The Journal of Experimental Medicine

Leddy, John P.; Bakemeier, Richard F.

doi: 10.1084/jem.121.1.1pmid: 14253485

The 7S γ-globulins causing erythrocyte autosensitization in 20 patients were isolated by elution and examined for homogeneity or heterogeneity of their L chain types and electrophoretic dispersion. The isolated erythrocyte autoantibodies from 12 patients contained only 1 detectable L chain type. Two of these "monotypic" populations showed appreciable restriction of electrophoretic dispersion, while 2 others more nearly resembled the electrophoretic heterogeneity of normal γ-globulins. The autoantibodies from the other 8 patients exhibited L chains of both types. The single "bitypic" population so tested was relatively polydisperse electrophoretically. As a comparison, anti-Rh o isoantibodies from 5 of 6 donors without known hematologic disease showed bitypic reactions, and 2 of these isoantibody populations were relatively polydisperse electrophoretically. One Rh isoantibody is described which contained only 1 demonstrable L chain type. The structural similarities to "paraproteins" observed in a significant proportion of these erythrocyte autoantibodies raise the possibility of their origin from a restricted population of antibody forming cells, and may have implications concerning the pathogenesis of erythrocyte autosensitization. Footnotes Submitted: 24 July 1964

Butler, Vincent P.; Tanenbaum, Stuart W.; Beiser, Sam M.

doi: 10.1084/jem.121.1.19pmid: 14253484

The specificity of the reaction of antipurin-6-oyl sera with thermally denatured DNA has been studied by means of hapten inhibition techniques. The relative order of effectiveness of various haptens as inhibitors of the complement fixation reaction between DNA and antipurin-6-oyl serum was found to be comparable to their relative order of effectiveness as inhibitors of the precipitin reaction between purin-6-oyl-protein conjugates and antipurin-6-oyl serum. Antipurin-6-oyl antibody has been purified and has been shown to be capable of reacting with thermally denatured DNA. It is concluded that the reactivity of these purine-specific antisera with DNA is a property of antibody with specificity for the purin-6-oyl moiety. In addition, the reaction between antipurinoyl sera and DNA has been demonstrated by the techniques of radioimmunoelectrophoresis and passive cutaneous anaphylaxis. Footnotes Submitted: 19 August 1964

Fedorko, Martha E.; Morse, Stephen I.

doi: 10.1084/jem.121.1.39pmid: 14253486

Acid mucopolysaccharides have been extracted from whole rabbit polymorphonuclear leucocytes and from the cytoplasmic granules of these cells. The leucocyte acid mucopolysaccharides can be separated into two fractions by the solubility of their CPC complexes in solutions of differing salt concentration. One of these fractions appears to be identical with hyaluronic acid; the other appears to be an atypical chondroitin sulfate. On both a dry weight and total protein basis the polymorphonuclear leucocyte granule contains approximately 2.6 times as much acid mucopolysaccharide as does the whole cell. Hyaluronic acid is concentrated in the granules in particular; its function is unknown. These results do not indicate that all lysosomes contain abundant acid mucopolysaccharides, for no detectable carbohydrate of this class could be extracted from lysosome-rich alveolar macrophages. Footnotes Submitted: 17 August 1964

Morse, Stephen I.

doi: 10.1084/jem.121.1.49pmid: 14253487

1. Intravenous injection into mice of phase I Bordetella pertussis vaccine resulted in a striking hyperleucocytosis with a predominating lymphocytosis. Intraperitoneal inoculation was less effective, and subcutaneous administration was inactive. 2. Active immunization prevented the hyperleucocytosis; passive immunization was less effective. 3. Reticuloendothelial blockage reduced the effect of the vaccine. 4. Extirpation of the spleen or thymus did not alter the leucocyte response. 5. Histologic studies suggested that the increase in circulating lymphocytes resulted from release of cells from lymphoid organs, including the thymus. Footnotes Submitted: 24 September 1964

Campbell, Charles H.

doi: 10.1084/jem.121.1.69pmid: 14253488

Multiplication of foot-and-mouth disease virus (FMDV) was compared in kidney cells from 7- to 35-day-old mice representing various degrees of age resistance to this virus. Three types of cell preparations were used: primary monolayer cultures, suspensions of dispersed cells, and suspensions of minced tissue. Virus multiplication in the two types of cell suspensions was related to the age of the donors both in regard to time when multiplication first became evident and to the amount of virus produced. While adsorption rates were similar in the cells from all age groups, virus multiplication began earlier in cells from younger mice and more virus was produced by these cells than by cells from older animals. There was no significant difference in the virus growth rates in the primary monolayer cultures of cells. The results indicate that kidney cells from mice 7 to 35 days old vary in their ability to produce virus in relation to the degree of susceptibility of the cell donors. After propagation of the cells in primary monolayer cultures, however, this difference no longer exists probably because of cell selection under the cultural conditions. Footnotes Submitted: 19 August 1964

Chapman, Niles D.; Dutton, Richard W.

doi: 10.1084/jem.121.1.85pmid: 14253489

An early proliferative response follows the mixing of homologous spleen or lymph node suspensions obtained from two unrelated rabbits. The rate of incorporation of radioactive thymidine has been used as a quantitative measure of this response. Thymus cells do not respond to homologous cell suspensions but may on occasion serve to stimulate the response in homologous spleen or lymph node cells. Homologous erythrocytes or autologous tissues do not stimulate a response. No response occurs if the two cell populations are separated by a Millipore membrane. Autoradiographic studies have established that 1 to 2 per cent of the intial cell population is involved in the response and they are large undifferentiated cells by the time they can first be identified as responders. There was no morphological evidence of any cellular interaction and the viability of mixed suspensions was not measurably different from that observed in separate suspension. Simultaneous additive responses could be obtained to homologous cells and to antigen when cell suspensions from immunized rabbits were used. The interaction of the cell populations from the two rabbits did not appear to suppress the response of each to antigen. The speed and magnitude of the response were in everyway comparable with the secondary response of cell suspensions from immunized rabbits exposed to the immunizing antigen. No evidence was obtained of any enhancement of the response to homologous cells by prior immunization with homologous tissues, but the possibility that it had occurred was not rigidly excluded. Footnotes Submitted: 24 August 1964

Tomasi, Thomas B.; Tan, Eng M.; Solomon, Alan; Prendergast, Robert A.

doi: 10.1084/jem.121.1.101pmid: 14253478

The γ 1 A present in saliva and colostrum exists largely in the form of higher polymers, the major component of which has a sedimentation coefficient of 11S. The 11S γ 1 A in these fluids differs from the polymers found in normal and myeloma sera both immunologically and by the fact that their sedimentation coefficients are unaffected by disulfide bond reduction in the absence of urea. However, like other γ-globulins the 11S γ 1 A molecules consist of multiple polypeptide chains linked by disulfide bonds. Local synthesis of γ 1 A in the salivary gland has been shown by fluorescent and autoradiographic studies, although the fraction of the total salivary γ 1 A which is derived from local production is uncertain. No evidence of transport of intravenously administered I 131 -labeled 7S γ 1 A from serum to saliva was obtained. Immunological specificity has been demonstrated in the salivary and colostral γ 1 A. Whether that portion of the γ 1 A which is immunologically specific is a piece incorporated during the local synthesis of γ 1 A in the gland or is added by the epithelial cell in the process of transport remains to be determined. Antibody activity (isohemagglutinins) have been demonstrated in saliva and colostrum and have been shown to be of the γ 1 A-type. In both of these fluids activity is associated primarily with γ 1 A-polymers of 11S and 18S sizes. There appears to be an immunological system which is characteristic of certain external secretions. Its properties including the local production of a distinctive type of antibody separate it from the "systemic" system responsible for the production of circulating antibody. This system may play a significant role in the body's defense mechanisms against allergens and microorganisms. Footnotes Submitted: 11 September 1964

Cohen, Edward P.; Talmage, David W.

doi: 10.1084/jem.121.1.125pmid: 14253479

Within 5 hours after the intravenous injection of particulate antigen into the primarily immunized mouse, precursors of antibody-forming cells began DNA synthesis as shown by the incorporation of tritium-labeled thymidine. DNA synthesis continued for at least 24 hours and essentially stopped by 48 hours. No DNA synthesis in antibody-forming precursor cells occurred before the injection of antigen. Footnotes Submitted: 17 September 1964

Jesaitis, Margeris A.

doi: 10.1084/jem.121.1.133pmid: 14253480

A study of the immunological properties of phage strains derived from T2 x T6 crosses revealed that the majority of the progeny differ serologically from the parental viruses. Some hybrids were found to contain head membranes having the serological specificity of both T2 and T6 phages, while others contained tail sheaths of the former and the tail fibers of the latter. Since the immunological properties of all hybrids were heritable, it has been concluded that the serological specificity of the head proteins of T2 and T6 is controlled by at least two genetic determinants, and that the specificities of the fiber and sheath proteins may be governed by single genes. Furthermore it was found that nucleic acids of hybrids had similar proportions of unsubstituted, mono- and diglucosylated hydroxymethylcytosine nucleotides to the nucleic acid of either T2 or T6 phage. Since the parental and hybrid viruses having chemically similar nucleic acids contained in some instances serologically different proteins, it has also been concluded that the extent of glucosylation of the hydroxymethylcytosine component of viral nucleic acids and the immunological properties of viral proteins are independently heritable traits of T2 and T6 bacteriophages. Footnotes Submitted: 25 September 1964

Cohn, Zanvil A.; Benson, Belinda

doi: 10.1084/jem.121.1.153pmid: 14253481

The in vitro differentiation of homogeneous populations of monocyte-like cells from the unstimulated mouse peritoneal cavity is described. Under the conditions employed, a progressive increase in cell size occurs without significant cell division. This process is characterized morphologically by the accumulation of phase-dense and neutral red-positive granules, mitochondria, and lipid droplets. The phase-dense granules react strongly for acid phosphatase. Biochemical determinations indicate marked increases in the total content and specific activity of acid phosphatase, cathepsin, and ß-glucuronidase. The production of acid phosphatase is more rapid and extensive than that of the other two hydrolases. From these data it appears that the conversion of a monocyte-like cell to a mature macrophage is accompanied by the formation of increased numbers of lysosome-like cytoplasmic organelles. Mouse peritoneal phagocytes stimulated in vivo with a bacterial lipopolysaccharide undergo a similar series of morphological and biochemical events. Footnotes Submitted: 27 August 1964