bluuebunny/crossref_metadata_2025 huggingface dataset:
A STUDY OF THE PASSIVE TRANSFER OF DELAYED HYPERSENSITIVITY TO DNP-POLY-L-LYSINE AND DNP-GL IN RESPONDER AND NONRESPONDER GUINEA PIGS
Authors: Green Ira , Paul William E. , Benacerraf Baruj
Published: June 2004

A study of the passive transfer of delayed hypersensitivity to DNP-poly-L-lysine and to DNP-GL was performed in Hartley guinea pigs. Delayed hypersensitivity to DNP-PLL and DNP-GL could be transferred successfully only by means of sensitized cells from genetic responder guinea pigs and in most cases, only into those guinea pigs genetically capable of responding to PLL.

The inability to transfer delayed hypersensitivity to DNP-PLL or DNP-GL to genetic nonresponder guinea pigs is not the result of the early destruction of the transferred cells by an incompatible host, since it was shown that delayed hypersensitivity to ovalbumin could be successfully transferred from guinea pigs with the PLL gene into genetic nonresponder animals.

The requirement of active participation of specific and genetically controlled host mechanisms in the successful passive transfer of delayed sensitivity to DNP-PLL and DNP-GL has been demonstrated.
COMPLEMENT AS A MEDIATOR OF INFLAMMATION
Authors: da Silva W. Dias , Eisele John W. , Lepow Irwin H.
Published: June 2004

Purified preparations of human C'1 esterase, C'4, C'2, C'3, and C'5 were labeled with 125I. Reaction mixtures were prepared containing a single labeled component and other unlabled components. After incubation at 37°C for 10 min at pH 7.4 in the presence of 5 x 10–4 M Mg2+, they were adjusted to pH 3.5 and subjected to sucrose density gradient ultracentrifugation and gel filtration at pH 3.5. In all cases, an activity capable of contracting guinea pig ileum with tachyphylaxis was obtained in low molecular weight fractions. However, these fractions were labeled only when 125I-C'3 was employed, indicating that biological activity was associated with a cleavage product of C'3. This fragment has been designated F(a)C'3 in a nomenclature consistent with that of immunoglobulin degradation products. The much larger, residual portion of the C'3 molecule has been designated F(b)C'3.

The biochemical characteristics of generation of F(a)C'3 were consistent with a mechanism involving action of C'1 esterase on C'4 and C'2, activation of C'2, and cleavage of C'3.

F(a)C'3 had a molecular weight by gel filtration techniques of 6800 or less. It was thermostable and susceptible to inactivation by endo- and exopeptidases. The isolated fragment possessed all of the biological properties of unfractionated mixtures of C'1 esterase, C'4, C'2, and C'3. In addition to contraction of guinea pig ileum, these included failure to contract rat uterus, enhancement of vascular permeability in guinea pig skin, degranulation of mast cells in guinea pig mesentery, and release of histamine from rat peritoneal mast cells. F(a)C'3 did not cross-desensitize guinea pig ileum to rat agar anaphylatoxin and vice versa.

The existence of different protein fragments with anaphylatoxin properties has been discussed. Distinctive characteristics of F(a)C'3 from classical anaphylatoxin generated by treatment of fresh rat serum with agar have been indicated.
IODINATION OF BACTERIA: A BACTERICIDAL MECHANISM
Authors: Klebanoff Seymour J.
Published: June 2004

Myeloperoxidase, iodide, and H2O2 have a bactericidal effect on Escherichia coli. Myeloperoxidase can be replaced in this system by lactoperoxidase or by a guinea pig leukocyte particulate preparation, H2O2 by an H2O2-generating system such as glucose and glucose oxidase, and iodide by thyroxine or triiodothyronine. The bactericidal effect was high at pH 5.0 and fell as the pH was increased.

Preincubation of myeloperoxidase, iodide, and H2O2 for 30 min before the addition of the bacteria largely prevented the bactericidal effect. Thus, the organisms must be present in the reaction mixture during iodide oxidation for maximum killing, which suggests the involvement of labile intermediates of iodide oxidation rather than the more stable end products of oxidation such as iodine.

Iodination of the bacteria by the myeloperoxidase-iodide-H2O2 system was demonstrated chemically and radioautographically. Iodination and the bactericidal effect were similarly affected by changes in experimental conditions in all the parameters tested (effect of preincubation, pH, and inhibitors).

Phagocytosis of bacteria by guinea pig leukocytes was associated with the conversion of iodide to a trichloroacetic acid-precipitable form. Iodide was localized radioautographically in the cytoplasm of human leukocytes which contained ingested bacteria. Iodide fixation was not observed in the absence of phagocytosis or in the presence of Tapazole.
FURTHER OBSERVATIONS ON THE MECHANISM OF RETICULOENDOTHELIAL BLOCKADE
Authors: Drutz David J. , Koenig M. Glenn , Rogers David E.
Published: June 2004

The mechanism of the reticuloendothelial "blockade" which followed injection of large quantities of chromic phosphate without exogenous stabilizing material was investigated in Wistar rats.

The RE blockade observed for several hours after induction appeared related to the continuing circulation of the chromic phosphate-blockading dose, and a reduction in the size of the particles used enhanced blockade.

RE blockade appeared to be particles specific and was not related to a generalized depression of RE-phagocytic cell function.

Studies in isolated perfused rat livers appeared to eliminate saturation of particle-specific macrophage clones as a likely explanation of blockade, and blockade could not be explained on the basis of depletion of serum opsonins.

In the system employed, it is postulated that blockade occurs when large numbers of circulating particles saturate specific macrophage cell membrane-binding sites rather than from physical stuffing of RE-phagocytic cells.
STUDIES ON CIRCULATING IMMUNE COMPLEXES
Authors: Cochrane Charles G. , Hawkins David
Published: June 2004

The relationship between certain physicochemical properties of circulating immune complexes and their ability to localize in vessel walls during a state of increased permeability was studied. The ability to become deposited was related to the large size of complexes, rather than to their net charge or to a specific affinity between complexes and structures of vessel walls.

Soluble complexes with sedimentation rates greater than 19S were capable of being entrapped along the vessel wall membranes, while complexes smaller than this were not. These large complexes were removed rapidly from the circulation, while smaller complexes persisted. Minimal levels of total complexes in the circulation necessary for detectable vascular localization were found to be as low as 15 µg antibody N/ml plasma.

In experimental serum sickness, a disease known to be induced by circulating immune complexes, the development of vascular and glomerular lesions occurred almost exclusively in rabbits having large (greater than 19S) circulating immune complexes. Animals with smaller complexes did not show deposition of complexes in glomeruli or development of glomerulonephritis. Their incidence of vasculitis was markedly reduced.
SUPPRESSION OF ADJUVANT DISEASE IN THE RAT BY HETEROLOGOUS ANTILYMPHOCYTE GLOBULIN
Authors: Currey H. L. F. , Ziff Morris
Published: June 2004

The effect of antilymphocyte globulin (ALG) on adjuvant arthritis, an immunologically induced disease in the rat, was studied. ALG was prepared from the serum of rabbits immunized against rat lymphocytes.

Adjuvant arthritis was induced in rats by a single intracutaneous injection of Freund's complete adjuvant; after 9 to 12 days, all control rats developed polyarthritis. Administration of antilymphocyte globulin about the time of adjuvant injection produced marked inhibition of clinical disease. Some suppression was apparent even when ALG was started after the onset of arthritis. Rats receiving ALG remained conspicuously healthy compared to controls. Urinary findings and renal histology showed no evidence of nephritis. The results of serum corticosterone determinations made it unlikely that adrenal stimulation contributed to the actions of ALG.

Antilymphocyte preparations lowered peripheral lymphocyte counts and suppressed primary antibody responses to sheep erythrocytes, but had little effect on the skin reaction to PPD, even in rats protected from arthritis.

All rats given ALG developed antibodies to rabbit globulin; there was no evidence that ALG inhibited the appearance of antibodies to itself, and prior hyperimmunization of rats with rabbit globulin did not interfere with the biological activity of ALG subsequently injected.

Antisera produced separately against lymph node and thymus cells had identical properties with regard to agglutination of lymphocytes and thymus cells. Administered to rats, these preparations were equally potent in lowering lymphocyte counts and suppressing both adjuvant arthritis and the primary antibody response to sheep erythrocytes.

It is concluded that ALG, as used in these experiments, is a potent immunosuppressive agent without obvious toxic effects.
HEMOPOIETIC COLONY STUDIES
Authors: Wolf N. S. , Trentin J. J.
Published: June 2004

In heavily irradiated mice, bone marrow regeneration of either endogenous or exogenous origin was shown to occur in discrete foci comparable to the more intensively studied spleen colonies. The number of endogenous bone marrow colonies was inversely related to dose of whole body X-irradiation. Endogenous marrow colonies were found after higher doses of irradiation than were endogenous spleen colonies. Most of them were granulocytic in nature.

Exogenous bone marrow colonies in lethally irradiated mice injected with bone marrow cells were proportional in number to the dose of cells injected, appeared at a time comparable to spleen colonies like which, at 7 or 8 days, they were of single differentiated cell line, either granuloid or erythroid or megakaryocytic, with a small percentage of "mixed" colonies.

Whereas erythroid colonies outnumber granuloid colonies in spleen, either in situ or subcutaneously transplanted (E:G colony ratio of about 3.5), granuloid colonies outnumber erythroid in bone marrow (E:G colony ratio of about 0.7). The characteristic E:G colony ratios of spleen and marrow appear more likely to be the result of a hemopoietic organ stromal influence on pluripotent colony forming units (CFU's) than of selective lodgment of committed (unipotent) granuloid and erythroid CFU's in bone marrow and spleen, respectively, as indicated by the following.

Bone marrow stem cells (CFU) which had reseeded the marrow cavity of irradiated primary recipients 18–24 hr earlier, were reharvested and retransplanted intravenously into irradiated secondary hosts. The E:G colony ratio of the colonies formed in the spleen of the secondary hosts was typical of primary spleen colonies (2.8), that of the colonies formed in the marrow cavity was typical of bone marrow colonies (0.6). Pieces of marrow stroma containing reseeded CPU's from the contralateral femur of these same primary recipients were implanted by trocar directly into the spleens of other irradiated secondary recipients. Those CPU's that developed in the intrasplenic-implanted marrow stroma yielded an. E:G colony ratio of 0.1. Those that migrated into the contiguous and remote portions of the spleen gave E:G colony ratios of 2.9 and 2.4, respectively.

Irradiated marrow stroma and normal spleen CPU's (a 1 mm cube of spleen) were loaded into the same trocar and implanted directly into the spleens of irradiated mice. The spleen CFU's that migrated into the implanted marrow stroma yielded five granuloid and two mixed colonies. The larger number that developed in the host spleen yielded an E:G colony ratio of 2.9 or higher.

Of those 19 mixed colonies that bridged the junction of spleen and implanted marrow stroma in each of the above two experiments, in every case, the erythroid portion of the colony was in the splenic stroma, the granuloid portion was in the marrow stroma.
SPECIFICITY OF CELLULAR IMMUNE RESPONSES
Authors: Paul William E. , Siskind Gregory W. , Benacerraf Baruj
Published: June 2004

In vitro antigen stimulation of DNA synthesis in lymph node cultures from immunized guinea pigs can be obtained with very low (10–4 µg/ml) antigen concentrations in the culture fluid. Immunization with low doses of DNP-GPA leads to a cell population capable of being stimulated, on the average, by low concentration of antigen whereas immunization with large antigen doses results in a sensitive cell population requiring, on the average, high antigen concentrations for stimulation. These findings correlate well with the affinity for hapten of the serum antibodies produced by these guinea pigs.

Both delayed reactions in vivo and DNA synthesis in vitro can be stimulated by hapten conjugated to proteins different from that used in primary immunization. However the immunizing conjugate is much more effective in terms of antigen concentration required for a given response.

These results can be understood in terms of a thermodynamically driven interaction of antigen (or "processed" antigen) with cell-associated antibody.
ADENO-ASSOCIATED SATELLITE VIRUS INTERFERENCE WITH THE REPLICATION OF ITS HELPER ADENOVIRUS
Authors: Parks Wade P. , Casazza Anna M. , Alcott Judith , Melnick Joseph L.
Published: June 2004

Adeno-associated satellite virus type 4 interferes with the replication of its helper adenovirus. No interferon-like soluble substance could be detected in satellite-infected cultures and other DNA- and RNA-containing viruses were not inhibited by coinfection with satellite virus under conditions which reduced adenovirus yields by more than 90% in monkey cells. Altering the concentration of adenovirus in the presence of constant amounts of satellite resulted in a constant degree of interference over a wide range of adenovirus inocula and suggested that adenovirus concentration was not a significant factor in the observed interference. The interference with adenovirus replication was abolished by pretreating satellite preparations with specific antiserum, ultraviolet light or heating at 80°C for 30 min. This suggested that infectious satellite virus mediated the interference. Satellite virus concentration was found to be a determinant of interference and studies indicated that the amount of interference with adenovirus was directly proportional to the concentration of satellite virus.

8 hr after adenovirus infection, the replication of adenovirus was no longer sensitive to satellite interference. This was true even though the satellite virus was enhanced as effectively as if the cells were infected simultaneously with both viruses.

Interference with adenovirus infectivity was accompanied by reduced yields of complement-fixing antigen and of virus particles which suggested that satellite virus interfered with the formation and not the function of adenovirus products. When cells were infected either with adenovirus alone or with adenovirus plus satellite, the same proportion of cells plated as adenovirus infectious centers. However, the number of plaque-forming units of adenovirus formed per cell in the satellite-infected cultures was reduced by approximately 90%, the same magnitude of reduction noted in whole cultures coinfected with satellite and adenovirus. This suggested that all cells infected with the two viruses were producing a reduced quantity of adenovirus.
ANTIGENS IN IMMUNITY
Authors: Nossal G. J. V. , Abbot A. , Mitchell Judith
Published: June 2004

Details of antigen trapping and processing in the rat lymph node have been investigated by the technique of high resolution radioautography. A series of 24 adult rats was injected with 20 µg of 125I-labeled Salmonella adelaide flagella, given as either a primary or a secondary stimulus into one hind foot-pad. At intervals ranging from 3 min to 3 wk, rats were killed and the popliteal nodes were processed for electron microscopic radioautography using Kodak NTE emulsion. The present paper deals with events in the lymph node medulla, and an accompanying report describes the radically different behavior of antigen in the cortical follicles.

In the medulla, lightly labeled granulocytes were transiently encountered, but by far the greatest bulk of antigen was in macrophages. Antigen entered these cells in two ways: by direct penetration of the plasma membrane; and by pinocytosis. In either case, the antigen rapidly became surrounded by tiny vesicles which may have represented Golgi-derived "protolysosomes." Vacuolar fusion ensued and a series of progressively larger and more complex antigen-containing "phagolysosomes" was formed. Substantial amounts of antigen could be detected in such bodies for at least 3 wk.

The antigen injection, as expected, caused extensive plasma-cytopoiesis. No evidence of label in plasma cells was obtained. No special anatomic relationship between plasma cells and antigen depot sites was discovered. These results are briefly discussed in relation to current theories of immune induction.
REGULATION OF CELLULAR ANTIBODY SYNTHESIS
Authors: Möller Göran
Published: June 2004

Transfer of spleen cells from mice immunized against sheep red blood cells (SRBC) into irradiated (600 R) nonimmune, syngeneic mice in the presence of antigen resulted in excessive cellular 7S production 7 days later. The number of 7S plaque-forming cells usually exceeded 106 per spleen and the mean proportion varied between 1 and 70%. In occasional animals all spleen cells were producing antibodies to SRBC. Serum antibody synthesis was also excessively increased, the titers in agglutination after 2-ME treatment and in hemolysis varying between 215 and 225. The generation time of the 7S PFC was found to be 9.6 hr in the secondary hosts.

It seemed possible that the excessive production of 7S PFC and antibodies in the irradiated nonimmune recipients was caused by the absence of feedback inhibition of the immune response by antibody, a mechanism which would normally function to restrict antibody synthesis. This conclusion was strengthened by the demonstration that transfer of antigen-stimulated immune cells into actively or passively immunized irradiated recipients resulted in a marked suppression of cellular 7S synthesis. Serial transfers of antigen-stimulated immune cell populations in irradiated hosts resulted in an equally high number of 7S PFC during the first four transfer generations. However, after the fifth to seventh transfer generation the number of 7S PFC rapidly declined and disappeared within one to three passages. Serum antibodies and 7S PFC declined in parallel during the last transfer generations. Further passages of antigen-stimulated spleen cells lacking 7S PFC did not lead to reappearance of PFC. Thus, antigen-sensitive cells have a limited lifespan and/or multiplication capacity. From the hypothesis that the 7S PFC developed by division from antigen-sensitive precursors it was calculated that 38–40 divisions occurred, Thus, one antigen-sensitive precursor has the potential to give rise to 1012 7S PFC.
THE PROVOCATION OF LATENT LYMPHOCYTIC CHORIOMENINGITIS VIRUS INFECTIONS IN MICE BY TREATMENT WITH ANTILYMPHOCYTIC SERUM
Authors: Volkert Mogens , Lundstedt Claus
Published: June 2004

The hypothesis that treatment with antilymphocytic serum (ALS) can provoke latent virus infections has been investigated.

In adult mice infections with sublethal doses of LCM virus usually result in the development of immunity to the virus and at the same time to a prolonged latent infection.

In the experiments described an intensive treatment with large doses of ALS was given to mice which had recovered from LCM virus infection. At the beginning of the treatment the mice had high titers of complement-fixing antibodies in their blood and no detectable virus. The data presented show that in spite of the immunity the ALS treatment provoked the occult virus and led to the development of viremia in all the treated mice. In some, very high virus titers were demonstrable. When the ALS treatment was discontinued the viremia disappeard again.

In most of the mice the ALS did not suppress the complement-fixing antibody titers and in some there was even a considerable increase in titer. In such cases the increases in virus titers and in antibody titers were closely related to one another. These results demonstrate once again that the complement-fixing antibodies to the LCM virus in mice probably do not influence the virus.
PAPAIN DIGESTION FRAGMENTS OF HUMAN IGM GLOBULINS
Authors: Mihaesco Constantin , Seligmann Maxime
Published: June 2004

Papain digestion of two Waldenström IgM globulins produced a high amount of small peptides and resulted in the formation of two end products, the Fabµ and Fcµ fragments.

The Fcµ fragment is characterized by a fast electrophoretic mobility, a high content in carbohydrate, and a high molecular weight. It was demonstrated that this fragment is made of heavy chain pieces belonging to several disulfide-linked monomeric subunits, presumably representing the carboxy-terminal end of the µ-chains. Fc fragments from the two macroglobulins could not be distinguished immunologically. An appreciable proportion of IgM molecules apparently underwent degradation without the formation of a stable Fc fragment. An Fc-like fragment, analogous to the reduced Fc fragment, was obtained at early stages of papain digestion of the IgM subunits.

The Fabµ fragment, with slow and individually distinct electrophoretic mobility, bears many physicochemical and immunological similarities to the Fabγ fragment. It consists of one light chain and one Fd piece, both of which were isolated. The interaction of these two constituents was demonstrated by gel diffusion studies. Fab fragments of both IgM globulins were resolved into two subpopulations with different electric charges.

In addition to these fragments, intermediary split products were observed at early stages of the degradation process, together with a high yield of small peptides mainly derived from the papain-sensitive region of the heavy chains. Immunologic data strongly suggested that this segment of µ-chains is situated between the Fd piece and the portion included in the Fc fragment.

Several experiments indicated the importance of conformational antigenic specificity in both Fab and Fc regions of the IgM globulins.
PATHOGENESIS OF THE GLOMERULONEPHRITIS OF NZB/W MICE
Authors: Lambert P. H. , Dixon Frank J.
Published: June 2004

The development of glomerulonephritis in NZB/W mice is closely related to the formation of antinuclear, particularly anti-DNA, antibodies. The developing inflammatory glomerular lesions are characterized by the deposition of γG- and ß1C-globulins plus DNA and possibly other nuclear antigens, presumably as complexes, in a granular to lumpy pattern along the capillary walls and in the mesangia. Elution studies revealed the γG-globulin in the glomeruli to be largely γG2A-type antibody to soluble nuclear antigens. Enhancement of the antinuclear antibody response by active immunization of young NZB/W mice with DNA-methylated BSA hastens the development and increases the severity of the glomerulonephritis. Similarly, injections of soluble DNA into NZB/W mice with circulating anti-DNA antibodies but with as yet little nephritis causes rapid progression of nephritis.
THE RESPONSE OF CULTURED MAMMALIAN CELLS TO DIPHTHERIA TOXIN
Authors: Moehring Joan M. , Moehring Thomas J.
Published: June 2004

We have investigated the response of the resistant mouse L cell to diphtheria toxin. Intact cells and cell-free systems were studied. It was determined that the cell-free system is as sensitive to toxin as those from sensitive reticulocyte, HeLa, and KB cells previously studied. Poly-L-ornithine, reported to stimulate macromolecular uptake, enhanced toxin activity under conditions which also stimulated acid phosphatase activity. Resistance, in the L cell, appears to be linked to the cell membrane and the process of macromolecular uptake, and not to any intrinsic property of either the polyribosomes or associated protein synthesizing factors. Because of the unique way in which diphtheria toxin inhibits protein synthesis, the ability of poly-L-ornthine to enhance this action provides convincing evidence that poly-L-ornithine stimulates a true absorption of macromolecules and not just increased surface adsorption.
HETEROGENEITY OF RABBIT IGM ANTIBODY AS DETECTED BY C'1a FIXATION
Authors: Hoyer Leon W. , Borsos Tibor , Rapp Herbert J. , Vannier Wilton E.
Published: June 2004

The C'1a-fixing properties of purified rabbit IgM anti-benzenearsonate antibody were determined. When tested with sheep erythrocytes to which hapten had been coupled by diazo linkage, the number of C'1a molecules fixed was 21% of the number of IgM antibody molecules bound to the erythrocyte surface. This was not due to loss of C'1a-fixing capacity during the purification procedure. Preparative electrophoresis of the antibody concentrated C'1a-fixing molecules in the anodal region so that antibody fractions with greater C'1a-fixing capacity were obtained. The demonstration that C'1a fixation is a property of a subpopulation of IgM molecules provides evidence for previously unrecognized µ-chain heterogeneity.
ANTIBODY PRODUCTION IN VITRO
Authors: Harris Gilmour
Published: June 2004

The effect of high specific activity thymidme-3H on proliferation and antibody production, using the hemolytic plaque-forming technique, by spleen cell suspensions in vitro from rabbits killed after a boost of SRC's has been studied. High specific activity thymidine-3H inhibited the proliferative ay well as the antibody response to antigen, and it was conduded that this was the result of the incorporation of radioactive 3H into the nuclei of dividing cells which were synthesizing antibody in these cultures.

The stimulation of the rate of DNA synthesis by specific antigen could be correlated with the ability of antigen to maintain antibody production, as measured by the specific hemolytic plaque-forming technique, above levels found in control cultures, incubated without antigen.

Radioautographic studies of PFC's in vitro showed that the majority of the cells arose from the DNA-synthesizing population of cells in these cultures, confirming the conclusions from the results of the inhibitory effects of high specific-activity thymidine-3H on PFC's. It was found that these PFC's, labeling with thymidine-14C, formed only a small proportion of all the cells labeled in this way in these cultures. The postulation was made that antigen, in vitro, provided a stimulation for cell proliferation in the responsive population of rabbit spleen cells, but that only a small proportion of this population could be induced by antigen to synthesize antibody.
ANTIBODY PRODUCTION IN VITRO
Authors: Harris Gilmour
Published: June 2004

The effects of actinomycin D and puromycin on spleen cell suspensions from rabbits immunized to SRC's were studied. These inhibitors, in high concentration, suppressed PFC's when added initially to recently isolated cells. When such cells were incubated for several days in the presence of both antigen and inhibitor, both actinomycin D and puromycin produced an increase in PFC's after the initial suppression. This recovery effect was best seen with cells from rabbits killed 3 days after boosting with SRC's, and was usually absent when cells were taken from rabbits killed 2 days after boosting. When actinomycin D or puromycin was added after several days in culture in the presence of SRC's, surviving PFC's were found to be not only resistant to these inhibitors, but there was also an increased number of PFC's compared to similar cultures incubated without these agents.

Radioautographic studies showed that PFC's stimulated by the presence of actinomycin D or puromycin were not incorporating precursors for RNA or protein synthesis. In view of the known mode of action of these inhibitors, it was postulated that they were stimulating antibody production by PFC's in vitro either by interfering with represser mechanisms or stimulating the completion of antibody molecules, perhaps by causing the release of preformed antibody chains from ribosomes.

Since the presence of specific antigens in vitro were necessary for these observed stimulatory effects on PFC's, and since antigens were producing an effect on antibody production on cells which were being suppressed by these inhibitors, added initially, it was further suggested that one role of antigen in the immune response was concerned with the completion of antibody synthesis on the ribosomes, perhaps by acting as an inducer as has been suggested previously (1).
THE EFFECT OF HETEROLOGOUS ANTI-LYMPHOCYTE SERUM ON MOUSE HEMOPOIETIC STEM CELLS
Authors: DeMeester Tom R. , Anderson Norman D. , Shaffer Charles F.
Published: June 2004

The present study has demonstrated that rabbit anti-mouse lymphocyte serum (RAMLS) has the capability of destroying bone marrow cells and suppressing hemopoietic stem cell function. The in vitro incubation of bone marrow suspensions with RAMLS caused extensive cell lysis with an apparent preferential destruction of lymphoid, erythroid, and blastoid elements. Using the spleen colony assay, the number of functional hemopoietic stem cells was found to be markedly reduced in bone marrow populations exposed to RAMLS in vitro. Further, this loss of stem cell function could be produced by exposing marrow suspensions to small concentrations of antiserum which did not produce detectable cytotoxic effects on the general marrow population.

A similar effect of RAMLS upon hemopoietic stem cells was found in vivo. The intravenous injection of RAMLS into lethally irradiated mice immediately after the infusion of isogeneic marrow cells reduced the number of spleen colonies formed, indicating that the antiserum could exhibit a deleterious effect upon stem cells in the bloodstream of the intact animal. Normal animals treated with daily subcutaneous injections of RAMLS for 3 wk had a significantly reduced marrow content of functional hemopoietic stem cells, suggesting that RAMLS can affect stem cells located in situ in the bone marrow. The experiments indicate that RAMLS possesses potential marrow toxicity.
IMMUNOPATHOLOGY OF LYMPHOCYTIC CHORIOMENINGITIS VIRUS INFECTION OF NEWBORN MICE
Authors: Hirsch Martin S. , Murphy Frederick A. , Hicklin Martin D.
Published: June 2004

Antithymocyte serum, when administered neonatally to mice, delayed the maturation of the lymphoid system, permitting development of cellular tolerance to LCM virus at an older age than is ordinarily possible. Humoral antibody formation was not prevented and the animals exhibited the paradox of high titers of both circulating virus and antibody. This, in turn, was followed by a chronic immunopathologic glomerulonephritis in most animals. Some animals developed wasting disease between 1 and 2 months of age, characterized by reticular cell hyperplasia and widespread infiltration into tissues and organs.
THE SERUM OPACITY REACTION OF STREPTOCOCCUS PYOGENES
Authors: Top Franklin H. , Wannamaker Lewis W.
Published: June 2004

Investigation into the antigenicity of streptococcal lipoproteinase has revealed the existence of multiple, immunologically distinct enzymes. Each lipoproteinase identified was found to be strain specific in that it was found only in strains of a particular T-agglutination pattern. In some T patterns, all streptococci of that T pattern which were examined shared a common lipoproteinase antigen. In other T patterns, strains which produced antigenically different lipoproteinases were identified. Evidence that the lipoproteinase antigen is distinct from other well-established cellular antigens of Group A streptococci has been presented. The production of this strain-specific enzyme by strains currently difficult to type by the standard M precipitin method may facilitate more precise identification of these strains and a better assessment of their role in the pathogenesis of Group A streptococcal infections and their sequelae.
PERSISTENCE OF IMMUNOGENICITY OF ANTIGEN AFTER UPTAKE BY MACROPHAGES
Authors: Unanue Emil R. , Askonas Brigitte A.
Published: June 2004

Peritoneal macrophages were cultured for several hours after uptake of 131I-hemocyanin. The cells degraded most of the 131I-labeled protein within 2–5 hr. Their ability to prime lymphocytes of syngeneic mice for a secondary immune challenge remained unchanged for long periods of time despite the loss of more than 90% of the original content of antigen. The persistence of immunogenicity was associated with a small percentage of antigen retained by the cell in a form which was protected from rapid breakdown and elimination.
ARGININE-RICH PROTEINS OF POLYMORPHONUCLEAR LEUKOCYTE LYSOSOMES
Authors: Zeya H. I. , Spitznagel J. K.
Published: June 2004

The cationic antibacterial proteins of rabbit PMN lysosomes have been resolved into at least five subfractions. Each of these showed substantial selectivity in its antibacterial action against several pathogenic bacteria, including two smooth and two rough Escherichia coli strains, three Staphylococcus aureus strains, one S. albus, three proteus species and four different cultures of streptococcus.

Each of the subfractions possesses a different electrophoretic mobility. Amino acid analyses of the three most cationic components revealed high contents of arginine consistent with their relative electrophoretic mobilities and very high arginine to lysine ratios. Aromatic amino acids were present in very low concentrations in these proteins and their light absorption at 2800 A was correspondingly weak.

The evidence of antibacterial specificity, along with marked differences in the arginine-lysine ratios, shows that the cationic antibacterial components of rabbit PMN lysosomes are biologically and chemically heterogeneous.
HEMATOPOIETIC ORIGIN OF MACROPHAGES AS STUDIED BY CHROMOSOME MARKERS IN MICE
Authors: Virolainen Martti
Published: June 2004

The origin of macrophages was studied in mouse radiation chimeras by chromosome marker technique. Macrophage cultures were established from peritoneal exudate, from lung washings, and from organ cultures of bone marrow, spleen, lymph node, thymus, and lung. Cultured macrophages were induced to divide by adding conditioned medium from L cell cultures.

In chimeras which were lethally irradiated and given injections of bone marrow or spleen cells, dividing macrophages were of donor type, independent of the source of the macrophages. When chimeras were established by injections of a mixture of bone marrow cells and cells from other hematopoietic tissues of two genetically different donors, the ratio of cells with different genotypes was approximately the same in bone marrow cells and in macrophage cultures. Thymus, lymph node, and peritoneal exudate cells were not found to contain precursor cells for macrophages. Precursor cells for macrophages and for bone marrow cells appeared to be equally sensitive to sublethal irradiation.

The results indicate that macrophages from different sources can all be derived from hematopoietic tissues, and suggest that only hematopoietic tissues contain precursor cells for macrophages which are capable of in vitro division. The close relationship between the source of cells in bone marrow and in macrophage cultures suggests that, at the maturation level at which the irradiated host is repopulated, the precursor or stem cells for macrophages may be identical with those for myeloid and erythroid series of cells.
KINETICS OF THE REACTION OF RENIN WITH NINE SYNTHETIC PEPTIDE SUBSTRATES
Authors: Skeggs Leonard T. , Lentz Kenneth E. , Kahn Joseph R. , Hochstrasser Harry
Published: June 2004

A number of peptides have been synthesized which represent portions of the tetradecapeptide renin substrate molecule, and which contain the hydrolyzable leu-leu bond. An automatic chemical method for determination of the velocity of the reaction of renin with these compounds was developed. Application of the method at several levels of substrate concentration permitted construction of Lineweaver-Burk plots, and calculation of Michaelis constants (Km) and maximal velocities (Vmax). The results show that the maximum affinity of the enzyme (lowest Km) for substrate is achieved only with the full tetradecapeptide molecule (asp1-arg2-val3-tyr4-ileu5-his6-pro7-phe8-his9-leu10-leu11-val12-tyr13-ser14). Removal of asp1 and arg2 from the N-terminal increases the Km eight-fold. Further, moderate increase in Km occurs when the next amino acids, val3, tyr4 and ileu5, are removed. The further removal of his6 results in a marked reduction in the Vmax. Removal of ser14 from the C-terminal of the nonapeptide his6-pro7-phe8-his9-leu10-leu11-val12-tyr13-ser14 does not greatly affect the Km nor the Vmax. Further removal of tyr13 from this compound results in complete loss of substrate activity. It is suggested that the compounds his6-pro7-phe8-his9-leu10-leu11-val12-tyr13-ser14 or his6-pro7-phe8-his9-leu10-leu11-val12-tyr13 might be used as substrates for the chemical assay and standardization of renin.
COMPETITION OF 19S AND 7S ANTIGEN RECEPTORS IN THE REGULATION OF THE PRIMARY IMMUNE RESPONSE
Authors: Henry Claudia , Jerne Niels K.
Published: June 2004

Prior to sheep red cells (SRC) mice were given 7S or 19S anti-SRC antibodies or mixtures of both. All 7S preparations suppressed the immune response. All 19S preparations enhanced the primary response, as measured by an up to 15-fold increase in the number of PFC per spleen. Results obtained with mixtures showed that 7S and 19S antibodies are competitive in their effect. The kinetics of the appearance of PFC in the mouse spleen after injection of SRC suggest that the depressive effect of 7S antibody simulates a reduction in SRC dose, whereas the enhancing effect of 19S antibody appears as a temporary increase in the rate at which PFC appear. Antibodies from one animal species are quite effective in another species.
THE ROLE OF AFFERENT LYMPHATICS IN THE REJECTION OF SKIN HOMOGRAFTS
Authors: Barker Clyde F. , Billingham R. E.
Published: June 2004

Experiments have been carried out on guinea pigs of two isogenic strains to elucidate the role of afferent lymphatic vessels in the rejection of orthotopic skin homografts. Graft beds were prepared in partially isolated skin flaps with an intact sustaining vascular "umbilical cord" in which a lymphatic connection with the host could be retained or abolished at will.

In the absence of demonstrable lymphatic connections between flap and host, intra-flap homografts long outlived similar grafts transplanted to conventional sites in intact skin and, rather than being specifically rejected, died as a consequence of ischemic necrosis of the flap. When lymphatic drainage was retained, intra-flap homografts were rejected in the usual manner. Hosts of long-term intra-flap homografts did not develop sensitivity, as evidenced by the "first set" type rejection of subsequent test grafts, or by the long-term survival of a second skin graft transplanted to a new flap raised on the opposite side of the host's body.

Intra-flap skin homografts were rejected if (a) the hosts had been presensitized, (b) they were grafted concomitantly with a skin homograft placed in a conventional site, or inoculated with a suspension of donor lymphoid cells, or (c) if the lymphatic drainage was restored by reimplantation of the hitherto partially isolated flap to an appropriate vascular bed. These findings and others indicate that an intact lymphatic drainage in its bed is essential for an orthotopic skin homograft to sensitize its host.

Various experiments were carried out in which intra-flap homografts were used as "indicators" for the acquisition of specific active or adoptive immunity by their hosts.

By transplanting skin homografts to conventional beds concomitantly with intra-flap grafts and then excising the former at various intervals, it has been found that a graft must be in residence for a minimum period of 4 days to evoke the development of a detectable level of sensitivity in the host. Furthermore, by replacing either freshly prepared or long-term skin flaps bearing skin homografts in vascular beds on the trunk and determining the subsequent survival times of the homografts, evidence has been obtained suggesting that reestablishment of a functional lymphatic system in a free skin graft may take as long as 9 days.

Using intra-flap homografts as indicators of adoptive immunization of the host, we found that as few as 50 x 106 isologous peripheral blood leukocytes from a specifically sensitized animal will transfer an effective level of sensitivity. We also found that hyperimmune serum, in relatively large amount, exerts a weak but definite adverse effect upon either freshly or recently transplanted intra-flap grafts.
THE INDUCTION OF GRAFT VERSUS HOST DISEASE IN MICE TREATED WITH CYCLOPHOSPHAMIDE
Authors: Owens Albert H. , Santos George W.
Published: June 2004

In these studies adult mice treated with cyclophosphamide and foreign immunologically competent cells developed a graft versus host disease which outwardly resembled that encountered in other experimental systems. Progressively larger doses of cyclophosphamide produced an increasingly severe disease whereas comparable doses of mechlorethamine were ineffective. Increasingly larger cell inocula from parental, allogeneic, and xenogeneic donors resulted in a correspondingly more severe disease. Nucleated cells obtained from the peripheral blood were found to be the most potent inducers of this syndrome, while cells from the spleen, bone marrow, and thymus displayed lesser degrees of reactivity in that order. No such graft versus host disease occurred in mice given saline, lysed, or heat-killed cells in place of viable foreign cells. Neither did the disorder develop when comparable inocula of isogeneic cells were used.
CELL WALL COMPOSITION AND VIRULENCE IN ESCHERICHIA COLI
Authors: Medearis Donald N. , Camitta Bruce M. , Heath Edward C.
Published: June 2004

Uridine diphosphate galactose 4-epimerase and phosphomannose isomerase-deficient mutants of Escherichia coli O111:B4 were studied to test the hypothesis that in E. coli a specific relationship exists between O antigenicity, virulence, and capacity to resist phagocytosis. The first mutant, designated J-5, produces a cell wall lipopolysaccharide, the side chains of which do not contain galactose, glucose, N-acetylglucosamine, or colitose. The second mutant produces a cell wall lipopolysaccharide which lacks only colitose. The capacity of these various organisms to kill mice was strikingly different. E. coli O111 was 1000 times as virulent as J-5, and 100 times as virulent as L-2.

The capacity of the organisms to kill mice was correlated with their ability to resist phagocytosis and to persist in the peritoneal cavity. The parent strain of O111 resisted phagocytosis by macrophages in vivo and polymorphonuclear leukocytes in vitro. The mutants did not, and the organism most deficient in the saccharide component of its LPS was most susceptible to phagocytosis and least virulent. These results were corroborated by growing the mutants in appropriately supplemented media which permitted the synthesis of complete LPS, reversed the susceptibility to phagocytosis, and restored virulence. Finally, serological reactivity was consistent with previous observations which had demonstrated that the O antigenicity of E. coli is determined by the saccharide composition of its cell wall lipopolysaccharide. Despite the difference in the capacity of the various log-phase organisms to kill mice when injected intraperitoneally, purified lipopolysaccharides extracted from them did not differ significantly in their capacity to kill or produce fever. Thus virulence was shown to be independent of endotoxin activity which in turn seemed to be unrelated to the saccharide composition of the cell wall LPS.

Collectively, these data provide at least a partial molecular definition of virulence in E. coli by demonstrating that the presence or absence of specific sugars in its cell wall lipopolysaccharide is a determinant of its antiphagocytic capacity and its virulence.
THE ORIGIN AND KINETICS OF MONONUCLEAR PHAGOCYTES
Authors: van Furth Ralph , Cohn Zanvil A.
Published: June 2004

The origin and turnover of efferent populations of mouse mononuclear phagocytes has been described. Mononuclear phagocytes were defined as mononuclear cells which are able to adhere to glass and phagocytize.

In vitro labeling studies with thymidine-3H showed that monocytes in the peripheral blood and peritoneal macrophages do not multiply and can be considered end cells in a normal, steady state situation. However, the mononuclear phagocytes of the bone marrow appear to be rapidly dividing cells.

This conclusion was supported by in vivo labeling experiments. A peak of labeled mononuclear phagocytes of the bone marrow was found 24 hr after a pulse of thymidine-3H. This was followed, 24 hr later, by a peak of labeled monocytes in the peripheral blood. From these experiments it was concluded that the rapidly dividing mononuclear phagocytes of the bone marrow, called promonocytes, are the progenitor cells of the monocytes. Labeling studies after splenectomy and after X-irradiation excluded other organs as a major source of the monocytes.

Peak labeling of both the blood monocyte and peritoneal macrophages occurred at the same time. A rapid entry of monocytes from the blood into the peritoneal cavity was observed, after a sterile inflammation was evoked by an injection of newborn calf serum. These data have led to the conclusion that monocytes give rise to peritoneal macrophages.

No indications have been obtained that mononuclear phagocytes originate from lymphocytes.

In the normal steady state the monocytes leave the circulation by a random process, with a half-time of 22 hr. The average blood transit time of the monocytes has been calculated to be 32 hr. The turnover rate of peritoneal macrophages was low and estimated at about 0.1% per hour.

On the basis of these studies the life history of mouse mononuclear phagocytes was formulated to be: promonocytes in the bone marrow, → monocytes in the peripheral blood, → macrophages in the tissue.
CATABOLIC ORIGIN OF A BENCE JONES PROTEIN FRAGMENT
Authors: Cioli D. , Baglioni C.
Published: June 2004

Gel filtration analysis of the urinary proteins of some patients with myeloma has shown the presence of "fragments" of Bence Jones proteins which correspond to the variable half of these proteins. Experiments have been carried out to establish the origin of a "fragment" observed in a patient who excreted a large amount of this protein. Labeled homologous Bence Jones protein has been injected into this and other control patients. Excretion of labeled "fragment" has been observed in all. Analysis by peptide mapping and radio-autography of this labeled "fragment" isolated from the urine showed that the invariable half of the Bence Jones protein was not excreted; it seemed thus likely that the invariable half was metabolized to small peptides and free amino acids.

A labeled Bence Jones protein which was excreted without any accompanying "fragment" was injected into the patient who excreted large amounts of "fragment." No excretion of labeled "fragment" was observed. It was thus concluded that the property of being degraded to "fragment" is characteristic of some "fragile" Bence Jones proteins and is not determined by the patient. Incubation with serum or urine of the "fragile" Bence Jones protein failed to produce any "fragment." "Fragments" of Bence Jones proteins are thus most likely formed during excretion of these proteins through the kidney and are products of the catabolism of Bence Jones proteins.