shows that the vaccine plays a part in immunity enhancing on the basis of the BCG (bacille calmette guerin) immunity, so that the protective effect of the BCG can be improved and prolonged; and the vaccine has no obvious side effects.

An inactivated Polio Vaccine derived from Sabin strain for safe and effective immunization against Poliomyelitis is provided. A process of preparation for such vaccine and formulations thereof are also provided. Administration of the vaccine of the present invention along with other antigens provides immunization not only against polio infection but also against other pathogens causing Hepatitis C. Hepatitis D. Hepatitis E. Meningitis A. Meningitis B. Meningitis C. Meningitis W. Meningitis Y. Pnemococcal (23 valent or more). Smallpox, Typhoid, Bacille Calmette Guerin, Tuberculosis. Human Immunodeficiency Virus. Anthrax or the like, to which children or adults not immunized earlier are susceptible, particularly to which children are susceptible.

The invention provides construction of TRAIL (Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand) recombinant bacille calmette guerin (rBCG), and relates to a shuttle expression vector comprising a signal peptide fragment of a major secretory antigen Ag85B of BCG and a gene fragment of a TRAIL and a construction method thereof. The obtained shuttle expression vector pMV261-Ag85B-TRAIL is used for constructing rBCGTRAIL, and can be applied to preparation of TRAIL rBCG for treating superficial bladder tumors, preventing postoperative recurrence thereof and preventing tuberculosis. The rBCG has dual functions of TRAIL and BCG, so that cooperative and synergistic actions of the TRAIL and BCG can be better brought into play; rBCG-TRAIL can secrete TRAIL, and the using amount of the rBCG-TRAIL can be lower than that of the BCG under the condition that the same or better immune effect is achieved, so that the toxic or side effect is reduced; and the rBCG-TRAIL can directly secrete TRAIL efficiently on a certain part, so that tumor cells can be killed in cooperation with the rBCG-TRAIL, and high cost caused by the use of a foreign cell factor is avoided.

The present invention describes an immunogenic formulation to be used in mammals against the respiratory syncytial virus (RSV), consisting in the Calmette-Guérin bacillus (BCG) strain or other attenuated Mycobacterium strain that expresses heterologously at least one protein or immunogenic fragment of the RSV subtype A or RSV subtype B strains, originated from proteins NS1, NS2, N, P, M, SH, M2 (ORF1), M2 (ORF2), L, F or G. The genetic material that encodes for these proteins or immunogenic fragments is inserted into the BCG genome or extrachromosomally in one or several copies, which expression is regulated by endogenous or exogenous BCG promoters, either constitutive or inducible. The viral proteins or immunogenic fragments can be expressed by BCG as cytoplasmic-soluble, extracellularly-secreted or cell membrane-bound proteins. The preparation can further contain combinations of previously described formulations. The formulation can be stabilized by freeze-drying (conservation range from 4° C. and 25° C.) or through low temperatures (−80° C.) in a buffered saline solution to be conserved prior to its use.

The invention relates to a preparation method of a diphasic quick differential medium of mycobacterium tuberculosis, a method for quickly differentiating the mycobacterium tuberculosis by using the medium, a method for testing the drug resistance on antituberculous drugs caused by the mycobacterium tuberculosis and a method for quickly culturing a bacillus Calmette and Guerin vaccine by using a liquid medium in the medium. The diphasic medium prepared by adopting the method can be used for quickly culturing the mycobacterium tuberculosis, the diphasic quick culture medium is capable of differentiating the mycobacterium tuberculosis within 2-5 days, the differentiated mycobacterium tuberculosis can be subjected to drug resistance detection of the antituberculous drugs such as rifampicin, isoniazide and streptomycin by virtue of a reverse transcription PCR (polymerase chain reaction) method, and the liquid medium prepared by adopting the method is capable of finishing the culture of the bacillus Calmette and Guerin vaccine within 6-9 days.

The invention provides antituberculous recombinant bacillus calmette-guerin (BCG delta mazG) with a mazG gene deleted. The mazG gene in a genome of the antituberculous recombinant bacillus calmette-guerin with the mazG gene deleted is replaced with a hygromycin resistant gene. The invention further provides a preparation method of the BCG delta mazG and application thereof in preparing a recombinant vaccine for preventing tuberculosis. After the BCG delta mazG is inoculated subcutaneously into a mouse, a T cell immune response is obviously improved, after the immunized mouse is infected with a mycobacterium tuberculosis standard strain H37Rv, the bacterial load of the infected animal lung and spleen is significantly decreased, the pathological degree is relieved, and after the immunized mouse is infected for five weeks, a T cell secondary immune response is significantly improved. It is found through bacterial in-vivo survival tests that after high-dose intravenous inoculation is conducted, the BCG delta mazG can survive on viscera of the spleen, the lung, the liver and the kidney of the mouse continuously for 20 w or above, and the amount of bacteria of a BCG delta mazG recombinant strain is obviously increased compared with a wild strain. It is proved through experiments that the BCG delta mazG recombinant strain has antituberculous immunogenicity, protective efficacy and in-vivo persistence activity which are significantly improved, and the antituberculous recombinant bacillus calmette-guerin (BCG delta mazG) with the mazG gene deleted is expected to become one of the novel candidate vaccines for preventing tuberculosis infection.

The present invention describes an immunogenic formulation to be used in mammals against the respiratory syncytial virus (RSV), consisting in the Calmette-Guérin bacillus (BCG) strain or other attenuated Mycobacterium strain that expresses heterologously at least one protein or immunogenic fragment of the RSV subtype A or RSV subtype B strains, originated from proteins NS1, NS2, N, P, M, SH, M2 (ORF1), M2 (ORF2), L, F or G. The genetic material that encodes for these proteins or immunogenic fragments is inserted into the BCG genome or extrachromosomally in one or several copies, which expression is regulated by endogenous or exogenous BCG promoters, either constitutive or inducible. The viral proteins or immunogenic fragments can be expressed by BCG as cytoplasmic-soluble, extracellularly-secreted or cell membrane-bound proteins. The preparation can further contain combinations of previously described formulations. The formulation can be stabilized by freeze-drying (conservation range from 4° C. and 25° C.) or through low temperatures (−80° C.) in a buffered saline solution to be conserved prior to its use.

The invention discloses a construction method of a drug evaluation model for dermal pathology of a tuberculosis rabbit. The method comprises: selecting a rabbit, injecting an immune drug into rabbit, conducting BCG (Bacillus Calmette Guerin) vaccine immune injection on the 15th to 20th day of drug intervention, and carrying out bacterial and pathological examination so as to construct a pathological model. The method of the invention establishes a pathological model of drug intervention on drug cutaneous tuberculosis with obvious symptoms so as to conduct visual research on tuberculosis pathology and bacterial pathogenicity, thus providing a visual animal model for vaccines and screening of drugs treating a tuberculosis necrotic liquefied cavity. And the model is stable and repeatable. The invention establishes a drug intervention procedure and observation indexes for research of drug intervention on a tuberculosis granuloma liquefaction process, and also provides a research basis for probing an immune mechanism about the formation of a tuberculosis liquefied cavity.

The invention discloses a modified new coronavirus SARS-CoV-2S protein, a recombinant plasmid and a recombinant bacillus calmette guerin vaccine constructed by the modified new coronavirus SARS-CoV-2S protein, and application of the recombinant bacillus calmette guerin vaccine, and belongs to the technical field of biological agents. Through modifying a partial sequence of gene promoter region and a partial sequence of gene terminator region, the modified new coronavirus S gene complete sequence can be smoothly expressed in an escherichia coli-mycobacterium tuberculosis shuttle plasmid pMV261 to obtain a recombinant plasmid pMVS; the recombinant plasmid pMVS is introduced into BCG through electric transformation, and recombinant BCG (rBCG) is obtained; the rBCG can successfully express S protein, arouse human immune response and induce antibody generation to prevent virus invasion, and the constructed rBCG is a subunit vaccine, has a better protection effect than parent BCG, can prevent or treat new coronavirus and mycobacterium tuberculosis, and has huge social benefits.

The invention discloses a method for preparing a pure protein derivative of tubercle bacillus by using a mycobacterium tuberculosis attenuated strain H37Ra, and belongs to the technical field of biological protein preparation. The mycobacterium tuberculosis attenuated strain CMCC93020 (H37Ra) is used as a strain, and a finished product of the pure protein derivative of the tubercle bacillus is obtained through amplification, inactivation, salting-out precipitation, degerming filtration, inspection, dilution and subpackaging. The defects in the prior art that strains have great harm to operators and the environment and have high requirements for production conditions (P3 production workshops) are overcome, and finally the protein which can be used for clinical diagnosis of tuberculosis, screening of bacillus calmette-guerin vaccine inoculation objects, monitoring of organism immune responses after bacillus calmette-guerin vaccine inoculation and epidemiological monitoring is obtained.

The invention provides a technical method for identifying human mycobacterium tuberculosis, mycobacterium bovis and bacillus calmette-guerin (BCG). Five target genes, i.e., 16S rRNA, IS6110, Rv2659c,ESAT6 and Rv1510 are selected for multiple PCR amplification, the mycobacterium tuberculosis can be rapidly and accurately identified, and interference caused by the mycobacterium bovis and BCG low-virulent strains (BCG) is effectively removed.

The present invention relates to specific skin reagent for diagnosing tuberculous infection and active tuberculosis, and belongs to the field of medical immunological diagnosis technology. The specific skin reagent consists of diluent liquid and dissolved tuberculous allergen, and the tuberculous allergen is tuberculous mycobacterium ESAT6 protein. The present invention also proposes applying tuberculous mycobacterium ESAT6 protein for human skin test as tuberculous allergen to detect tuberculous infecting person and tuberculosis patient and identify the specific reaction of inoculated Calmette-Guerin bacillus vaccine. The present invention can induce the immune response of tuberculous infecting person. The reagent can induce delayed allergic reaction of tuberculous infecting person and can identify BCG inoculation from the allergic reaction of non-tuberculous mycobacterium and tuberculous mycobacterium infection.

The invention discloses a medical immunity diagnose technique, especially providing a bacillus tubercle mycobacterium gold mark diagnose agent based on combined protein. Wherein, it uses B cell antigen determinants of bacillus tubercle mycobacterium main excrete proteins as ESAT6, MPT64, PstS-1, Ag85B to combine a new group of protein as antigen, as the specific test antibody in the clinic serology diagnosis of phthisis. The invention combines the B cell antigen determinants of bacillus tubercle mycobacterium main excrete proteins to be displayed by gene project, and purified to attain one new protein, and via gold mark filter method to test the serum of tuberculosis patient. The invention has high sensitivity, simple operation, and high speed, which can identify the sensitized condition caused by contacting the non- bacillus tubercle mycobacterium or inoculating beg vaccine, and the real bacillus tubercle mycobacterium. It can be used in the clinic serology diagnosis of clinic tuberculosis.

Embodiments of the invention relate to methods and systems for the detection of Mycobacterium tuberculosis. Mycobacterium tuberculosis kills more than one million people each year. To better understand why M. tuberculosis is virulent and to discover chemical markers of this pathogen, we compared its lipid profile to that of the attenuated but related mycobacterium, Mycobacterium bovis Bacille Calmette Guerin (BCG). This strategy identified previously unknown compounds that are specific to M. tuberculosis, e.g. 1-tuberculosinyladenosine, N6-tuberculosinyladenosine, and various tuberculosinyladenosines having mycolic acids, produced by the Rv3378c enzyme.

By taking guide functions of specific cellular immunity and monoclonal antibody immunity generated in vaccine inoculation as theoretical basis, the effective component of bacillus calmette Guerin (BCG), namely muramyl dipeptide (MDP), and a monoclonal antibody Rituximab of lymphoma cell membrane resistant antigen CD20 are combined to prepare a muramyl dipeptide-anti-CD20 immune conjugate, namely MDP-Rituximab, wherein in-vivo and in-vitro anti-tumour effects of immunocompetent cells subjected to BCG immunity under mediation of MDP-Rituximab are known. The conjugate disclosed by the invention keeps the immunogenicity of MDP, stimulates organisms to generate specific T cell immune response, and brings MDP to be around residual lymphoma cells with the help of immune guide function that a Rituximab antibody is combined with lymphoma cell membrane CD20 molecules; the final purpose of eliminating residual tumours is achieved through immune response mediated by MDP induced T cells; therefore, due to the method, the drug resistance caused by exclusive use of Rituximab is overcome; furthermore, the use amount of the antibody is reduced; and the application prospect of the muramyl dipeptide-anti-CD20 immune conjugate in malignant lymphoma is predicted.

This invention involves reorganization BCG vaccine for secretion of people interferon- alpha2a, and structuring method. Utilize gene engineering to clone BCG vaccine Ag85B signal peptide part and the genes of people IFN - alpha 2a playing a secreting role to pMV261 to receive the BCG vaccine shuttling expression carrier pMSIFN - alpha 2a. Adopt electric boring technology to lead pMSIFN - alpha 2a into BCG to recombinate rBGGIFN - alpha2a. Depending on rBGGIFN - alpha 2a duplicating in BCG and the secretion function of the signal peptide, can secrete IFN - alpha2a efficiently.

The invention relates to a construction and an application of a novel PhoPR gene overexpressed BCG (Bacillus Calmette-Guerin) mutant strain. The construction comprises the following steps: firstly, converting a carrier containing a BCG overexpressed PhoPR gene into BCG, screening, detecting and confirming a successfully converted strain as the novel PhoPR gene overexpressed BCG mutant strain. Theinvention relates to the novel PhoPR gene overexpressed BCG mutant strain, namely, BCG: PhoPR strain. Such a strain is mainly used for researching the functions of BCG PhoPR gene overexpressed associated protein PhoPR at the aspects of growth of mycobacterium tuberculosis in host, drug tolerance, permeability regulation, acid resistance, pathogenicity, and the like, and is used for developing andresearching novel vaccines for interfering and treating tuberculosis.

The invention belongs to the biomedical field, in particular to the use of Bacteroides fragilis in the preparation of drugs for the treatment and prevention of tuberculosis. The invention also provides the application of Bacteroides fragilis in the preparation of pharmaceutical compositions, foods, health products and food additives for treating and / or preventing tuberculosis. The Bacteroides fragilis provided by the invention can effectively treat and / or prevent tuberculosis and enhance the immune protection effect of tuberculosis bacillus Calmette-Guerin vaccine (BCG), and has wide application prospect.

The invention discloses 2-amino-2-deoxy-D-glucose and an application of hydrochloride, sulfate and myo-inositol thereof in preparing a mycobacterium tuberculosis culture medium or a bacillus calmette guerin vaccine mycobacterium culture medium. The 2-amino-2-deoxy-D-glucose and the hydrochloride, sulfate and myo-inositol thereof serving as a growth factor of mycobacterium tuberculosis have good application prospects in developing a novel mycobacterium tuberculosis rapid culture medium, and can be applied to the rapid diagnosis of tuberculosis, type identification and drug sensitivity detection; and by adding one of the four compounds or the mixture of several ones of the four compounds into the culture medium, the flora density in the liquid culture medium is far higher than that of a blank control group, the macroscopic bacterial colony appears one week earlier in the solid culture medium than in the blank control group, and the bacterial plaque is obviously larger than that of the blank control group.

The invention discloses a BCG (bacillus calmette-guerin) vaccine complex combined with a nano drug carrier. The BCG vaccine complex is of a complex structure formed by combining the nano drug carrierand BCG vaccine thalli by virtue of an acting force between avidin and biotin protein; the nano drug carrier is nano particles which are coated with a polylactic acid-glycolic acid copolymer, subjected to surface modification by the avidin and have the particle size of 120nm to 180nm; and the surface of each BCG vaccine thallus is combined with one or more nano particles. The BCG vaccine complex disclosed by the invention has very good adhesion and absorption effects on the epithelial layer of the bladder, drug particles are loaded to the surfaces of the BCG thalli, and the BCG is used as thecarrier, so that the uptake of the nano drug carrier in the bladder and tumor targeting are enhanced, and the tumor killing effect of a drug can be remarkably improved. Meanwhile, the drug-loaded BCGcan stimulate strong immune response in part of the bladder and the whole body, so that a combined treatment mode combining immunotherapy and chemotherapy is realized.

The invention relates to a live recombinant Mycobacterium bovis-BCG strain comprising a nucleic acid capable of expression, the nucleic acid encoding at least one protein or polypeptide that exhibits alanine dehydrogenase activity, glutamine synthetase activity, or serine dehydratase activity.