The Journal of Experimental Medicine

Gustafsson, Bengt E.; Norman, Arne

doi: 10.1084/jem.116.3.273pmid: 13903130

In a colony of germfree rats 50 per cent of the males had urinary calculi composed of calcium citrate and calcium oxalate. Genetically closely related conventional animals on the same sterilized diet did not present a single case of stone formation. The tendency to calculus formation disappeared when germfree animals were contaminated with the intestinal flora from conventional rats. The calculus formation can readily be explained by the high calcium, high citrate, and high pH of the urine. This pattern was changed to that of conventional rats when the germfree rats were infected with intestinal microorganisms. Footnotes Submitted: 22 April 1962

Sanford, Jay P.; Hunter, Betty W.; Donaldson, Paul

doi: 10.1084/jem.116.3.285pmid: 14496913

1. Hematogenous E. coli pyelonephritis was produced in rats. The localization of the organisms and the persistence of bacterial antigens was followed by fluorescent antibody techniques as well as by standard histological and bacteriological methods. 2. Salient sequential features were as follows: Single organisms passed through vessel walls into the renal interstitium and began multiplication and subsequently evoked a leucocytic response. Bacterial multiplication did not occur in glomeruli or renal tubular cells. Bacteria did not appear within renal tubular lumina until microabscesses were well developed in the renal interstitium. Bacteriuria appeared late and represented secondary invasion rather than filtration of organisms. The infection healed spontaneously but, while sterile, the parenchymal scars contained large amounts of residual bacterial antigen. The persistence of bacterial antigen did not result in continuing inflammatory changes or progressive scarring. 3. The persistence of bacterial antigens is postulated to constitute a major antigenic stimulus responsible for active immunity in experimental hematogenous pyelonephritis. Footnotes Submitted: 26 April 1962

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

doi: 10.1084/jem.116.3.295pmid: 13920994

Antibody formation in vitro by red and white pulp of the spleen and by bone marrow tissue was studied at various days after an intravenous booster injection of soluble antigens such as ovalbumin and bovine gamma globulin (BGG). When the booster injection of antigen was given early (10 days) after an intravenous primary injection, high antibody formation could be demonstrated in the spleen primarily 2 to 3 days after the injection, but much less afterwards. When the booster injection was given later (1 month) after the primary, the antibody production by the spleen lasted longer and higher serum titers were obtained. The bone marrow formed antibody in both cases but, particularly with the short interval between injections, its response was delayed as compared to the spleen. It was also shown that during antibody formation the production of gamma globulin in vitro was enhanced. Histologically the antibody production was always correlated to immature plasma cell proliferation, located at the border of red and white pulp and in the red pulp of the spleen. When endotoxin had been injected at the time of a primary BGG injection, and a second antigen injection was given 5 to 10 days later, a booster response could be elicited which was sometimes limited to the white pulp on day 1, and on day 2 was divided between "red" and "white" pulp. The response induced at day 10, at the peak of secondary nodule proliferation, lasted very long and was accompanied by an enormous plasma cellular proliferation in and around the periarteriolar lymphoid areas of the spleen. The possible importance of the secondary nodules of the white pulp in the preparation for a secondary response is discussed. Footnotes Submitted: 1 April 1962

Hoyer, Leon W.; Good, Robert A.; Condie, Richard M.

doi: 10.1084/jem.116.3.311pmid: 14449435

1. 6-Mercaptopurine (6-MP) prevents experimental allergic encephalomyelitis (EAE) during the period of drug administration in both rabbits and guinea pigs. The disease is suppressed even when treatment is started as late as the 5th day after antigenic stimulation in guinea pigs and the 12th day in rabbits. 2. After discontinuation of 6-MP treatment, there is a latent period before the disease is noted. The length of this latent period is not modified by the duration of 6-MP treatment. 3. The effect of 6-MP on EAE is not the result of leukopenia, non-specific toxicity and debilitation, anti-inflammatory activity, or mere masking of clinical signs of the disease. It is, rather, the result of 6-MP's specific anti-immunologic activity. 4. The effects of 6-MP on antibody production, delayed hypersensitivity, and EAE are compared. This provides indirect evidence for the importance of circulating antibody in the pathogenesis of EAE. 5. The important considerations in the use of 6-MP are discussed and the possible usefulness of 6-MP in human neurologic diseases is considered. Footnotes Submitted: 14 May 1962

Morgan, Herbert R.; Andrese, Angelo P.

doi: 10.1084/jem.116.3.329pmid: 19867215

Chick embryo fibroblasts infected with Rous sarcoma virus in vitro are rendered malignant for such cells produce typical Rous sarcomas when injected into susceptible chicks since the tumors produced predominantly retain the sex chromatin patterns of the donor cells when such cells are injected into a recipient of the opposite sex. However, examination of the sex chromatin of cells at the periphery of the tumor shows presence of recipient cells though the bulk of the tumor is clearly of donor cell origin. Such tumors grow and cause death of the recipient. Injection of RSV induces tumors of the sex of the recipient as also does the injection of transformed cells rendered incapable of multiplication by x-rays. Following their injection into susceptible chicks, the cells transformed in vitro by virus behave in the same manner as tumor cells obtained from tumors induced by virus in vivo and cultivated in the same conditions in vitro . When such tumors induced by transformed cells are serially transferred in recipients of the opposite sex, they gradually convert to the sex of the recipient indicating that the tumors are not indefinitely transplantable. These chick embryo fibroblasts transformed in vitro show the same neo-plastic properties as tumor cells when they are introduced into the cheek pouch of the hamster or the eye of the guinea pig. Footnotes Submitted: 17 May 1962

Rothbard, Sidney; Watson, Robert F.

doi: 10.1084/jem.116.3.337pmid: 14494413

Antibody to rat collagen, prepared in rabbits and injected into the circulation of normal or adjuvant-prepared rats, becomes fixed to its antigen and can then be identified in tissue sections under ultraviolet light by its fluorescence after application of fluorescein-conjugated anti-rabbit globulin. In heart, lung, liver, spleen, adrenal, kidney, jejunum, lymph node, thymus, joint synovia, peripheral nerve, aorta, skeletal muscle, eye, and brain, the antibody was found at all sites where collagen and reticulin are normally present, but except for the kidneys of the adjuvant-prepared rats, no pathological abnormalities were demonstrated. It was not found within cells. Specific fluorescence was absent from tissues of rats injected with normal rabbit serum or rabbit anti-fish collagen serum or rabbit anti-rat collagen serum after absorption with rat collagen, but was present when the anti-rat collagen serum had been absorbed with fish collagen. The reaction could be blocked by pretreatment of sections with unlabeled anti-rabbit globulin and did not occur with heterologous labeled anti-duck globulin. After serial treatment in vitro with homologous antibody to collagen and the conjugated anti-rabbit globulin, purified reconstituted collagen fibers showed the same fluorescence as fibers in the tissues; no fluorescence of the fibers occurred when heterologous antibody to collagen was applied. These findings indicate that the antibody to rat collagen is directed toward an antigen present in both collagen and reticulin. Footnotes Submitted: 24 May 1962

Friedman, Robert M.; Baron, Samuel; Buckler, Charles E.; Steinmuller, Robert I.

doi: 10.1084/jem.116.3.347pmid: 13895287

In guinea pigs, methotrexate and x-radiation have been employed to block the development of delayed hypersensitivity and of antibody to vaccinia virus during experimental infection. Animals whose specific responses were blocked by x-radiation and methotrexate recovered from vaccinia virus infection as rapidly as non-treated control animals suggesting that these responses were not essential in recovery of the infected animals. A virus-inhibiting substance with the properties of interferon was present in skin of untreated and methotrexate- and x-radiation-treated animals infected with vaccinia virus. Footnotes Submitted: 3 May 1962

Henson, James B.; Gorham, John R.; Leader, Robert W.; Wagner, Bernard M.

doi: 10.1084/jem.116.3.357pmid: 13906569

Hypergammaglobulinemia in mink was produced by the injection of crude tissue suspensions from mink with spontaneous Aleutian disease. The initiating factor was found to be resistant to 0.3 per cent formalin for 2 weeks but not 40 weeks at 5°C. Foreign antigens as well as formalinized normal mink tissue from homologous and heterologous genotypes did not cause a detectable change in the serum protein values. Mink homozygous recessive for the Aleutian gene were found to be significantly more susceptible to the experimental disease. Possible pathogenetic mechanisms as well as similarities between the mink disease and certain immunologic and connective tissue diseases of man are discussed. Footnotes Submitted: 2 May 1962

Weissmann, Gerald; Fell, Honor B.

doi: 10.1084/jem.116.3.365pmid: 14005935

1. The effect of hydrocortisone on the development of fetal rat skin in organ culture, and on its repair after exposure to a mixed beam from a mercury lamp, are described. 2. The addition of hydrocortisone (7.5 µg/ml) to the culture medium (HC medium) caused accelerated differentiation and keratinisation of the epidermis followed by atrophic changes as in vivo . 3. Explants were grown for 2 days in either normal or HC medium and then irradiated with light from an Hanovia lamp. 4. Irradiation of the control explants produced severe necrosis in both epidermis and dermis and much disorganisation of the dermal intercellular material; 2 days after exposure the s. corneum with adherent cellular debris had become either completely detached from the denuded dermis, or raised to form a fluid-filled blister. Epidermal regeneration had begun by the 4th day after irradiation and was usually complete by the 6th day. 5. Hydrocortisone modified the response to irradiation as follows: (1) reduced and retarded cellular breakdown, (2) prevented vesication, (3) preserved the organisation of the dermal intercellular material, (4) hastened epithelialisation, (5) accelerated the differentiation of the new epidermis. Effects (2), (3), and (4) were probably secondary to (1). 6. Experiments with various light filters showed that the effective wavelengths for producing lesions in the skin explants were those below 3000 A. 7. It is suggested that the beneficial effect of hydrocortisone on the repair of irradiated skin explants might be due, at least in part, to a reduced proteolytic activity in the damaged tissue through a stabilising action of the hormone on the lysosomes. 8. The relationship of these findings to clinical observations is discussed. Footnotes Submitted: 21 May 1962

Mackaness, G. B.

doi: 10.1084/jem.116.3.381pmid: 14467923

The mouse was found to be natively susceptible to Listeria monocytogenes . Its susceptibility was attributed to the capacity of the organism to survive and multiplying in host macrophages. During the first 3 days of a primary infection the bacterial populations of spleen and liver were found to increase at a constant rate. On the 4th day of infection the host became hypersensitive to Listeria antigens and at the same time bacterial growth ceased. A rapid inactivation of the organism ensued. Convalescent mice were resistant to challenge, but no protective factor could be found in their serum. Histological evidence suggested that acquired resistance was the result of a change occurring in the host's mononuclear phagocytes. When challenged in vitro , the macrophages of convalescent mice were found to resist infection with Listeria monocytogenes. Listeria -resistant cells appeared during the course of infection at a time which corresponded with the development of the antibacterial mechanism in the spleen. They persisted for as long as the antibacterial mechanism remained intact in this organ. This period of absolute resistance to Listeria lasted about 3 weeks. Thereafter, the host remained hypersensitive but unable to inactivate a challenge inoculum of Listeria . However, it remained capable of producing an accelerated response to reinfection. This was thought to depend upon an ability to generate a new population of resistant cells from a residuum of specifically sensitized macrophages or macrophage precursors still surviving in the tissues as a result of the immunological activation which occurred during the primary infection. Footnotes Submitted: 5 March 1962