Human Tubercle Bacilli Research Articles

Epidemiological trends of tuberculosis in low and high prevalence countries.

The study of the epidemiology of tuberculosis and the need for a rational approach to the problem of tuberculosis control require a sound knowledge of the risk of transmission of tuberculosis infection from host to host. This knowledge is required today both in countries with a high prevalence and in those with a low prevalence of the disease. The risk of transmission of tuberculosis infection in a given community during a particular period of time is most reliably expressed numerically in terms of a series of average annual infection risks in successive calender years.The infection risk indicates the proportion of the population which will be primarily infected, or reinfected, with tubercle bacilli in the course of one year. The risk is usually expressed as a percentage or as a rate. The annual trend (decrease) in the risk of infection is even more important than the actual level of this risk.Tuberculin surveys in schoolchildren, which are repeated, say every five years, in the same area provide us with the necessary information to make estimates for the annual infection risk. In areas where infections with the socalled atypical mycobacteriae are frequent, simultaneous testing with two "Sensitins", one prepared from human tubercle bacilli and one from atypical mycobacteriae will be of value for estimating the proportion of those infected with virulent bacilli and those infected with other mycobacteriae.

Zn- and Mg- Containing Tricalcium Phosphates-Based Adjuvants for Cancer Immunotherapy

Zn-, and Mg-containing tricalcium phosphates (TCPs) loaded with a hydrothermal extract of a human tubercle bacillus (HTB) were prepared by immersing Zn-TCP and Mg-TCP in HTB-containing supersaturated calcium phosphate solutions. The in vitro and in vivo immunogenic activities of the HTB-loaded Zn-, and Mg-TCPs (Zn-Ap-HTB and Mg-Ap-HTB, respectively) were evaluated as potential immunopotentiating adjuvants for cancer immunotherapy. The Zn-Ap-HTB and Mg-Ap-HTB adjuvants showed no obvious cytotoxicity and more effectively stimulated granulocyte-macrophage colony-stimulating factor (GM-CSF) secretion by macrophage-like cells than unprocessed HTB or HTB-loaded TCP (T-Ap-HTB) in vitro. Zn-Ap-HTB and Mg-Ap-HTB mixed with liquid-nitrogen-treated tumor tissue markedly inhibited the in vivo development of rechallenged Lewis lung carcinoma (LLC) cells compared with T-Ap-HTB and the unprocessed HTB mixed liquid-nitrogen-treated tumor tissue. Zn-Ap-HTB and Mg-Ap-HTB contributed to eliciting potent systemic antitumor immunity in vivo.

Novel Apatite-Pathogen-Associated Molecular Patterns Adjuvants for Cancer Immune Therapy

An apatite-pathogen-associated molecular patterns (PAMPs) adjuvant was prepared by coprecipitation of apatite and PAMPs using a supersaturated calcium phosphate solution supplemented with PAMPs. In this study, we used a hydrothermal extract of human tubercle bacillus (HTB) as PAMPs. The effects of coprecipitation conditions on immunogenic activity were studied. The adjuvants prepared using extract of at 10 and 20 μg/mL showed higher in vitro immunogenic activity than that at 2 μg/mL. The adjuvants prepared under stir at speed of 300 rpm showed better in vitro immunogenic activity than those under still. The adjuvants prepared using 10 μg/mL of extract of HTB under stir at speed of 300 rpm may be promising for cancer immune therapy.

Human pulmonary infections with bovine and environment (atypical) mycobacteria in jos, Nigeria.

To examine sputum specimens from patients with persistent bronchopulmonary disorders for mycobacterium species and to characterize the recovered isolates with a view to determining the extent of involvement of environmental mycobacteria in pulmonary infections. Analytical study using standard microscopy, culture and biochemical test for the identification of mycobacterium species. Jos University Teaching Hospital (JUTH) and 2 referral hospitals: Plateau Specialist Hospital and Evangelical Churches of West Africa (ECWA) Evangel Hospital in Jos, Nigeria. Three hundred and twenty nine (329) volunteer new patients seen at the chest clinic and general out patient departments with broncho-pulmonary disorders. Patients already on anti-tuberculosis were excluded from the study. Subjects were administered antituberculosis drugs and or other treatment regimes after proper diagnosis Sixty-five (65) mycobacterial isolates were obtained and differentiated into human tubercle bacilli, bovine and or environmental (atypical) mycobacteria on the basis of nine identification tests. Of the 65 mycobacterial isolates subjected to the tests, 40 (61.54%) were identified as mycobacterium tuberculosis, 10 (15.38%) as M. bovis and 15 (23.08%) as environmental mycobacteria. Among the environmental group, 9 (20.69%) were classified as M. avium 3 (3.45%) each as M. kansasi and M. fortuitum. The study confirms the involvement of bovine and environmental mycobacteria in pulmonary infections. This may be related to the rising prevalence of HIV/AIDS globally. The need for adequate bacteriological analysis in current-day diagnosis of pulmonary tuberculosis in indicated.

Synthetic Peptides Identify Promiscuous Human Th1 Cell Epitopes of the Secreted Mycobacterial Antigen MPB70

MPB70 is a secreted protein of Mycobacterium bovis and Mycobacterium tuberculosis which stimulates both cellular and humoral immune responses during infection with bovine and human tubercle bacilli. In addition, vaccination with MPB70 has been shown to induce Th1 cell responses and protection in animal models of tuberculosis. The present study was carried out to map the dominant human Th1 cell epitopes of MPB70 in relation to major histocompatibility complex (MHC) class II restriction in healthy subjects showing strong T-cell responses to complex mycobacterial antigens. Peripheral blood mononuclear cells (PBMC) from HLA-DR-typed donors were tested with complex mycobacterial antigens (whole-cell M. tuberculosis and M. tuberculosis culture filtrates), with MPB70 purified from the culture filtrate of M. bovis BCG Tokyo, and with 13 synthetic peptides (25-mers overlapping by 10 residues) covering the sequence of MPB70. The donors that responded to the complex antigens and MPB70 also responded to the cocktail of synthetic MPB70 peptides. Testing of PBMC with individual peptides showed that peptides p5 (amino acids [aa] 61 to 85), p6 (aa 76 to 100), p8 (aa 106 to 130), p12 (aa 166 to 190), and p13 (aa 181 to 193) were most frequently recognized in proliferation and gamma interferon (IFN-gamma) assays. Testing of antigen-specific CD4(+) T-cell lines with the individual peptides of MPB70 confirmed that peptides p8, p12, and p13 contain immunodominant Th1 cell epitopes of MPB70. MHC restriction analysis with HLA-typed donors showed that MPB70 and its immunodominant peptides were presented to T cells promiscuously. The T-cell lines responding to MPB70 and peptides p8, p12, and p13 in IFN-gamma assays mediated antigen-peptide-specific cytotoxic activity against monocytes/macrophages pulsed with the whole-protein antigen or the peptides. In conclusion, the promiscuous recognition of MPB70 and its immunodominant peptide defined epitopes (aa 106 to 130 and 166 to 193) by IFN-gamma-producing Th1 cells supports possible application of this secreted antigen to subunit vaccine design.

Evolution and clonal traits of Mycobacterium tuberculosis complex in Guinea-Bissau.

Two hundred twenty-nine consecutive isolates of Mycobacterium tuberculosis complex from patients with pulmonary tuberculosis in Guinea-Bissau, which is located in West Africa, were analyzed for clonal origin by biochemical typing and DNA fingerprinting. By using four biochemical tests (resistance to thiophene-2-carboxylic acid hydrazide, niacin production, nitrate reductase test, and pyrazinamidase test), the isolates could be assigned to five different biovars. The characteristics of four strains conformed fully with the biochemical criteria for M. bovis, while those of 85 isolates agreed with the biochemical criteria for M. tuberculosis. The remaining 140 isolates could be allocated into one of three biovars (biovars 2 to 4) representing a spectrum between the classical bovine (biovar 1) and human (biovar 5) tubercle bacilli. By using two genotyping methods, restriction fragment length polymorphism analysis with IS6110 (IS6110 RFLP analysis) and spoligotyping, the isolates could be separated into three groups (groups A to C) of the M. tuberculosis complex. Group A (n = 95), which contained the majority of classical human M. tuberculosis isolates, had large numbers of copies of IS6110 elements (mean number of copies, 9) and a distinctive spoligotyping pattern that lacked spacers 33 to 36. Isolates of the major group, group B (n = 119), had fewer IS6110 copies (mean copy number, 5) and a spoligotyping pattern that lacked spacers 7 to 9 and 39 and mainly comprised isolates of biovars 1 to 4. Group C isolates (n = 15) had one to three IS6110 copies, had a spoligotyping pattern that lacked spacers 29 to 34, and represented biovar 3 to 5 isolates. Four isolates whose biochemical characteristics conformed with those of M. bovis clustered with the group B isolates and had spoligotype patterns that differed from those previously reported for M. bovis, in that they possessed spacers 40 to 43. Interestingly, isolates of group B and, to a certain extent, also isolates of group C showed a high degree of variability in biochemical traits, despite genotypic identity in terms of IS6110 RFLP and spoligotype patterns. We hypothesize that isolates of groups B and C have their evolutionary origin in West Africa, while group A isolates are of European descent.

Rapid differentiation of bovine and human tubercle bacilli based on a characteristic mutation in the bovine pyrazinamidase gene.

Bovine tuberculosis (TB) caused by Mycobacterium bovis is an important veterinary disease that can also afflict humans. Although M. bovis shares an almost identical genome with M. tuberculosis, subtle differences in host specificity and several biochemical parameters can be used to distinguish the two closely related species. The current method for distinguishing M. bovis from M. tuberculosis relies on tedious testing of biochemical parameters, including natural resistance to pyrazinamide and defective pyrazinamidase (PZase) activity of M. bovis strains. In this study, we report the development of a rapid PCR-single-strand conformation polymorphism (SSCP) assay to differentiate M. bovis from M. tuberculosis strains, based on the detection of a single characteristic point mutation in the PZase gene (pncA) of M. bovis. Eighty-seven of 89 M. bovis strains could be distinguished from M. tuberculosis strains. Surprisingly, two animal isolates which were initially identified as M. bovis were shown to be M. africanum because they had a wild-type pncA sequence with positive PZase. These two M. africanum strains contain multiple (three and six) copies of insertion sequence IS6110, a feature they have in common with M. tuberculosis. The implication of this finding for the taxonomy of M. tuberculosis complex is discussed in relation to host preference and epidemiology. The development of a rapid PCR-SSCP test for distinguishing M. bovis from M. tuberculosis will be useful for monitoring the spread of bovine TB to humans in areas where bovine TB is endemic and for directing the treatment of human TB caused by M. bovis.

ＨＢｅ抗原陽性Ｂ型慢性肝炎に対する人型結核菌体抽出物質（ＳＳＭ）の効果

The effect on the HBeAg/anti-HBe system of SSM (specific substance Maruyama), an extract from human tubercle bacilli, was evaluated in patients with HBeAg positive type B chronic hepatitis. Twenty-five (25) HBeAg positive patients with biopsy-proven chronic hepatitis were injected subcutaneously with SSM solution twice a week for two years. Fifteen (15) comparable patients served as untreated controls for one year. HBeAg and anti-HBe were measured by the RIA method. (1) In the group receiving SSM injections, HBeAg disappeared in 8 out of 25 patients (32.0%) one year after the treatment and in 15 out of 23 (65.2%) within two years. Seroconversion from HBeAg to anti-HBe occurred in 5 of the 25 patients (20.0%) after one year and in 7 out of 23 (30.4%) within two years. Of the 15 untreated controls, HBeAg disappeared in 3 patients (20%), and seroconversion from HBeAg to anti-HBe occurred in 1 (6.7%) within one year. (2) In the SSM group, the HBeAg cut off index decreased significantly (p less than 0.01), from 5.2 +/- 1.9 during the pretreatment period to 2.7 +/- 2.3 twelve months after the treatment. (3) In the SSM group, a transient rise in sGOT and sGPT was observed in some patients three to nine months after starting the treatment. HBeAg disappeared in all these patients. (4) No notable side effects were observed in any of the patients treated with SSM. These results indicate that SSM treatment improves the HBeAg/anti-HBe system in patients with HBeAg positive chronic hepatitis.

Surgery for Crohn's disease in 28 children with the presence of acid-fast bacilli

Obstruction, abscesses, fistulae, treatment failure, and growth retardation are not always absolute indications for surgery in Crohn's disease. At the Children's Hospital of Eastern Ontario we have advocated earlier surgery since 1974. Forty-eight cases are reviewed, 28 surgical (17 ileocecocolic, 2 ileocolic with normal rectum, 3 colorectal, 1 jejunal, 5 perineal). Ileocecocolic resection with primary anastomosis produced remission for at least 3.2 years. Remission in colectomy with primary ileocolic anastomosis was at least 1 year. Ileostomy and colorectal resection produced remissions for at least 1.66 years. After jejunal resection remission was 1.58 years. Significant height and weight gains in over 80% of cases encourage earlier surgery. Adequate medical treatment has not often influenced the disease process. Surgery does not remove useful, medically restorable to normal bowel. Most postoperative cases required no medications and none jave required steroids. Permanent cure of anorectal disease alone was not achieved. Surgical mortality was 0%. The average hospital stay was 11.6 days. Postoperative fistulae occurred in 1 case and closed spontaneously. Mycobacterium tuberculosis was found in 2 resected specimens and in 1 human tubercle bacilli were cultured after 5 months. Both cases healed spontaneously, and both were otherwise undistinguishable from the other cases of Crohn's disease. Both were white, native-born Canadians. On examination of all the other resected specimens in our series, no acid-fast bacilli were seen.

Effects of the prolongation of survival time on immunotherapy with an extract from human tubercle bacilli (SSM) in gastric cancer in cases of non-curative resection. Statistical analysis of the results of long-term observation

A retrospective study on the effects of postoperative adjuvant immunotherapy for gastric cancer was made. In order to evaluate the efficacy of immunotherapy, this study was undertaken from a different point of view, that is, immunotherapy was put on a basic treatment and immunochemotherapy was used as a control.Six hundred and three patients subjected to non-curative resection were selected from 699 patients having been operated palliatively and recorded in the registration form of The Research Institute of Vaccine Therapy for Tumors and Infectious Diseases, Nippon Medical School from April 1979 to March 1980. 460 patients were excluded from this study because they were not satisfied with the criteria; e.g., 208 out of 460 excluded patients from this study gave no reply for the questionnarie, and other 252 cases had not been confirmed histologically the cancer and/or the initial therapy for some of these 252 cases was not SSM. Therefore, this report is based on 143 patients. In 69 cases immunotherapy with an extract from human tubercle bacilli (SSM) as sole treatment was carried out (immuno group), and combined therapy with SSM plus chemotherapeutic agents, mainly Mitomycin C, 5-Fluorouracil, FT-207 and Cytosine arabinoside, was given to 74 patients (combi group).There were no significant differences in the back ground factors of the two groups. The survival rate was higher in the immuno group, 3 and 5 year survival rate in the immuno group was 32.4% and 27.7%, and that of the combi group was 15.9% and 8.9% respectively. It showed that the survival rate of the immuno group was significantly higher than that of the combi group.In addition, stratifying the back ground factors caused non-curative resection, the influence for the prognosis was studied by means of statistical analysis. In comparison between the two groups, all survival curves in the immuno group were surperior to those in the combi group. Especially, those on H factor positive cases were exceedingly.These results suggested that immunotherapy with SSM was effective in patients undergone a non-curative resection for gastric cancer, and it was emphasized that the establishment of sole treatment group with immunotherapy was significant for the evaluation of effectiveness of immunotherapy.

Subdivision ofMycobacterium tuberculosisfor epidemiological purposes: a seven year study of the ‘Classical’ and ‘Asian’ types of the human tubercle bacillus in South-East England

Human strains of Mycobacterium tuberculosis were divided into the 'Classical' and 'Asian' types according to their sensitivity to thiophen-2-carboxylic acid hydrazide. The isolation of these two types in South-East England was studied during a seven-year period (1977-1983). The 'Asian' type was more prevalent among ethnic Asian patients than among ethnic Europeans. Among Europeans there was a decline in the isolation rate of 'Classical' strains and a small but significant increase in 'Asian' strains during the study period, so that the proportion of the latter type in this group is increasing. The type of bacillus was unrelated to the site of isolation except that the incidence of lymphadenitis due to the 'Asian' type among European females was significantly higher than expected. In general, European patients tended to be older than Asian patients, and the differences in age distribution according to site of isolation and type of bacillus in each ethnic group were small. A notable exception occurred with European females infected with the 'Asian' type, whose age distribution was similar to the Asians. In view of the differences in behaviour of the two types of human tubercle bacilli in this country there is a need to continue a bacteriological surveillance and also to determine whether the nature of the host-pathogen interaction varies according to the type of bacillus.

Statistical analysis on the postoperative survivals of cancer patients in the treatment of SSM (an extract from human tubercle bacilli, so called Maruyama vaccine)

Statistical analysis on the postoperative survivals of cancer patients in the treatment of SSM (an extract from human tubercle bacilli, so called Maruyama vaccine)

Evidence for host-dependent modification and restriction of bacteriophage DNA in Mycobacterium tuberculosis.

Wild isolates of Mycobacterium tuberculosis may be divided into the three internationally recognized phage types on the basis of susceptibility to mycobacteriophages DS6A, BK1 and D34. Strains of type A are lysed at high efficiency by DS6A only; type B is lysed by BK1 grown on Mycobacterium smegmatis ATCC607 and DS6A, while type C is lysed additionally by D34 grown on atypical Mycobacterium F130. Propagation of D34 on a C-strain (D34-C) or BK1 on a B-strain (BK1-B) has no effect on viral host-range. D34-C has an efficiency of plating (e.o.p.) of 10(-5) on type B strains and 10(-7) on A strains. BK1-B plates on A strains at an e.o.p. of 10(-5). BK1 recovered from and repropagated on an A strain (BK1-A) has an e.o.p. of 1-0 on strains of all classes. D34-B has an e.o.p. of 1-0 on strains of type B and C, while D34-A plates with high efficiency on types B and C and displayed an e.o.p. of 10(-4) on type A. Repropagation of these viruses on the M. tuberculosis strains originally lysed by them results in the restoration of their previous host range. Variations in plating efficiency cannot be explained by differences in viral absorption alone. These findings suggest that the three phage types of human tubercle bacilli are related by a hierarchical pattern of DNA restriction and modification in which the C pattern is included in the B, and both patterns are included in A-modified DNA. Viruses such as DS6A which are equally virulent for strains of all classes are not susceptible to host dependent restriction.

Studies of tuberculosis in an in relation to infection in cattle

A study has been made in Sweden to investigate whether the risk of tuberculous infection and its trend with time in man in different areas were related to varying prevalences of tuberculous infection in cattle. It was found that the level of the infection risk in man was related to the prevalence of tuberculous infection in cattle, varying from 3·4 per cent in 1935 at age 15 in counties with less than 2 per cent infected cattle to 5·8 per cent in counties with 20 per cent infected cattle or more. However, the downward trend of the risk of infection with time was not found to be associated with the prevalence of tuberculous infection in cattle. The correlation between the risk of infection in man and the prevalence of infection in cattle in the 24 Swedish counties was positive and highly significant (+0·79). The relationship between cattle tuberculosis and tuberculosis in man was further studied by correlating infection in cattle with tuberculosis mortality and morbidity in man. The correlation with mortality was negative and highly significant (−0·77), i.e. counties with little cattle tuberculosis had a high tuberculosis mortality in man, and vice versa. There were similar large negative correlations with measures of tuberculosis incidence. When variations between the counties in relevant environmental factors, namely capital, urbanization and overcrowding, were taken into account, a strong positive association remains between the prevalence of infected cattle and the risk of tuberculous infection in man, but the associations with tuberculous mortality and morbidity, though they remain negative, become weaker. It is suggested that the probable explanation of these findings is the long-term protection against adult infection with human tubercle bacilli conferred by bovine infection in childhood.

Isolation and characterization of toxic glycolipids from the firmly bound lipids of human tubercle bacilli

Isolation and characterization of toxic glycolipids from the firmly bound lipids of human tubercle bacilli

Der bakteriologisch-serologische Kurs

This is an eminently practical book. It provides a good guide for the technician who wants to learn how to use the bacteriologic and serologic laboratory. It leads him from dealing with infectious material through all the techniques until the final microscopical diagnosis. Technically, it goes clearly beyond the requirements of the course work in most American schools. It provides a clear and practical description of the pitfalls and the frequent mistakes in technique and interpretation of bacteriologic examinations. As the basis for scientific work and training, however, it is clearly not sufficient. It does not have a bibliography, which makes the usefulness of some of the described techniques rather questionable. To give one example, the description of the test to differentiate atypical from human tubercle bacilli is insufficient for the worker who really wants to follow it, and without a bibliography it is, therefore, useless.

Isolation of tuberculin active peptides from cell wall fraction of human tubercle bacillus strain Aoyama B.

Isolation of tuberculin active peptides from cell wall fraction of human tubercle bacillus strain Aoyama B.

Studies on chemotherapeutics for Mycobacterium tuberculosis. 23. Synthesis and antibacterial activity on Mycobacterium tuberculosis of 3-methoxy-4-phenoxybenzaldehyde and 3-hydroxy-4-phenoxybenzaldehyde derivatives

Forty-two kinds of compounds mainly related to 3-methoxy-4-phenoxybenzaldehyde thiosemicarbazone and 22 kinds of compounds similar to its demethylated compound, 3-hydroxy-4-phenoxybenzaldehyde, were synthesized and their antibacterial action against human tubercle bacilli, H37Rv sensitive strain, was tested, using a Tween-albumin medium. Against the control growth-inhibitory values of 0.0313 μg/ml for isonicotinoyl hydrazide (INH) and 0.625 μg/ml for tibione (TBI), 3-methoxy-4-phenoxybenzaldehyde (p-tolyl) thiosemicarbazone and 4-(o-methoxyphenyl) thiosemicarbazone showed the value of 3.13 μg/ml, and 4-allyl-, 4-propyl-, 4-butyl-, 4-(p-methoxyphenyl)-, 4-(o-ethoxyphenyl)-, and 4-(p-iso-butoxyphenyl)-thiosemicarbazones and amidinohydrazone hydrochloride all showed growth inhibition in a concentration of 6.25 μg/ml. 3-Hydroxy-4-phenoxybenzaldehyde derivatives all failed to show any antibacterial activity and this fact indicated that the presence of a methoxyl group was significant in the appearance of antibacterial activity against this strain of tubercle bacilli. 3-Methoxy-4-phenoxybenzaldehyde isonicotinoylhydrazone inhibited the growth of tubercle bacilli at a concentration of 0.078 μg/ml, but it was found to have cross-resistance with INH.

結核菌に対する化学療法剤の研究(第22報) : N<SUP>2</SUP>-(2-Pyridyl)-3-p-ethoxyphenyl-pseudothiohydantoinおよびN<SUP>2</SUP>-(2-Benzothiazoly)-3-allylpseudothiohydantoin誘導体の合成ならびに抗菌作用について

Twenty-five derivatives of N2-(2-pyridyl)-3-p-ethoxyphenylpseudothiohydantoin and 27 derivatives of N2-(2-benzothiazolyl)-3-allylpseudothiohydantoin were synthesized and their antibacterial action against human tubercle bacilli, H37Rv strain, was tested in a Tween-albumin medium. None of the compounds tested showed any marked effect except that two of these, 5-(5-methyl-2-furfurylidene) and 5-(8-hydroxyquinolyl-5-methylene)-N2-(2-benzothiazolyl)-3-allypseudothiohydantoins inhibited the growth in a concentration of 12.5 μg/ml.

Diagnostic and Prognostic Significance of the Quantitative Tuberculin Tests

Excerpt Fifty years ago practically everyone over 20 years of age in the industrialized areas of Europe and the United States had been infected with human tubercle bacilli and gave strong positive ...