To compare the effects of polysaccharide nucleic acid fraction of bacillus calmette guerin (BCG-PSN) and thymopeptides on T-lymphocytes of normal and immunosuppressed mice, CD4+ and CD8+ T-lymphocyte subsets of single nucleic cell in thymus, spleen and peripheral blood were detected successively by flow cytometry after application of BCG-PSN and thymopeptides. Meanwhile, CD4+/CD8+ ratio was also calculated. The results showed that both BCG-PSN and thymopeptides could decrease the proportion of CD4+ CD8+ T-lymphocyte subsets in the thymus, at the same time increase CD4+ T-lymphocyte, CD8+ T-lymphocyte proportion in the three tissues. The fluctuation in amplitude was greater in thymopeptides group than that in BCG-PSN group. It is concluded that acting location of thymopeptides is in thymus, its stimulating action is stronger than that of BCG-PSN, while BCG-PSN not only accelerates the differentiation in thymus, but also has some direct stimulation to peripheral CD4+ T-lymphocytes, and can maintain CD4+/CD8+ ratio within normal range. So, BCG-PSN is safer.
Adjuvants, Immunologic/*pharmacology ; Immunocompromised Host ; Mycobacterium bovis/*chemistry ; Nucleic Acids/pharmacology ; Peptide Fragments/*pharmacology ; Polysaccharides, Bacterial/*pharmacology ; T-Lymphocyte Subsets/*drug effects ; Thymus Gland/chemistry

A case of tuberculosis is reported in an eight-year-old, male, elk (Cervus elaphus nelsoni). The elk showed severe coughing, respiratory distress, abdominal breathing, anorexia, and severe progressive emaciation in the elk farm. At necropsy, the elk appeared in poor body condition. Mild enlargement of retropharyngeal and submandibular lymph node was observed in the head. Diffuse fibrinous pleuritis and purple red lobar pneumonia were found in the thorax. Well demarcated numerous dark yellow discrete or confluent nodules from 0.3 to 2 cm in diameter were scattered in the whole lung. Bronchial and mediastinal lymph nodes were also enlarged. Histopathologically, lungs had typical classical tuberculous granulomas, multiple abscesses, and numerous macrophages and Langhans giant cells infiltration in alveolar lumen. In the lymph nodes, there were small clusters of necrosis and infiltration of numerous macrophages, epithelioid cells, and Langhans giant cells. With the acid-fast staining, numerous mycobacteria were revealed in the lung and lymph nodes. According to this study, there are differences of the histopathologic lesions and the numbers of acid-fast bacilli in the lesions between this elk and cattle. Mycobacterium bovis was confirmed as a causative agent in this elk using bacterial isolation, biochemical characteristics, and PCR technique. The isolate was negative for niacin test, nitrate reductase, and pyrazinamidase. This is a first report for bovine tuberculosis of farmed elk in Asia.
Animals ; DNA, Bacterial/chemistry/genetics ; Deer/*microbiology ; Fatal Outcome ; Korea ; Lung/microbiology/pathology ; Male ; Mycobacterium bovis/genetics/*isolation&purification ; Polymerase Chain Reaction ; Tuberculosis/microbiology/*veterinary

To compare the effects of polysaccharide nucleic acid fraction of bacillus calmette guerin (BCG-PSN) and thymopeptides on T-lymphocytes of normal and immunosuppressed mice, CD4+ and CD8+ T-lymphocyte subsets of single nucleic cell in thymus, spleen and peripheral blood were detected successively by flow cytometry after application of BCG-PSN and thymopeptides. Meanwhile, CD4+/CD8+ ratio was also calculated. The results showed that both BCG-PSN and thymopeptides could decrease the proportion of CD4+ CD8+ T-lymphocyte subsets in the thymus, at the same time increase CD4+ T-lymphocyte, CD8+ T-lymphocyte proportion in the three tissues. The fluctuation in amplitude was greater in thymopeptides group than that in BCG-PSN group. It is concluded that acting location of thymopeptides is in thymus, its stimulating action is stronger than that of BCG-PSN, while BCG-PSN not only accelerates the differentiation in thymus, but also has some direct stimulation to peripheral CD4+ T-lymphocytes, and can maintain CD4+/CD8+ ratio within normal range. So, BCG-PSN is safer.
Adjuvants, Immunologic ; pharmacology ; Animals ; Female ; Immunocompromised Host ; Male ; Mice ; Mycobacterium bovis ; chemistry ; Nucleic Acids ; pharmacology ; Peptide Fragments ; pharmacology ; Polysaccharides, Bacterial ; pharmacology ; T-Lymphocyte Subsets ; drug effects ; Thymus Gland ; chemistry

New anti-tubercular drugs and drug targets are urgently needed to reduce the time for treatment and also to identify agents that will be effective against Mycobacterium tuberculosis persisting intracellularly. Mycobacteria have a unique cell wall. Deletion of the gene for arylamine N-acetyltransferase (NAT) decreases mycobacterial cell wall lipids, particularly the distinctive mycolates, and also increases antibiotic susceptibility and killing within macrophage of Mycobacterium bovis BCG. The nat gene and its associated gene cluster are almost identical in sequence in M. bovis BCG and M. tuberculosis. The gene cluster is essential for intracellular survival of mycobacteria. We have therefore used pure NAT protein for high-throughput screening to identify several classes of small molecules that inhibit NAT activity. Here, we characterize one class of such molecules-triazoles-in relation to its effects on the target enzyme and on both M. bovis BCG and M. tuberculosis. The most potent triazole mimics the effects of deletion of the nat gene on growth, lipid disruption and intracellular survival. We also present the structure-activity relationship between NAT inhibition and effects on mycobacterial growth, and use ligand-protein analysis to give further insight into the structure-activity relationships. We conclude that screening a chemical library with NAT protein yields compounds that have high potential as anti-tubercular agents and that the inhibitors will allow further exploration of the biochemical pathway in which NAT is involved.
Antitubercular Agents ; chemistry ; isolation & purification ; pharmacology ; Arylamine N-Acetyltransferase ; antagonists & inhibitors ; chemistry ; Enzyme Inhibitors ; chemistry ; isolation & purification ; pharmacology ; High-Throughput Screening Assays ; Humans ; Mycobacterium bovis ; drug effects ; enzymology ; genetics ; Mycobacterium tuberculosis ; drug effects ; enzymology ; genetics ; Protein Conformation ; Structure-Activity Relationship ; Triazoles ; chemistry ; isolation & purification ; pharmacology

Cancer is the main cause of death in developed countries. However, in underdeveloped countries infections and parasitic diseases are the main causes of death. There are raising scientific evidences indicating that parasitic infections induce antitumor activity against certain types of cancers. In this study, the effects of Toxoplasma gondii and Toxocara canis egg antigens in comparison with Bacillus Calmette Guerin (BCG) (known to have anticancer distinctive) on WEHI-164 fibosarcoma transplanted to BALB/c mice was investigated. Groups of 6 male BALB/c mice injected with T. gondii antigen, BCG, or T. canis egg antigen as case groups and alum alone as control groups. All mice were then challenged with WEHI-164 fibrosarcoma cells. The mice were examined for growth of the solid tumor and the tumor sizes were measured every other day up to 4 wk. The mean tumor area in T. gondii, BCG, or alum alone injected mice in 4 different days of measurements was 25 mm2, 23 mm2, and 186 mm2 respectively. Also the mean tumor area in T. canis injected mice in 4 different days was 25.5 mm2 compared to the control group (alum treated) which was 155 mm2. T. gondii parasites and T. canis egg antigens induced inhibition of the tumor growth in the fibrosarcoma mouse model. We need further study to clarify the mechanisms of anti-cancer effects.
Animals ; Antigens, Helminth/isolation & purification/pharmacology/*therapeutic use ; Antigens, Protozoan/isolation & purification/pharmacology/*therapeutic use ; Antineoplastic Agents/isolation & purification/pharmacology/*therapeutic use ; Chemoprevention/*methods ; Female ; Fibrosarcoma/pathology/*prevention & control ; Male ; Mice ; Mice, Inbred BALB C ; Mycobacterium bovis ; Toxocara canis/*chemistry ; Toxoplasma/*chemistry

OBJECTIVETo investigate the protective effects of Hongbeiyegen (HBYG) against immunological liver injury induced by bacille Calmette-Guerin (BCG) and lipopolysaccharide (LPS).

METHODSImmunological liver injury was induced in rats by BCG and LPS injected via the tail vein. The liver index, thymus index and spleen index were calculated and the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and nitric oxide (NO) and liver homogenate contents of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) were determined.

RESULTSHBYG significantly improved the liver index, thymus index and spleen index, and reduced the serum levels of ALT, AST and NO, and as the liver homogenate contents of TNF-alpha and IL-1beta.

CONCLUSIONHBYG offers obvious protective effects against immunological injury liver in mice.

Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Chemical and Drug Induced Liver Injury ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Euphorbiaceae ; chemistry ; Female ; Interleukin-1beta ; metabolism ; Lipopolysaccharides ; Liver ; drug effects ; metabolism ; pathology ; Liver Diseases ; immunology ; prevention & control ; Male ; Mice ; Mice, Inbred Strains ; Mycobacterium bovis ; Nitric Oxide ; blood ; Phytotherapy ; Plant Roots ; chemistry ; Treatment Outcome ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo observe the effect of polysaccharide ATPS-2 from Armillariella tabescens on the immunological liver injury in mice induced by BCG plus LPS.

METHODBCG and LPS were adopted to establish BCG plus LPS liver injury model in mice. The content of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and NO, the activity of superoxide dismutase (SOD) and malondiadehyde (MDA) content of liver homogenate in mice were measured by colorimetric method. The content of tumor necrosis factor-alpha (TNF-alpha), interleukin-1 (IL-1), on serum were measured by enzyme linked immunosorbent assay (ELISA) , and T- and B-lymphocyte proliferation were measured by MTT. Index of liver, spleen and thymus were calculated after treatment.

RESULTPolysaccharide ATPS-2 from A. tabescens (25, 50, 100 mg x kg(-1)) could obviously reduce the high level of ALT, AST, NO and TNF-alpha, IL-1 on serum, inhibit the high level of MDA, increase the low activity of SOD in liver homogenate and enhance T-and B-lymphocyte proliferation, elevate the spleen, thymic index and decrease liver index of the mice to different extent.

CONCLUSIONPolysaccharide ATPS-2 from A. tabescens had apparently protective effects in the immunological liver injury mice induced by BCG plus LPS.

Agaricales ; chemistry ; Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; B-Lymphocytes ; cytology ; Cell Proliferation ; drug effects ; Chemical and Drug Induced Liver Injury ; Interleukin-1 ; blood ; Lipopolysaccharides ; Liver ; metabolism ; pathology ; Liver Diseases ; immunology ; metabolism ; pathology ; Male ; Malondialdehyde ; metabolism ; Mice ; Mycobacterium bovis ; Nitric Oxide ; blood ; Polysaccharides ; isolation & purification ; pharmacology ; Protective Agents ; isolation & purification ; pharmacology ; Random Allocation ; Superoxide Dismutase ; metabolism ; T-Lymphocytes ; cytology ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo detect the effects of Polyporus polysaccharide (PPS), Bacillus Calmette-Guerin (BCG), and their combination on the nuclear factor kappa B (NF-κB) signaling pathway associated-gene expression and investigate the molecular mechanisms of the toxic-reducing effect of PPS in coordination with BCG against bladder cancer.

METHODSAfter T739 cells were treated with PPS, BCG and their combination, the changes in mRNA and protein expression of inhibitor of kappa B kinase beta (IKKβ), NF-κB subunit p65 (NF-κB p65), intracellular adhesion molecule 1 (ICAM1) and chemokine (C-c motif) ligand 2 (CCL2) in bladder cancer cell line T739 were determined by relative quantitative real-time PCR, Western blot, and flow cytometry (FCM). NF-κB p65 DNA-binding activity in T739 cell was detected by biotinylated probe-ELISA, and NF-κB p65 nuclear expression in T739 cell was observed by immunohistochemistry.

RESULTSCompared with the T739 control group, the mRNA expression of IKBKB (IKKβ), Rel A (NF-κB p65), ICAM1 and CCL2 in T739 cells treated with BCG were increased obviously (Ratio>2.0), as well as the expression of IKKβ, CCL2 and ICAM1 proteins. Meanwhile, NF-κB p65 DNA-binding activity and NF-κB p65 nuclear expression in T739 cells treated with BCG were up-regulated significantly (P<0.05). Compared with the control, the increased expression in T739 cells were simultaneously down-regulated after PPS treatment, except for ICAM1 protein expression. With cells treated with a combination of BCG and PPS, the expression of genes associated with the NF-κB signaling pathway, such as IKBKB, ICAM1 and CCL2, were all down-regulated compared to the BCG group, as well as Rel A mRNA expression, NF-κB p65 DNA-binding activity and NF-κB p65 nuclear expression.

CONCLUSIONSPPS could inhibit the over-activation of the NF-κB signaling pathway induced by BCG in bladder cancer cells and accordingly attenuate the adverse reactions to BCG therapy.

Cell Line, Tumor ; Cell Nucleus ; drug effects ; metabolism ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Mycobacterium bovis ; drug effects ; NF-kappa B ; metabolism ; Neoplasm Proteins ; genetics ; metabolism ; Polyporus ; chemistry ; Polysaccharides ; pharmacology ; Signal Transduction ; drug effects ; Urinary Bladder Neoplasms ; genetics ; microbiology ; pathology

BACKGROUND: BCG, a potent inducer of Th1 immune response, has been suggested to suppress Th2 response which is known to mediate IgE-mediated allergic disorders, in particular allergic asthma. Schultz-Dale reaction is known to be a model of IgE-mediated hypersensitivity. This study was done to investigate whether BCG infection suppresses the Schultz-Dale reaction by inhibiting Th2 response and allergen-specific IgE production. METHODS: Twenty-four Sprague-Dawley rats were sensitized and provoked with ovalbumin (OVA). A pretreatment of 6 x 10(4) colony forming units of BCG or saline was done 7 days before sensitization. The Schultz-Dale reaction was represented as tracheal smooth muscle contractions to 50 micrograms/mL OVA challenge in vitro. Serum OVA-specific IgE levels and IFN-gamma and IL-4 concentrations in bronchoalveolar lavage fluid (BALF) were measured. RESULTS: The Schultz-Dale reaction and serum OVA-specific IgE levels were significantly decreased in BCG infected and OVA sensitized rats compared with only sensitized rats (p < 0.01 and p < 0.05, respectively). As compared with only sensitized rats, IL-4 concentration and a ratio of IFN-gamma:IL-4 in BCG infected and OVA sensitized rats were significantly decreased (p < 0.001) and increased (p < 0.05), respectively. The Schultz-Dale reaction was correlated with OVA-specific IgE levels (r = 0.50, p < 0.05), IL-4 concentration (r = 0.69, p < 0.001), and ratio of IFN-:IL-4 (r = -0.44, p < 0.05). OVA-specific IgE levels were correlated with IL-4 concentration (r = 0.61, p < 0.01) and ratio of IFN-gamma:IL-4 (r = -0.48, p < 0.05). CONCLUSION: These findings suggest that BCG infection prior to allergen sensitization may inhibit Schultz-Dale reaction developed in the sensitized rat tracheal smooth muscle via the suppressive effects of Th2 immune response and allergen-specific IgE production.
Animal ; Bronchial Provocation Tests ; Bronchoalveolar Lavage Fluid/*chemistry ; Comparative Study ; Cytokines/analysis ; Disease Models, Animal ; Hypersensitivity, Immediate/*immunology ; Immunization ; Immunoglobulin E/*analysis ; Male ; Mycobacterium bovis/*immunology ; Probability ; Rats ; Rats, Sprague-Dawley ; Reference Values ; Sensitivity and Specificity ; T-Lymphocytes/*immunology ; Tuberculosis/*immunology/veterinary