bluuebunny/crossref_metadata_2025 huggingface dataset:
RAPIDLY LABELED NUCLEAR AND CYTOPLASMIC RENAL RNA
Authors: Malt Ronald A.
Published: June 2004

Much of the RNA in mouse kidneys is not recovered when they are extracted with phenol-SDS at 60°C. Better recovery is achieved by extracting in hypertonic buffer with phenol-SDS, chloroform, and isoamyl alcohol after treatment with DNase.

1. Among the polydisperse RNA in the renal nuclei are small amounts of 45S RNA and large amounts of 32S RNA; the latter first appear when 18S RNA is detectable in the cytoplasm 10 min after administration of undine-H3.

2. In the cytoplasm rapidly labeled 18S RNA is transported as part of a 45S particle to which mRNA is attached.

3. Compared to HeLa cells in culture, normal mouse kidney is poor in the polydisperse nuclear precursors of rRNA and rich in the cytoplasmic polydisperse RNA that sediments like mRNA.
STRUCTURAL STUDIES OF HUMAN γG-MYELOMA PROTEINS OF DIFFERENT ANTIGENIC SUBGROUPS AND GENETIC SPECIFICITIES
Authors: Frangione B. , Franklin E. C. , Fudenberg H. H. , Koshland M. E.
Published: June 2004

1. Peptide maps of Fc fragments or heavy chains of 36 G myeloma proteins and two "heavy chain disease" proteins belonging to the four γ-chain subgroups revealed very striking similarities between them. However differences in a few peptides were noted. This was most pronounced for the Ge(γ2d) subgroup which lacked three peptides characteristic of the other three subgroups. While Fc fragments from different proteins belonging to the same subgroup appeared very similar, minor differences in addition to those based on currently recognized Gm factors were occasionally noted.

2. Fc fragments from Gm(a+) We(γ2b) proteins had a peptide previously shown to be characteristic of normal Gm(a+) γG-globulins. Fc fragments from Gm(a-) molecules belonging to the We(γ2b), Vi(γ2c), or Ne(γ2a) subgroups, whether Gm(b+), Gm(f+), or Gm(-), had the peptide previously identified in Gm(b+f+) normal γG-globulin. This "non-a" peptide was absent in peptide maps from Gm(-) molecules of the Ge(γ2d) subgroup which contained instead another peptide with the same electrophoretic mobility but migrating slightly further on chromatography.

3. Both the "a" and "non-a" peptides were pentapeptides having three amino acids in common, and differing in the other two. The "a" peptide contained one residue of lysine, aspartic acid, threonine, leucine, and glutamic acid. The "non-a" peptides prepared from Gm(b+), Gm(f+), and Gm(-) proteins were identical and contained one residue of lysine, threonine, and methionine sulfone, and two residues of glutamic acid.

4. Several possible mechanisms for the origin of these differences, and their possible role in serologic specificity are discussed.
ANTIGENS OF LEUKEMIAS INDUCED BY NATURALLY OCCURRING MURINE LEUKEMIA VIRUS: THEIR RELATION TO THE ANTIGENS OF GROSS VIRUS AND OTHER MURINE LEUKEMIA VIRUSES
Authors: Geering Gayla , Old Lloyd J. , Boyse Edward A.
Published: June 2004

Leukemias can be induced in W/Fu inbred rats by neonatal inoculation of normal thymus cells of C58 mice. These leukemias are not transplantable to C58 mice or to adult W/Fu rats, but they can be kept in passage in W/Fu rats aged 0 to 7 days. Adult W/Fu rats inoculated repeatedly with these isogenic leukemias produce cytotoxic and precipitating antibodies. These antisera are of particular value in the analysis of the antigens of leukemia cells and of leukemia viruses because their mode of preparation precludes the formation of antibody against any normal constituents of the cell.

Analysis based on the cytotoxic test indicates the presence of 2 distinct cell surface antigens in leukemias induced by Passage A Gross virus or occurring spontaneously in mice of high-incidence strains. All leukemias and other tissues known to contain G (Gross) leukemia antigen have both determinants, but certain leukemias of low-incidence strains have only 1 of them and so were previously classified G-.

Immunoprecipitation with these antisera reveals the presence of a cellular antigen common to G+ cells and absent from G- cells; the same antigen can be demonstrated in ether-treated Gross virus, but not in intact virus. This antigen is present also in ether-treated preparations of the Friend, Moloney, and Rauscher leukemia viruses, but not in Bittner (mammary tumor) virus. Thus it may be regarded as a group-specific antigen of murine leukemia viruses, in contrast to the type-specific cellular antigens demonstrable by the cytotoxic test. Four additional antigens associated with leukemias induced by wild-type Gross virus have been demonstrated by immunoprecipitation, but their relation to viral and cellular antigens has not been determined.
FUNCTIONAL ANATOMY OF THE LYMPHOCYTE IN IMMUNOLOGICAL REACTIONS IN VITRO
Authors: McFarland William , Heilman Dorothy H. , Moorhead John F.
Published: June 2004

The motile lymphocyte in vitro has a prominent "tail" that becomes a means of "attachment" to other cells and debris during interaction. The term "uropod" is proposed to designate this specialized cytoplasmic projection which appears totally different, anatomically and functionally, from the pseudopods. Observations of lymphoblasts during mitosis indicate that the uropod is formed immediately following mitosis at the point of final cytoplasmic connection between daughter cells, a fact that may prove significant as lymphocyte function is better understood.

In the mixed leukocyte reaction the lymphocyte interacts with macrophages, cell debris, and lymphoblasts via the uropod, suggesting that stimulatory material may be acquired through this specialized appendage. Lymphoblast-lymphocyte interaction is noteworthy and implies that immunologically committed cells may be mustered through horizontal as well as vertical processes: horizontally by lymphoblast-lymphocyte interaction and vertically by mitosis of transformed lymphoblasts. The possible relevance of these in vitro observations to lymphocyte functions in vivo is discussed.
ALLOANTIBODY PRODUCTION MEASURED BY PLAQUE ASSAY IN RELATION TO STRONG AND WEAK HISTOINCOMPATIBILITY
Authors: Hildemann W. H. , Pinkerton W.
Published: June 2004

A plaque technique which provides a sensitive and quantitative determination of allohemolysin (isohemolysin) production by lymphoid cells of mice and Syrian hamsters is described. Much ongoing attention has been given to minimizing sources of spurious plaques while improving the reproducibility of the technique. Primary intravenous immunization with allogeneic blood cells led to the early appearance of plaque-forming spleen cells responding to both strong and weak histocompatibility antigens in inbred strains of mice. The strongest histoincompatibility (A/J → BL/6), characterized by H-2 plus non-H-2 differences and a median skin allograft survival time of 8.2 ± 0.5 days, gave a peak average plaque production of 29.6 per 10 x 106 viable spleen cells after 6 days. At the other extreme with an average allograft survival time of 91 days, C3H(H-1a) → C3H.K(H-1b) showed a feeble production of plaque-forming cells with a peak response of 3.7 per 10 x 106 viable spleen cells at 9 days. Histocompatibility antigens determined by genes at the H-1, H-Y, and H-X loci were demonstrated to be erythrocyte antigens as well. Peak plaque production occurred earlier and reached higher levels across the stronger histocompatibility barriers (H-2 plus non-H-2 or H-2 alone) in temporal accord with the rapidity of skin allograft rejection. Weak H-Y or H-1 differences leading to prolonged allograft survival were associated with moderate, but not commensurate delays in the initial occurrence and recruitment of plaque-forming cells. With one exception (C57BL/10 → B10.A), the average plaque counts found at the peak of the primary responses were inversely proportional to the median survival times of skin allografts in the same strain combinations. Nevertheless, in light of the disparity between the times of allograft rejection and the time course of appearance of plaque-forming cells with the "weaker" congenic combinations, the early 19S antibodies do not appear to be directly involved in allograft rejection.

There appears to be no limitation in the weakness of histocompatibility antigens capable of evoking antibodies detectible by plaque assay. The extent to which molecular species of mouse antibodies other than 19S hemolysins are induced and detected under the conditions employed remains to be determined. The responses in strain combinations involving stronger histocompatibility barriers were characterized by exponential increases in plaque-forming cells, for 2 to 4 days until maximal levels were reached. Repeated immunization in several trials led to substantial increases over the maximum primary response in the number of demonstrable plaque-forming cells. Further preliminary results are reported concerning alloimmune plaque production in three strains of Syrian hamsters exhibiting intermediate to weak histocompatibility differences.
AN ANTIVIRAL SUBSTANCE FROM PENICILLIUM FUNICULOSUM
Authors: Shope Richard E.
Published: June 2004

Helenine, NDV, and statolon, all known inducers of interferon in mice, all exerted a marked antiviral effect against Semliki Forest virus. This AV effect was, so far as can be demonstrated, mediated through, the induced interferon.

The same three materials also exerted a marked antipassive immunity effect. All the evidence that can be brought to bear indicates that this API effect like the AV effect is mediated through interferon known to be induced by the three materials.

If the API effect does indeed have interferon as its basis, this represents a new and totally unsuspected action of interferon.
ANTIBODY SYNTHESIS AT THE CELLULAR LEVEL
Authors: Wigzell Hans
Published: June 2004

The specific suppressing activity of passively administered antibody on 7S antibody synthesis against sheep and chicken red blood cells has been investigated at the cellular level using the indirect hemolytic agar-plaque technique. 7S antibody production was found to be sensitive to antibody-induced suppression. No inhibitory effect of transferred antibody was seen until 48 to 72 hr after administration. This indicates that the action of antibody is not by direct suppression of synthesis of already committed cells but rather by removal from the system of the stimulus for maintenance of 7S synthesis. The sensitivity of the 7S system to inhibition decreases with time after immunization but significant specific suppression could still be obtained if transfer of antibody was delayed until 40 days after immunization. The present findings emphasize the role of antibody as a feedback factor during a substantial postpeak period of 7S antibody synthesis and suggest an important role of antigen in stabilizing the 7S antibody production.
STUDIES OF PPLO INFECTION
Authors: Thomas Lewis , Aleu Fernando , Bitensky Mark W. , Davidson Morton , Gesner Bertram
Published: June 2004

Rolling disease has been produced and studied in rats and mice, using the exotoxin of the A strain of Mycoplasma neurolyticum.

The primary lesion of the brain consists of spongiform degeneration, associated with vesicle formation in the cortex and underlying white matter of the cerebral hemispheres, and in the molecular layer of the cerebellum. The brains of animals surviving 2 days or longer show extensive necrotizing lesions resembling ischemic necrosis, in both cerebral hemispheres.

The brains of rats and mice with rolling disease become deeply stained by intraperitoneally injected trypan blue, indicating early disruption of the blood brain barrier.

The toxin appears to be a thermolabile protein with a molecular weight exceeding 200,000. It is only active when injected by vein, and causes no disease when injected intracerebrally, intraperitoneally or subcutaneously, suggesting the existence of specific receptors within the vascular bed of the central nervous system.

Protection is afforded by rabbit antibody against the toxin, but only when antibody is injected within less than 3 min after intravenous injection of toxin, indicating rapid fixation to receptors in the brain.

The toxin is inactivated by incubation for 10 min at 37°C with suspensions of the sedimentable component of normal brain. The inactivating factor in brain sediment is very thermostable, not affected by trypsin, and eliminated by treatment with periodate. Similar inactivation of toxin is demonstrable with water-soluble gangliosides of brain.

A theoretical concept to explain the action of the toxin is proposed.
STUDIES OF PPLO INFECTION
Authors: Thomas Lewis , Bitensky Mark W.
Published: June 2004

Concentrated suspensions of washed Mycoplasma neurolyticum produce rolling disease in mice and rats, with neurological manifestations and pathological lesions similar to those seen with the exotoxin of this organism.

Pretreatment of animals with tetracycline protects completely against the toxic effects of washed suspensions of mycoplasmas, while tetracycline affords no protection against the exotoxin. Freeze-thawing disruption of mycoplasma suspensions eliminates their neurotoxicity, while the same treatment does not affect exotoxin. The toxicity of intact organisms is not affected by exposure to the sedimentable component of brain, nor to ganglioside. These observations are interpreted to indicate that the neurotoxicity of living mycoplasmas must be due to their production of toxin after they have been injected into the animal.

Resting mycoplasmas, suspended in Ringer's solution in dialysis sacs submerged in PPLO broth) produce considerable amounts of toxin within 15 min of incubation at 37°C. Toxin is also produced, although in somewhat less amount, by washed organisms suspended in phosphate buffer containing glucose. The formation of toxin is prevented by the presence of puromycin, but not by the aminonucleoside analogue of puromycin, indicating that active protein synthesis is involved in the elaboration of toxin.

The similarities between the neurotoxicity of the intact organisms of M. neurolyticum and Mycoplasma gallisepticum are discussed.
ON THE NATURE OF THE "LIPOVIRUS"
Authors: Chang R. Shihman , Pan I-Hung , Rosenau Barbara J.
Published: June 2004

Experiments designed to elucidate the nature of the "lipovirus" are described. The development of characteristic nuclear lesions in human cells in vitro depended on the presence of an ameboid cell in the inoculum. The spatial separation of the ameboid cells from the human cells by a membrane filter 150 µ in thickness was sufficient to prevent the development of nuclear lesions. Nuclear lesions appeared to be the primary change of the affected human cells. This development of nuclear lesions was partially suppressed by FUDR and the suppression was reversed by thymidine.

Time-lapse microcinematography showed that a 30 min intermittent contact between an ameboid cell and a human cell resulted in the retraction of both progenies of the human cell after a lapse of about 36 hr. Other human cells not in contact with the ameboid cell remained polygonal and continued to divide.

Radioautography of the ameboid cell revealed the presence in the cytoplasm of thymidine-containing DNAse-sensitive materials.

The development of antigens related to the ameboid cell within the cytoplasm of the human cell is described in the accompanying report (4).
EXPERIMENTAL ALLERGIC GLOMERULONEPHRITIS INDUCED IN THE RABBIT WITH HOMOLOGOUS RENAL ANTIGENS
Authors: Unanue Emil R. , Dixon Frank J. , Feldman Joseph D.
Published: June 2004

Rabbits immunized to several homologous renal antigens developed a variety of autoantikidney antibodies. Some of these antibodies reacted with the host's glomeruli and appeared to cause glomerulonephritis. Passive transfer of sera from some of these nephritic rabbits into normal, unilaterally nephrectomized rabbits led to the induction of nephritis. The production of autoantibody to glomeruli was transitory in most instances in spite of continued immunization. In some rabbits immunized to whole kidney homogenate, extracts or sediment, antibodies were found fixed to renal tubular basement membranes where an ultrastructural lesion was demonstrated. Rabbits also produced antikidney antibody apparently to tubular cytoplasmic components which did not fix to kidney in vivo and were of no pathogenetic significance.

Immunization with autologous renal basement membranes induced a small autoantibody response in half the rabbits. This response was not associated with detectable renal injury.
EQUINE ANTIHAPTEN ANTIBODY
Authors: Rockey John H.
Published: June 2004

Eight antigenically unique immunoglobulins have been identified in purified equine anti-p-azophenyl-ß-lactoside (Lac) antibody isolated from a single horse. The Fc fragments of the γGa-, γGb-, γGc-, and -γA-globulins have been shown to possess unique antigenic determinants. Common γG- and γA-Fc fragment antigenic determinants, which were absent from the 10Sγ1- and γM-globulins, have also been observed. All antibody populations share two antigenically distinct light (B, L) chain variants.

The association of anti-Lac antibody with the hapten p-(p-dimethylamino-benzeneazo)-phenyl-ß-lactoside has been measured by equilibrium dialysis and by fluorescence quenching. A variation in the affinity of anti-Lac antibody for hapten has been observed. The affinity of antibody was unaltered by enzymatic removal of the Fc fragments by peptic digestion or dissociation of the two combining sites on the papain 3.5S Fab fragments, indicating that the observed heterogeneity of affinities was not a direct function of the heterogeneity in structure of the Fc fragments. Isolated heavy (A, H) chains of γA-anti-Lac antibody have been shown to have retamed affinity for Lac dye by equilibrium dialysis and by analytical ultracentrifugation, employing a combination of schlieren and absorption optics.

The heavy (A, H) chains from two physically separable, antigenically distinct antibody populations, isolated from the same animal and having affinity for the same haptenic determinant, have been found to differ in their amino acid composition. Anti-Lac antibody light (B, L) chains have also been shown to be chemically heterogeneous, and contained populations of polypeptide chains possessing, and populations lacking methionine.

The relevance of the observed structural heterogeneity of equine anti-Lac antibody to the problem of defining the mechanism of acquisition of immunological specificity is briefly discussed.
STUDIES ON RABBIT LYMPHOCYTES IN VITRO
Authors: Sell Stewart
Published: June 2004

The addition of sheep antisera to rabbit IgG or to rabbit IgG subunits (L chain, H chain, Fab piece, Fc piece) to in vitro cultures of rabbit peripheral lymphocytes may induce up to 80–90% of the small lymphocytes to transform into immature "blast" cells. As many as 1 x 107 molecules of antibody may be required to stimulate each cell. It is concluded that each peripheral lymphocyte carries the antigenic specificities of the entire IgG molecule and thus could carry, and may produce specific antibody molecules. Such a conclusion supports the theory that the small lymphocyte may be the site of specific recognition of antigens in the primary response.
BACTERIAL INTERFERENCE IN EXPERIMENTAL BURNS
Authors: Anthony Bascom F. , Wannamaker Lewis W.
Published: June 2004

A standardized, full thickness, dermal burn in rabbits was used to study interference between strains of Staph. aureus inoculated on the wound surface. Several strains appeared equally capable of colonizing lesions and of preventing superinfection by other staphylococci inoculated at a later time. In addition, cross-infection between rabbits colonized by different strains (502A and Q461) and placed together in cages was prevented, presumably by the same mechanism. Interference appeared to be a strictly local phenomenon, since it did not occur when an animal was colonized by strain 502A at one burn site and subsequently challenged with strain Q461 at a separate lesion.

For interference to occur, a minimal time interval (9 hr) was required between inoculation of the interfering strain and inoculation of the challenge strain. In vivo growth rates indicated rapid growth in the first 24 hr by the interfering strain but no detectable multiplication by the challenge strain. Heat-killed staphylococci, even in large numbers, were incapable of producing interference. Penicillin treatment of animals colonized by strain 502A (penicillin-sensitive) abolished interference with strain Q461 (penicillin-resistant). These findings indicate that bacterial multiplication by the interfering strain is an essential feature of this phenomenon.

The mechanism of interference between strains of Staph. aureus remains obscure. There was no evidence in these studies for direct bacterial antagonism in vitro or in vivo between most of the strains examined; yet, all were capable of producing interference. Attempts to identify antistaphylococcal activity in passively transferred tissue homogenates and serum collected from infected animals were also negative. The ability of large inocula of staphylococci grown in broth to superinfect colonized lesions indicates that the numerical superiority of the interfering strain over the challenge strain is an important aspect of interference. The observation that in vivo-grown organisms may superinfect in significantly smaller quantities is suggestive of a qualitative advantage as well.
ONTOGENY OF MACROPHAGE RESISTANCE TO MOUSE HEPATITIS IN VIVO AND IN VITRO
Authors: Gallily Ruth , Warwick Anne , Bang Frederik B.
Published: June 2004

Adult or weanling C3H mice were found to be genetically resistant to a strain of mouse hepatitis virus. Infant C3H mice, however, developed infection and died from mouse hepatitis virus when minimal infectious doses of virus were given to them. There was a delay in the time of death compared to that of the genetically susceptible strain, and the virus recovered from these mice had increased pathogenicity for C3H mice.

The ontogeny of resistance to hepatitis in the C3H mice thus progresses from delayed susceptibility to complete resistance as the age of the host increases. It is reflected in increased resistance of macrophages derived in vitro from liver cultures of infant mice of different ages. This increase in resistance with age was reduced by maintaining the cultures for a longer period of time before inoculation, or by increasing the number of explants in a given culture. Resistant cells were uniformly furnished by mice age 16 days, or more. It is concluded that a process of maturation of resistance of the cells takes place after the mice are born, but that this does not continue under in vitro conditions, and that it may be modified by the environment of the cells.
RABBIT MACROPHAGE INTERFERONS
Authors: Smith Thomas J. , Wagner Robert R.
Published: June 2004

Antiviral factors present in cultures of rabbit peritoneal macrophages or rabbit kidney (RK) cells infected with Newcastle disease virus (NDV) and those in cultures of uninfected macrophages all fulfilled the biological and physicochemical criteria for classification as interferons. Virus-induced macrophage and RK interferons were slightly more stable to heat or acid than "spontaneously produced" or endotoxin-induced macrophage interferon. Interferon activity in serum of NDV-infected rabbits was decidedly more labile than NDV-induced macrophage interferon. However, these differences in lability were too slight to serve as a useful basis for distinguishing one rabbit interferon from another.

Rabbit interferons from various sources could be differentiated by filtration through Sephadex G-100 and their molecular weights estimated by comparison with elution profiles of a series of marker proteins of known molecular weight. Each of four different preparations of rabbit interferons was found to contain more than one molecular component. Elution peaks for three NDV-induced interferons were equivalent to the following molecular weights: RK ≃44,000–45,000 and > 134,000 (variable and < 1% when present); macrophage ≃37,000, 44,000–45,000, and > 134,000 (variable and <1% when present); and serum ≃50,000–52,000 and > 134,000 (∼10% and heat labile). NDV-induced serum interferon may also contain another molecular component of mol wt ≃45,000 represented by a trailing shoulder from the major 51,000 mol wt peak.

Endotoxin-induced macrophage interferon proved to be polydisperse. Sephadex filtration of this interferon did not reveal clear and consistent elution patterns, partially owing to its low initial titer and lability. However, variable peaks of biological activity could be detected in Sephadex fractions equivalent to approximate molecular weight values of > 134,000, 72,000–78,000, 33,000–38,000, 28,000–30,000, and possibly a component of 42,000–45,000. A major component of mol wt ≃37,000 was present in all samples of endotoxin-induced macrophage interferon. The other constituents may be biologically active subunits or polymers.

These data indicate that rabbit macrophages produce two primary kinds of interferon: (a) an RK-like component of mol wt ≃45,000 that is synthesized in greatest amount after viral induction, and (b) a different species of mol wt ≃ 37,000 that can also be synthesized in the absence of viral induction. The presence of major interferon constituents of mol wt ≃51,000 and > 134,000 in rabbit serum after viral induction suggests that macrophages are not the principal interferon-producing cells that respond to intravenous injection of NDV.
BIOLOGICAL ACTIVITY OF THE CLEAVAGE PRODUCT OF HUMAN IMMUNOGLOBULIN G WITH CYANOGEN BROMIDE
Authors: Lahav Mira , Arnon Ruth , Sela Michael
Published: June 2004

Treatment of human IgG with cyanogen bromide in 0.05 M HCl under specified conditions resulted in the cleavage of about half of its methionyl peptide bonds. A major fragment of about 5S was isolated from the reaction mixture by gel filtration in quantitative yield. The CNBr fragment reacted fully with goat antiserum against human light chain, but its reaction with anti-heavy chain was markedly decreased. The treatment with CNBr caused a drastic decrease in the following biological activities of IgG: complement fixing, skin binding, reaction with antiglobulin factors, and reaction with specific anti-Gm(12) serum. On the other hand, the reaction with serum of anti-Gm(1) or anti-Gm(4) specificity was not impaired and antibody activity, namely antistreptolysin and isohemagglutinin, was retained after the treatment with CNBr.

It is concluded that the CNBr cleaves preferentially the methionyl bonds in the Fc portion of IgG, and that the major fragment obtained, denoted F(ab'')2, has still the combining properties of a divalent antibody. The possible therapeutic uses of F(ab'')2 are discussed.
THE EFFECT OF TOLERANCE ON THE SPECIFICITY OF THE ANTIBODY RESPONSE AND ON IMMUNOGENICITY
Authors: St. Rose J. E. M. , Cinader B.
Published: June 2004

Animals were rendered tolerant to human albumin and were then immunized with azo derivatives of human albumin which differed in the number of hapten groups per molecule and in the extent of conformational change. The incidence and specificity of the resulting antibody response was studied and the presence of antibody to azo groups and to conformationally altered protein determinants was demonstrated. Reactivity with the tolerance-inducing antigen was shown to be due to antibodies directed against conformationally altered protein determinants. The difference in the response of tolerant animals to hapten-poor and hapten-rich derivatives was attributed to the extent of conformational alteration. A genetic factor appeared to be implicated in the capacity of tolerant animals to respond to an antigen which cross-reacts with tolerance-inducing macromolecules.
IN VITRO DEMONSTRATION OF HUMORAL AND CELL-BOUND IMMUNITY AGAINST COMMON SPECIFIC TRANSPLANTATION ANTIGEN(S) OF ADENOVIRUS 12-INDUCED MOUSE AND HAMSTER TUMORS
Authors: Hellström Ingegerd , Sjögren Hans O.
Published: June 2004

A colony inhibition (CI) technique was used to demonstrate in vitro tumor-specific antigenicity common for neoplasms induced by the adenovirus type 12. Immunization of mice against syngeneic mouse adeno 12 tumor cells evoked immunity demonstrable against other adeno 12 neoplasms of mice and of hamster as well. The immunity was detected as antibody activity in the sera of specifically immunized mice and as a similar activity of the lymph node cells of these animals. The finding in transplantation experiments that infection with adenovirus types 12 and 7 induces an isograft immunity against adeno 12 tumors was confirmed by CI tests on the sera of virus-infected mice. The probable identity of the antigen detected by the CI technique and the tumor-specific transplantation antigen is discussed.
PERSISTENCE OF GROUP A STREPTOCOCCAL CELL WALLS RELATED TO CHRONIC INFLAMMATION OF RABBIT DERMAL CONNECTIVE TISSUE
Authors: Ohanian Sarkis H. , Schwab John H.
Published: June 2004

Antibodies specific for the mucopeptide and group-specific C polysaccharide antigens of Group A streptococcal cell walls were prepared by acid dissociation of immune precipitates, and labeled with either fluorescein or 125I.

Employing both fluorescent and radioautographic procedures the persistence of the antigens was followed in skin sites injected with cell wall fragments. Both antigens persisted within macrophages for at least 54 days in those animals which developed no chronic tissue response. In animals which did develop chronic nodular lesions the concentration of antigen decreased as the inflammatory process subsided. Lesion activity was thus associated with the presence of cell wall material.

The fate of these antigens was also determined following the intradermal injection of intact Group A streptococcal cells. Cell wall antigens persisted in the tissue site considerably longer than morphologically identifiable streptococci, indicating that cell wall fragments are released during dismantling of streptococci in phagocytic cells.
GENERATION OF CHEMOTACTIC ACTIVITY IN RABBIT SERUM BY PLASMINOGEN-STREPTOKINASE MIXTURES
Authors: Taylor Fletcher B. , Ward Peter A.
Published: June 2004

The addition of plasminogen and streptokinase to rabbit serum results in appearance of chemotactic activity which is relatively heat-labile and dialyzable. Generation of this activity requires heat-labile factors in serum, but not the sixth component of complement. The optimal mole ratio for generation of chemotactic activity is 1:1 to 1:3 for plasminogen to streptokinase. Neither material alone is sufficient to generate the activity in fresh serum. Generation of chemotactic activity can be blocked by the presence of ϵ-aminocaproic acid. This plasmin-generated chemotactic factor sediments slowly in a sucrose density gradient during ultracentrifugation.

The addition of plasminogen and streptokinase to rabbit serum containing the preformed chemotactic factor (activated trimolecular complex of C'5, C'6, C'7) causes destruction of the chemotactic factor and loss of hemolytic activity of the sixth component of complement. The same mole ratio of reactants for optimal expression of the phenomenon holds as for plasmin generation of chemotactic activity in untreated serum.
IMMUNOPATHOLOGY OF NZB/BL MICE
Authors: Mellors Robert C. , Huang Chen Ya
Published: June 2004

Hemolytic disease characterized by slight anemia, reticulocytosis, hemosiderosis, extramedullary hematopoiesis, and positive indirect antiglobulin (Coombs') tests and renal disease with proteinuria, hypoalbuminemia, and glomerular lesions were produced in Swiss mice by neonatal intraperitoneal inoculation of cell-free filtrates prepared from the spleens of old NZB/Bl mice. Lymphoid cell and plasma cell hyperplasia as well as hypergammaglobulinemia occurred in some of these inoculated mice.

Type "C" murine oncogenic virus-like particles, indistinguishable from those previously described (1), were shown by electron microscopic study to be present in distinctive locations, notably in the basal foldings of convoluted tubules in the kidneys of a newborn NZB/Bl mouse, old NZB/Bl mice, a CBA x NZB F1 hybrid mouse, and a Swiss mouse inoculated with NZB/Bl spleen cell-free filtrate.

These observations point to two (perhaps related) circumstances which may be requisites for the pathogenic action of this newly discovered virus within and outside the strain NZB: infection of newborn, or infant, mice; persistent, possibly tolerant, infection of adult mice.
HYBRID FORMATION BETWEEN BACTERIAL ENDOTOXINS
Authors: Rudbach Jon A. , Milner Kelsey C. , Ribi Edgar
Published: June 2004

When endotoxins extracted from enteric bacteria were mixed in the presence of sodium deoxycholate, and the bile salt was subsequently removed by dialysis or by extraction with ethanol, a new type of endotoxin was formed. The latter material was as biologically active as the original endotoxins and possessed a combination of antigenic determinants that were previously unique to each of the individual endotoxins in the mixture. This hybrid formation between endotoxins was detected by immunodiffusion and radioautography and by quantitative precipitation procedures.
VIRAL ONCOLYSIS: INCREASED IMMUNOGENICITY OF HOST CELL ANTIGEN ASSOCIATED WITH INFLUENZA VIRUS
Authors: Lindenmann Jean , Klein Paul A.
Published: June 2004

A2G mice could be solidly immunized against the Ehrlich ascites tumor by single intraperitoneal injections of homogenized and lyophilized tumor cells which had been infected with oncolytic strains of influenza A virus. Similar homogenates from noninfected tumor cells were not immunogenic, even when mixed with egg-grown virus.

The immunizing principle in viral oncolysates could not be separated from the oncolytic virus by differential centrifugation or adsorption to and elution from red cells. It could be inhibited by antibody raised in rabbits against the egg-grown oncolytic virus. This reaction showed serologic specificity. Thus, the immunogenicity of an oncolysate produced with the WSA strain of neurotropic influenza virus could be inhibited by rabbit anti-WSA, but not by rabbit antibody to the TUR strain of fowl plague virus. Conversely, the immunogenicity of an oncolysate prepared with the TUR strain could be inhibited by rabbit anti-TUR, but not by anti-WSA.

When mice were preimmunized (primed) with egg-grown WSA virus, their antitumor response to a later injection of WSA oncolysate was of the anamnestic type. Priming with egg-grown influenza B virus had no such effect.

It was concluded that the immunogenicity of certain host cell components was greatly increased by incorporation into the makeup of the oncolytic virus.
STUDIES ON PNEUMONIA VIRUS OF MICE (PVM) IN CELL CULTURE
Authors: Compans Richard W. , Harter Donald H. , Choppin Purnell W.
Published: June 2004

Pneumonia virus of mice (PVM) particles are spheres 80–120 mµ in diameter, or filaments of similar diameter with lengths up to 3 µ. The particles possess an outer spike-covered envelope and helical internal component 120–150 A in diameter. Virus particles acquire their envelope by a budding process at the cell membrane; mature particles are seen only extracellularly.

Dense inclusions are prominent in the cytoplasm of PVM-infected BHK21 cells by 48 hr after inoculation. The inclusions appear to consist of aggregates of the internal component of PVM, and the helical component has been isolated in a cesium chloride gradient from extracts of osmotically shocked cells.

Murine erythrocytes, which are agglutinated by PVM, adsorb to the surface of infected cells and to budding and extracellular PVM particles.

On the basis of its structure and morphogenesis, PVM appears to be a myxovirus; however, it does not fit into either of the established subgroups of myxoviruses. The 120–150 A diameter of the PVM internal component differs from the diameters of the internal components of the two established subgroups of myxoviruses, and suggests that a third subgroup of these viruses may exist.
CELLS AS ANTIGEN CARRIERS AND AS IMMUNOGLOBULIN PRODUCERS
Authors: Chou Chi-Tao , Dubiski S. , Cinader Bernhard
Published: June 2004

The injection into newborn rabbits of a small quantity of human albumin, associated with red blood corpuscles or nucleated rabbit cells, induces an antibody response in the majority of animals, whereas the same quantity of antigen in solution fails to stimulate antibody formation or induces tolerance. The promoting capacity of the cells depends on attachment of antigen to them.

The antibody produced after the injection of albumin, associated with nucleated cells, is of recipient origin. However, immunoglobulin carrying the marker of donor cells can be demonstrated in the recipient animals, and may reach serum concentrations similar to those normally present in animals which are heterozygous with respect to the marker. It appears that the antibody-promoting function and the synthetic capacity for allotype are quite distinct and that the period required for allotype formation is very short with mononuclear peritoneal exudate cells and is very much longer with cells from the thymus. The capacity of cells from lymph nodes for sustained allotype formation is less than that of thymus cells but greater than that of mononuclear peritoneal exudate cells.
PRIMARY STIMULATION AND MEASUREMENT OF ANTIBODY PRODUCTION TO SHEEP RED BLOOD CELLS IN VITRO
Authors: Robinson W. A. , Marbrook J. , Diener E.
Published: June 2004

A method for the primary stimulation and measurement of antibody production to sheep red blood cells in vitro has been described. In this system when mouse spleen cells are incubated with sheep erythrocytes large complement-dependent plaques are formed. The number and size of plaques increases from day 1 to day 3 of incubation with an average of 4.4 plaque areas per 1 x 106 cells plated at day 3. There is a linear relationship between the number of spleen cells plated and the number of plaques formed. Plaque formation is inhibited by colchicine, actinomycin D, and rabbit anti-mouse globulin. This system offers a possible means for the direct in vitro measurement of the number of cells in a population susceptible to antigenic stimulation by sheep erythrocytes.
IMMUNIZATION OF DISSOCIATED SPLEEN CELL CULTURES FROM NORMAL MICE
Authors: Mishell Robert I. , Dutton Richard W.
Published: June 2004

A culture system for cell suspensions from mouse spleens has been described. The system provides adequate conditions for in vitro immunization on initial exposure to heterologous erythrocytes. The in vitro response closely parallels that observed in vivo with respect to size, early kinetics, antigen dose, and the inhibitory effect of passive antibody.

The response of cultured cells differs in two respects from that seen in vivo. There is an increase in the ability to discriminate between different varieties of homologous erythrocytes and the in vitro response does not appear to be limited by whatever mechanisms regulate the in vivo response.
CELL POPULATIONS AND CELL PROLIFERATION IN THE IN VITRO RESPONSE OF NORMAL MOUSE SPLEEN TO HETEROLOGOUS ERYTHROCYTES
Authors: Dutton Richard W. , Mishell Robert I.
Published: June 2004

The role of proliferation in the development of 19S antibody-forming cells in the primary response has been investigated in an in vitro system.

Spleen cell suspensions from normal, unimmunized mice were cultured in vitro in the presence of mammalian erythrocytes and the number of 19S hemolytic plaque-forming cells that arose 4 days later was measured. The hot pulse technique for the selective irradiation of those cells which synthesize DNA during a defined period of time has been described.

The effect of such hot pulses administered at various times after the addition of antigen on the subsequent appearance of antibody-forming cells was determined.

The results established that: (a) the onset of DNA synthesis does not start for approximately 24–32 hr after the addition of antigen, (b) essentially all the antibody-forming cells arise by cell division, and (c) different cell populations are involved in the response to two non-cross-reacting antigens.
STREPTOCOCCAL NUCLEASES
Authors: Wannamaker Lewis W. , Hayes Barbara , Yasmineh Walid
Published: June 2004

Preparations of streptococcal DNAse D with high specific activity and free of other streptococcal nucleases have been obtained by zone electrophoresis and column chromatography. Antisera prepared by injecting rabbits with such preparations specifically neutralize the activity of this enzyme.

As with DNAse B, preparations of DNAse D regularly exhibit ribonuclease activity. For both B and D enzymes, the order of substrate preference is thymus DNA, yeast RNA, bacterial RNA; but the specific activity of the D enzyme is higher than that of the B enzyme with respect to thymus DNA and lower with respect to bacterial RNA.

Both the deoxyribonuclease and the ribonuclease activities exhibited by preparations of both enzymes are inhibited by bacterial RNA, but approximately 100-fold greater concentrations of bacterial RNA are required to achieve inhibition of the deoxyribonuclease activity of the D enzyme equivalent to the inhibition of the B enzyme. The deoxyribonuclease activity of the D enzyme is also inhibited by yeast RNA, but even larger amounts are required.

These observations indicate that the D enzyme is immunologically distinct from the other streptococcal nucleases and that it differs quantitatively from the B enzyme with respect to relative specific activities on different substrates and behavior in the presence of the bacterial ribonucleic acid inhibitor.
HURLER'S SYNDROME: A GENETIC STUDY OF CLONES IN CELL CULTURE WITH PARTICULAR REFERENCE TO THE LYON HYPOTHESIS
Authors: Danes B. Shannon , Bearn Alexander G.
Published: June 2004

Clones of skin fibroblasts from normal individuals, patients with different mucopolysaccharidoses, and certain of their relatives have been examined for cellular metachromasia and cellular uronic acid.

All the clones derived from affected individuals and heterozygous carriers in families with the autosomal forms of Hurler's syndrome showed marked metachromasia and increased cellular uronic acid. Since only one cell population was demonstrated in clones derived from heterozygous carriers, no evidence for autosomal inactivation was obtained.

Clones derived from affected individuals with the X-linked recessive form of Hurler's syndrome contained uniform populations of metachromatic staining cells which demonstrated increased cellular uronic acid. Clones derived from the noncarrier fathers showed no cellular metachromasia or increased cellular uronic acid. Clones derived from the heterozygous mothers and sisters showed two populations both qualitatively and quantitatively. On the average, 72% of these clones were metachromatic and demonstrated an increased uronic acid content; 28% of the clones showed no metachromasia and the uronic acid content was similar to that found in normal individuals.

The appearance of two distinct cell populations in clones derived from females heterozygous for the X-linked recessive form of Hurler's syndrome provides evidence in favor of the Lyon hypothesis.
EFFECTS OF CHRONIC EXCESS SALT INGESTION
Authors: Dahl Lewis K. , Knudsen Knud D. , Heine Martha , Leitl George
Published: June 2004

Parabiosis has been found to modify the expected blood pressure response of rats from two strains with opposite genetic propensities for experimental hypertension. When a member from one strain was united in parabiosis with a member from the other and both were maintained on high NaCl diet, the rat from the strain ordinarily resistant to it rapidly developed hypertension, in contrast to appropriate controls from this strain. The development of hypertension in this resistant animal preceded that in its mate from the strain highly sensitive to hypertension. In the latter, both the level of hypertension and mortality were significantly less than in its control. It seems likely that the hypertension observed is the resistant parabiont was initiated in its partner from the sensitive strain. This modification in blood pressures was not observed in the absence of a high NaCl diet.

Parabiosis between animals from the same strain did not alter their response. Thus, as in earlier experiences (1–4) the interaction of a nongenetic factor (NaCl) with the appropriate genetic substrate appeared to be necessary for the development of hypertension.

The findings are interpreted as evidence that a transmittable humoral influence plays an important role in the pathogenesis of rat hypertension. The presence of this agent is genetically determined but, under the conditions of these experiments, it took the added stimulus of dietary NaCl to demonstrate its existence.
ANTIHAPTEN ANTIBODY SPECIFICITY AND L CHAIN TYPE
Authors: Nussenzweig Victor , Benacerraf Baruj
Published: June 2004

Two types of light polypeptide chain (κ and λ) are present in guinea pig immunoglobulins. The ratios of K/L molecules in anti-hapten antibodies differ sometimes markedly from that found in normal serum. Anti-DNP antibodies and anti-pipsyl antibodies have, respectively, a higher and a lower than normal K/L ratio. The possibility that the L chain type affects the range of configurations which the antibody-combining site may assume was investigated. Fractional precipitation of anti-DNP antibodies from serum of guinea pigs immunized with DNP44-BSA was performed using limiting amounts of antigen. Antibody fractions were purified from each precipitate, their affinities for ϵ-DNP-L-lysine measured by fluorescence quenching (K0) and the K/L ratio estimated by precipitation with specific antisera. Increasing concentrations of L molecules were found in fractions with decreasing K0. In other experiments, fractional precipitation and purification of antibodies which cross-react with DNP was performed in serum of animals immunized with pipsyl-BGG. The K/L ratio in antibodies isolated from these fractions was much higher than in fractions which do not cross-react with DNP. These results show that K molecules are better adapted to react with the DNP hapten than L molecules. The K/L ratio in anti-DNP antibody was shown to increase in the course of immunization, at the same time that the K0 is increasing. This rise in K0 is markedly delayed when larger doses of antigen are employed.
STUDIES ON THE MODE OF ACTION OF DIPHTHERIA TOXIN
Authors: Goor Ronald S. , Pappenheimer A. M. , Ames Elizabeth
Published: June 2004

Inhibition of soluble transferase II activity in cell-free systems by diphtheria toxin and NAD can be prevented or reversed in the presence of a sufficient concentration of nicotinamide. Quantitative studies on inhibition of peptide bond formation in cell-free extracts by toxin and NAD have indicated that two successive reversible reactions are involved. First, toxin and NAD interact mole for mole to form a relatively dissociable complex. This toxin-NAD complex then reacts with transferase II to form an enzymatically inactive product that is but slightly dissociated. In the presence of sufficient nicotinamide, however, the latter complex can be broken down to yield active transferase II once more.

Based on the above model, an equation has been derived that accurately predicts the per cent inhibition of amino acid incorporation in cell-free systems at any given toxin and NAD level. The observed inhibition appears to be independent of the sensitivity to toxin of the cell species from which the extracts were derived, and depends only on the toxin and NAD concentrations.

Although the model satisfactorily explains inhibition of peptide bond formation by toxin in cell-free systems, further assumptions are needed to explain how still lower concentrations of toxin are able to arrest protein synthesis completely in the living cell.