The Journal of Experimental Medicine

Broberger, Ove; Perlmann, Peter

doi: 10.1084/jem.110.5.657pmid: 13804543

Sera from 30 children, suffering from ulcerative colitis, were examined for the presence of antibodies capable of reacting with antigens of normal human tissue. It was possible to demonstrate that most of the sera contained a precipitating and hemagglutinating factor, reacting with a constituent of human colonic tissue. This constituent was obtained, within 1 hour after death, from colonic tissue of newborn babies who had died without feeding. It could also be prepared from fetal tissue. The antigen can be extracted with phenol-water at 65°C. and seems to be a polysaccharide. The precipitating factor in the sera of the patients behaves electrophoretically as a γ-globulin. Phenol-water extracts from liver and from kidney also reacted positively with the sera from certain patients. There are indications which suggest that the antigen obtained from these tissues is identical with that from colon. Sera from 38 healthy children did not give any reactions with the extracts used. In additional controls, the sera of 32 children with various diseases, all of suspected autoimmune origin, were also tested. A few of these reacted positively with the phenol-water extracts from the organs mentioned above. Most likely, the antigen reacting in these cases is different from that reacting with the antibodies in the sera from patients with ulcerative colitis. The possible role of the antibodies in the pathogenesis of ulcerative colitis and the mechanism of their formation are discussed. Footnotes Submitted: 15 July 1959

Gustafsson, Bengt E.; Laurell, Carl-Bertil

doi: 10.1084/jem.110.5.675pmid: 13830427

The earlier observed pronounced hypogammaglobulinemia in germfree rats of different ages has been confirmed. Using an immunologic technique the concentration of immunologic gamma globulins were found to vary between 10 and 15 per cent of the values observed in ordinary rats. Upon contamination of germfree rats with the normal microbial flora a pronounced lag phase was noted before the gamma globulin level became normal. This lag phase was most pronounced in growing rats. Newborn rats seem to start gamma globulin production more rapidly than older germfree rats. The response with regard to gamma globulin production on contamination of germfree rats with different types of bacterial cells through the natural routes is not identical. Footnotes Submitted: 20 July 1959

Beeson, Paul B.; Rowley, D.

doi: 10.1084/jem.110.5.685pmid: 13798253

In studying the problem of the peculiar susceptibility of the kidney to coliform bacterial infection it was found that kidney tissue, unlike that of other organs, interferes with the ability of normal serum to destroy these organisms. The effect was attributable to strong anticomplementary activity, 5 to 15 times greater than that of other organs. Inactivation of complement by kidney tissue was found to have characteristics of a chemical reaction, the active principle being heat-labile, non-dialysable, and difficult to separate from tissue particles. Attempts to purify it or to obtain it in a soluble form usually resulted in great loss in activity. The component of complement affected was the fourth; i.e ., that which is characterized by susceptibility to injury by ammonia. Similarities were found to exist between the conditions of ammonia formation and complement inactivation by kidney homogenates, the most notable being enhancement by phosphate and glutamine. The possibility is suggested that these findings may help to explain the vulnerability of the kidney to certain infections, especially those due to bacteria which are destroyed by the combined action of complement and antibody. Footnotes Submitted: 1 July 1959

Green, H.; Barrow, P.; Goldberg, B.

doi: 10.1084/jem.110.5.699pmid: 13851485

Rabbit antibody + complement alters the permeability properties of mouse Krebs ascites tumor cells and erythrocytes. When antibody + C' acts on ascites tumor cells in a low protein medium, intracellular K + is lost from the cells at a rate far greater than the normal leak rate. At the same time the cells lose amino acids and ribonucleotides and become fully permeable to the Na + of the medium. When antibody + C' acts in a low protein medium, the cells swell extensively and lose most of their macromolecules to the medium (hemoglobin from erythrocytes, protein and RNA from the ascites tumor cells). If the antibody + C' acts in a medium containing protein in sufficient concentration to balance the colloid osmotic pressure of the cells, the swelling is prevented; no macromolecules are then lost from the cells, but the loss of K + and entrance of Na + are not altered, and the loss of amino acids and ribonucleotides is only slightly affected. It therefore appears that the action of antibody + C' is to produce functional "holes" in the animal cell membrane which permit the equilibration of cations and small molecules between cell and medium. This leads to an increase in the osmotic pressure of the cell and a rapid influx of water. The cell membrane and its "holes" are thereby stretched, permitting macromolecules to escape from the cell. Footnotes Submitted: 1 July 1959

Gelzer, Justus; Suter, Emanuel

doi: 10.1084/jem.110.5.715pmid: 13827241

The effect of antibody on the fate of Salmonella typhimurium within mononuclear phagocytes (MN) of rabbits was studied in vitro . Monocytes and bacteria were incubated either in absence or presence of antibody. After 45 minutes during which phagocytosis occurred infected cells were washed to remove extracellular bacilli and free antibody. The cells were then reincubated in a medium without addition of antibody, and the interaction between the MN and bacteria was followed, correlating bacterial viability and the morphology of the mixture. The following results were obtained. The anti- Salmonella antibody was not bactericidal even in presence of complement and did not enhance phagocytosis. Regardless of whether antibody was present or absent during phagocytosis, the bacteria appeared to multiply within the cells. When no antibody was present during phagocytosis the infected cells were severely damaged within a few hours of incubation, and extensive extracellular multiplication was dominating. When antibody was present during phagocytosis MN packed with bacteria persisted for a long time. Little or no extracellular growth occurred. It was possible to demonstrate the presence of the antibody within the infected MN, using the fluorescent antibody technique. The antibody appeared as a coat around the bacteria. Antibody entered the cells only during phagocytosis, presumably attached to the bacteria. The active factor of the immune serum was found in the gamma globulin fraction and reacted specifically with the somatic antigen of Salmonella typhimurium . The antiflagellar portion of the antiserum was not involved in the phenomenon described. It is concluded that this antibody protects monocytes against the effect of intracellularly located Salmonella . Footnotes Submitted: 2 July 1959

Landy, Maurice; Trapani, Robert-John; Shear, Murray J.

doi: 10.1084/jem.110.5.731pmid: 14413831

The immunological properties of typhoid endotoxin were investigated after incubating it with serum under conditions in which EDC (endotoxin-detoxifying component) was operative. Brief interaction of endotoxin in vitro with this system in serum (human, rabbit, mouse) abolished its antigenicity for rabbits. Suitable modification of the host permitted separate elicitation of toxic and antigenic responses to typhoid endotoxin. Endotoxin altered by EDC was still capable of precipitating specific antibody; moreover, it did so in a manner which resembled that of the haptenic polysaccharide. Footnotes Submitted: 13 July 1959

Rowley, Donald A.; Chutkow, Jerry; Attig, Charles

doi: 10.1084/jem.110.5.751pmid: 14439721

Hemophilus pertussis vaccine injected into normal rat skin produced local edema lasting several days. Four to 6 days later the injected site became severely inflamed. When uninjected skin was challenged 5 to 28 days after the initial injection, severe inflammation developed at the site of challenge within 12 to 24 hours. This secondary hypersensitive response was elicited by a dose of vaccine which produced little or no initial or delayed inflammation in a normal rat. Specific cutaneous hypersensitivity to a particulate antigen ( i.e . typhoid vaccine) or a soluble antigen (human or rabbit gamma globulin) developed when rats were injected with a mixture of the antigen and pertussis vaccine. Pertussis vaccine mixed with typhoid vaccine did not enhance circulating agglutinin formation to typhoid vaccine. Cutaneous hypersensitivity to pertussis vaccine was passively transferred to normal rats by lymph node cells but not with serum from hypersensitive rats. Sensitization with pertussis vaccine did not enhance the edema-producing activity of histamine, serotonin, or 48/80 given subcutaneously. Mast cells in areas of hypersensitive inflammation were not damaged appreciably. The hypersensitive inflammation was not inhibited by treatment with anti-serotonin and antihistaminic drugs. Hypersensitive rats "depleted of mast cells" responded to challenge with pertussis vaccine with severe inflammation though their response to 48/80 was depressed. Hypersensitive rats treated with x-irradiation showed decreased hypersensitive inflammation though they responded normally to 48/80 and histamine. These studies failed to demonstrate a role for circulating antibody in the cutaneous hypersensitive inflammation produced in the rat by pertussis vaccine. Furthermore, the findings indicated that the cutaneous hypersensitive inflammation is not mediated by tissue serotonin and/or histamine. Footnotes Submitted: 10 July 1959

Levene, Charles I.; Gross, Jerome

doi: 10.1084/jem.110.5.771pmid: 14416144

The lathyrogenic agents, ß-aminopropionitrile and semicarbizide, when applied to the chorio-allantoic membrane of the chick embryo produced a dramatic increase in fragility of the embryo. This alteration was not associated with a change in the concentration of collagen, except in aorta, but was accompanied by a sharp increase in the amount of collagen extractible in cold 1 M NaCl from skin, bone, and aorta. Increase in fragility and extractible collagen began within 3 hours after introduction of the agent and rose steadily for at least 72 hours. Essentially no collagen could be extracted from tissues of normal chick embryos. Both fragility and amount of extractible collagen were dosage- and time-dependent. It is concluded that the extractible collagen in lathyrism consists of a large proportion of dissolved fibers previously insoluble and formed prior to administration of the agent. The data also suggest that the "lathyritic" collagen in vivo is not in molecular dispersion but in an aggregate or fibrillar form. It is dispersed by cooling. The extracted collagen could be reconstituted to typical striated fibrils in vitro and the molecule appeared to be normal in the gross, with regard to asymmetry ratio and intramolecular helical structure. The evidence at hand suggests that at least one of the defects induced by lathyrogenic agents is an interference with the normal intermolecular cross-linking within the collagen fibril. Footnotes Submitted: 20 July 1959

Bradley, G. Mary; Spink, Wesley W.

doi: 10.1084/jem.110.5.791pmid: 13803714

When small numbers of Brucella melitensis were inoculated into ABC mice, occasional hepatic granulomas without necrosis were demonstrated. The greatest multiplication of brucellae was detected in the spleens. Because it had been previously observed that ACTH or cortisone markedly accelerated the multiplication of brucellae in the livers of infected mice with destruction of liver cells, it was considered that triiodothyronine might likewise exaggerate a brucella infection by stimulating endogenous adrenal secretion. Although adrenal hypertrophy was produced, infection of mice treated with triiodothyronine resulted in severe hepatic necrosis or infarcts without the multiplication of brucellae in either the livers or spleens. The lesions were not encountered in untreated infected mice or in control mice treated with triiodothyronine. The necrosis was associated with minimal inflammatory reaction. The necrosis was not induced in mice treated with triiodothyronine and given brucella endotoxin. The precise genesis of the acute hepatic necrosis cited in these experiments remains undefined. Triiodothyronine did not cause deaths in mice infected with Br. melitensis . The infection was neither enhanced nor suppressed. Footnotes Submitted: 30 June 1959

Vogel, F. Stephen

doi: 10.1084/jem.110.5.801pmid: 19867166

Goldfish kept in water containing ionized copper and a detergent added with the aim of decreasing coagulation of the mucus on the gills, took in and retained this metal in their brains, livers, and kidneys, in concentrations comparable to those that occur naturally in Wilson's disease, as chemical assays disclosed. Histochemical studies made it clear that much copper had accumulated within the large neurons, principally in those of the telencephalon and anterior horn region of the spinal cord and in the tubular epithelial cells of the kidneys, the nuclei of the parenchymal cells of the liver, the sarcoplasm of the skeletal muscle, and in the epithelial covering of the gills. The intraneuronal deposition of copper was regularly associated after a time with conspicuous cytologic changes, notably contraction and hyperchromaticity of the nerve cells with tortuosity and fragmentation of the axis cylinders and lysis and loss of neurons. The accumulation of metal in the renal epithelium was frequently accompanied by necrosis and was regularly associated with hyperplasia and calcification of the epithelial cells of the larger renal tubules in all goldfish kept for prolonged periods in copper-rich water. The deposition of copper in the liver was not accompanied by consistent cytologic changes. The similarity of the cytologic alterations induced in the central nervous systems by copper and those that occur naturally in hepatolenticular degeneration in human beings provides evidence that copper itself plays an important role in the pathologic alterations of the brain in Wilson's disease. Footnotes Submitted: 15 July 1959