Heat-killed Tubercle Bacilli Research Articles

The black scourge? Race and the Rockefeller Foundation's tuberculosis commission in interwar Jamaica.

From 1927 to 1942, the Rockefeller Foundation ran a tuberculosis commission in Jamaica that researched the epidemiology of the disease, examined the efficacy of a vaccine with heat-killed tubercle bacilli, and offered basic treatment to tuberculosis sufferers. Drawing upon diaries and scientific writings by the staff employed by the commission, among other sources, this article explores the role that race played in the tuberculosis commission. It assesses how race shaped the research conducted by the commission, how it informed staff interactions and staff/patient relations, and the clash and/or confluence of "imported" and local racial ideas in the commission's work.

Modifications in tissue kallikrein activity with indomethacin and prednisolone treatment in arthritic rats

This investigation was conducted to evaluate the tissue kallikrein activity in the synovial and paw tissues of control, non-treated adjuvant arthritic and adjuvant arthritic rats treated either with indomethacin or prednisolone. Adjuvant arthritis was induced in male Sprague-Dawley rats in the right knee by injecting 0.05 ml of a fine suspension of heat-killed Mycobacterium tubercle bacilli in liquid paraffin (5 mg/ml). Indomethacin (2.5 mg/kg given orally) and prednisolone (3.0 mg/kg given orally) treatment for 9 days caused a significant reduction (P < 0.001) in knee swelling. In non-treated adjuvant rats, synovial tissue kallikrein levels were raised (P < 0.01) than the synovial tissue kallikrein obtained from control normal rats. Prednisolone treatment resulted in reduction (P < 0.05) and indomethacin treatment produced raise (P < 0.01) in synovial tissue kallikrein levels. In non-treated arthritic rats, the paw tissue kallikrein levels were lower (P < 0.001) than the control rats; whereas, rats treated with prednisolone and indomethacine showed higher levels (P < 0.05) of paw tissue kallikrein. These results may suggest that the prednisone and indomethacin differ in their actions in inflamed synovial tissue kallikrein and have a similar effect in non-inflamed paw tissues.

The tuberculin skin test in relation to immunological in vitro reactions in BCG-vaccinated healthcare workers.

The aim was to study the tuberculin skin test in relation to immunological in vitro reactions in bacille Calmette-Guerin (BCG)-vaccinated healthcare workers. The present study was performed in Sweden, a country with a low incidence of tuberculosis, a high BCG vaccination efficacy and high tuberculin conversion rates. BCG-vaccinated healthcare workers (n=381) were tuberculin skin tested. From these, 11 subjects with negative tuberculin reactions (<6 mm) were matched for age and sex with 11 subjects with large positive reactions (> or = 15 mm). Lymphocyte transformation and the production of interferon-gamma (IFN-gamma) were analysed after stimulation in vitro of peripheral blood mononuclear cells with tuberculin purified protein derivative, heat-killed tubercle bacilli and a culture filtrate from tubercle bacilli. In the tuberculin-positive group the lymphocyte transformation response was 2-3 times larger, and IFN-gamma production was 7-10 times larger, than in the tuberculin-negative group (p<0.001). The present results suggest that a positive tuberculin skin test in bacille Calmette-Guerin-vaccinated subjects indicates a stronger immune response of the protective T-helper 1-type than does a negative test. In similar settings, the study supports the traditional practice of regarding the tuberculin skin test in bacille Calmette-Guerin-vaccinated subjects as an indicator of a protective immune response against tuberculosis.

Relationship between macrophage infiltration and epidermopoiesis in delayed-type hypersensitivity.

The relationship between epidermopoiesis and macrophage infiltration was studied in delayed-type hypersensitivity (DTH) skin lesions in guinea pigs that had been sensitized with heat-killed tubercle bacilli or bovine serum albumin (BSA). Macrophages were identified with acid-phosphatase and nonspecific esterase stains, and the epidermal proliferative response was studied at DTH challenge sites by autoradiography. The number of macrophages in the sensitized animals was higher than that in the nonsensitized animals 48-72 h following challenge injections, when labelling indices were also elevated in the former group. Soluble factor(s) from cultured macrophages transiently enhanced the DNA synthesis of epidermal cells in cultures and in the sites injected with the factor(s). These results suggest that macrophages retained in the DTH lesion may play a role in an acceleration of epidermal proliferation, thus leading to acanthosis and lichenification.

Potentiation of the rat delayed-type hypersensitivity reaction by the Fc portion of human IgG 1

Fc fragments derived from a human IgG 1 myeloma protein potentiate the rat delayed-type hypersensitivity (DTH) reaction to antigen challenge. Lewis rats immunized with heat-killed tubercle bacilli give augmented DTH reactions to the purified protein derivative of tuberculin when Fc fragments are included in the challenge dose. Similar potentiation of DTH by pFc′ fragments indicates that the active site is located in the CH 3 domain of IgG 1. Histologic evaluation of the augmented reaction sites revealed predominantly mononuclear cell infiltrates characteristic of DTH reactions. Skin tests of tubercle bacilli-sensitized rats with an unrelated antigen and/or Fc fragments fail to elicit significant reactions. Augmentation of the DTH reaction to purified protein derivative is restricted to the Fc or pFc′ region fragments since intact monomeric IgG 1, Fab fragments, and bovine serum albumin were all shown not to be active potentiators. The DTH reaction of ovalbumin-sensitized rats was similarly augmented when Fc fragments were included with a challenge dose of ovalbumin, thus supporting the general nature of the phenomenon. These results support the concept of Ig molecules as multifunctional proteins that can not only serve effector functions but also participate in the regulation of immune responses.

Effect of mycobacterial water-soluble adjuvant (MAF3) on the immune response of guinea pigs to the hapten-carrier conjugates.

To clarify the complex mechanisms of the immune responses, many adjuvants have been used as immunologic tools. A classical example of such materials is Freund's complete adjuvant (FCA). It is well known that the heat-killed tubercle bacilli in mineral oil are the most important constituents for the adjuvant activity of FCA. Recently, it has been demonstrated that the tubercle bacilli in FCA can be replaced with water-soluble fractions from mycobacteria (1, 7, 10-13) and other Gram-positive bacteria (8). These water-soluble fractions probably share common chemical constituents. MAF3, which was prepared from the delipidated cells of Mycobacterium tuberculosis strain Aoyama B by hydrogenolysis and gel filtration (10), was found to be one of such water-soluble components responsible for the adjuvant activity. Although adjuvant activity of MAF3 on the antibody response, especially of the IgG2 class, and on the induction of delayed-type hypersensitivity (DTH) reaction to ovalbumin (OA) have been observed in guinea pigs, its effect on the immune response to other types of antigens such as hapten-carrier conjugates are not fully investigated yet. In this paper, we describe the adjuvant effect of MAF3 on the antibody response and the induction of DTH reaction in guinea pigs immunized with 2, 4-dinitrophenyl (DNP)-conjugates, DNP-bovine serum albumin (BSA), DNP-dextran or DNP-guinea pig serum albumin (GPA) emulsified with Freund's incomplete adjuvant (FIA) which lacks mycobacterial components. Female Hartley/s guinea pigs, weighing about 300 to 500 g, were used in all experiments. BSA (fraction V, Seikagaku-Kogyo, Tokyo), OA ( •~ 5 crystalline, ICN Pharmaceuticals Inc., Ohio) and GPA (Cohn's fraction V from normal guinea

Elicitation of endotoxemic effects in C3H/HeJ mice with glucocorticoid antagonizing factor and partial characterization of the factor

C3H/HeJ mice were used to study the origin and nature of endotoxin-induced glucocorticoid antagonizing factor (GAF). In conventional mice GAF is believed to be responsible for a variety of effects that occur as a result of an injection of endotoxin, including the inhibition of hormonal induction of hepatic phosphoenolpyruvate carboxykinase and of glyconeogenesis. Responses in such animals are seen whether the endotoxin is extracted with phenol-water or with trichloroacetic acid. C3H/HeJ mice do not respond (or produce GAF?) after an intravenous injection of phenol-water lipopolysaccharide, but they react normally (produce GAF?) when given a trichloroacetic acid preparation. They also behave the same as conventional animals when injected with serum from poisoned normal mice, especially when the reticuloendothelial system of the donors has been activated by prior injections of Zymosan or heat-killed tubercle bacilli. The C3H/HeJ mice have been used, therefore, as assay animals to establish that peak levels of GAF appear in donor serum about 2 h after an injection of lipopolysaccharide, and it is produced intraperitoneally in C3H/HeJ mice given a mixture of endotoxin and peritoneal exudate cells derived from responder mice. GAF elutes from Sephadex G-200 along with markers of known molecular weight in the region of 100,000 to 200,000. It is inactivated by trypsin and by heating at 75 degrees C for 1 h.

In vitro studies on the mechanism of acquired resistance to tuberculous infection. I. The relationship between lymphocytes and macrophages in cellular immunity to tuberculous infection.

The relationship between lymphocytes and macrophages in cellular immunity against tuberculous infection was studied by means of an in vitro cell culture system without addition of streptomycin. The peritoneal macrophages were obtained from normal mice or mice immunized with heat-killed tubercle bacilli in paraffin oil, boosted with live BCG and infected with H37Rv cells in vitro. The infected monolayers of macrophages were cultivated for 48 hr with immune lymphoid cells obtained from immunized mice. The intracellular growth of H37Rv cells 3,5 and 7 days after infection was examined by counting tubercle bacilli within infected macrophages under a microscope. 1) The increase of bacilli within macrophages derived from immunized mice was slightly smaller than that in normal macrophages. 2) The addition of immune lymph node cells to the macrophage monolayers resulted in a marked decrease in the number of bacilli within both normal and "immune" macrophages. Conversely, normal lymph node cells exhibited an enhancing effect on the intracellular bacillary growth. 3) Immune lymph node cells showed a higher capacity to cause macrophages to suppress intracellular growth of bacilli than that of splenic lymphoid cells or thyrmocytes after addition to macrophage monolayers. 4) The treatment of lymphoid cells with inhibitors of protein synthesis, cycloheximide or streptovitacin A, resulted in a remarkable reduction of the ability of sensitized lymphocytes to cause macrophages to suppress multiplication of intracellular bacilli.

Evaluation of the animal assay of tuberculin intended for use in humans: II. Usefulness

The reactions of guinea pigs sensitized with heat-killed tubercle bacilli in oil to tests containing serial dilutions of tuberculins of different manufacture in diluents containing 5 parts/ million of Tween-80 were compared with their reactions to a single dilution and to serial dilutions of the international standard of mammalian tuberculin PPD in phosphate buffer not containing Tween. The studies were done double blind in two laboratories, each using 24 animals for each comparison. An analysis of the data indicates that the potency ratio between products calculated from a six or an eight point parallel line assay design, while providing a reasonably accurate estimate of the true potency ratio between products in animals, may be very different from the potency ratio between these products in humans. The apparent dependency of the potency ratio between tuberculins on the animal species in which it is estimated makes the animal assay system less useful in the quality control of tuberculin intended for use in humans. Various aspects of this problem and potential solutions are discussed.

Skin reaction, inhibition of macrophage migration, and lymphocyte transformation with tuberculin active peptide (TAP) and arabinogalactan obtained from tubercle bacilli.

Arabinogalactan purified from heat-killed tubercle bacilli failed to elicit a delayed type of skin reaction and had no ability to induce in vitro lymphocyte blast formation in sensitized guinea pigs. It was, however, inhibitory to migration of bronchoalveolar washing cells by the indirect test, but not by the direct test, and capable of eliciting an immediate type of skin reaction and anaphylaxis when injected into sensitized guinea pigs. Tuberculin active peptide (TAP) was active in all of in vitro and in vivo cell-mediated immune responses, but not in immediate responses.

Prevention of tuberculous cavity formation by desensitization with tuberculin-active peptide.

Repeated injections of tuberculin-active peptide without toxicity prevented cavity formation in rabbit lungs that had been injected with heat-killed tubercle bacilli. This desensitization procedure also suppressed the delayed type skin reaction and the in vitro migration inhibition of alveolar cells with tuberculins, but it had no significant influence upon the passive hemagglutination titer of antipolysaccharide antibody. The correlation between cavity formation and delayed hypersensitivity is discussed.

Pyrazinamide deamidase in tuberculous infection.

Pyrazinamide deamidase of mouse liver was suppressed by tuberculous infection. Such decrease of enzyme activity was also induced by the injection of heat-killed tubercle bacilli or defatted residues. Not only virulent bacilli, but also avirulent Mycobacterium tuberculosis, attenuated Mycobacterium bovis, and nonpathogenic acid-fast bacilli induced enzyme suppression. Prior vaccination with BCG or neonatal thymectomy did not prevent the decrease of enzyme activity induced by the subsequent challenge infection. Phagocytosis of particulate substances did not cause the phenomenon. Tests of various fractions of tubercle bacilli showed that cord factor was the active principle responsible for the suppression of pyrazinamide deamidase in tuberculous infection.

The relationship of delayed hypersensitivity to acquired antituberculous immunity: II. Effect of adjuvant on the allergenicity and immunogenicity of heat-killed tubercle bacilli

Mice were injected subcutaneously on two occasions with a heat-killed, lyophilized suspension of M. tuberculosis H 37R v dispersed in a water-in-oil emulsion or as a saline suspension. The levels of tuberculin sensitivity were followed for 30 days, at which time the mice were infected intravenously with approximately 10 6 living BCG (Montreal). At intervals thereafter, groups of mice were challenged with 10 LD 50's Listeria monocytogenes so that the development of nonspecific resistance could be followed in relation to the fate of BCG and the state of hypersensitivity in the different treatment groups. Nonvaccinated controls, and mice receiving adjuvant alone, required 10–12 days to respond to the eliciting dose of BCG with the development of detectable tuberculin sensitivity and cellular resistance to the Listeria challenge. They were also unable to limit the growth of BCG in the lung. Mice receiving H 37R v in saline behaved like the normal controls; but animals receiving H 37R v in adjuvant quickly developed high levels of tuberculin sensitivity, and their resistance to Listeria and BCG both appeared much sooner. Seven months later, mice of this group showed a marked rise in tuberculin sensitivity and an accelerated onset of cellular immunity in response to the BCG infection. The significance of these findings is discussed in relation to the mechanism of antituberculous immunity and the evaluation of nonliving vaccines.

Cellular Reactivity to Tuberculin in Immune and Serologically-Unresponsive Guinea Pigs

Abstract We have reported that the serologic reactivity of young guinea pigs to mycobacterial antigens decreased proportionately as the sensitizing dose of heat-killed tubercle bacilli was increased (1). Using animals from 0 to 4 weeks old, we found that approximately 67% of those that received doses of 0.5 mg (dry weight) or less exhibited precipitating and complement-fixing antibodies against an unheated mycobacterial culture filtrate when tested 4 weeks after injection, whereas only 34% of the animals that received doses greater than 0.5 mg reacted serologically when similarly tested. Serologic unresponsiveness was induced quite effectively by injecting newborn animals with a 5-mg dose; only 1 of 14 animals sensitized in this manner exhibited anti-mycobacterial precipitins when tested 4 to 5 weeks later. Hypersensitivity to tuberculin purified protein derivative (PPD) in the serologically-unresponsive animals, however, appeared to be unaffected. Animals injected with the 5-mg dose at birth exhibited unequivocal skin reactivity to PPD 5 weeks later, but the reactions were smaller than those of adults that received the same sensitizing dose (1).

19-S- and 7S- Antibody Activities of Middlebrook-Dobus Indirect Hemagglutination and Hemolysis Reactions in Guinea Pigs Sensitized with Heat Killed Tubercle Bacilli

19-S- and 7S- Antibody Activities of Middlebrook-Dobus Indirect Hemagglutination and Hemolysis Reactions in Guinea Pigs Sensitized with Heat Killed Tubercle Bacilli

Studies on Antibody-holding Cells of Animals Sensitized with Heat-killed Tubercle Bacilli by Fluorescent Antibody Technique

Studies on Antibody-holding Cells of Animals Sensitized with Heat-killed Tubercle Bacilli by Fluorescent Antibody Technique

ツベルクリン反応の受身伝達に関する研究 : 第1報 加熱死結核菌流パラ浮遊液注射によるラットへの影響について

ツベルクリン反応の受身伝達に関する研究 : 第1報 加熱死結核菌流パラ浮遊液注射によるラットへの影響について

Experimental Studies on Passive Transfer of Tuberculin Allergy : Report II. Immunological studies on peritoneal cells of sensitized with heat-killed tubercle bacilli

Experimental Studies on Passive Transfer of Tuberculin Allergy : Report II. Immunological studies on peritoneal cells of sensitized with heat-killed tubercle bacilli

Reversion and Reconversion Rate of Tuberculin Skin Reactions in Correlation with the Use of Prednisone

The cutaneous tuberculin sensitivity found in healthy persons who have had a tuberculous infection, or in sufferers from tuberculosis, or in experimental animals previously sensitized with BCG or heat-killed tubercle bacilli, is usually inhibited or diminished by certain agents including adrenocorticotrophic hormone and adrenal corticosteroids, administered either systemically or locally.’” Our present study is based on a sizable group of natural tuberculin converters who received prednisone at random or as a trial treatment for certain forms of tuberculosis for a varying period until at least the reversion of skin sensitivity was definitely obtained. The primary purpose is to record the exact time of inhibition of tuberculin sensitivity with the use of corticosteroid in the same dosage and also the time of reconversion of the reaction after withdrawal of treatment.

加熱結核死菌免疫における流動パラフィンadjuvantの免疫増強機序に関する実験的検討

加熱結核死菌免疫における流動パラフィンadjuvantの免疫増強機序に関する実験的検討