bluuebunny/crossref_metadata_2025 huggingface dataset:
ENZYMATICALLY PRODUCED SUBUNITS OF PROTEINS FORMED BY PLASMA CELLS IN MICE
Authors: Fahey J. L. , Askonas Brigitte A.
Published: June 2004

Gamma globulin and antibody obtained from inbred C3H mice are split by papain and cysteine into fragments roughly one-third the size of the original Molecule (S20,w = 3.5S). The papain digests were characterized by starch gel electrophoresis and immunological methods. The highly heterogeneous fragments could be divided into two groups with distinct antigenic determinants (S and F), which were separated by DEAE ion-exchange cellulose chromatography. Approximately two-thirds of the fragments had S antigenic groupings and one-third had F antigenic groupings. These data are consistent with the view that mouse gamma globulin is split by papain and cysteine into three major fragments, two of which are of the S type and one of the F type.

Antibody activity of the original molecule was present in the S fragments. Although the S fragments did not precipitate the antigen (hemocyanin) they were shown to bind antigen specifically in the manner of univalent antibodies.

The S fragments of normal γ-globulin were very heterogeneous with a broad spectrum of electrophoretic mobilities. Comparison of S fragments from slow and fast migrating globulins showed that the mobilities of the original γ-globulin samples were largely reflected in the mobilities of their S fragments. Additional observations indicated that the F fragments also may help to determine the electrophoretic mobility of intact γ-globulin molecules.

S fragments of differing electrophoretic mobility were shown to have the same antigenic determinants, indicating that the structural differences responsible for the electrophoretic mobility differences were not involved in the antigenic groupings identified with rabbit antisera.

The F fragments of normal γ-globulin migrated more rapidly than the S fragments, were less heterogeneous, and showed several bands on starch gel electrophoresis. The F fragments differed antigenically from the S fragments, and had no antibody activity. Two groups of F fragments (F and F') were detected with some antisera.

The γ-myeloma protein (5563) formed in a C3H plasma cell tumor and similarly fragmented by treatment with papain and cysteine, produced much more discrete S and F components than were found in the normal γ-globulin digest. The electrophoretic properties of the myeloma protein fragments were within the range observed for normal γ-globulin fragments. Although the γ-myeloma protein shares antigenic determinants with normal γ-globulins it lacks some of the antigenic groupings present in the γ-globulin preparation. Both S and F fragments from the myeloma protein share antigenic determinants with the corresponding fragments from normal γ-globulin. In addition, both S and F fragments of normal γ-globulin possess antigenic groupings not present in fragments of the γ-myeloma protein, accounting for the antigenic deficiency observed on comparison of the γ-myeloma protein with normal γ-globulins.
ENZYMATICALLY PRODUCED SUBUNITS OF PROTEINS FORMED BY PLASMA CELLS IN MICE
Authors: Askonas Brigitte A. , Fahey J. L.
Published: June 2004

The relationship of Bence Jones protein (mol wt = 45,000) to a ß2A-myeloma protein (mol wt = 160,000) formed by the same mouse plasma cell tumor (MPC-2) was investigated. The ß2A-myeloma protein was split by treatment with papain and cysteine into fragments (S20,w = 3.7S), similar in size to the Bence Jones protein (S20,w = 3.6S). Two types of fragments with distinct antigenic groupings designated S and F, were present in the MPC-2 myeloma protein digest. These were partially separated by DEAE-cellulose chromatography.

The Bence Jones protein was found to share antigenic determinants with S fragments from the MPC-2 ß2A-myeloma protein and with S fragments from γ-globulins. Physicochemical observations indicated, however, that the Bence Jones protein was not identical to the globulin fragments produced by treatment with papain and cysteine.

Comparison of the S and F fragments from ß2A- and γ-globulins revealed that the antigenic features shared by the various globulins derived from plasma cells (γ- and ß2A-myeloma proteins, the range of normal γ-globulins) are largely properties of the S fragments, whereas the distinctive antigenic differences between the γ- and ß2A-myeloma proteins were properties which appeared in the F fragments of the molecules.
HYPERSENSITIVITY IN NEWBORN GUINEA PIGS
Authors: Salvin S. B. , Gregg M. B. , Smith R. F.
Published: June 2004

Neonatal guinea pigs during the first 2 weeks of life did not indicate the presence of delayed hypersensitivity intradermally, after sensitization with purified soluble antigens in dose levels that induced detectable delayed hypersensitivity in the skin of adults. Although Arthus type allergy was detectable in newborns, circulating antibody frequently preceded its appearance by several days. Passive Arthus reactions were not produced in newborns as readily as in adults. Contact hypersensitivity and allergic encephalomyelitis were induced in newborns, but corneal reactions were not. Total body irradiation with 200 r inhibited antibody formation in newborns, as in adults. In addition, the induction period for anamnestic responses in newborns and the antigen elimination rate were the same as in adults. Passive transfer of delayed hypersensitivity from sensitized newborns to normal adults was accomplished.
AN ANTIGENIC BASIS FOR VIRULENCE IN STRAINS OF SALMONELLA TYPHIMURIUM
Authors: Jenkin Charles R.
Published: June 2004

A study has been made of the antigenic relationships between a virulent and an avirulent strain of Salmonella typhimurium. Evidence is presented which supports the hypothesis that the susceptibility of the mouse to infection by S. typhimurium is due to an antigenic relationship between host and parasite. The antigen shared between host and parasite, appears to be absent or is masked in the avirulent strain.
THE PREPARATION AND SOME PROPERTIES OF PENICILLENIC ACID DERIVATIVES RELEVANT TO PENICILLIN HYPERSENSITIVITY
Authors: Parser Charles W. , deWeck Alain L. , Kern Milton , Eisen Herman N.
Published: June 2004

A number of penicillenate and penicilloyl derivatives potentially useful in the study of penicillin hypersensitivity have been prepared and some of their properties described.
A TRANSIENT STAGE OF SUSPECTED DELAYED SENSITIVITY DURING THE EARLY INDUCTION PHASE OF IMMEDIATE CORNEAL SENSITIVITY
Authors: Parks James J. , Leibowitz Howard M. I. , Maumenee A. Edward
Published: June 2004

1. If quantities of bovine serum albumin or bovine gamma globulin in the range of 2 to 4 mg are inoculated intracorneally into rabbits, a biphasic reaction occurs in the cornea.

2. The primary reaction, which becomes manifest approximately 3 days after inoculation and lasts several days, is characterized by a diffuse clouding of the cornea. During this period, no antibody can be demonstrated either by serological or histological techniques. The animals react with a delayed type of skin reaction and the sensitivity can be passively transferred to normal guinea pigs with the cells of pooled lymph nodes or buffy coats. The corneal reaction is therefore considered a manifestation of delayed sensitivity.

3. The secondary reaction in the cornea, usually occurring about the 14th day after inoculation is the "Wessely phenomenon." This reaction is characterized by a precipitation of immune complexes in the cornea resulting in a visible annular corneal opacity. Circulating antibody can be readily demonstrated by both serological and histological techniques and the animals demonstrate typical Arthus skin sensitivity.

4. Generally intervening between these two phases of corneal activity is a stage in which the inoculated eye appears essentially normal. This stage is eliminated if both corneas are inoculated initially.
THE EFFECT OF TRACHOMA VIRUS VACCINE ON THE COURSE OF EXPERIMENTAL TRACHOMA INFECTION IN BLIND HUMAN VOLUNTEERS
Authors: Grayston J. Thomas , Wang San-Pin , Yang Yen-Fei , Woolridge Robert L.
Published: June 2004

The TRIC-Taiwan-1-1958 strain of elementary body virus isolated from a trachoma patient on Taiwan has been proven capable of reproducing trachoma by experimental inoculation of six human volunteers. Virus material derived from the seventh passage in embryonated hen eggs caused the clinical picture of trachoma in every inoculation, even at the dilution of 10–4 of infected yolk sacs (approximately 1 EID50). There was a similar clinical picture with each inoculation beginning with an acute follicular conjunctivitis which progressed for 4 months and then persisted with chronic changes until 9 months when treatment was begun. The illness was generally more acute than would be expected in natural trachoma. That trachoma was reproduced was shown by the involvement of the cornea with epithelial keratitis and pannus, and by the occurrence of gelatinous follicles and eventual cicatrization of the conjunctiva. These clinical findings were supported by repeated demonstrations of typical inclusion bodies of Halberstaedter-Prowazek from conjunctival and even corneal cells, by repeated reisolation of elementary body virus in egg yolk sacs, and by the development of complement-fixing antibody with a "specific" trachoma antigen in each volunteer. Control inoculations with adenovirus type 4 and normal yolk sac showed different clinical and laboratory findings.

Experimental trachoma vaccine was given to three of the volunteers to study its effect on the course of illness. An antibody response to the vaccine was demonstrated and there was a modification of disease in the volunteers receiving vaccine. While the three volunteers who received placebo each developed cross-infection of their uninoculated eye and had an acute reactivation of the bilateral disease after 1 to 2 months of antibiotic eye ointment therapy, the vaccinated volunteers remained free of infection in uninoculated eyes and showed no relapse after ointment therapy.

Treatment with sulfamethoxypyridazine, a sulfa drug with prolonged action, proved to be an effective and relatively simple method of therapy for experimental trachoma.
THE CORRELATION OF A BIPHASIC METABOLIC RESPONSE WITH A BIPHASIC RESPONSE IN RESISTANCE TO TUBERCULOSIS IN RABBITS
Authors: Allison Marvin J. , Zappasodi Peter , Lurie Max B.
Published: June 2004

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.
ELECTRON MICROSCOPIC STUDIES OF EXPERIMENTAL NEPHRITIS WITH FERRITIN-CONJUGATED ANTIBODY
Authors: Andres Giuseppe A. , Morgan Councilman , Hsu Konrad C. , Rifkind Richard A. , Seegal Beatrice C.
Published: June 2004

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.
THE BIOLOGICAL, IMMUNOLOGICAL, AND PHYSICOCHEMICAL CHARACTERIZATION OF A TRANSMISSIBLE AGENT CAPABLE OF INDUCING DNA AND THYMINE DEGRADATION IN CULTURED HUMAN CELLS
Authors: Chang R. Shihman , Humes Maryellen
Published: June 2004

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.
TRANSFORMATION OF ADULT ALLOGENEIC SMALL LYMPHOCYTES AFTER TRANSFUSION INTO NEWBORN RATS
Authors: Porter K. A. , Cooper E. H.
Published: June 2004

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.
FACTORS INVOLVED IN THE INDUCTION OF NON-SPECIFIC RESISTANCE TO STREPTOCOCCAL INFECTION IN MICE BY ENDOTOXIN
Authors: Michael J. Gabriel , Massell Benedict F.
Published: June 2004

Endotoxins derived from several species of Gram-negative bacteria, while inducing non-specific resistance to typhoid bacilli in mice, failed to increase the resistance of these animals to infection with virulent strains of Group A streptococci. However, if administration of endotoxin was followed by injection of minute amounts of type-specific antiserum, a substantial degree of protection against the streptococcal pathogen was obtained. The same amount of type-specific antiserum given to the animals by itself did not have any effect on the outcome of the streptococcal infection. Fresh rabbit blood, obtained from animals pretreated with endotoxin, together with anti-M protein immune serum, was found strongly bactericidal for streptococci. These observations suggest that, at least with regard to streptococcal infection, both humoral and cellular factors are required for induction of non-specific resistance.
THE POPULATION-DEPENDENT REQUIREMENT BY CULTURED MAMMALIAN CELLS FOR METABOLITES WHICH THEY CAN SYNTHESIZE
Authors: Eagle Harry , Piez Karl
Published: June 2004

At least seven compounds synthesized by cultured cells in amounts which should suffice for sustained growth have nevertheless proved under certain conditions necessary for survival (asparagine, cystine, glutamine, homocystine inositol, pyruvate, serine). In every instance so far examined, that requirement has been population-dependent, disappearing at cell densities sufficiently large to bring the concentration in the medium and in the cellular pool to metabolically effective levels before the cells died of the specific deficiency.

At population densities of less than 100 cells per ml, serine was required by all cultured cells so far studied. With more exigent strains, such as the RT6 strain of rabbit fibroblast and the P388 mouse leukemia, the serine requirement disappeared only at populations in excess of 50,000 and 150,000 cells per ml, respectively. The requirement for pyruvate by the latter cell as an alternative to serine also disappeared at that population density.

In a cystine-free medium there were population-dependent requirements for cystine, homocystine, or serine, depending on the experimental conditions. With methionine and glucose as cystine precursors, the critical population density permitting cellular survival and growth was in excess of 200,000 cells/ml. The provision of homocystine as an intermediate reduced the critical population density to 10,000 to 60,000 cells/ml; with the further provision of serine, the critical cell concentration permitting growth was reduced to 50 to 500 cells/ml.

Cells adapted to glutamic acid, and capable of utilizing it as a substitute for glutamine, nevertheless required exogeneous glutamine at cellular densities of less than 50,000 cells per ml. In some experiments, the provision of asparagine reduced the critical population density to 10,000 cells/ml, presumably because of its glutamine-sparing action.

Inositol is required by most cell lines, despite their demonstrated capacity to synthesize it from glucose. With at least one cell line (HeLa), sustained growth was occasionally achieved in an inositol-free medium if the population density was maintained in excess of 240,000 cells/ml.

The possible implications of these findings with respect to the loss of specific organ functions in dispersed cell culture are discussed.
COLICINE K
Authors: Rüde Erwin , Goebel Walther F.
Published: June 2004

The somatic antigen of the non-colicinogenic bacillus E. coli K235 L-OC- has been isolated, and its chemical and serological properties have been compared with those of colicine K. The antigen of the non-colicinogenic bacillus has a protein content significantly lower than that of the C+ antigen, a difference which might be related to the antibacterial activity of the latter.

The lipocarbohydrate components of the two antigens are chemically very similar; both contain the same proportions of galactose, glucose, heptose, rhamnose, glucosamine, and mannosamine.

When tested by agar diffusion, the two antigens are indistinguishable, as are their lipocarbohydrate components.

Our studies indicate that the bactericidal activity of colicine K does not reside in its lipocarbohydrate but in its protein component.
ROLE OF THE THYMUS IN IMMUNE REACTIONS IN RATS
Authors: Arnason Barry G. , Janković Branislav D. , Waksman Byron H. , Wennersten Christine
Published: June 2004

In rats thymectomized at birth and tested in adult life, ability to develop autoallergic encephalomyelitis was completely suppressed, there was a marked diminution in the degree of tuberculin sensitization appearing after a single injection of mycobacteria in oil, rejection of skin homografts was markedly delayed, and adjuvant arthritis was not appreciably affected. At the same time there was a striking decrease in the circulating level of small lymphocytes.
STUDIES ON THE CHEMISTRY AND IMMUNOCHEMISTRY OF CELL WALLS OF STAPHYLOCOCCUS AUREUS
Authors: Morse Stephen I.
Published: June 2004

The cell walls of an 80/81 strain of Staphylococcus aureus (NYH-6) contain alanine, glycine, glutamic acid, lysine, muramic acid, glucosamine, and ribitol phosphate.

94 per cent of the phosphorus and 41 per cent of the glucosamine are removed by extraction of the cell walls with hot 5 per cent TCA, but significant amounts of the other constituents are not extracted by this procedure.

The residue after hot TCA extraction (mucopeptide) is susceptible to lysozyme whereas the intact cell walls are resistant.

Staphylococcus aureus cell walls are agglutinated by S. aureus antisera. Agglutination of the cell walls of one S. aureus strain is inhibited by absorption of antisera with cell walls of other S. aureus strains but not by absorption with S. albus cell walls.

The ribitol teichoic acid can be isolated from cold TCA extracts of the cell walls. This compound consists almost entirely of ribitol phosphate and glucosamine.

The isolated teichoic acid of strain NYH-6 is readily fixed to tanned sheep erythrocytes and these sensitized cells are agglutinated by S. aureus antisera. Cold TCA extracts of cell walls of other strains of S. aureus inhibit hemagglutination whereas extracts of S. albus walls do not.

Studies on the inhibition of both hemagglutination and precipitation indicate that the antigenic determinant of S. aureus NYH-6 teichoic acid is ß-N-acetylglucosamine.
STUDIES ON THE INTERACTIONS BETWEEN COMPONENTS OF STAPHYLOCOCCUS AUREUS AND STAPHYLOCOCCUS BACTERIOPHAGE
Authors: Morse Stephen I.
Published: June 2004

The cell walls of Staphylococcus aureus are capable of inactivating S. aureus bacteriophage. Furthermore, the cell walls isolated from S. aureus of a given phage type inactivate a variety of different staphylococcal bacteriophages. Under the conditions employed neither the isolated mucopeptide nor teichoic acid components of the cell walls act as bacteriophage receptor.
EXPERIMENTAL ALLERGIC ENCEPHALOMYELITIS: THE EFFECT OF 6-MERCAPTOPURINE
Authors: Hoyer Leon W. , Good Robert A. , Condie Richard M.
Published: June 2004

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.
COMPARATIVE STUDIES IN ROUS SARCOMA WITH VIRUS, TUMOR CELLS AND CHICK EMBRYO CELLS TRANSFORMED IN VITRO BY VIRUS
Authors: Morgan Herbert R. , Andrese Angelo P.
Published: June 2004

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.
ANTIGENICITY OF RAT COLLAGEN
Authors: Rothbard Sidney , Watson Robert F.
Published: June 2004

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.
THE INTRACELLULAR DISTRIBUTION OF GAMMA GLOBULIN IN A MOUSE PLASMA CELL TUMOR (X5563) AS REVEALED BY FLUORESCENCE AND ELECTRON MICROSCOPY
Authors: Rifkind Richard A. , Osserman Elliott F. , Hsu Konrad C. , Morgan Councilman
Published: June 2004

Ferritin- and fluorescein-conjugated antibody staining has been applied to a study of a mouse plasma cell tumor. The presence of myeloma globulin within cisternae of the endoplasmic reticulum was observed at a stage of the secretory process when the remainder of the cytoplasm was essentially free of labeled globulin. The distribution of ferritin suggested a functional heterogeneity among units of the endoplasmic reticulum. Apparently, progressive accumulation of globulin results in distension of the endoplasmic reticulum and, occasionally, in the appearance of considerable quantities of this secretory protein in the extracisternal cytoplasmic matrix. Participation of the Golgi apparatus in the packaging and release of small quantitites of globulin seems likely. In addition, however, fragmentation of the peripheral cytoplasm with rupture of distended ergastoplasmic vesicles appeared to be another pathway whereby globulin is secreted.
AUTOSENSITIZATION REACTION IN VITRO
Authors: Koprowski Hilary , Fernandes Mario V.
Published: June 2004

Lymph node cells were obtained from an inbred strain of Lewis rats injected with guinea pig cord tissue in Freund's adjuvant. These cells, when added to tissue culture monolayers of puppy brain, aggregated on or around the glial elements. This reaction, called contactual agglutination, was followed by the specific destruction of glial cells, leaving cultures consisting only of fibroblasts. No such reaction was noted when lymph node cells obtained either from normal rats or those injected with adjuvant alone were used. Absorption of serum obtained from rats injected with guinea pig cord tissue by non-sensitized lymph node cells made them reactive in brain tissue culture. The contactual agglutination test seems to provide an opportunity for investigation of sensitization reaction in tissue culture systems.
SPECIFIC FRACTIONATION OF A POPULATION OF ANTIDEXTRAN MOLECULES WITH COMBINING SITES OF VARIOUS SIZES
Authors: Schlossman Stuart F. , Kabat Elvin A.
Published: June 2004

Two human antidextran sera were each fractionated into two populations of antibody by specific absorption of the antidextran on an insoluble dextran (sephadex), washing away non-specific protein, eluting the first fraction of antibody with isomaltose or isomaltotriose, and the second fraction with isomaltohexaose. The differences in behavior of the purified antibody fractions alone, or reconstituted in serum, in quantitative inhibition studies with the isomaltose series of oligosaccharides, or in quantitative precipitin studies with NRC dextrans of graded molecular weight, could be ascribed to differences in the sizes of their combining sites. It was shown that the antibody fractions eluted with isomaltose or isomaltotriose were made up largely of antibodies inhibited readily by the smaller oligosaccharides and therefore having a higher proportion of molecules with smaller-sized combining sites; whereas those fractions eluted with isomaltohexaose contained primarily antibodies readily inhibitable only with the larger oligosaccharides, and hence were considered to have a higher proportion of larger-sized combining sites. The purified antibody fractions were shown to be only fast gamma globulin,—to possess the same mobility in immunoelectrophoresis; in addition the antibody fractions from one individual in double diffusion in agar gave lines which fused with one another, with gamma globulin Fr. II, and with a purified antidextran solution containing the entire population of antidextran antibodies. Evidence was also presented which indicated little or no change in the populations of antibody molecules produced in these two individuals for a period as long as 10 years following immunization. It is felt that these data offer substantial support for the hypothesis that the antidextran produced in a single individual consists of a heterogeneous population of antibody molecules with antibody-combining sites of various sizes.
GENETIC CHARACTERS OF HUMAN γ-GLOBULINS IN MYELOMA PROTEINS
Authors: Harboe M. , Osterland C. K. , Mannik M. , Kunkel H. G.
Published: June 2004

The genetic factors Gm(a), Gm(b), Gm(x), and Inv(a), Inv(b) described for normal human γ-globulin were all found in different myeloma proteins. A single myeloma protein never contained more than one product of alternate alleles even in heterozygous individuals. However, factors determined by the two different loci were often found in the same myeloma protein. The Gm(a) character of the myeloma protein parallelled that of the normal γ-globulin of the same serum in most cases. In contrast, the Gm(b) character was usually absent in the myeloma protein when it was directly demonstrable in the normal γ-globulin. The myeloma proteins from six Negroes were Gm(a+b-), whereas the normal γ-globulin was Gm(a+b+). This indicates that the effect of gene Gmb is similar in Negroes and whites, even though its relation to gene Gma is different in the two races.

Gm factors were found only in the 7S γ-globulin type myelomas and not in other products of plasma cell tumors. Inv characters were, however, present in all four types of proteins studied, namely 7S and 19S γ-globulins, ß2A-globulins, and Bence Jones proteins. In two instances, genetic heterogeneity of the protein products was demonstrated suggesting the proliferation of more than one clone of plasma cells in some multiple myeloma patients.

The accumulated evidence obtained in this study strongly suggested that the presence and absence of genetic characters was compatible with the concept that myeloma proteins were closely analogous to individual moieties in the spectrum of normal γ-globulins rather than truly abnormal proteins. Their study offered evidence of a heterogeneity of genetic characters among the normal γ-globulins in a given individual. It also appears probable that in normal individuals single plasma cells have a restricted capacity to express genetic information in their protein product.
A CARRIER STATE OF MUMPS VIRUS IN HUMAN CONJUNCTIVA CELLS
Authors: Walker Duard L. , Hinze Harry C.
Published: June 2004

Mumps virus produced a carrier state in human conjunctiva cells that was maintained for more than 100 subcultures over a period of 3 years. Antiserum in the medium was not required. The virus had little apparent effect on the cells which grew at a rate similar to uninfected control cells. Mumps virus was regularly found in the culture medium at levels about 0.9 log higher than the cell-associated virus. When first tested after 30 subcultures, the virus was found to have lost its cytopathogenicity for cells ordinarily susceptible to mumps virus, but was identifiable as mumps virus by neutralization with specific antiserum. Use of fluorescein-labeled antiserum revealed that 80 to 95 per cent of cells in the carrier cultures contained mumps virus antigen. The antigen was concentrated in a few sharply circumscribed, discrete masses in the cell cytoplasm rather than in many granules throughout the cytoplasm as is characteristic of cell infection by cytopathogenic mumps virus. The carrier cultures were resistant to the destructive effect of a cytopathogenic line of mumps virus, but showed little resistance to the cytopathogenic effect of vesicular stomatitis, Sendai, or Newcastle disease viruses.
EFFECTS OF BACTERIAL ENDOTOXINS ON METABOLISM
Authors: Berry L. Joe , Smythe Dorothy S. , Kolbye Susannah McC.
Published: June 2004

The greater susceptibility to the lethal effects of bacterial endotoxin (heat-killed Salmonella typhimurium or Escherichia coli lipopolysaccharide, in mice infected with an attenuated strain of Mycobacterium tuberculosis (BCG) was confirmed. It reached a maximum at 2 weeks postinfection and gradually diminished for an additional 6 weeks.

At the time of maximum susceptibility several metabolic and physiological differences became apparent. BCG-infected mice die sooner (4 to 12 hours) and without the diarrhea, conjunctivitis, and general symptomatology associated with endotoxin deaths of normal animals. Reticuloendothelial blockade results in only a small change in reactivity to endotoxin, in contrast to normal mice. Subcutaneous injection of 2 units of ACTH is followed by no significant increase in urinary nitrogen excretion while in control animals it more than doubles. Plasma clearance of intravenously administered inulin is approximately normal in BCG-infected mice 17 hours after an LD50 dose of endotoxin but control mice similarly treated show renal impairment. In line with this result is the absence of elevated carcass non-protein nitrogen (NPN) following endotoxin poisoning or at the moment of death from endotoxemia in the hyperreactive animals in contrast to the two- to threefold increase in carcass NPN in normal mice under similar conditions. Body carbohydrate is at a minimum and becomes depleted to a level approximating that found at death more rapidly in BCG-infected mice given endotoxin than in controls. There is also a lower ratio of carbohydrate anabolized to protein catabolized following cortisone administration to BCG-infected mice than in control mice. This is found in adrenalectomized mice and in stressed animals and is reported elsewhere.

Some of the differences just described can be attributed to a refractory adrenal cortex. There is less depletion of adrenal cholesterol in vivo and lower corticoid synthesis in vitro than in normal mice yet this is not fundamentally responsible for the greater susceptibility of BCG-infected animals to endotoxin since adrenalectomized mice, which are even more susceptible, are metabolically and physiologically more comparable to normal mice than to BCG-infected mice. One can conclude, therefore, that the hyperreactivity of BCG-infected mice is more than an intensification of the normal response to endotoxin.
THE MODE OF ACTION OF ANTIBIOTICS IN STIMULATING GROWTH OF CHICKS
Authors: Eyssen H. , de Somer P.
Published: June 2004

A study was made of the effect of antibiotics on growth of chicks and on intestinal absorption of fats and carbohydrates.

Around the 8th day of life, chicks fed an antibiotic-free casein-sucrose diet developed a transitory syndrome of malabsorption of fats and carbohydrates, associated with disturbance of the efficiency of feed utilization and poor weight increase. Administration of virginiamycin, at a level of 20 ppm, suppressed this period of malabsorption and resulted in improved feed conversion and increased weight gain.

The temporary growth depression and malabsorption were not observed in disinfected rooms in new quarters. Under these conditions virginiamycin did not stimulate growth nor was the efficiency of feed utilization improved by the antibiotic. However, the growth-depressing flora could be introduced to the new quarters by feeding each bird 50 mg of fresh feces collected from chicks in old quarters.

Both the intestinal absorption and the growth-promoting effect of virginiamycin were influenced by the type of carbohydrate in the basal diet, and have been found to be most pronounced when sucrose was fed as the sole source of carbohydrate. The malabsorption was less obvious when cornstarch was substituted for sucrose. In this case virginiamycin had only a limited effect on growth and on feed conversion.

The present investigations suggest that antibiotics stimulate growth of chicks by their antibacterial action against Gram-positive microorganisms which interfere with the absorption of nutrients. Furthermore, the growth-promoting effect seems to be most pronounced during a limited period of a few days around the 8th day of life.
THE MECHANISM OF BLOCKADE OF THE RETICULOENDOTHELIAL SYSTEM
Authors: Murray I. MacKay
Published: June 2004

Rats were injected intravenously with various particulate materials with and without the addition of various stabilizing agents. One hour after the injection of the blockading colloid, a tracer dose of similar or different colloidal material was injected. A half-time clearance rate of the tracer dose greater than 4 minutes was taken as evidence of blockade of the RES.

Blockade of the tracer dose occurred when the surface properties of the particles in both blockading and tracer doses were identical. Different particles stabilized by the same agent behaved as identical particles and identical particles stabilized by different agents behaved as different particles.

The opsonization of a tracer dose of gelatin-stabilized colloid in vitro by a specific globulin fraction obtained from heterologous and homologous plasma prevented its blockade, while opsonization of both blockading and tracer doses with the same proteins resulted in blockade. However, when both blockading and tracer doses were opsonized with isologous globulins, blockade did not occur.
THE IN VITRO PRODUCTION OF ANTIBODY BY DELAYED PHASE CORNEAL LIMBAL LYMPHOID FOCI
Authors: Leibowitz Howard M. I. , Parks James J.
Published: June 2004

The intracorneal inoculation of a sufficient quantity of a soluble protein antigen into the eye of a rabbit produces a biphasic allergic reaction in that cornea. The earlier stage, characterized by a diffuse corneal clouding, is a manifestation of delayed hypersensitivity and is accompanied by a limbal infiltrate composed predominantly of lymphocytic-mononuclear elements. The later response, known as the Wessely Phenomenon, is a ring of opacification in the cornea which is visible in the gross. This reaction is dependent upon the presence of specific circulating antibodies and is therefore classified among the immediate types of hypersensitivity. It is accompanied by a dense limbal infiltration of plasma cells. Intervening between the two reactions is a period of several days during which the eye appears relatively normal.

Explants containing large numbers of infiltrating lymphocytic-mononuclear elements were removed from the corneal-scleral junction of experimental eyes during the height of the delayed type hypersensitivity reaction and maintained in vitro in tissue culture. At a later date the overlay fluid in which the explants were maintained was shown to contain specific humoral antibodies, demonstrating the capability of cells present at a delayed reaction for the later production of antibodies. The possible linkage of the two modes of immune phenomena is discussed.
TWO MAJOR TYPES OF NORMAL 7S γ-GLOBULIN
Authors: Mannik Mart , Kunkel Henry G.
Published: June 2004

Normal 7S human γ-globulin was found to contain two fundamental antigenic groups of molecules. The group 1 molecules of normal γglobulin correspond antigenically to group 1 multiple myeloma proteins and Bence Jones proteins; and group 2 molecules of normal γ-globulin correspond antigenically to group 2 multiple myeloma proteins and Bence-Jones proteins. Among pooled human Fr II and several individual γ-globulin preparations, approximately 60 per cent of molecules belong to group 1 and approximately 30 per cent of molecules to group 2 in this classification. The possible existence of a third minor antigenic group, constituting about 10 per cent, is discussed. Antisera to Bence Jones proteins of antigenic group 1 and group 2, in conjunction with I-131-labeled 7S γ-globulin proved to be the most useful system for defining the antigenic groups of normal γ-globulin. The group-specific antigenic determinants of normal 7S γ-globulin molecules were located on the S fragments of these proteins.
THE EFFECT OF ANTIBACTERIAL DRUGS ON THE WEIGHT OF MICE
Authors: Dubos Rene , Schaedler Russell W. , Costello Richard L.
Published: June 2004

NCS mice gained weight rapidly when fed a gluten diet deficient in several amino acids, but their weight gain on the same regimen was very much retarded if they were given antibacterial drugs, even for a short period of time. This retardation of growth could not be entirely corrected by supplementing the gluten diet with lysine and threonine.

The decrease in growth rate brought about by antibacterial drugs could probably be traced to the alteration in intestinal flora resulting from drug treatment. The intensity and duration of both types of changes were related to the dose of drug administered, and to the length of the treatment period.

Whatever the nutritional regimen, treatment with penicillin caused a retardation of weight gain in NCS mice. The retardation was more pronounced, and longer lasting, when the animals were fed semisynthetic regimens (containing casein or gluten) than when they were fed crude diets (pellets) containing natural materials of ill defined composition. These differences probably had their origin in the fact that the changes in fecal flora induced by the drugs were profoundly influenced by the composition of the diet.

Antibacterial drugs which retarded weight gain of Swiss NCS mice, in contrast increased weight gain in ordinary Swiss mice raised under usual conditions. It is probable that this difference in response to the antibacterial drugs resulted from the fact that ordinary Swiss mice have a much more complex intestinal flora than NCS animals.
IN VITRO STUDIES OF MAMMALIAN SOMATIC CELL VARIATION
Authors: Cann Howard M. , Herzenberg Leonard A.
Published: June 2004

When long term cultures of mouse lymphoma cells, known to possess the isoantigenic phenotype determined by the H-2d allele, are incubated with anti H-2d isoantibody and guinea pig complement, slightly more than 99 per cent of cells are killed under optimal conditions. Growth in mass culture and colony formation by single cells after incubation with isoantibody and complement are employed to assess the cytotoxic effect. The cytotoxic action of isoantibody is complement-dependent, for viability of cells exposed to antibody alone is unaltered. When excess isoantibody and optimum concentrations of complement are used, killing begins as soon as these reagents are mixed with the cells, and no further killing occurs after 5 to 15 minutes at 37°C. About 80 per cent of cells are killed with an isoantiserum containing antibody to two isoantigenic components of the H-2d complex. That the cytotoxic action is mediated through the H-2 isoantigen is shown by (a) isoantiserum containing only anti H-2d antibody produces maximal cell killing, and (b) isoantiserum from which anti H-2d antibody has been removed by absorption loses all cytotoxic activity.

Variant cells resistant to the cytotoxic action of anti H-2d isoantibody were isolated from lymphoma cell populations surviving multiple exposures to isoantibody and complement. These variants can be distinguished morphologically from the isoantibody-sensitive parent cell line. Although variants are resistant to anti H-2d isoantibody, these cells possess H-2d isoantigen but in a lower concentration than found in cells of the parent line. The basis for resistance to cytotoxic isoantibody is discussed.
LYSINE DEFICIENCY AND HOST RESISTANCE TO ANTHRAX
Authors: Gray Irving
Published: June 2004

1. Lysine deficiency has been produced in rats by placing the weanling animals on an experimental diet in which gluten replaced the casein of the control diet. Both diets were complete in all other known requirements.

2. The resistance of the deficient animals to a subcutaneous challenge of several concentrations of B. anthracis spores, was decreased. Deaths occurred within 2 days post inoculation.

3. When lysine was added to the gluten diet to bring the total concentration to that of the control diet, the growth rate of the animals was maintained but a decrease in resistance remained, although not as great as on the gluten diet.

4. Changes in the tissues associated with lysine deficiency are reported.

5. It is not unreasonable to state that within the time frame of our experiments, the decreased ability of the RES of the host to clear the invading organism from the tissues and subsequently to break down the organism is a major factor of the decreased resistance of the lysine-deficient rats to anthrax.