scholarly journals MULTIPLICATION OF TUBERCLE BACILLI WITHIN MONONUCLEAR PHAGOCYTES IN TISSUE CULTURES DERIVED FROM NORMAL ANIMALS AND ANIMALS VACCINATED WITH BCG

1953 ◽  
Vol 97 (2) ◽  
pp. 235-245 ◽  
Author(s):  
Emanuel Suter
Keyword(s):  
Tissue Culture ◽  
Guinea Pig ◽  
Guinea Pigs ◽  
Culture Medium ◽  
Normal Animal ◽  
Mononuclear Phagocytes ◽  
Tissue Cultures ◽  
Inhibition Of Growth ◽  
Intracellular Multiplication

When monocytes derived from normal guinea pigs or rabbits were infected with tubercle bacilli and cultivated in vitro, the bacilli multiplied abundantly within the cytoplasm of these cells. By contrast, intracellular multiplication of the bacilli was retarded or completely inhibited within the monocytes of rabbits or guinea pigs vaccinated with BCG. This inhibition of growth was observed with both virulent or attenuated strains of tubercle bacilli. Under the conditions used in the present study, the ability of monocytes to inhibit bacillary proliferation was the same whether serum from a normal animal or from vaccinated animals was used in the tissue culture medium. Moreover, the serum of vaccinated animals did not inhibit multiplication of tubercle bacilli within monocytes derived from a normal animal. The ability of guinea pig monocytes to interfere with intracellular bacillary proliferation was first perceptible 8 days after vaccination.

scholarly journals THE MULTIPLICATION OF TUBERCLE BACILLI WITHIN NORMAL PHAGOCYTES IN TISSUE CULTURE

1952 ◽  
Vol 96 (2) ◽  
pp. 137-150 ◽  
Author(s):  
Emanuel Suter
Keyword(s):  
Tissue Culture ◽  
Culture Medium ◽  
Mononuclear Cells ◽  
Virulent Strain ◽  
Host Cells ◽  
Avirulent Strain ◽  
Glass Slides ◽  
Intracellular Multiplication ◽  
Cultivation In Vitro

A technique has been described for the cultivation in vitro of normal mononuclear cells on glass slides in a liquid medium. Under these conditions the monocytes transformed into macrophages which proliferated as in ordinary tissue culture. These cultures of monocytes could be infected with tubercle bacilli. The numbers of stainable tubercle bacilli within the monocytes increased steadily in cultures infected with virulent or attenuated strains. Evidence is given to support the view that this increase in numbers of bacilli was due to intracellular multiplication. There was no evidence of intracellular bacillary multiplication in cultures infected with an avirulent strain. Tubercle bacilli multiplying within phagocytes in vitro exert a damaging effect upon the host cells. The damage was most obvious in cells infected with a virulent strain. Tubercle bacilli within phagocytes were protected against the bacteriostatic effect of streptomycin added in a concentration of 5 γ per ml. of culture medium. This permitted the use of streptomycin in infected cultures to prevent extracellular multiplication of the bacilli.

scholarly journals FURTHER STUDIES ON TYPHUS FEVER

1936 ◽  
Vol 64 (5) ◽  
pp. 673-687 ◽  
Author(s):  
Hans Zinsser ◽  
Attilio Macchiavello
Keyword(s):  
Tissue Culture ◽  
Guinea Pig ◽  
Guinea Pigs ◽  
Horse Serum ◽  
Tissue Cultures ◽  
Practical Application ◽  
Typhus Fever ◽  
Culture Virus ◽  
Pig Serum

1. Guinea pigs can be actively immunized against European typhus fever with homologous formalinized Rickettsia tissue cultures, provided sufficient amounts are injected. The method is suggested for practical application in man. 2. Serovaccination against European typhus fever can be successfully applied to guinea pigs by a variety of methods, the simplest of which consists of the injection of mixtures of virulent defibrinated guinea pig blood and convalescent guinea pig serum taken from 3 to 5 days after defervescence. Similar results can be obtained with mixtures in which tissue culture virus, either with convalescent guinea pig serum or with antimurine horse serum, is used. There is no indication so far that such animals become carriers. Possible application of these methods to typhus epidemics is discussed.

scholarly journals THE NEUTRALIZATION OR DESTRUCTION OF DIPHTHERIA TOXIN BY TISSUE

1931 ◽  
Vol 53 (6) ◽  
pp. 821-826 ◽  
Author(s):  
Augustus Wadsworth ◽  
Ella N. Hoppe
Keyword(s):  
Guinea Pig ◽  
Cardiac Muscle ◽  
Muscle Tissue ◽  
Guinea Pigs ◽  
Diphtheria Toxin ◽  
Cardiac Tissue ◽  
Tissue Cultures ◽  
Adult Heart ◽  
Embryo Extract

As determined by the intracutaneous test in guinea pigs, diphtheria toxin is not altered in the presence of cardiac tissue obtained from the fetal or from the adult heart of the guinea pig. Tissue cultures were apparently uninjured by the presence of the toxin in the dilutions used in these experiments, and, when washed with embryo extract after removal of the diluted toxin, continued to grow. Embryonic guinea pig cardiac muscle tissue growing in cultures in vitro possesses the power of neutralizing, binding, or destroying diphtheria toxin so that it is no longer toxic for normal guinea pigs. Such neutralization takes place through the intervention of growing tissue and is a property which is lacking in similar surviving tissue not in a state of cultivation. Thus, it appears that the living, growing cells of the tissues neutralize or destroy limited quantities of toxin; only when the quantity of toxin exceeds a certain limit is its action injurious.

scholarly journals THE ESSENTIAL PARTICIPATION OF AN ENZYME IN THE INHIBITION OF GROWTH OF TUBERCLE BACILLI BY SPERMINE

1953 ◽  
Vol 97 (3) ◽  
pp. 327-343 ◽  
Author(s):  
James G. Hirsch
Keyword(s):  
Guinea Pig ◽  
Culture Medium ◽  
Culture Media ◽  
Enzymatic Reaction ◽  
Rabbit Serum ◽  
Inhibition Of Growth ◽  
Plasma Fraction ◽  
Bovine Plasma ◽  
Chemical Contaminant

Spermine inhibits the multiplication of tubercle bacilli in vitro only if another tissue substance is present in the culture medium. This activating substance occurs as a chemical contaminant of the albumin in commercial bovine plasma fraction V ordinarily added to culture media; it is also found in whole bovine and sheep serum and in aqueous extracts of the guinea pig kidney. No detectable spermine activator is contained in whole human, guinea pig, or rabbit serum. The serum constituent which renders spermine inhibitory for the growth of acid-fast bacteria has the properties of a protein of the alpha globulin classification. This protein is an enzyme which acts on spermine. Presumably, some product of the enzymatic reaction exerts a toxic effect on mammalian tubercle bacilli.

Multiplication of Mycobacterium tuberculosis Within Normal and “Immune” Mouse Macrophages Cultivated With and Without Streptomycin

1970 ◽  
Vol 1 (1) ◽  
pp. 30-40
Author(s):  
Ronald J. Patterson ◽  
Guy P. Youmans
Keyword(s):  
Tissue Culture ◽  
Mycobacterium Tuberculosis ◽  
Culture Medium ◽  
Peritoneal Macrophages ◽  
Mononuclear Phagocytes ◽  
Immune Mouse ◽  
Mouse Macrophages ◽  
Intracellular Multiplication ◽  
Mycobacterial Growth ◽  
Mouse Peritoneal Macrophages

Acquired cellular immunity to infection with Mycobacterium tuberculosis is believed to reside in the capacity of mononuclear phagocytes of immunized animals to inhibit intracellular multiplication of the parasite. However, in macrophage tissue culture systems, it has been customary to employ streptomycin in the medium for the purpose of restricting extracellular, but not intracellular, growth of M. tuberculosis. In contrast, our data show that small amounts of streptomycin markedly inhibit intracellular as well as extracellular growth of M. tuberculosis in normal mouse peritoneal macrophages, and that the degree of this inhibition is directly proportional to the concentration of streptomycin used. In the absence of streptomycin, virulent tubercle bacilli grew as rapidly in “immune” macrophages as in normal macrophages. “Immune” macrophages, however, were slightly more resistant to destruction by the intracellularly multiplying mycobacteria. In the presence of streptomycin, however, intracellular mycobacterial growth was inhibited more in “immune” macrophages than in normal macrophages, and this effect also was directly proportional to the concentration of streptomycin used. Virulent mycobacteria grew somewhat more slowly within mouse peritoneal macrophages obtained after induction of a peritoneal exudate with glycogen than in noninduced cells. The rate of multiplication, though, was the same within normal and “immune” induced peritoneal cells except in the presence of streptomycin. As with noninduced macrophages, this drug inhibited the intracellular multiplication of virulent tubercle bacilli more effectively within “immune” induced than within normal induced cells. It would appear, therefore, that the greater inhibition of intracellular multiplication of virulent tubercle bacilli in “immune” macrophages in tissue culture noted by a number of investigators in the past may have been an artifact created by the use of streptomycin in the tissue culture medium.

scholarly journals STUDIES ON THE MECHANISM OF IMMUNITY IN TUBERCULOSIS

1939 ◽  
Vol 69 (4) ◽  
pp. 579-599 ◽  
Author(s):  
Max B. Lurie
Keyword(s):  
Guinea Pigs ◽  
Physiological Activity ◽  
Normal Animal ◽  
Mononuclear Phagocytes ◽  
Ion Concentration ◽  
Phagocytic Capacity ◽  
Hydrogen Ion Concentration ◽  
Carbon Particles ◽  
Mesenchyme Cells

1. Tuberculous and vaccinated rabbits and guinea pigs mobilize mononuclear phagocytes at the site of a non-specific inflammation with greater rapidity than do normal animals, just as they respond to tubercle bacilli. 2. The succession of cells that characterizes inflammation in general is accelerated in allergic rabbits and guinea pigs in response to non specific irritants. 3. The pH at the site of reinfection with tubercie bacffli in immun ized rabbits and guinea pigs and at the site of a non-specific inflammation in the latter is slightly lower than in a similar site in a normal animal. 4. No constant relation was found between the mobifization of mononuclears and the hydrogen ion concentration at the site of inflammation. 5. The rate of mitotic and amitotic division of mononuclears in allergic rabbits and guinea pigs in response to non-specific irritants is greater than in normal animals. 6. Mononuclears derived from actively tuberculous or vaccinated guinea pigs exhibit greater in vitro phagocytic capacity for carbon particles than mononuclears obtained from normal animals. 7. Mononuclears of tuberculous rabbits ingest more staphylococci than the phagocytes of the same type originating from normal animals. 8. Mononuclears originating from actively tuberculous rabbits and guinea pigs exhibit greater in vitro phagocytic capacity for tuberde bacilli than mononuclears obtained from normal animals. 9. The enhancement of the phagocytic capacity for tubercie bacilli afforded mononuclears by vaccination with a bacillus of low virulence is lower, and of questionable significance. 10. The increased phagocytic activity of mononuclears derived from tuberculous or vaccinated rabbits and guinea pigs for tubercie bacilli and for non-specific particulate matter occurs in media containing sera derived from normal and from tuberculous individuals. 11. The more rapid mobilization of mononuclears by immunized animals in response to specific as well as non-specific irritants is associated with their increased physiological activity. The significance of this enhanced activity conferred by the tuberculous process on the mesenchyme cells is discussed in relation to the mechanism of immunity to tuberculosis and other phenomena.

Inhibition of Human and Animal Platelet Adhesiveness to Glass Bead Columns by Adenosine, Dipyridamole, Chlorpromazine and Acetylsalicylic Acid

1976 ◽  
Vol 36 (02) ◽  
pp. 401-410 ◽  
Author(s):  
Buichi Fujttani ◽  
Toshimichi Tsuboi ◽  
Kazuko Takeno ◽  
Kouichi Yoshida ◽  
Masanao Shimizu
Keyword(s):  
Guinea Pig ◽  
Acetylsalicylic Acid ◽  
Guinea Pigs ◽  
Glass Bead ◽  
Animal Species ◽  
Inhibitory Effect ◽  
Rabbit Platelet ◽  
Platelet Adhesiveness

SummaryThe differences among human, rabbit and guinea-pig platelet adhesiveness as for inhibitions by adenosine, dipyridamole, chlorpromazine and acetylsalicylic acid are described, and the influence of measurement conditions on platelet adhesiveness is also reported. Platelet adhesiveness of human and animal species decreased with an increase of heparin concentrations and an increase of flow rate of blood passing through a glass bead column. Human and rabbit platelet adhesiveness was inhibited in vitro by adenosine, dipyridamole and chlorpromazine, but not by acetylsalicylic acid. On the other hand, guinea-pig platelet adhesiveness was inhibited by the four drugs including acetylsalicylic acid. In in vivo study, adenosine, dipyridamole and chlorpromazine inhibited platelet adhesiveness in rabbits and guinea-pigs. Acetylsalicylic acid showed the inhibitory effect in guinea-pigs, but not in rabbits.

scholarly journals In Vitro Production of Erythropoietin by Kidneys Perfused With a Serum-free Solution

Blood ◽  
1974 ◽  
Vol 44 (1) ◽  
pp. 77-85 ◽  
Author(s):  
Allan J. Erslev
Keyword(s):  
Tissue Culture ◽  
Atmospheric Pressure ◽  
Culture Medium ◽  
Kidney Tissue ◽  
In Vitro Production ◽  
In Vitro Perfusion ◽  
Serum Free ◽  
Enzymatic Activation ◽  
Vitro Production

Abstract Normal rabbits exposed to 0.4 atmospheric pressure for 3 hr will generate about 40-60 U of erythropoietin during a subsequent 3-hr period. If the kidneys were removed from 3-hr hypoxic animals, washed carefully, and perfused for 3 hr by recirculation with a serum-tissue culture mixture, each kidney generated about 14 U of erythropoietin in vitro. Perfusion of normal kidneys did not result in the production of erythropoietin, and only small amounts were generated if the perfusate contained Puromycin. Three-hour hypoxic kidneys perfused for 3 hr with a serum-free tissue culture medium were found to generate about 8 U of erythropoietin per kidney and similar kidneys perfused with saline about 1 U. These results indicate that erythropoietin is synthesized by kidney tissue and not produced by enzymatic activation of a plasma substrate.

scholarly journals FURTHER STUDIES ON THE ROLE OF PROTEASES IN THE ALLERGIC REACTION

1961 ◽  
Vol 113 (2) ◽  
pp. 359-380 ◽  
Author(s):  
Georges Ungar ◽  
Takuso Yamura ◽  
Jacqueline B. Isola ◽  
Sidney Kobrin
Keyword(s):  
Amino Acid ◽  
Guinea Pig ◽  
Proteolytic Activity ◽  
Guinea Pigs ◽  
Protease Activity ◽  
Amino Acid Esters ◽  
Protein Substrates ◽  
Protease Activation ◽  
Acid Esters

Protease activity was measured through the hydrolysis of synthetic amino acid esters in body fluids and tissues of guinea pigs, rats, mice, and humans. Significant in vitro activation was observed in serum and lung slices of sensitized guinea pigs on addition of the specific antigen. Increased proteolytic activity was also seen in reverse anaphylaxis. More marked activation occurred when guinea pig serum was treated with peptone and guinea pig or rat serum was treated with agar. Protease activation was demonstrated in specimens of human skin under the influence of a poison ivy extract or croton oil added in vitro. Urinary protease activity of guinea pigs increased significantly during the first hours of anaphylactic shock and very markedly in peptone shock. Peptone shock, elicited in mice pretreated with H. pertussis, was accompanied by a considerable increase in protease activity in the peritoneal fluid as compared with non-pretreated mice which were insensitive to peptone. Proteolytic activity resulting from the activation procedures was due to a number of proteases. The dominant substrate affinity and inhibition patterns suggest that serum and urine proteases are similar to but not identical with plasmin. Anaphylactic activation exhibited patterns different from those resulting from the action of anaphylactoid agents. Tissue enzymes are either of cathepsin- or chymotrypsin-type or mixtures of both. Some of the activated enzymes, although remarkably effective in hydrolyzing amino acid esters, show no activity on protein substrates. This does not justify, however, their designation as "esterases." They probably belong to the class of specific proteases acting only on a single or a small number of functionally significant protein substrates. There is at present sufficient evidence to prove not only that protease activation does occur in anaphylaxis and anaphylactoid conditions but also that it is an important component of the chain of reactions leading to the allergic response.

The effects of furosemide and bumetanide on lung liquid production by in vitro lungs from fetal guinea pigs

1990 ◽  
Vol 68 (8) ◽  
pp. 1131-1135 ◽  
Author(s):  
J. Thom ◽  
A. M. Perks
Keyword(s):  
Body Weight ◽  
Guinea Pig ◽  
Guinea Pigs ◽  
Loop Diuretics ◽  
Dilution Technique ◽  
Blue Dextran ◽  
Lung Fluid ◽  
Lung Liquid ◽  
Secretion Rates

Lungs from fetal guinea pigs of 61 ± 3 days of gestation were supported in vitro for 3 h, and lung liquid secretion rates were measured by a dye dilution technique based on Blue Dextran 2000. Ten preparations that had received no treatment showed an average secretion rate of 1.12 ± 0.28 mL∙kg−1 body weight∙h−1 during the first hour, and there were no significant changes over the following 2 h. In studies of 54 fetal lungs, furosemide, bumetanide, control ethanol carrier, or saline alone were placed in the supporting medium during the middle hour of the 3-h incubations (ABA design). Furosemide at 10−3 M reduced secretion 83.4 ± 16.8%; at 10−4 and 10−5 M it produced smaller reductions. Bumetanide at 10−3 M usually produced reabsorption (129.9 ± 23.0% reduction), at 10−4 M it reduced secretion 30.9 ± 11.8%, but at 10−5 M it was ineffective. Control carrier and saline were without effect. The ability of the loop diuretics to produce reabsorption of fluid in some preparations suggests the unmasking of an active reabsorptive process. The results also suggest that lung liquid secretion in the fetal guinea pig, as in the sheep, is dependent on a Na+ and Cl− cotransport system.Key words: fetus, lung fluid, bumetanide, furosemide.

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