The Journal of Experimental Medicine

Ceppellini, Ruggero; Landy, Maurice

doi: 10.1084/jem.117.3.321pmid: 14019651

Erythrocytes coated with bacterial capsular polysaccharides, notably the Vi antigen, were no longer agglutinated by antibodies directed against the various antigens native to the red cell surface. These effects could not be attributed to prevention of antibody uptake even though in some systems the uptake of antibody was diminished. In fact, agglutination by Rh-incomplete antibody was brought back to the original titer only after the sensitized Vi-coated cells had been subjected to ten alternating exposures to globulin and antiglobulin. Hemagglutination by Newcastle, mumps, and influenza viruses was also suppressed. Erythrocytes coated with Vi polysaccharide assumed the distinctive physicochemical attributes of this acidic polymer which results in a stabilization of the erythrocyte suspension as manifested by increased electrophoretic mobility and a striking decrease in the rate of sedimentation. Among the possible models for explaining the nature of the Vi effect on immune agglutination, the data favor interference with lattice formation. Footnotes Submitted: 14 September 1962

Pierce, G. B.; Midgley, A. R.; Ram, J. Sri

doi: 10.1084/jem.117.3.339pmid: 13943400

A parietal yolk sac carcinoma of the mouse that secretes large quantities of basement membrane-like material has been used to study the formation of basement membranes. Suitably characterized fluorescein-labeled antibodies against this material stained basement membranes of epithelial structures and vessels, as well as reticulin. When absorbed with reticulin and vascular basement membranes of the spleen until these structures no longer fluoresced, the antibody still stained the basement membrane-like material of the tumor, its normal embryonic counterpart (Reichert's membrane), and the basement membranes at the bases of epithelial cells. The observation made previously that parietal yolk sac cells secreted, in the absence of connective tissue and reticulin, the basement membrane (Reichert's membrane) upon which they rested has been confirmed through the localization of ferritin-labeled antibody to the endoplasmic reticulin of the secreting cells. Since a basement membrane proven to be an epithelial secretion is antigenically similar to basement membranes at the bases of all epithelial cells studied but antigenically different from connective tissue elements, it is postulated that the basement membranes at the bases of epithelial cells in general are an epithelial secretion, and are not a condensation of ground substance as is commonly believed. Footnotes Submitted: 22 October 1962

Bornstein, Donald L.; Bredenberg, Carl; Wood, W. Barry

doi: 10.1084/jem.117.3.349pmid: 14014022

Although the absolute febrile responses of trained individual rabbits injected intravenously with small to moderate doses of leucocytic pyrogen vary over an appreciable range, the relative responses of each rabbit to changes in dosage are satisfactorily reproducible. The quantitative dose-response relationship is characterized by a hyperthermic ceiling at which the intensity of the febrile reaction is relatively constant over a wide dosage range. Only at lower dose levels, where the dose-response curve is reasonably steep, is the magnitude of the fever produced proportional to the amount of pyrogen injected. When sufficiently large doses of LP are injected, the hyperthermic ceiling is exceeded. The fevers thus induced are biphasic in character and, in this way, resemble the usual response to bacterial endotoxin. Similar biphasic fevers result from continuous infusions of relatively low concentrations of LP at a constant rate. Repeated intermittent injections of moderate doses of LP likewise cause prolonged biphasic fevers, but, once the fever has become established, the reaction to each individual injection becomes markedly depressed. When large doses of LP are injected at daily intervals, the characteristic biphasic response occurs only following the first injection. Thereafter a state of tolerance intervenes in which the late secondary rise in temperature fails to occur. This form of tolerance lasts as long as the daily injections are continued but subsides within a few days after the injections are stopped. During the transient tolerance the rabbit's responsiveness to small doses of LP (in the sensitive range of the dose response curve) is depressed. In addition, the amount of endogenous pyrogen mobilized from the tissues by a large dose of LP is not as great as that generated in a normal rabbit. The relations of these findings to biphasic fevers, tolerance, and the accuracy of the conventional method of pyrogen assay are briefly discussed. Footnotes Submitted: 27 November 1962

Lee, Leung

doi: 10.1084/jem.117.3.365pmid: 13929108

In the presence of reticuloendothelial blockade, the intravenous injection of a protein antigen into specifically immunized rabbits or the infusion of soluble immune complexes into normal animals has been shown to result in the production of bilateral renal cortical necrosis. The similarity in the pathogenesis of this lesion and that seen in the classical generalized Shwartzman reaction produced by bacterial endotoxins is indicated by ( a ) the failure of both lesions to develop in animals pretreated with large doses of heparin, ( b ) by the finding of "heparin-precipitable fibrinogen" in the circulation, and ( c ) by the presence of massive fibrin deposits within the glomerular capillaries. These findings indicate that antigen-antibody reactions in vivo are capable of activating the blood coagulation system and that the mode of action of bacterial endotoxins may have an immunological basis. Footnotes Submitted: 4 November 1962

Freimer, Earl H.

doi: 10.1084/jem.117.3.377pmid: 13959531

Intact bacterial membranes have been isolated from protoplasts prepared from Group A streptococci by a cell wall-dissolving enzyme. A membrane fraction with identical chemical and serological properties has been obtained by differential centrifugation of mechanically disrupted streptococci. The membrane is chemically distinct from the cell wall and is composed of 72 per cent protein, 26 per cent lipid, and 2 per cent carbohydrate. Capillary precipitin tests and analysis by microdiffusion have demonstrated that the membrane contains antigens distinct from those of the cell wall and from those of the cytoplasm which it envelops. Evidence is presented which demonstrates that this antigenic material is common to the membranes of Group A streptococci, and that it can be distinguished by immunodiffusion from related antigenic substances present in membranes of several other serological groups of hemolytic streptococci. This antigenic material does not cross-react with the membrane antigens of other Gram-positive cocci. Footnotes Submitted: 17 October 1962

Coe, John E.; Salvin, S. B.

doi: 10.1084/jem.117.3.401pmid: 14021926

"Gastric feeding" of adult guinea pigs with dinitrochlorobenzene (DCB) resulted in a specific unresponsiveness to sensitization with the specific contact hapten. The more DCB gastric-fed to a guinea pig, the more complete the unresponsiveness to the hapten. When mycobacteria were incorporated into the sensitizing emulsion, the state of unresponsiveness to the dinitrophenyl (DNP) group was less apparent. When animals gastric-fed with DCB were later sensitized with an in vitro conjugate of the hapten combined with a heterologous protein such as dinitrophenyl-hen egg albumin (DNP·HEA), an immune response similar to that in the controls occurred both to the hapten and to the protein carrier. However, when the tolerant animals were sensitized with a conjugate containing a homologous protein carrier such as dinitrophenyl guinea pig serum (DNP·GPS), they showed diminished immune responses in comparison with those in the non-tolerant controls. The presence of circulating anti-DNP antibodies from sensitization with DNP-HEA did not affect the unresponsiveness to the specific contact hapten, regardless of whether these antibodies are present before or after induction of tolerance. Sensitization with picryl chloride (PiCl) (a cross-reacting hapten), either before or after gastric feeding of DCB, did not affect the state of unresponsiveness to DNP. Similarly when the DNP-tolerant animal was sensitized with PiCl, the subsequent immune response was similar to that in the controls; cross-reactions with the DNP group both in the contact and circulating antibody phase occurred at a rate similar to that in the controls. The foregoing relationships can be explained by presuming that, upon the gastric feeding of DCB, an in vivo conjugate is formed with a somatic protein, which determines the basic specificity of the tolerance. Acquired tolerance seems to manifest an immunologic specificity similar to that of delayed hypersensitivity, a relationship not unexpected if delayed hypersensitivity is an early phase of the immune response. Footnotes Submitted: 15 October 1962

Opie, Eugene L.

doi: 10.1084/jem.117.3.425pmid: 13940208

Changes characteristic of an inflammatory reaction caused by the injection of an injurious agent into the peritoneal cavity were measurable by the quantity of peritoneal fluid, by the number of leucocytes that entered the cavity, and by the abundance of exuded protein. Inflammation increased in severity along with ion dissociation and increase of the valence of electrolytes. With chlorides, sulfates, and nitrates it increased in each instance with the valence of their basic ions. The reactions caused by sodium chloride, sodium sulfate, and sodium citrate increased with the valence of their acid ions. Salts of heavy metals which fix and precipitate proteins caused more active inflammation than did other electrolytes. Histamine and arginine caused active inflammatory reactions with similar characteristics. The amino compounds, urea, citrulline, and creatinine, glycine, alanine, histidine, arginine, and histamine, produced inflammatory reaction in the order of severity with which, in foregoing experiments, they had caused necrosis when injected into the dermis. The acids and alkalis that were tested caused active inflammation when their acidity approached pH 1 or their alkalinity pH 11 respectively, but with approximate neutrality the inflammatory reaction became scant. These changes were in accord with the extent of necrosis when acid or alkalis, in foregoing experiments, were injected into the dermis. When histamine or arginine combines with hydrochloric acid to form histamine dihydrochloride or arginine monohydrochloride alkalinity is lost and inflammatory reactions caused by the hydrochlorides are relatively mild. The characteristic changes which accompany histamine and arginine do not occur. The combination of histamine with phosphoric acid to form histamine acid phosphate has similar relation to histamine. Inflammatory reactions caused by monobasic, dibasic, and tribasic sodium phosphate increased in activity in accord with increasing alkalinity referable to the basic ions of these salts. The activity of inflammatory reactions has been found to vary in accord with the chemical constitution of the injurious agents that have been tested. Footnotes Submitted: 18 November 1962

Najarian, John S.; Feldman, Joseph D.

doi: 10.1084/jem.117.3.449pmid: 13937305

Passive transfer of transplantation immunity was accomplished in inbred guinea pigs with tritiated thymidine-labeled lymphoid cells sensitized to homologous tissues. Autoradiographs of the homologous skin graft sites disclosed the presence of relatively few or no labeled cells at the site of rejection. Passive transfer of transplantation immunity was also accomplished with sensitized lymphoid cells enclosed in cell-impenetrable Millipore chambers. Previous studies with passive transfer of tuberculin sensitivity in guinea pigs revealed that the specifically sensitized cells could be easily found at the site of challenge in the presence of specific antigen and were ineffective when enclosed in Millipore chambers. It appeared, then, that the homograft reaction and delayed sensitivity of tuberculin type were achieved by different immunologic mechanisms within the same species. Footnotes Submitted: 7 November 1962

Uhr, Jonathan W.; Finkelstein, Martin S.

doi: 10.1084/jem.117.3.457pmid: 13995245

Injection of a sufficient dose of bacteriophage ϕX 174 into guinea pigs results in the formation of rapidly sedimenting antibody molecules (19S), and later, slowly sedimenting molecules (7S). Above a threshold dose of antigen, the relative rate of 19S formation is maximal and dose-independent; below this dose, slower relative rates are obtained. The time for doubling the serum 19S level is as short as 6 to 8 hours, suggesting that the absolute rate of antibody formation per cell is increasing in addition to proliferation of antibody-producing cells. Synthesis of 19S after injection of 10 10 ϕX virtually ceases at 10 days after which 19S antibody activity disappears from the circulation with a half-life of approximately 24 hours. A second injection of ϕX on day 5 or 9 prolongs 19S synthesis, indicating that antigen not only can regulate the relative rate, but also is essential for continued synthesis of 19S. 19S synthesis is also prolonged in guinea pigs by injection of ϕX with endotoxin or by 400 r whole body x-irradiation 24 hours after injection of phage into rabbits. The primary 7S response is not detected until approximately 1 week after immunization and relative rates are antigen-dependent. Primary 7S synthesis can continue for many months and leads to preparation for a secondary antibody response (immunological memory) during which only 7S is detected. In contrast, in animals that form precipitating 19S without detectable 7S, a second injection of phage 1 month later results in a second 19S response which closely resembles the first. These findings have led to the suggestion that formation of 19S does not lead to persisting immunological memory. Footnotes Submitted: 19 November 1962

Bellanti, Joseph A.; Eitzman, Donald V.; Robbins, John B.; Smith, Richard T.

doi: 10.1084/jem.117.3.479pmid: 13970471

By intensive stimulation with large amounts of Salmonella flagellar antigen, newborn rabbits were induced to form high titer flagellar agglutinins usually by the 7th to 10th day of life. Characterization of the agglutinins at various times during the first 30 days of life revealed that the earliest antibody which appeared was a gamma-1 macroglobulin, and that 7S gamma-2 globulins did not appear until the 4th or 5th week of life. In contrast, the adult animals produced macroglobulin antibodies for only 3 to 5 days before the lower molecular weight variety appeared. The infant macroglobulin appears to be similar in all respects to adult macroglobulin antibodies. These data are interpreted to indicate that the newborn and adult rabbit differ in their response to this type of stimulus not in timing of macroglobulin antibody production, but chiefly in the prolonged interval, which precedes the development of the capacity for the 7S type response in the newborn animal. Footnotes Submitted: 8 November 1962