The Journal of Experimental Medicine

Möller, Göran

doi: 10.1084/jem.114.4.415pmid: 19867192

The fluorescent antibody technique has been applied for the demonstration of mouse isoantigens at the cellular level. Specific reactions were obtained by the indirect or "sandwich" technique with a variety of living normal and neoplastic cells. Isoantigens of the H-2 system and of other systems could be demonstrated as well and appeared to be localized at the cell membrane. As far as the H-2 system was concerned, the membrane localization could be confirmed on histological sections. Different types of non-specific staining reactions have been identified and described. Pinocytosis and cell injury led to such reactions that were morphologically distinguishable from the specific "ring" reaction and as far as pinocytosis is concerned, could be easily avoided by reducing the incubation time. In addition, a non-specific staining reaction morphologically indistinguishable from the specific "ring" reaction could be seen in a small proportion of bone marrow and lymph node cells but in no other cell type studied. The possible nature of this reaction is discussed. Footnotes Submitted: 15 March 1961

Erichsen, Stian; Eng, Jan; Morgan, Herbert R.

doi: 10.1084/jem.114.4.435pmid: 19867193

The chick embryo fibroblast infected by Rous sarcoma virus in vitro acquires the capacity to produce the acid mucopolysaccharides which are found in the tumors caused by this virus and which are also produced by tumor cells in vitro . The transformed cell acquires synthetic as well as morphologic, metabolic, and proliferative properties characteristic of Rous sarcoma tumor cells in vivo and in vitro and the transformed cell may be analogous to the tumor cell produced by virus infection in vivo . Footnotes Submitted: 15 May 1961

Magill, Thomas P.

doi: 10.1084/jem.114.4.441pmid: 19867194

During passage in mice which had been vaccinated with the homologous, and with closely related strains of influenza virus, the passage strain developed a lessened susceptibility to the deleterious effects of the "immune" environment, concommittant with which was a developed capacity to evoke antibodies which reacted with earlier strains of virus—a capacity which was inapparent in the parent strain. However, the parent strain exhibited a relatively broad range of surface reactivity which was not apparent in the derived strain. The data are interpreted to mean that the hereditary change resulted from spatial rearrangement and quantitative redistribution of antigens in the virus particle (in which the surface is viewed as being distinct from the inner bulk), and are viewed as enhancing the idea that influenza virus variation ( i.e ., "mutation") may result from a rearrangement of existing hereditary elements. Footnotes Submitted: 9 May 1961

Hochwald, G. M.; Thorbecke, G. J.; Asofsky, R.

doi: 10.1084/jem.114.4.459pmid: 19867195

The development of a new method for the determination of the sites of serum protein formation has been described. The method involves the incorporation of C 14 -labeled amino acids by tissues cultured in vitro , and subsequent autoradiography of immunoelectrophoretic patterns prepared from a mixture of culture fluids and carrier serum with an antiserum against the carrier serum. This technique has been used to demonstrate formation of γ-globulin, of ß 2 -macroglobulin, and of a component of C' 3 by mouse spleen tissue, and of various other serum proteins by liver tissue. The specificity and sensitivity of this method have been discussed, and some of its advantages and pitfalls were mentioned. In addition, a rabbit antimouse serum was prepared, and the immunoelectrophoretic patterns obtained with mouse serum were compared with those described in the literature. Footnotes Submitted: 9 May 1961

Asofsky, R.; Thorbecke, G. J.

doi: 10.1084/jem.114.4.471pmid: 19867196

Human, and monkey tissues were cultured in the presence of labeled amino acid. The culture fluids were then examined for labeled serum proteins by means of autoradiography of immunoelectrophoretic patterns prepared from mixtures of the fluids and carrier sera. Gamma globulin, ß 2A -globulin, ß 2M globulin, and ß 1C -globulin were found to be formed in human lymph node, bone marrow, and ileum. Gamma globulin, ß 2M -globulin, and ß 1C -globulin were found to be formed in monkey spleen, lymph node, and bone marrow. Formation of several other serum proteins was observed in the monkey liver. Footnotes Submitted: 9 May 1961

Ito, Yohei; Evans, Charles A.

doi: 10.1084/jem.114.4.485pmid: 19867197

A deoxyribonucleic acid preparation which showed infectivity and tumorigenic activity in domestic rabbits was isolated from the papillomatous tissue of wild cottontail rabbits by phenolic deproteinization procedure. The activity of the preparation could be completely abolished by its exposure to a minute amount (0.02 µg/ml) of DNAase. Antisera against Shope papilloma virus did not block the tumorigenic activity of the preparation, and trypsin and chymotrypsin had no effect on it. The extraction with phenol of a partially purified virus preparation also yielded extracts with tumorigenic potency. Extracts obtained from the domestic rabbit papilloma and submitted to phenolic deproteinization also proved infective and tumorigenic in rabbits of this sort, although the level of "tumorigenicity" was much lower than that of the cottontail preparations. Tests for intact virus, carried out with half of the extracts yielded wholly negative findings. Footnotes Submitted: 19 May 1961

Spink, Wesley W.; Vick, James

doi: 10.1084/jem.114.4.501pmid: 19867198

Peripheral vascular failure caused by endotoxin in the dog has an initial stage of vasoconstriction. Preliminary studies in vitro demonstrated that the constriction was due to the interaction of endotoxin with a heat-labile serum or plasma factor and platelets, resulting in the liberation of histamine. Further studies on the intact dog support and extend this concept. A standardized dose of Escherichia coli endotoxin produced fatal shock in control adult mongrel dogs within 28 hours. The characteristic pattern of changes included progressive hypotension, oliguria and anuria, hemoconcentration, and acidosis. Normal dogs were protected against endotoxin by transfusions of blood in which the essential serum factor was depleted in one of two ways. First, plasma separated from the blood of normal animals was heated at 56°C for 30 minutes, and the infused reconstituted whole blood protected normal dogs. Protection was not afforded by unheated reconstituted blood. Second, blood from immune dogs obtained within 24 hours after a second lethal dose of endotoxin protected recipient dogs. However, protection was not demonstrated with blood collected 72 hours after a second injection of endotoxin. The nature of the serum factor essential for endotoxin activity is not known. It is postulated that an enzyme or enzyme system is involved, and the possible role of complement is discussed. Footnotes Submitted: 1 June 1961

Kretschmer, Roberto R.; Peréz-Tamayo, Ruy

doi: 10.1084/jem.114.4.509pmid: 19867199

Gross and microscopic observations of skin homograft rejection carried out in cortisone-conditioned and non-conditioned rabbits seem to indicate that humoral antibodies play an important role in the phenomenon. Thus, local administration of isoimmune serum to animals bearing skin homografts resulted in a significantly earlier rejection of that particular test graft without modifying the course of a neighboring control-skin graft. This result appears to support the idea that homograft rejection is not only due to cellular antibodies but to a combination of both humoral and cellular immune responses, which should not be regarded as completely unrelated. Footnotes Submitted: 31 May 1961

Franklin, Edward C.; Stanworth, Denis R.

doi: 10.1084/jem.114.4.521pmid: 19867200

The antigenic properties of normal 19S γ-globulin, pathologic macroglobulins, ß 2A -myeloma proteins, and Bence Jones proteins have been compared with 7S γ-globulin and the small 3.5S units derived from it by gel diffusion precipitin techniques. These studies demonstrate that the determinant groups on the 7S γ-globulin molecule responsible for the cross-reaction with each of the other proteins are associated with the two fragments of 7S γ-globulin which have the antibody-combining sites. The antigenic specificity of the 7S γ-globulin which distinguishes it from each of these proteins is associated primarily with the fragment that is richest in hexose and can not combine with antigen. However when compared with certain of the paraproteins additional antigenic specificity was also found to reside in the fragments with antibody-combining activity. The finding of similar antigenic relationships in rabbit γ-globulins suggests that some of the biological properties associated only with the 7S γ-globulins and not with the other immune globulins may reside in the fragment which also carries the antigenic specificity of the protein. Footnotes Submitted: 9 May 1961

Allison, Fred; Lancaster, Margie G.

doi: 10.1084/jem.114.4.535pmid: 19867201

Vigorous anticoagulation with heparin sodium and sodium warfarin singly and in combination did not prevent the margination and endothelial sticking reaction of leucocytes in rabbit ear chambers damaged by heat. The general inflammatory reaction observed in this preparation was similarly uninfluenced by the anticoagulants. An unexpected finding after administration of heparin was the enhanced formation of platelet and fibrin-like thrombi within damaged ear chambers. Sodium warfarin did not induce or prevent this heparin effect. Production of these heparin-associated thrombi was minimized in animals subjected to defibrinogenation in vivo whereas leucocytic sticking was not modified. Although defibrinogenation was not absolute, these experiments represent additional proof that the sticking of white blood cells to vascular endothelium is not causally related to the fibrinogen-fibrin system. Footnotes Submitted: 12 May 1961