The Journal of Experimental Medicine

Cochrane, Charles G.; Weigle, William O.

doi: 10.1084/jem.108.5.591pmid: 13587844

The in vivo activity of soluble antigen-antibody complexes was tested by a single intradermal injection in rabbits. Skin reactions were obtained marked by erythema, induration, and occasionally hemorrhage and necrosis. Microscopically, diffuse inflammation and occasional vascular necrosis could be found at all dosages. This indicates that soluble antigen-antibody complexes are phlogogenic and provides support for the suggestion that complexes are responsible for the lesions seen in serum sickness. The reactions were similar in severity to local passive Arthus (LPA) reactions at equal dosages of antibody in the dosage range studied. BSA antigen could be found in large concentrations in affected vessel walls of both reverse passive Arthus (RPA) and active or classical Arthus reactions. It is suggested that this predominantly vascular localization of antigen might bring about the relative severity of the RPA and active Arthus reactions, as contrasted to the complex and LPA reactions. The finding of affected vessels in the complex and LPA reactions containing little or no antigen and antibody, while these components were present in adjacent areas, suggests that the antigen-antibody combination may cause vascular reaction and damage by the release of physiologically active mediators from the tissue or tissue fluid. Footnotes Submitted: 18 June 1958

Dalldorf, Gilbert; Weigand, Heribert

doi: 10.1084/jem.108.5.605pmid: 13587845

Young cynomolgus monkeys inoculated intracerebrally with an attenuated Type 1 polio virus and, after 5 days, with a monkey adapted Coxsackie A-14 virus frequently became paralyzed. Neither virus alone was capable of inducing paralysis. Similar results were observed when the AB IV strain of Coxsackie A-7 was substituted for the A-14 virus. In this case the 2nd inoculation was made intramuscularly. Paralytic poliomyelitis may at times represent the summation of two infections, the total of motor neuron destruction by two independent and noninterfering enteroviruses. Footnotes Submitted: 19 June 1958

Bader, John P.; Morgan, Herbert R.

doi: 10.1084/jem.108.5.617pmid: 13587846

Mouse fibroblasts (L cells) fail to support the growth of psittacosis virus (6BC strain) if they are maintained on a medium containing only inorganic salts and glucose for 2 days prior to infection. Virus propagation can be stimulated by the addition of a synthetic medium containing amino acids, water-soluble vitamins, glutamine, glucose, and inorganic salts. By omitting single amino acids from the complete synthetic medium, tyrosine, threonine, methionine, isoleucine, phenylalanine, tryptophan, leucine, valine, and cysteine or cystine were found to be essential for stimulation, while lysine, arginine, histidine, hydroxyproline, proline, glutamic acid, aspartic acid, serine, alanine, and glycine were not essential. The cells on deficient media showed varying degrees of degenerative changes, but there was little correlation between ability to support psittacosis virus growth and morphologic condition of the cells. Glucose is also an essential component of the medium for viral growth, but the absence of glutamine had no effect on stimulation of virus propagation. L cell cultures maintained on media deficient in phenylalanine or tryptophan for 2 days before infection were also found to be incapable of supporting virus growth. The implications of this study in latent viral infections are discussed. Footnotes Submitted: 26 June 1958

Irving Lieberman; Peter Ove

doi: 10.1084/jem.108.5.631pmid: 13587847

Norman, Philip S.; Hill, Betsy M.

doi: 10.1084/jem.108.5.639pmid: 13587848

There are two inhibitors of plasmin in the plasma proteins of the human. α1-antiplasmin is heat-labile and migrates as an α1-globulin on electrophoresis. It combines non-dissociably with plasmin at a rate that depends on temperature. α2-antiplasmin is relatively more heat-stable and migrates as an α2-globulin on electrophoresis. It combines dissociably with plasmin independently of temperature. Footnotes Submitted: 2 July 1958

Padgett, Billie L.; Walker, Duard L.

doi: 10.1084/jem.108.5.651pmid: 13587849

The rate of elution of the variant virus from chicken RBC is progressively decreased as the temperature of incubation is increased above 25°C. The activity of the parent virus, on the other hand, is increased as the temperature is increased up to 40°C. The major cause of the decreased activity of the variant at temperatures above 2S°C. is an inhibition of the variant enzyme rather than its inactivation. The activity of the variant enzyme is stimulated in the presence of strontium, calcium, and barium ions. Manganese has only a slight effect, and magnesium has no stimulatory effect on the elution of the variant virus. Elution of the variant at 37°C. is progressively inhibited at pH values above 7, while the parent virus is still active at pH 8. In the absence of calcium the variant enzyme, hemagglutinin, and infectivity are more heat labile than those of the parent virus. The addition of calcium increases the heat stability of all three properties of the variant, and in the presence of calcium the infectivity of the variant is as stable as that of the parent virus. Footnotes Submitted: 14 July 1958

Kidd, John G.

doi: 10.1084/jem.108.5.665pmid: 13587850

Conjugates made by coupling diazotized arsanilic acid with one or another of a variety of proteins regularly brought about the complete regression of established 6C3HED lymphomas in living mice without perceptibly harming the latter, while untreated control animals regularly died with lymphomatosis. Histologic studies made plain that the lymphoma cells promptly die in mice treated with the arsenic-azoproteins, while those in untreated control animals continue to proliferate. Various inorganic and organic arsenicals (including arsanilic acid and 4-arsonophenyldiazotate) were essentially devoid of effect on the lymphoma cells in vivo , and this proved true as well of the proteins employed (serum albumins and globulins procured from several species, casein, and ovalbumin). Mixtures of arsanilic acid and the several proteins, various sulfur-azoproteins, and a number of other substances— viz ., amethopterin, chlorambucil, 6-mercaptopurine, 8-azaguanine, azaserine, 6-azauracil, 5-fluorouracil, thioTEPA, and DON, each given in maximal tolerated amounts—also failed to influence notably the course of established 6C3HED lymphomas in vivo . Although readily overcoming Lymphoma E9514 cells growing in the subcutaneous tissues of susceptible mice, the arsenic-azoproteins had little or no effect once these cells had reached the livers and spleens of susceptible hosts. Furthermore the arsenic-azoproteins had little or no effect in vivo on the cells of Lymphoma AKRL1, L1210, and L4946. The findings were considered in relationship to the respective susceptibilities of several types of lymphoma cells to other anti-lymphoma agents—notably guinea pig serum, immune serums prepared in rabbits with mouse lymphoma cells as antigens, and a variety of chemical compounds. Taken together, the observations provide proof that lymphoma cells of various types, although resembling one another quite closely in growth characteristics following transplantation in susceptible hosts, and in morphology as disclosed by ordinary microscopy, differ notably in susceptibility to the effects of the several anti-lymphoma agents. Footnotes Submitted: 1 July 1958

Skarnes, Robert C.; Rosen, Fred S.; Shear, Murray J.; Landy, Maurice

doi: 10.1084/jem.108.5.685pmid: 13587851

A humoral substance which inactivates endotoxin in vitro has been shown to be clearly distinguishable from complement, properdin, and specific antibody. For the present, it is designated "endotoxin-detoxifying component" or EDC. Animal species could be grouped in three categories with regard to the EDC activity of their sera; rat serum was highly potent; chimpanzee, dog, horse, and guinea pig sera were much less active; mouse, rabbit, and sheep sera exhibited no activity. The EDC potency of human sera varied widely, ranging from high to barely discernible activity. In contrast to the variations of EDC potency in serum, citrated plasma from all species manifested high potency of about the same magnitude. The influence of time, temperature, pH, and concentration of reactants on the inactivation of endotoxin by EDC was examined. EDC activity in plasma and serum was found to be labile to beating at 56°C. for 1 hour. Bacterial endotoxins, derived by different isolation procedures from smooth and rough Gram-negative species, varied considerably in susceptibility to EDC action. Footnotes Submitted: 9 July 1958

Rosen, Fred S.; Skarnes, Robert C.; Landy, Maurice; Shear, Murray J.

doi: 10.1084/jem.108.5.701pmid: 13587852

The uniformly high potency of citrated plasma as compared with the limited capacity of serum to inactivate endotoxin in vitro was found to be a consequence of the anticoagulant employed in collecting the plasma. Addition of calcium to plasma suppressed the activity of its endotoxm detoxifying component (EDC) whereas the addition of calcium-binding anticoagulants rendered serum comparable to plasma. Dialysis of plasma resulted in a marked reduction of its EDC activity despite the concommittant elimination of calcium. EDC activity could then be fully restored upon the addition of calcium-binding anticoagulants. Resin-treated plasma, without added anticoagulant, had EDC activity equal to plasma obtained with calcium-binding anticoagulants. Following dialysis, resin-treated plasma also sustained a marked reduction in EDC activity which could be fully restored by calcium-binding anticoagulants. Restoration was also obtained with the dialysate even after ashing. These findings indicated that the suppression of EDC activity by calcium is not direct but is mediated through its effects on an anionic component of plasma which is required for inactivation of endotoxin by EDC. Footnotes Submitted: 9 July 1958

Rowe, Wallace P.; Hartley, Janet W.; Roizman, Bernard; Levy, Hilton B.

doi: 10.1084/jem.108.5.713pmid: 13587853

Infectious tissue culture fluids of the majority of serotypes of adenovirus at low dilutions detach HeLa or KB cells from glass surfaces within a few hours after inoculation. A reproducible method for testing cell detachment was devised. The factor present in infectious tissue culture fluids and responsible for cell detachment is trypsin-sensitive and non-dialyzable; it is smaller and more resistant to the effect of heat or ultraviolet light than the infectious virus particle. Cell detachment activity was found to be temperature-dependent, and the cell-detaching titer of infectious tissue culture fluids was not affected by repeated exposure to HeLa cells. Inhibition of cell detachment by human or rabbit sera was observed only when other antibodies to adenovirus antigens were also present, but the antibody inhibiting cell detachment could not be correlated quantitatively with complement-fixing or homologous neutralizing antibody. Footnotes Submitted: 15 July 1958