Water soluble palmitic acid in media for cultivation of leprosy derived psychrophilic mycobacteria from Mycobacterium leprae infected tissues

Cultivation trials for Mycobacterium leprae resulted in growth of Mycobacterium psychrophilum (L). Media were inoculated with host grown Mycobacterium leprae cells from armadillo tissues, Nu mice foot pads or human lepromata. Cultures were obtained in liquid and on semisolid multifactoria 1 media containing water soluble palmitic acid or its salts. Ammonium thioglycolate and Napalmitate served as carbon and energy sources. The water soluble palmitic acid remained in perfect solution following sterilization in the autoclave, thus easily accessible to the cells. The cyclodextrin-Fe complex served as a siderophore to grow the obtained leprosy derived psychrophilic cells. The leprosy derived cultures and subcultures grew opimally at+10 degrees Celssius but deteriorated rapidly at + 32 degrees Celsius, in the multifactorial media. No growth occurred in 7H9 media. Cultures were not identified for classification.

Soulier, savon et balai contre un parasite intra-cellulaire Mycobaterium leprae

les suspensiones purifiées de Mycobacterium leprae ont été isolées à partir des lépromes humains. Les extraits bacillaires avaient une activité élevée en ribulose-diphosphate carboxylase (RuDC). A cause de l`absence d`apha-cétoglutarate deshedrogenase, le cycle tricarboxylique était imcomplet. L NADH-oxydade était présente seulement à l`état de traces. Ce profil métabolique montre que Mycobacterium leprae possède génétiquement des caracteristiques autotrophiques similaires à celles des bacilles du sol. La présente du RuDC montre que Mycobacterium leprae pourrait utilisier le dioxide de carbone comme source unique de carbone

Reticuloendothelial response in murine leprosy

The capillary-endothelial and reticuloendothelial responses in rats with murine leprosy were compared with responses in uninfected controls. Experiments were performed on normal and serotonin-depleted rats, 20, 70 and 120 days after inoculation. Capillary reactions were provoked with dextran and with histamine, then visualized with India ink. A definite fluctuation in capillary response is demonstrable during the progression of the disease. In the early stage of the infection there is a stimulated systemic capillary response, but in the advanced stage this capillary alertness is no longer present. Similar stimulated endothelial response is present in the incipient and slowly progressing lesions of inoculation, whereas in the late necrotic stage there is no endothelial activation in the granuloma. Apart from the altered capillary response there is no change in the phagocytic function of the reticuloendothelial system as shown by the India ink test made 20 and 70 days after inoculation.

Experimental transmission of murine leprosy to the Guinea-pig by means of induced macrophage exudate and suppression of the natural defence mechanism

A method is report for the experimental transmission of rat leprosy to a normally resistant host, the guinea-pig, by (1) inducing a macrophage exudate in the peritoneal cavity before intraperitoneal injection of the infectious tissue homogenizate, and (2) simultaneous suppression of the natural resistance with an antihistamine (mepyramine maleate) or a complement-inhibiting compound (sodium polyanethol sulphonate). Large granulomas developed in 8 weeks in the peritoneal cavities of the guinea-pis so treated. Using these granulomas, the infection was transmissable to subgeneration of the guinea-pig, and could be transmitted back again into the rat. In other words, M. leprae murium retained its infectiousness in the resistant host. Histologic examinations have shown that the granuloma of the guinea-pig is structurally similar o that of the rat. The tissue gives a strongly positive reaction to the tetrazolium violet test. The method described permits the parasite to escape the damaging action of the extracellular inhibitors which Hanks considers to be the barriers to experimental transmission.

Action of histamine and antihistamine on the ingestion of murine leprosy bacilli by macrophages of the rat and the Guinea-pig

Phagocytosis of the tubercle bacillus of a BCG culture, and of the murine leprosy bacillus of a leproma suspension, by macrophages of glycogen-induced peritoneal exudates of guinea-pigs and albino rats was studied to determine the influence of histamine and an antihistamine (mepyramine maleate). Ingestion of BCG by the cells of both species was stimulated by histamine and inhibited by the antihistamine. On the other hand, phagocytosis of the murine bacillus was not influenced by either of the two substances, this contrary result being obtained with both kinds of exudate cells. A raw, bacillus-free, aqueous extract of the rat leproma inhibited phagocytosis of BCG by the guinea-pig monocytes. The active substance is considered responsible for the monopolization of the monocytes by M. leprae murium. The rat leproma extract and the antihistamine induce similar morphological and functional alterations of the monocytes; absence of pseudopodia and vacuoles, shrinkage of the cell, and a passive behavior with regard to BCG.

Studies on the effects of phagocytic stimulation on microbial disease. XI. Action of chaulmoogra derivatives on endothelial cells of skin vessels

Chaulmoogra oil and certain chaulmoogra derivatives have been found to induce the same intense phagocytic activity of the endothelial cells of the skin capillaries as is induced by histamine. The phenomenon could be inhibited in either case by previous administration of a synthetic anti-histaminic substance. From these facts it is supposed that chaulmoogra preparations induce the phenomenon by liberating latent histamine. It is therefore concluded that: 1- By the administration of active chaulmoogra derivatives, a large number of inactive endothelial cels may be stimulated to help the cellular defense mechanism, thus increasing the capacity of the reticuloendothelial system. 2- The experiments suggest that histamine activation is involved in this mechanism, but this needs further direct experimental evidence. 3- If histamine activiation is the mechanism of the action of chaulmoogra derivatives, a cycle of defense in which histamine plays an important role may be supposed. Since the phagocytic cells that participate in the pathology of leprosy are paralyzed in their function of destroying the parasite, the active chaulmoogra derivatives could stimulate this section of the defense mechanism, which interferes with bacteriolytic activity.

Studies on the effects of phagocytic stimulation on microbial disease. XII. Action of chaulmoogra oil on the reticuloendothelial system

Four groups of albino rats were injected intravenously with india ink after the parenteral administration of various substances. In the control, the ink disappeared from the blood stream in 39 to 68 minutes. The administration of synthetic antihistamine slowed the rapidity of the phagocytosis of the ink particles by the reticuloendothelial system, whereas the administration of histamine, and also of chaulmoogra oil, increased the rapidity of phagocytosis. The increased phagocytic activity thus induced by chaulmoogra oil was found to be inhibited by the simultaneous administration of an antihistamine substance. These experiments have brought further evidence about the mechanism of action of chaulmoogra oil in the treatment of leprosy, namely, that it stimulates the cellular defense mechanism of the host to increased activity.