bluuebunny/crossref_metadata_2025 huggingface dataset:
IMMUNE RESPONSE TO CHEMICALLY MODIFIED FLAGELLIN
Authors: Parish C. R.
Published: June 2004

Flagellin (mol. wt. 40,000) from S. adelaide organisms was acetoacetylated to varying extents with diketene (acetoacetic anhydride). Chemical studies demonstrated that the amino groups of flagellin were more readily acetoacetylated than the hydroxyl groups. Several antigenic tests revealed that as flagellin was acetoacetylated to increasing extents there was a steady decline in the affinity of the molecule for anti-flagellin antibodies.

Loss in antigenic activity following acetoacetylation was not related to the number of acetoacetyl groups attached but was determined by the type of residue substituted. Reactive lysine residues were much less important anti-genically than easily substituted hydroxyl groups.

Acetoacetylation very readily destroyed the antibody-forming capacity of flagellin in rats. This fall in immunogenicity was related to the antigenic activity of the preparations. In fact, only a 40% reduction in the antigenic activity of flagellin produced a 90–95% reduction in primary antibody formation. The more heavily acetoacetylated flagellins produced no detectable antibody and, in fact, rendered adult rats tolerant (in terms of antibody formation) to a subsequent challenge of flagellin. Tolerance was induced by acetoacetylated flagellins which had drastically reduced affinities for anti-flagellin antibodies.

These results were interpreted as indicating that the affinity of antigen for receptors on cells appears to be of crucial importance in determining whether antibody formation or immunological tolerance (antibody suppression) occurs.
THE ANTIGENS AND AUTOANTIGENS OF THE SEMINAL VESICLE
Authors: Orsini Frank , Shulman Sidney
Published: June 2004

Guinea pig vesicular fluid was characterized both biochemically and immunologically. Biochemical analyses showed this fluid to be homogeneous by ultracentrifugal analyses, revealing a single boundary with a sedimentation coeflicient of 1.5 S. In contrast, electrophoretic separation methods revealed six components, of which three were major components, of approximately equal proportions. They were termed I, II, and III. One of these components (II) was shown to be strongly antigenic in heteroimmunization, whereas components I and III failed to show any antigenicity, even after diverse attempts. This antigen (component II) was found to be highly tissue specific and species specific.

Through procedures of isoimmunization, component II was also found to be immunogenic, giving rise (in male animals) to autoantibodies, A high proportion of injected guinea pigs showed positive skin tests and many revealed tissue lesions when the seminal vesicles were examined histologically. It is therefore concluded that experimental autoimmune disease of the seminal vesicle has been induced.
CONTRIBUTION OF BONE MARROW CELLS AND LACK OF EXPRESSION OF THYMOCYTES IN GENETIC CONTROLS OF IMMUNE RESPONSES FOR TWO IMMUNOPOTENT REGIONS WITHIN POLY-(PHE,GLU)-POLY-PRO--POLY-LYS IN INBRED MOUSE STRAINS
Authors: Mozes Edna , Shearer G. M.
Published: June 2004

Previous cellular studies on the genetic regulation of immunological responsiveness for two immunopotent regions within the branched chain synthetic polypeptide (Phe, G)-Pro--L demonstrated a direct correlation between the number of detectable immunocompetent splenic precursor cells and the response patterns of SJL, DBA/1, and F1 mice (21). In order to establish the cellular origin(s) of the genetic defect, the present study first demonstrated that thymus and bone marrow cell cooperation was required for (Phe, G)- and Pro--L-specific immune responses. Secondly, limiting dilution experiments, in which several graded and limiting inocula of marrow cells were mixed with a non-limiting number of 108 thymocytes and injected into irradiated, syngeneic recipients, indicated that the low responsiveness of the SJL and DBA/1 strains to the (Phe, G) and Pro--L specificities, respectively, could be attributed to a reduced number of precursor cells found in bone marrow. About five times more marrow precursors were detected in SJL mice for Pro--L than for (Phe, G), whereas about five times as many precursor cells were estimated for (Phe, G) as for Pro--L in the DBA/1 strain. These differences are similar to those obtained using spleen cells from unimmunized SJL and DBA/1 donors (21), and indicate that these genetically determined variations in responsiveness can be accounted for by differences in the frequencies of monospecific populations of immunocompetent cells present in bone marrow.

In contrast, limiting dilution transfers of thymocytes or thymus-derived cells with an excess of syngeneic marrow cells resulted in equally frequent (Phe, G) and Pro--L responses for both SJL ad DBA/1 strains. This finding in conjunction with the observation that the generation of (Phe, G)- and Pro--L-specific responses were associated in individual recipients injected with limiting inocula of thymocytes indicated that a single population of thymocytes was stimulated by (Phe,G)-Pro--L. Therefore, it is improbable that the thymic population of immunocompetent cells contributes to expression of these genetically controlled defects.
PEYER'S PATCHES: AN ENRICHED SOURCE OF PRECURSORS FOR IGA-PRODUCING IMMUNOCYTES IN THE RABBIT
Authors: Craig Susan W. , Cebra John J.
Published: June 2004

The proliferative and differentiative potential of Peyer's patch, peripheral blood, and popliteal lymph node cells was assessed by allogeneic cell transfer followed by quantitation of donor immunocytes by immunofluorescence. It was found that Peyer's patches are a highly enriched source of cells which have the potential to proliferate and differentiate into IgA-producing immunocytes and that the Peyer's patch cells are far more efficient in seeding the gut of irradiated recipient rabbits with donor cells that give rise to immunoglobulin-producing cells than cells from peripheral blood or popliteal lymph nodes.
CHARACTERIZATION OF IMMUNOGLOBULIN STRUCTURES FROM THE SURFACE OF CHRONIC LYMPHOCYTIC LEUKEMIA CELLS
Authors: Eskeland Trond , Klein Eva , Inoue Masaharu , Johansson Bo
Published: June 2004

Chronic lymphocytic leukemia cells with relatively large amounts of mu and kappa immunoglobulin structures on the surface, and apparently very small amounts intracellularly, were subjected to homogenization or washing after freezing and thawing. After a light centrifugation, which sedimented the nuclei and unbroken cells, most of the immunoglobulin structures were found in the supernatant. Ultracentrifugation, which was performed to remove the membranes from the supernatant, sedimented only half the amount of the immunoglobulin structures.

By sucrose density gradient ultracentrifugation and Sephadex G-200 filtration, the unsedimented immunoglobulin structures were shown to consist of 7S IgM and free kappa chains. About 80,000 7S IgM molecules were calculated to be present on each cell. The amount of kappa chains not associated with IgM was estimated to be equal to the amount of kappa chains in IgM. Inhibition of passive hemagglutination was used to detect and quantitate the immunoglobulin structures.
CELL-TO-CELL INTERACTION IN THE IMMUNE RESPONSE
Authors: Miller J. F. A. P. , Sprent J.
Published: June 2004

Collaboration between thymus-derived lymphocytes and nonthymus-derived antibody-forming cell precursors occurs in the primary antibody response of mice to heterologous erythrocytes and serum proteins. The purpose of the experiments reported here was to determine whether collaboration took place in an adoptive secondary antibody response.

A chimeric population of lymphocytes was produced by reconstituting neonatally thymectomized CBA mice soon after birth with (CBA x C57BL)F1 thymus lymphocytes. These mice could be effectively primed to fowl immunoglobulin G (FγG) and their thoracic duct lymphocytes adoptively transferred memory responses to irradiated mice. The activity of these cells was impaired markedly by preincubation with CBA anti-C57BL serum and to a lesser extent by anti-θ-serum. Reversal of this deficiency was obtained by adding T cells in the form of thoracic duct cells from normal CBA mice. Cells from FγG-primed mice were at least 10 times as effective as cells from normal mice or from CBA mice primed to horse erythrocytes. These results were considered to support the concept that memory resides in the T cell population and that collaboration between T and B cells is necessary for an optimal secondary antibody response.

Poor antibody responses were obtained in irradiated mice given mixtures of thoracic duct cells from primed mice and of B cells from unprimed mice (in the form of spleen or thoracic duct cells from thymectomized donors). In contrast to the situation with T cells, the deficiency in the B cell population could not be reversed by adding B cells from unprimed mice. It was considered that memory resides in B cells as well as in T cells and that priming probably entails a change in the B cell population which is fundamentally different from that produced in the T cell population.
CELL-MEDIATED IMMUNITY AND ANTIBODY RESPONSES IN THE RESPIRATORY TRACT AFTER LOCAL AND SYSTEMIC IMMUNIZATION
Authors: Waldman Robert H. , Henney Christopher S.
Published: June 2004

The cell-mediated immune response on the secretory surface of the lower respiratory tract was investigated and compared to the systemic cell-mediated immune response and to the secretory and systemic antibody responses. Guinea pigs were immunized either parenterally or locally with dinitrophenylated human IgG. The effect of rendering animals tolerant on the local cell-mediated response and on the local antibody response was also studied. The antibody response in the bronchial washing fluids was predominantly of the IgA class. In the animals rendered tolerant by administering large doses of the antigen intravenously, the secretory antibody response was significantly reduced to about 40% of the levels in the animals which were not tolerant. Cellular immunity was assayed using the inhibition of macrophage migration. In animals which were immunized subcutaneously, splenic lymphocytes strongly inhibited the migration of normal guinea pig macrophages; however cells obtained from bronchial washings from the same animals exhibited little or no inhibition of macrophage migration in the presence of antigen. In the animals which were immunized by nose drops, splenic lymphocytes showed virtually no inhibition of macrophage migration; however, bronchial washing lymphocytes strongly inhibited the migration of normal macrophages. Thus, nose drop immunization gave rise to a significantly greater local cellular immune response than did parenteral immunization. In those animals which were immunized by nose drops after a tolerizing dose of antigen, there was virtually no inhibition of macrophage migration by either the cells from the spleens or bronchial washings. Studies were done which demonstrated that a substance similar to macrophage-inhibition factor was liberated by the bronchial washing cells on incubation with the antigen. Studies were also done which indicated that secretory antibody played no role in the inhibition of macrophage migration when bronchial washing lymphocytes were used. Lymphocytes were purified from bronchial washings and were found to inhibit normal macrophage migration in the presence of antigen to the same extent that the mixed population of lymphocytes and pulmonary macrophages did. Thus this study supports the concept that local cellular immunity can be initiated independently of systemic cellular immunity.
ABNORMAL DEVELOPMENT OF THE THYMUS IN "NUDE" MICE
Authors: Wortis Henry H. , Nehlsen Sandra , Owen John J.
Published: June 2004

Nude mice bearing grafts of normal thymus reject skin grafts and have low, but higher than usual, lymphocyte counts. Nude bone marrow can successfully repopulate the thymus and thymus-derived areas of lethelly irradiated recipient mice. Attempts to reconstitute nude mice with normal fetal liver failed. The so-called thymic rudiment of nude mice when grafted to normal mice did not develop thymocytes. These experiments show that nude mice suffer from a defect of the epithelial portion of the thymus rather than of the precursors of thymocytes.
ANTIGENIC PROMOTION
Authors: Wu Chin-Yu , Cinader Bernard
Published: June 2004

Rabbits were immunized with p-azobenzene arsonic acid derivatives of human serum albumin (HA-As) or of dissociated keyhole limpet hemocyanin. The IgM response to the hapten was evaluated in terms of the number of hapten-specific plaque-forming cells in the lymph node draining the injection site. In some experiments, antibody was measured by agglutination of tanned and sensitized erythrocytes.

The hapten response of animals immunized with HA-As was increased (promoting effect) when the animals were injected with one of several structurally unrelated macromolecules: keyhole limpet hemocyanin (KLH), horse spleen ferritin (HSF), lysozyme (Lys), alum-precipitated human gamma globulin (alum-precipitated HGG). Different macromolecules differed in the magnitude of the promoting effect they induced, e.g., promotion by the associated form of KLH was greater than that by the dissociated form; alum-precipitated HGG was a better promoter than was soluble HGG. The relative magnitude of promotion by different macromolecules (associated vs. dissociated KLH, alum-precipitated vs. soluble HGG) correlated with the relative magnitude of the carrier effect, as judged by the hapten response induced by p-azobenzene arsonic acid conjugated to various proteins. Promotion was detected by agglutination assay of circulating antibody, by plaque assay of cells from the popliteal lymph node draining the site of preinjection, but not by plaque assay of cells from the contralateral lymph node. Promotion was dependent on the dose of the promoting macromolecule and on the dose of the hapten-protein conjugate. It was not observed in animals tolerant to the promoting macromolecule. Inhibition (i.e. antigenic competition), rather than promotion, was observed upon a secondary response to the preinjected macromolecule or when the hapten-protein conjugate was incorporated in Freund's adjuvant.
RESPONSE OF CULTURED MACROPHAGES TO MYCOBACTERIUM TUBERCULOSIS, WITH OBSERVATIONS ON FUSION OF LYSOSOMES WITH PHAGOSOMES
Authors: Armstrong J. A. , Hart P. D'Arcy
Published: June 2004

The cytological response to the ingestion of tubercle bacilli by cultured mouse peritoneal macrophages has been studied by electron microscopy. Methods included a quantitative assessment based on systematic surveying of cell profiles, and of phagosomes and their contained bacteria, encountered in thin sections; classification of the sectioned bacteria into visibly damaged and apparently intact categories; prelabeling of dense granules (secondary lysosomes) with ferritin as an aid to identifying the occurrence and frequency of phagosome-lysosome fusion; and monitoring of bacterial growth and viability by light microscopy and cultural counts. The situations studied were as follows: progressive infection with the multiplying virulent strain H37Rv; ingestion of the same strain previously inactivated by gamma radiation; infection with an attenuated strain (BCG); and a stabilized virulent infection induced by the surfactant Macrocyclon.

In the bacterial suspensions used routinely for inoculation, about half the bacilli were viable, matching closely the proportions of intact and damaged organisms identified with the electron microscope. In the inoculated macrophages, some phagosomes containing intact bacilli and others containing damaged bacilli were always to be found; but the proportion of organisms scored as damaged increased, and that of intact organisms decreased, in situations where the population as a whole had been rendered nonviable before inoculation, or where they became so intracellularly as in the late stages of a BCG infection.

Evidence of fusion of ferritin-marked lysosomes with some bacterium-containing phagosomes was obtained in all experiments, but a significant difference was regularly observed according to whether the bacilli were damaged or intact. Virtually all phagosomes containing damaged bacilli showed signs of fusion; but when many phagosomes were present containing apparently intact organisms (as with actively multiplying strain H37Rv or with this strain held at a steady level of viability by Macrocyclon, and also with strain BCG at an early stage of that infection), signs of fusion of lysosomes with these phagosomes were infrequent.

From these findings it is inferred that intracellular survival of M. tuberculosis in cultured macrophages is associated with a tendency to nonfusion of dense granules with the phagosome, thus avoiding direct exposure of the bacilli to the contents of these organelles. It is suggested, further, that fusion of dense granules with the phagosome, leading to digestion, is determined by recognition of the bacillus as nonviable. The possibility is discussed that the cytological response to different mycobacterial infections may reflect differences of a basic nature between facultative and obligate intracellular parasitism.
ANTIBODY-MEDIATED ACTIVATION OF A DEFECTIVE ß-D-GALACTOSIDASE
Authors: Celada Franco , Ellis John , Bodlund Kerstin , Rotman Boris
Published: June 2004

Two closely related protein antigens were used to study immunogenic competition. Namely, normal ß-D-galactosidase of Escherichia coli (Z) and a genetically defective ß-D-galactosidase (AMEF) which seems to differ from the normal in one amino acid substitution. A unique characteristic of this pair of antigens is that, although they are indistinguishable in precipitation and absorption tests with antibodies, the enzymatic activity of AMEF is specifically increased several-hundredfold in the presence of antibodies directed against Z. The following results show that Z and AMEF also differ in their immunogenic ability: (a) antibodies directed against Z activated AMEF; antibodies directed against AMEF did not activate, but competed specifically with activating antibodies. (b) Animals immunized with AMEF failed to produce activating antibodies when they were subsequently challenged with Z, although the presence of some cells primed to produce activating antibodies could be demonstrated by adoptive transfer. (c) Animals preimmunized with Z were stimulated in their production of activating antibodies by AMEF challenge, although not as efficiently as with Z.

A model explaining these observations by competition for the immunogenic site among antigen-sensitive cells carrying cross-reacting receptors is presented.
QUANTITATIVE INVESTIGATIONS OF IDIOTYPIC ANTIBODIES
Authors: Spring Susan B. , Schroeder Kenneth W. , Nisonoff Alfred
Published: June 2004

The effect of challenge by antigen on persistence of clones of antibody-producing cells and on the induction of new clones was investigated through quantitative measurements of idiotypic specificities.

In each of nine rabbits idiotypic specificities present in the earliest bleedings were completely replaced after a few months; subsequent changes occurred much more slowly. On a quantitative basis the population of molecules used as immunogen always reacted most effectively with the homologous anti-idiotypic antiserum.

Little effect of increased antigen dose on the rate of change of idiotype was observed. Even large amounts of antigen administered every 2 wk caused only gradual changes in idiotypic specificities. This was attributed either to more effective capture of antigen by memory cells, as compared to precursor cells, or to the induction of tolerance in those clones that were not expressed. In two of three rabbits on a monthly injection schedule, the idiotypic specificities identified underwent very slow changes over a period as long as 17 months. Changes occurred more rapidly when antigen was administered every 2 wk.

In each of four rabbits investigated, all idiotypic specificities identified before a 5 month rest period were still present afterwards, indicating the survival of essentially all clones of antibody-producing cells during that interval. Quantitative inhibition data indicated that some new clones of cells were initiated.
EFFECT OF PHAGOCYTOSIS ON MEMBRANE TRANSPORT OF NONELECTROLYTES
Authors: Tsan Min-Fu , Berlin Richard D.
Published: June 2004

The activities of specific transport systems were determined before and after large portions of the surface membrane had been interiorized by phagocytosis of inert particles. In five separate transport systems in rabbit polymorphonuclear leukocytes (adenosine and two adenine transport systems) and alveolar macrophages (adenosine and lysine transport systems), the rate of transport was unaffected even after an estimated 35–50% of the membrane had been internalized. Studies of the kinetics of lysine and adenosine transport, exchange diffusion of lysine transport in alveolar macrophages, and the specificities of adenine transport in polymorphonuclear leukocytes indicate that the nature of the membrane transport systems is not altered by phagocytosis. Therefore the constancy of transport indicates that the number of carriers remains the same before and after phagocytosis. It was also shown that this constancy of transport did not depend on the introduction into the surface of new transport sites during phagocytosis. Therefore transport sites are preserved on the surface during the internalization of membrane which accompanies phagocytosis. The results are best explained by the concept that the membrane is mosaic in character with geographically separate transport and phagocytic sites.
NONANTIGENICITY AND IMMUNOLOGIC TOLERANCE: THE ROLE OF THE CARRIER IN THE INDUCTION OF TOLERANCE TO THE HAPTEN
Authors: Golan Dov Theo , Borel Yves
Published: June 2004

Treatment of adult mice with dinitrophenyl (DNP) bound to isogeneic serum resulted in a specific inability to respond to DNP after challenge with DNP-keyhole limpet hemocyanin (KLH) in complete Freund's adjuvant. The unresponsiveness to the hapten had all the characteristics of immunologic tolerance: it had a definite induction time; it was transient but could be maintained by additional injections of the tolerogen; it was antigen specific and dose dependent. In addition, the induction of tolerance to DNP is dependent on the nature of the carrier.

Two main conclusions can be drawn from these data: DNP conjugates of three homopolymers of lysine were found to be nonimmunogenic in mice, yet tolerogenic. Thus, antigenicity is not necessary to induce tolerance.

Among the various carriers tested, isogeneic 7S immunoglobulin (IgG) was found to be the most effective to induce and maintain tolerance to the hapten. This suggests that IgG may have a function other than its usual role as an immunoglobulin.
THE DEVELOPMENT OF NEUTROPHILIC POLYMORPHONUCLEAR LEUKOCYTES IN HUMAN BONE MARROW
Authors: Bainton Dorothy Ford , Ullyot Joan L. , Farquhar Marilyn G.
Published: June 2004

Neutrophilic leukocytes (PMN) and their precursors from normal human marrow and blood were examined by histochemical staining and by electron microscopy and cytochemistry in order to determine the origin and nature of their cytoplasmic granules. Human neutrophils contain two basic types of granules, azurophils and specifics, which differ in morphology, contents, and time of origin. Azurophils are large and may be spherical or ellipsoid, the latter with a crystalline inclusion. They are produced in the first secretory stage (promyelocyte), contain peroxidase and various lysosomal enzymes, and thus correspond to modified primary lysosomes. Specifics are smaller, may be spherical or elongated, and are formed during a later secretory stage (myelocyte). They lack lysosomal enzymes and contain alkaline phosphatase and basic protein; their contents remain largely undetermined. Specifics outnumber azurophils in the mature PMN because of reduction in numbers of azurophils per cell by cell division in the myelocyte stage. The findings indicate that the situation is basically the same as described previously in the rabbit, insofar as the origin, enzymic activity, and persistence in the mature cell of the two types (azurophil and specific) of granules are concerned. The main difference between PMN of the two species is in the morphology (size, shape, and density) of the granules, especially the azurophils.
OBSERVATIONS ON TISSUE GRAFTS ESTABLISHED IN RABBIT EAR CHAMBERS
Authors: Hobbs J. B. , Cliff W. J.
Published: June 2004

An in vivo microscopic and ultrastructural study of tissues transferred to the transparent rabbit ear chamber is presented. Fragments of liver, kidney, thyroid, and myometrium were successfully auto- or allografted into the chamber from donors of all ages and allowed continuous in vivo observation of parenchymal structure and function, as well as of the graft vasculature which plays such an active role during rejection.

Circulation was quickly reestablished by anastomosis of graft vessels to those of the ear chamber membrane and only minor reversible parenchymal changes occurred. Both vessels and parenchyma retained the characteristics of the organs of origin on both light and electron microscopy and were observed functioning in vivo for periods up to 1 yr in the case of autografts, and until rejection occurred in allografts. In the latter case, rejection did not occur in tissues obtained from adult and neonatal donors for nearly 3 months, while tissues of fetal origin were generally rejected much earlier.

The kidney grafts provide a unique opportunity for a close comparative study of mammalian fetal and adult glomerular blood flow under varying rates of perfusion, and the tubular epithelium could be observed regenerating after initial acute tubular necrosis. Renal tubules from fetal, neonatal, and adult donors were all capable of metaplastic change to form a highly ciliated epithelium.

Grafts of these tissues will allow the fine detail of the processes of rejection to be studied continuously in vivo.
EFFECT OF GROWTH HORMONE ON THE METABOLISM OF THYMUS AND ON THE IMMUNE RESPONSE AGAINST SHEEP ERYTHROCYTES
Authors: Pandian M. R. , Talwar G. P.
Published: June 2004

The effect of pituitary growth hormone on the biosynthesis of DNA in the thymus and other lymphoid organs, as well as the ability of the rat to respond immunologically to sheep red blood cells, has been evaluated. There is a marked reduction in plaque-forming cells, hemagglutination titers, and DNA synthesis in animals when examined at 15 wk after hypophysectomy. Administration of bovine growth hormone (BGH) leads to the enhancement of DNA synthesis in lymphoid organs and recovery of the immune response. Similar effects of the hormone are observed in plateaued rats. Injection of rabbit anti-BGH globulins, in contrast to normal rabbit globulins, over 5 days causes a drop in the weight of the thymus and in the rate of DNA synthesis in this organ. The thymus is also the organ in which stimulation of DNA synthesis is observed at a time period earlier than the spleen and lymph nodes after a single injection of BGH. The hormone stimulates not only the incorporation of thymidine-3H into DNA in the cortical cells, but also the incorporation of sodium sulfate-35S into TCA-insoluble biopolymers reported to be elaborated in the medullary area of the thymus.

An in vitro system for the action of BGH on the thymus has been described. There is an obligatory requirement for calcium, but not for fetal calf serum in the medium for the hormone effect. An early action of the hormone is the enhanced incorporation of uridine-G-3H into RNA in thymocytes which is followed by a stimulation of the synthesis of proteins and DNA. The stimulatory action of growth hormone on RNA synthesis is not because of a facilitated uptake of the radioactive uridine by the cells under hormonal influence, a mechanism by which insulin is observed to increase RNA synthesis in thymocytes in vitro. The action of growth hormone on thymocytes is specific, since thyroid-stimulating hormone (TSH), luteinizing hormone (LH), and heat-inactivated growth hormone are not effective. BGH has also a beneficial action on the regeneration of the thymus and spleen in starved rats.
THE PREPARATION AND PROPERTIES OF MACROPHAGE-L CELL HYBRIDS
Authors: Gordon Saimon , Ripps Carolyn S. , Cohn Zanvil
Published: June 2004

The plasma membrane of the mouse peritoneal macrophage has specific receptors which enable the cell to bind IgG or complement-coated sheep red cells and is also rich in a divalent cation-dependent adenosine triphosphatase (ATPase) activity. L cells lack these macrophage membrane markers. The question of macrophage membrane receptor expression was investigated in DBA/2 mouse macrophage x mouse LMTK- cell hybrids produced with the aid of Sendai virus. Three independent clones and one mass culture were isolated by their ability to grow in hypoxanthine, aminopterin, and thymidine (HAT) selection medium. These hybrids retained 85–100% of the sum of two parent cells' chromosomes and expressed several genes derived from both parents, including glucose phosphate isomerase isozymes and H-2 antigens. The hybrids displayed ATPase activity which was intermediate between that of the macrophage and L cell. The macrophage specific receptors for antibody or complement-coated red cells could not be demonstrated on hybrid cells. The selective absence of these receptors is probably because of a failure in gene expression rather than to loss of genes.
CHANGE IN THE STRUCTURE OF SHOPE PAPILLOMA VIRUS-INDUCED ARGINASE ASSOCIATED WITH MUTATION OF THE VIRUS
Authors: Rogers Stanfield
Published: June 2004

The change in the state of the virus-induced enzyme associated with a mutation in the virus provides additional evidence that the enzyme is synthesized from virus rather than rabbit genetic information. This change in structure results in differences in stability of polymerization, degree of optical rotary dispersion (ORD) specific rotation, change in elution characteristics from carboxymethyl cellulose, and a reduction in specific activity of the arginase.

Liver arginase differs markedly in ORD characteristics from the virus-induced enzyme. In contrast to the virus-induced enzyme, it showed no negative Cotton effect at 233 nm until it was activated with manganese. Manganese had no influence on the ORD spectrum of virus-induced arginase. In addition, liver arginase is denatured by 4 M urea, while the virus-induced enzyme requires 10 M urea for denaturation.
MODULATION OF FUNCTION OF THE ACTIVATED FIRST COMPONENT OF COMPLEMENT BY A FRAGMENT DERIVED FROM SERUM
Authors: Gigli Irma , Kaplan Allen P. , Austen K. Frank
Published: June 2004

An activity designated Kf can be separated from human serum and shown to give a 100–300% enhancement in the hemolytic activity of fully activated, fractionated C1. The enhancement of C1 activity is not because of activation of precursor C1 and it is not attributable to an effect on C1 binding. EAC42 or EAC4 intermediates interacted with C1Kf exhibit a greater Tmax and shorter Zmax than when such intermediates are reacted with the same number of hemolytic units of C1. C3 consumption by the EAC1Kf42 intermediate greatly exceeds that of the EAC142 intermediate produced from the same EAC4 cells by comparable inputs of the other two complement components. Taken together, these findings suggest that Kf-treated C1 achieves more efficient utilization of C4 and C2 to create a larger number of 42 sites as appreciated on the intermediates by shorter Tmax and a greater Zmax, and an increased capacity to utilize C3. The capacity of Kf to enhance C1 upon introduction into whole serum of a patient with hereditary angioedema (HAE) in a manner comparable to its effect on fractionated C1 suggests that the effect of Kf may be pertinent to certain pathophysiologic conditions of man.
IDENTIFICATION OF MARROW-DERIVED AND THYMUS-DERIVED SMALL LYMPHOCYTES IN THE LYMPHOID TISSUE AND THORACIC DUCT LYMPH OF NORMAL RATS
Authors: Howard Jonathan C. , Hunt S. V. , Gowans J. L.
Published: June 2004

These experiments show that small lymphocytes from the thoracic duct of rats are normally a mixture of thymus-derived and marrow-derived cells, and define the traffic areas in lymphoid tissues through which the two populations recirculate.

Thoracic duct lymphocytes were labeled in vitro with uridine-3H and their histological distribution in the lymphoid tissues of normal recipients was demonstrated by radioautography. Labeled lymphocytes occupied two adjacent areas distinguished by a marked difference in the intensity of labeling; heavily labeled cells were found in thymus-dependent traffic areas of lymphocyte recirculation, while lightly labeled cells localized in the thymus-independent follicular areas around germinal centers.

A corresponding heterogeneity of uridine uptake among small lymphocytes from normal donors was demonstrated by sedimentation at 1 g; slowly sedimenting cells incorporated little uridine and localized in follicular areas after transfusion while rapidly sedimenting cells incorporated more uridine and localized in thymus-dependent areas after transfusion.

Experimentally prepared marrow-derived small lymphocytes behaved in sedimentation studies and after transfusion like a pure population of the lightly labeled small lymphocytes in normal lymph. Artificially reconstituted mixtures of marrow-derived and thymus-derived lymphocytes were qualitatively indistinguishable from normal lymphocyte populations.
AN ULTRASTRUCTURAL STUDY OF LYMPHOCYTES WITH SURFACE-BOUND IMMUNOGLOBULIN
Authors: Perkins William D. , Karnovsky Morris J. , Unanue Emil R.
Published: June 2004

This report is on a radioautographic study of lymphocytes exposed to 125I-labeled anti-Ig in an attempt to identify surface-bound Ig molecules. The results as studied by ultrastructural radioautography confirmed the presence of surface-bound Ig on a certain population of lymphocytes. The specificity of the anti-Ig was determined by using appropriate controls that included the use of an absorbed anti-Ig and anti-hemocyanin antibody. The labeling pattern resulting from the interaction of labeled anti-Ig and Ig was found to be specifically associated with the cell surface and random in its distribution. Morphological differences were not apparent between labeled and nonlabeled lymphocytes in the spleen and lymph nodes. In the thymus, most lymphocytes did not exhibit detectable Ig. The few thymic lymphocytes that were labeled had unique morphological characteristics that included fewer ribosomes, many of which were monoribosomes. Relative to the amount in their cytoplasmic organelles, plasma cells had surface Ig but to a lesser degree than lymphocytes. Finally, macrophages were nonspecifically labeled and contained antibody on their membranes as well as intracellularly.
THE PROLIFERATION OF PLASMA CELLS FROM MOUSE BONE MARROW IN VITRO
Authors: Nakamura Kunie
Published: June 2004

An in vitro proliferative system for immunoglobulin-G-forming plasma cells from the bone marrow of mice was established by the addition of antigenic protein and thymic cells or their homogenate to bone marrow cultures. The promoting activity of the thymus on plasma cells was independent from mouse strain, but it differed in strength with the variations of donor strains. Synthesis of immunoglobulin-G in proliferating plasma cells and its antibody reactivity against the administered antigen were demonstrated by immunocytological analyses.
ANALYSIS OF GRAFT-VERSUS-HOST DISEASE IN SYRIAN HAMSTERS
Authors: Streilein J. Wayne
Published: June 2004

The so-called refractory state, one sequela of acute graft-versus-host disease, has been studied in adult (CB x MHA)F1 hybrid Syrian hamsters inoculated with sublethal numbers of MHA-anti-CB lymphoid cells. Intracutaneous challenge of these animals with 200 million MHA-anti-CB lymphoid cells after the acute syndrome subsided failed to evoke epidermal necrolysis, whereas a similar challenge administered to normal F1 recipients invariably resulted in lethal epidermolysis. Moreover, the gradual attrition of lymphatic tissues in these hosts and their fading capacity to display adequately immune lymphocyte transfer reactions in the skin coincided with increasing evidence of host refractoriness, suggesting a causal interrelationship. It was possible to circumvent refractoriness by challenging these animals intracutaneously with MHA-anti-CB cells if: (a) the hosts had been lethally irradiated and reconstituted with F1 hematopoietic cells, or (b) the intracutaneous inocula contained admixed F1 lymphoid cells. This evidence provides additional support for the hypothesis that in GVH disease donor lymphocytes attack primarily host lymphoid cells bearing offending homologous antigens. The GVH process can continue so long as these lymphocyte-bound antigens persist within the host, and will abate only as the aggregate host lymphatic mass is effectively destroyed (hamsters) or its antigenic determinants are masked by isoantibodies (rats, mice, man?). At this point, insufficient target tissues remain for rechallenge to incite significant recrudescence of the disease.
RECEPTORS ON IMMUNOCOMPETENT CELLS
Authors: Davie Joseph M. , Paul William E.
Published: June 2004

The interaction of antigen with specific, cell-associated receptors was measured in thermodynamic terms. The binding of 125I-labeled 2,4-dinitrophenyl guinea pig albumin (DNP16GPA-125I) to lymphocytes from guinea pigs immunized to DNP16GPA is a temperature-dependent, reversible process. Measurement of association and dissociation rates of antigen-receptor complexes permits calculation of antigen-cell binding constants. These may also be calculated by equilibrium-binding techniques. Although differences in the constants calculated in these two ways exist, a clear increase in avidity of cell receptor for antigen occurs in the course of the immune response. This change in receptor avidity provides evidence that the time-dependent change in affinity of serum antibody (maturation) indeed has a cellular basis.

The magnitude of the equilibrium constant is, in part, due to binding of more than one DNP group per molecule of antigen. Thus, multivalent ligands bind more effectively to cell receptors than univalent or paucivalent ligands when measured by the number of antigen molecules bound, the dissociation rate of antigen-receptor complexes, and in the relative capacity to inhibit a standard multivalent ligand (DNP16GPA-125I) from binding.
CELL-TO-CELL INTERACTION IN THE IMMUNE RESPONSE
Authors: Anderson R. E. , Sprent J. , Miller J. F. A. P.
Published: June 2004

The helper function of carrier-primed T cells was found to be radiosensitive in vivo. The results could not be attributed to interference with the spleen-seeking properties of the irradiated cells. It is suggested that T cell division is essential for the induction of 7S antibody responses in vivo.
C3 PROACTIVATOR CONVERTASE AND ITS MODE OF ACTION
Authors: Müller-Eberhard Hans J. , Götze Otto
Published: June 2004

The activity in human serum which is responsible for conversion of C3 proactivator (C3PA) to C3 activator was shown to reside in a 3S α-globulin. The factor, called C3PA convertase (C3PAse), was obtained in partially purified form. For conversion of C3PA, C3PAse required participation of metal ions and of a C3 fragment, which in physicochemical and antigenic properties resembled C3b. Isolated, native C3 failed to substitute for the fragment, but did restore the impaired C3 activator system in hydrazine-treated serum. Unlike native C3, the C3 fragment initiated conversion of C3PA in whole serum. A hypothetical concept which envisions the C3 fragment as effector of C3PAse has been proposed.
INDUCTION OF IMMUNITY AND TOLERANCE IN VITRO BY HAPTEN PROTEIN CONJUGATES
Authors: Feldmann Marc
Published: June 2004

Of many dinitrophenylated (DNP) protein conjugates tested, only DNP conjugated to polymerized flagellin (DNP-POL) (or the structurally related bacterial flagella) elicited a primary anti-DNP response in vitro. Other DNP proteins, such as DNP-monomeric flagellin (DNP-MON), were capable of inducing secondary responses in vitro. The capacity of DNP-POL to immunize spleen cell suspensions devoid of thymus-derived cells was the reason for the greater immunogenicity of DNP-POL, since even large numbers of flagellin-reactive activated thymus cells did not increase the anti-DNP response of normal spleen cells immunized with DNP-POL, whereas the thymus-dependent response to DNP-MON was markedly increased.

The capacity of various batches of DNP-POL to immunize normal spleen cells in vitro varied markedly, depending on the number of DNP groups conjugated. DNP-POL with few DNP groups conjugated was immunogenic, but even at very high concentrations did not induce tolerance. In contrast, highly conjugated DNP-POL did not immunize, but readily induced tolerance. DNP-POL with intermediate degrees of conjugation were, like unconjugated polymeric flagellin, capable of inducing both immunity and tolerance. Since DNP-POL immunizes bone marrow-derived lymphocytes (B cells) directly the reduced response was not due to a masking of carrier determinants, necessary for cell collaboration. By using mixed DNP-5-(dimethylamino)-1-naphthalyl (dansyl)-POL conjugates, it was found that the inhibitory effect of a high degree of hapten conjugated was hapten specific. Depolymerization of DNP-POL to DNP-MON, which does not induce primary anti-DNP responses, was excluded by centrifugation analysis and electron microscopy.

The relationship of the degree of hapten conjugation on DNP-POL to the capacity to induce tolerance and immunity in B cells has clarified the mechanism of immunological triggering of these cells. A model of the mechanism of "signal" discrimination between immunity and tolerance in B cells, based on these findings, is proposed.
SEROLOGICALLY DEMONSTRABLE ALLOANTIGENS OF MOUSE EPIDERMAL CELLS
Authors: Scheid Margrit , Boyse Edward A. , Carswell Elizabeth A. , Old Lloyd J.
Published: June 2004

Single cells were prepared from mouse tail epidermis by a method which gives high viability counts and so permits their use in cytotoxicity tests.

According to tests with standard alloantisera, the antigen phenotype of mouse epidermal cells is H-2+θ+Sk+H-Y+TL-Ly-A-Ly-B,C-PC-.

The skin differentiation alloantigen Sk, which is responsible for homograft reactions directed selectively against skin, is expressed also on brain, but not on other cell types; it is present on the transplanted neuroblastoma C1300. Cytotoxicity tests with epidermal cells of H-2 congenic mouse stocks confirm that the Sk locus is not closely linked to H-2.

The lymphoid cell differentiation antigen θ also is present on both epidermal cells and brain. Mice frequently retain θ-incompatible or Sk-incompatible skin grafts although they have formed substantial titers of θ or Sk antibody in response to grafting.

Male (H-Y) antigen is demonstrable on epidermal cells by cytotoxicity tests with H-Y antibody, as it is also on one other type of cell, spermatozoa.
CELLULAR IMMUNOABSORBENTS IN TRANSPLANTATION IMMUNITY
Authors: Berke Gideon , Levey Raphael H.
Published: June 2004

Mouse lymphoid cells, sensitized against tumor allografts, can be deprived of the immunoreactive cells by in vitro absorption with specific fibroblast monolayers. Populations of lymphocytes so depleted are less effective in retarding tumor growth in vivo and in lysing tumor cells in vitro. Moreover, the adsorbed immunoreactive cells can be recovered specifically and are subsequently efficient in inhibiting tumor growth in vivo and in killing tumor cells in vitro.

Further evidence is presented for the suggestion that the destruction of target cells in vitro by sensitized lymphoid cells is truly representative of the mode of destruction of grafted cells in vivo.
LEUKOTACTIC FACTORS ELABORATED BY VIRUS-INFECTED TISSUES
Authors: Ward Peter A. , Cohen Stanley , Flanagan Thomas D.
Published: June 2004

Infection of chick embryos wih either Newcastle disease virus or mumps virus and infection of BGM cell cultures with mumps virus result in the elaboration of chemotactic activity for neutrophils and macrophages. These factors cannot be found in lysates of uninfected cells. They do not appear to be associated with the viral particles per se, but rather are present in virus-free supernates from infected fluids. Ultracentrifugal studies of the neutrophil chemotactic activity in allantoic fluid of embryos infected with the two different viruses indicate a similar biphasic distribution of activity, while fluid from the mammalian cell cultures shows a single zone of leukotactic activity, further suggesting that the infected cell, rather than the virus, is responsible for the leukotactic activity.

Virus-infected cells also release a substance(s) which is itself not leukotactic but which can interact with human C3 or C5 to generate such activity. This leukotactic factor-generating substance is similar to that reported in another virus-infected cell system.

It is postulated that the leukotactic factors elaborated as a result of virus infection of cells may play a protective role in vivo.
VARIATION IN ACCESSIBLE CELL SURFACE IMMUNOGLOBULIN AMONG ANTIBODY-FORMING CELLS
Authors: Nossal G. J. V. , Lewis Heather
Published: June 2004

Spleen cells from CBA mice that had been primarily or secondarily immunized with sheep red blood cells were reacted at 0°C with a 125I-labeled polyvalent rabbit anti-mouse globulin reagent. After suitable washing, the cells were placed in a plaque-revealing monolayer and warmed to 37°C. Plaques appeared within 10–20 min. Single plaque-forming cells (PFC) were taken from the middle of plaques, were washed by micromanipulation, and were singly dried on glass slides. The amount of attached antireceptor was assessed by quantitative radioautography.

Great variation in "receptor density" was encountered among the 258 single cells studied. However, early, immature PFC in both primary and secondary responses had statistically significantly more receptors than late, mature PFC. On any given day point, no difference was found between IgM- and IgG-forming cells. The results were consistent with the view that cells still able to be driven to further proliferation by antigen retain receptors, and conversely that cells, as they mature, lose both receptors and ability to be influenced by antigen.
EFFECT OF COLCHICINE AND VINBLASTINE ON THE TOPOGRAPHICAL SEPARATION OF MEMBRANE FUNCTIONS
Authors: Ukena Thomas E. , Berlin Richard D.
Published: June 2004

The topographical separation of membrane functions into phagocytic and transport areas, inferred from physiological studies, is not demonstrable in cells treated with colchicine and vinblastine, alkaloids which bind to microtubular proteins.
THE RESPONSE OF T CELLS TO HISTOCOMPATIBILITY-2 ANTIGENS
Authors: Gershon Richard K. , Liebhaber Stephen A.
Published: June 2004

The DNA synthetic response of a wide variety of parental thymocyte doses was studied in the lymphoid tissues of lethally irradiated F1 mice. The response curves of the thymocytes were strikingly similar in shape to those of T cells responding to antigens, such as sheep red cells, which are more labile than the histocompatibility antigens of the F1 host. The response was characterized by a dose-dependent latent period of 1–3 days, followed by a sharp increase in activity and a significant subsequent shutoff. Larger thymocyte doses tended to shorten the latent period. A comparison of the responses of different cell doses to one another indicated that the response usually simulated a suppressed first-order reaction. However, occasional multiple order reactions were observed. We have interpreted these data to indicate that both positive and negative interactions occur between thymocytes, without the mediation of B cell products such as conventional antibodies.