The Journal of Experimental Medicine

Allison, Marvin J.; Zappasodi, Peter; Lurie, Max B.

doi: 10.1084/jem.115.5.881pmid: 13860625

We have found a phase of susceptibility associated with a reduced metabolic activity on the part of peritoneal mononuclear phagocytes taken from BCG-vaccinated rabbits. A second stage of heightened resistance to infection was found to be associated with a heightened metabolic activity. The period of susceptibility in BCG vaccination is primarily concerned with initiation of the infection and not with the progression of the disease, which in both stages is increased. These reactions are discussed in relation to other conditions, such as nonspecific protein therapy and the administration of endotoxin, which also have similiar biphasic stages of resistance. Of incidental interest is the fact that rabbits who received 400 roentgen units are two years later still unable to respond to BCG vaccination with an increase in resistance. We conclude that there is a relationship between the level of certain metabolic activities of reticuloendothelial cells and resistance to tuberculosis. Footnotes Submitted: 3 December 1961

Mellors, Robert C.; Brzosko, Witold J.

doi: 10.1084/jem.115.5.891pmid: 14472423

After intravenous injection in mice, rabbit immune complexes, solubilized in antigen excess and containing fluorescent antigens (BSA* or OA*) or fluorescent antibody, or both, were promptly localized in reticuloendothelial cells, and polymorphonuclear leukocytes, of the sinusoids of liver and the red pulp of spleen; in glomeruli and elsewhere in kidney; in capillary endothelium of heart and lung; and in hepatic cells. Thereafter manifold processes occurred. Within 48 hours the immune complexes were scarcely detectable in liver and splenic red pulp but now were localized in the germinal centers of white pulp where heretofore they had been seen only in trace amounts. This new localization presumably was associated with the antibody-forming activity of the germinal centers, for the immune phase of antigen clearance from the blood had already begun. Although the immune complexes were localized in various regions of the nephrons and their appertaining blood vessels, the initial sites of predilection were the glomerular capillary walls and intercapillary spaces. After 48 hours the immune complexes were still detectable, although in diminished amounts, in the glomeruli but had by now essentially disappeared from other renal sites. The localization of immune complexes in the kidney was associated with proteinuria and with structural changes which closely simulated in some instances those of human membranous glomerulonephritis, of focal and diffuse types, and consisted mainly of eosinophilic swellings of the glomerular capillary walls, intercapillary spaces, and basement membranes. There was a close correspondence between the distributions of the eosinophilic swellings and the fluorescent immune complexes. The renal localization and persistence of fluorescent antigens (BSA* or OA*), after separate injections in mice, differed from that of fluorescent immune complexes in several respects. For example BSA* showed predilection for the glomerular basement membranes and was localized sparsely in the capillary walls and intercapillary spaces; OA* was localized only in minute amounts; and neither was detectable in more than trace amounts at 48 hours after injection. These fluorescent proteins (of low molecular weights, 40,000, 70,000) did not cause glomerulonephritis within the time interval studied, whereas fluorescent immune complexes, containing on the average two molecules of antigen to one of antibody (with minimum molecular weights of 240,000 to 300,000) produced glomerulonephritis in some instances, in confirmation of the observations of others. Since the localization of the immune complexes occurred immediately and without known immunologic relation to the kidney itself, the selective physical retention of proteins by structures comprising the glomerular ultrafilters appeared to be of pathogenic significance in this form of membranous glomerulonephritis in mice, as perhaps also in nephrotic glomerulonephritis in man. If after injection of fluorescent immune complex, homologous antiserum was also administered intravenously so as to produce acute anaphylactic death, coarse and occlusive depositions of immune precipitates occurred in pulmonary, myocardial, and renal capillaries, and in hepatic sinusoids. Footnotes Submitted: 5 December 1961

Hull, R. N.; Cherry, W. R.; Tritch, O. J.

doi: 10.1084/jem.115.5.903pmid: 14449901

The establishment of two strains of rhesus monkey kidney cells in continuous tissue culture, the development of a subline adapted to chemically defined medium, and the isolation of several clonal derivatives were described. Growth characteristics, chromosome numbers, malignant potentiality, and freeze storage data are presented. The cells were studied for their sensitivity to a large number of viruses and were extensively compared with primary cultures of monkey kidney cells for sensitivity to poliovirus. The cell strains were not sensitive to all the viruses which could be grown in primary cultures of the same tissue but were susceptible to most of them. In some instances an advantage to the use of the cell strain for certain viruses was noted. Footnotes Submitted: 10 December 1961

Germuth, Frederick G.; Maumenee, A. Edward; Senterfit, Laurence B.; Pollack, Abou D.

doi: 10.1084/jem.115.5.919pmid: 19867210

The injection of antigen into the center of the avascular cornea of homologously sensitized animals induced a ring of opacification between the center of the cornea and the limbus. This ring of opacification was composed of a line of deeply eosinophilic amorphous material in a matrix of swollen collagen fibers, palisaded by polymorphonuclear leucocytes. By use of fluor-tagged antigen, it was shown that the line of damage in the cornea coincided with the precipitation of antigen presumably by antibody entering the cornea from the limbal vessels. With the passage of time, the antigen-antibody precipitates were removed, at least in part by phagocytosis, and the ring of opacification was replaced by ingrowing blood vessels surrounded by plasma cells. Treatment of sensitized animals with nitrogen mustard showed that antigen-antibody interaction could injure the corneal stroma in the relative absence of polymorphonuclear leucocytes. Footnotes Submitted: 18 December 1961

Andres, Giuseppe A.; Morgan, Councilman; Hsu, Konrad C.; Rifkind, Richard A.; Seegal, Beatrice C.

doi: 10.1084/jem.115.5.929pmid: 13861458

Ferritin-conjugated antibody has been used to identify by electron microscopy the sites at which nephrotoxic globulins localize in rat kidney during acute experimental glomerulonephritis. Antibody was concentrated in the glomerular basement membrane and in basement membrane-like material contained in distended cisternae of the endoplasmic reticulum. These data confirm and amplify, at the ultrastructural level, the results of studies obtained with the fluorescent antibody technique, and are consistent with the hypothesis that the cisternae and capillary basement membrane possess common proteins. Footnotes Submitted: 18 January 1962

Chang, R. Shihman; Humes, Maryellen

doi: 10.1084/jem.115.5.937pmid: 13878102

Experiments designed to characterize an unidentified transmissible agent brought forth the following findings: The cytopathology consisted of the formation of intranuclear globules, collapse of the involved nuclei, and the extrusion of nuclear materials. The relatively dormant primary human amnion cells were less susceptible than the rapidly growing cell lines. Similarly, the slowly multiplying ribose variants were less susceptible than their corresponding parent cell lines. Interferon-like activity was released from infected cells. Infectivity was readily demonstrated following storage at 0–4°C for at least 8 months or at 37°C for at least 2 weeks. Freeze-thawing, however, markedly reduced or completely destroyed its infectivity. Infectivity was destroyed completely by ether and chloroform; partially by desoxycholate, and not affected by trypsin, papain, RNAse, DNAse, hyaluronidase, lysozyme, lecithinase, or pancreatic lipase. The rate of inactivation by 0.025 per cent formalin was much slower than that of vaccinia and herpes viruses. Its synthesis was suppressed by 5-fluorodeoxyuridine. This suppression was not reversed by thymidine and/or uracil. Heat-stable neutralizing antibody could not be demonstrated in 379 human and animal serums, in human gamma globulins, or in serums from animals "immunized" with this agent. Heat-labile inhibitors (lipoprotein-like) capable of inhibiting the infectivity of this agent were demonstrated in 154 of the 157 serums tested. Experimental evidence indicated the non-identity of this ubiquitous inhibitor and the properdin system. The non-infectious complex between this agent and the ubiquitous serum inhibitor may be dissociated (hence, become infectious) by simple dilution. Repeated attempts to reisolate a similar agent have not been successful. We have hypothesized that this agent is a virus consisting of DNA wrapped in a surface coat rich in lipid, and suggest that this virus be referred to tentatively as a lipovirus. Footnotes Submitted: 1 January 1962

Chang, R. Shihman; Geyer, Robert P.; Andrus, Stephen B.

doi: 10.1084/jem.115.5.959pmid: 13878101

The release of a toxic product from cultured human cells infected by a new cytopathic agent (the lipovirus) was described. This toxin was dissociable from the infectious particles. It induced sudanophilia of human and mouse cells, an increase in the total fatty acid content, and a change in the major constituent fatty acids. Similar toxin was not demonstrated in cultures infected by the vaccinia herpes simplex, adeno 3, polyoma, polio 1 and 2, Coxsackie B1, parainfluenza A, and Rous sarcoma viruses. Preliminary characterization indicated that this toxin was resistant to tryptic digestion and could not be dialyzed or neutralized by human gamma globulin. It was inactivated at 58°C for 30 minutes, but stable at 37°, 4°, and –60°C. The significance of these findings is discussed. Footnotes Submitted: 1 January 1962

Chang, R. Shihman; Liepins, H.

doi: 10.1084/jem.115.5.967pmid: 13878103

DNA and thymine degradation on cultured human, mouse, and chick cells were studied. Significant increase in DNA-degrading activity was demonstrated in human embryonic cells killed by freeze-thawing, liver cells killed with mitomycin C, mouse embryonic cells infected with encephalomyocarditis virus, and in all cells killed by the lipovirus. Twelve other viral agents, actinomycin D, and 5-fluorodeoxyuridine failed to produce a similar increase. Thymidine-2-C 14 -labeled cultures, either live, killed, or infected by 19 different physical-chemical and biological agents, did not release detectable quantity of C 14 C 2 . Following infection with the lipovirus 20 to 60 per cent of the total radioactivity of thymidine-2-C 14 -labeled cultures was liberated as C 14 O 2 . It was postulated that the lipovirus introduced into the host cells the missing genetic information necessary for the synthesis of one or more enzymes responsible for the reductive catabolism of thymine. Footnotes Submitted: 1 January 1962

Szulman, Aron E.

doi: 10.1084/jem.115.5.977pmid: 19867211

The H antigen was mapped out by immunofluorescence in human tissues (including those of fetuses from 15 cm crown-heel length) from individuals of the various groups within the ABO system, both secretors and non-secretors. The distribution of the antigen can be summarized under the following headings: Cell walls of endothelium : present throughout the cardiovascular system; Cell walls of stratified epithelia : in skin, non-cornifying squamous stratified membranes, transitional epithelia; Mucus : occurring wherever the latter is produced in secretor individuals and confined to a few special topographical areas in non-secretors; Secretions and excretions : the pancreatic and sudoriferous (independent of secretor status), and mammary and uterine (governed by the secretor makeup) all contain it. The distribution of the H antigen is most fully represented in tissues of group O. It follows an over-all universal pattern, characteristically modified in non-secretors, equally valid for antigens A and B described in a preceding study. Within this pattern, in tissues of the non-O groups, the complement of the H substance in its various forms wanes in a manner consistent with the hypothesis that it serves as a substrate for the A 1 , A 2 , B genes, exerting their action with different degrees of efficiency. The secretor:non-secretor phenomena can be most simply interpreted by viewing the non-secretor, recessive gene (in the homozygous, ss condition) as inhibiting the production of some of the water-soluble forms of the blood group substances. Since the gene was never found responsible for dissociation of the H and A, B antigens its inhibitory action is thought to be wrought at the point of formation of the basic H substance or its precursor. Footnotes Submitted: 12 December 1961

Porter, K. A.; Cooper, E. H.

doi: 10.1084/jem.115.5.997pmid: 14488083

Newborn rats of one inbred strain were given an intracardiac injection of adult thoracic duct lymphocytes from another inbred strain. It was found that although there was a direct relationship between the number of small lymphocytes injected and the incidence of fatal runt disease, there was no particular relationship between the large lymphocyte content of an inoculum and its runt-inducing potentiality. Using tritiated thymidine and an autoradiographic technique, the small lymphocytes in the inoculum were labelled and found to migrate in large numbers into the cortex of the lymph nodes and into the Peyer's patches of the host animal, and in smaller numbers into the white pulp of the spleen. Within 24 hours isotope, previously present in the DNA of small lymphocytes, appeared in a number of the large pyroninophilic cells which were a characteristic feature of the spleen and lymph nodes in this early phase of runt disease. When the large lymphocytes in the inoculum were labelled they were found to migrate to the red pulp of the spleen, medulla of the lymph nodes, and the Peyer's patches and the lamina propria of the small intestine. Later some labelled small lymphocytes appeared at these sites. These findings suggest that: ( 1 ) Some small lymphocytes are immunologically competent cells, and ( 2 ) After introduction into the circulation of a newborn rat, these same small lymphocytes are the first cells to react with the antigens of the host, and in the process they become transformed into large pyroninophilic cells capable of division. The large lymphocytes seem to play little part in this initiating of an immunological response, but do give rise to some small lymphocytes. Footnotes Submitted: 7 January 1962