bluuebunny/crossref_metadata_2025 huggingface dataset:
CELLULAR DIFFERENTIATION OF THE IMMUNE SYSTEM OF MICE
Authors: Shearer G. M. , Cudkowicz G.
Published: June 2004

Marrow cell suspensions of unprimed donor mice have been transplanted into X-irradiated syngeneic hosts. 5–46 days later, bone cavities and spleens contained regenerated cells of the immune system which required interaction with thymocytes (from intact donors) and antigen (SRBC) to form antigen-sensitive units (ASU) and to generate mature immunocytes. These cells were capable of differentiating either into direct or indirect hemolytic plaque-forming cells (PFC). The precursors of PFC regenerated earlier than the other cell type necessary for immunocompetence, the antigen-reactive cell (ARC). The latter was not found until 10 or more days after transplantation. Availability of ARC was inferred from PFC responses elicited by grafted mice challenged with SRBC at varying intervals.

In a second series of experiments, graded numbers of marrow cells (ranging from 107 to 5 x 107) were transplanted with 5 x 107 or 108 thymocytes into irradiated mice, and SRBC were given 18 hr later. After 9–12 days the recipient spleens contained all or some of the following immunocytes: direct and indirect PFC, and hemagglutinating cluster-forming cells. The frequency of each immune response varied independently of the others, but in relation to the number of marrow cells grafted. This was interpreted to indicate that ASU formed in irradiated mice by interaction of marrow and thymus cells were similar to those of intact mice. In particular, they were specialized for the molecular class (IgM or IgG) and function (lysis or agglutination) of the antibody to be secreted by their descendent immunocytes. Hence, class-differentiation appeared to be conferred upon ASU by their marrow-derived components.
REGULATION OF THE IMMUNE RESPONSE
Authors: Sinclair Nicholas R. StC.
Published: June 2004

The ability of 7S and F(ab')2 antibody fragments to suppress priming with low doses of antigen was compared. The 7S preparation was approximately 100–1000 times more potent than the F(ab')2 preparation when the agglutinin titers of the two preparations were the same. The presence of any ability to suppress priming in the F(ab')2 preparation may reflect an inherent capacity of the F(ab')2 antibody or contamination with small amounts of 7S antibody.

The difference between 7S and F(ab')2 antibody in ability to suppress priming is attributed to the lack of the Fc portion on the F(ab')2 antibody. The Fc portion may be needed to prevent rapid excretion of antibody from the body, to induce rapid phagocytosis of antigen-antibody complexes with consequent breakdown and elimination of antigen, or to inactivate or suppress the antigen-sensitive cells from reacting to antigenic determinants. More detailed studies will permit a better assessment of the importance of these three possible regulatory roles of the Fc portion of the immunoglobulin in the immune response.
INDIVIDUAL ANTIGENIC SPECIFICITY OF HUMAN MACROGLOBULINS
Authors: Harboe M. , Solheim B. G. , Deverill Johanna
Published: June 2004

The localization of individually specific antigenic determinants on the monoclonal γM Mö was studied in gel diffusion experiments with 19S γM Mö and its 7S subunit. Various types of such determinants were delineated. Some were demonstrable both on the intact macroglobulin molecule and on the subunit. The latter was often unable to precipitate with the corresponding antibody, but combined with it and inhibited precipitation. Immunization of rabbits with purified γMs Mö was the most useful way to obtain antisera demonstrating individually specific antigenic determinants on the subunit by direct precipitation. Still other determinants were apparently destroyed by mild reduction of the γM molecule.

Reduction and reassociation of mixtures of two monoclonal γM-globulins resulted in formation of hybrid molecules containing subunits of different origin. Individually specific antisera against the two components were useful for demonstrating hybrid formation. Serological activity, i.e., cold agglutinin activity, was present in the hybrid molecules.
ANTIBODY AND IMMUNOGLOBULIN PRODUCTION AT THE CELLULAR LEVEL IN BURSECTOMIZED-IRRADIATED CHICKENS
Authors: Alm Gunnar V. , Peterson Raymond D. A.
Published: June 2004

The effect of bursectomy combined with sublethal X-irradiation in the newly hatched chicken on the immunoglobulin and antibody producing capacity in later life was investigated. The previous findings of a significant incidence of hypogammaglobulinemia in such animals were confirmed. Spleen cells from severely hypogammaglobulinemic animals synthesized and secreted little or no immunoglobulin. Such spleen lymphoid cells contained fewer immunoglobulin antigenic determinants than spleen cells from irradiated control animals as evidenced by their relative inability to respond by an increased DNA synthesis after in vitro culture with rabbit antiserum to chicken immunoglobulin. Therefore, the deficiency in the immunoglobulin synthesis extends not only to actively secreting cells such as plasma cells, but to the entire lymphoid cell population.

As expected, most irradiated-bursectomized chickens, irrespective of plasma immunoglobulin levels failed to produce detectable amount of circulating antibodies to Brucella abortus antigen in the primary immune response. Severely hypogammaglobulinemic animals were completely unable to elaborate any plaque forming cells (PFC) in the primary response to sheep red blood cells (SRBC). The results of this investigation support the contention that in the severely hypogammaglobulinemic bursectomized-irradiated chicken the entire antibody producing and immunoglobulin producing cell line is absent. The possibility remains that precursor or stem cells are present but are not appropriately directed to antibody synthesis by other cell types.
THE ENTRY OF GRANULE-ASSOCIATED PEROXIDASE INTO THE PHAGOCYTIC VACUOLES OF EOSINOPHILS
Authors: Cotran Ramzi S. , Litt Mortimer
Published: June 2004

The ultrastructural localization of peroxidase was studied in guinea pig eosinophils which had phagocytized zymosan or Escherichia coli. After phagocytosis, the membrane of the granule was joined to the membrane of the phagocytic vacuole, and the enzyme, which is ordinarily restricted to the matrix of the granule, was seen in the vacuole surrounding the ingested particle.
BIOLOGICAL PROPERTIES OF CELL WALL MUCOPEPTIDE OF HEMOLYTIC STREPTOCOCCI
Authors: Rotta Jiři , Bednář Blahoslav
Published: June 2004

Several of the toxic properties of streptococcal mucopeptide have been studied in detail. Intravenous injection of as little as 1 µg of mucopeptide, solubilized by ultrasonic treatment, elicits a reproducible febrile response. Rabbits which are made tolerant to Escherichia coli endotoxin are only partially tolerant to the subsequent injection of streptococcal mucopeptide. Soluble mucopeptide was successfully employed to prepare and provoke the localized Shwartzman reaction.

Intravenous injection of 80 µg of solubilized mucopeptide leads to diffuse cellular infiltration as well as focal areas of myocardial necrosis, surrounded by inflammatory cells.
THE LOSS OF PHENOTYPIC TRAITS BY DIFFERENTIATED CELLS
Authors: Chacko S. , Abbott J. , Holtzer S. , Holtzer H.
Published: June 2004

A single, functional, mitotically quiescent chondrocyte may be induced to reenter the mitotic cyde, and produce a progeny of over 1011 cells. Sessile, adherent, polygonal cells deposit matrix, whereas amoeboid, dispersed, flattened fibroblastic cells do not. The prior synthetic history of a cell is of greater importance in determining whether the characteristic chondrogenic phenotype will be expressed, rather than growth in "permissive" or "nonpermissive" medium. Clonal conditions select for stem-like cells, some of whose progeny may become polygonal chondrocytes. The retention of the characteristic chondrogenic phenotype in vitro is favored by pruning the dedifferentiated chondrocytes which arise in these cultures. Dedifferentiated chondrocytes interfere with the deposition and synthesis of chondroitin sulfate by neighboring functional chondrocytes. Possible mechanisms are proposed to explain this type of cell-cell or cell exudate interference.

If the progeny of a single, genetically programmed chondrocyte may or may not synthesize chondroitin sulfate, then extragenic sites in the cytoplasm or cell surface must influence the decision as to which cluster of "luxur" molecules the cell will synthesize.
THE PATHOGENESIS OF ALEUTIAN DISEASE OF MINK
Authors: Porter David D. , Larsen Austin E. , Porter Helen G.
Published: June 2004

Mink inoculated with 1 x 105 ID50 of Aleutian disease virus revealed very high virus titers in the tissues 8–18 days later. The highest virus titers observed were 5 x 108 ID50 per g of spleen and 1 x 109 ID50 per g of liver 10 days after inoculation. Concomitant with the increase in infectious virus titers, viral antigen(s) was found in the cytoplasm of macrophages in the spleen and lymph nodes and in Kupffer cells in the liver. Antiviral antibody was assayed by indirect immunofluorescence, using sections of infected liver as the source of antigen. A few mink infected for 9 days and all those infected 10 days or more developed antibody to Aleutian disease virus antigen(s). By 60 days after infection, when hypergammaglobulinemia was marked, the mink had an exceptionally high mean antibody titer of 100,000.

The pathogenesis of the glomerulonephritis of Aleutian disease is apparently related to formation of viral antigen-antibody-complement complexes which lodge in glomerular capillaries. No evidence was found that viral infection of the kidney took place, and no autoimmune responses were found.

In this "slow-virus" disease the virus replicates rapidly and the morphologic and biochemical manifestations of disease are apparently due to the continuing interplay between a replicating antigen and the host immune response.
SPECIES SPECIFICITY OF LEUKOCYTIC PYROGENS
Authors: Bornstein Donald L. , Woods James W.
Published: June 2004

Polymorphonuclear neutrophilic leukocytes of the dog, cat, and goat release leukocytic pyrogen under the same conditions as the heterophile polymorphonuclear leukocytes of the rabbit. The characteristics of the febrile response to an intravenous injection of homologous leukocytic pyrogen in all four species are very similar: a brisk monophasic fever reaching a peak between 30 and 50 min with smooth defervescence to the baseline by 3 hr. Shivering, which is not obvious in the rabbit, is noted in the dog, cat, and goat during the first 30 min. Quantitative differences in response reveal the cat to be the most sensitive of of these species to homologous leukocytic pyrogen, followed by the rabbit, dog, and goat.

The response to heterologous pyrogen is in most cases markedly diminished compared to that after equal doses of homologous protein, suggesting the operation of species specificity, although canine and feline pyrogen behaved very similarly in all tests. Species specificity of leukocytic pyrogen is probably related to amino acid substitutions in different species of a common mammalian protein effector molecule.
STUDIES ON ANTIGENIC COMPETITION
Authors: Brody Neil I. , Siskind Gregory W.
Published: June 2004

Antigenic competition was studied in a haptenic system. It was found that: (a) The extent of competition is greater when larger doses of antigen are employed. (b) Antigenic competition appears to be independent of the carrier molecule. (c) The affinity of the antibody produced in antigenic competition is approximately equal to the affinity of antibody formed by animals immunized with only one antigen. (d) Antigenic competition only occurs when both antigens are injected so as to drain into the same regional lymph nodes.

The results suggest that antigenic competition occurs locally at the site of antigen stimulation and is not mediated by a circulating factor, by tolerance induction, or by suppression due to synthesis of cross-reacting antibodies.
DENSITY DISTRIBUTION ANALYSIS OF ANTIGEN-SENSITIVE CELLS IN THE RAT
Authors: Haskill J. Stephen
Published: June 2004

Analysis of the cell populations capable of initiating a response to sheep and horse erythrocyte antigens has been carried out by means of equilibrium density gradient centrifugation. The results indicate that there are at least six distinguishable AS cell populations for sheep erythrocytes, but only three for horse erythrocytes in the spleen of the Lewis rat. Evidence is presented for the existence of metabolic, physiological, and immunological differences among these populations. It is suggested that at least one population of AS cells responds only to the specific antigen and at least one other population is sensitive to stimulation by a broad range of antigens. It is assumed that the difference between these two AS cells results from a process of differentiation of AS cells primed into DNA synthesis by antigen stimulation.
REGULATION OF THE SECONDARY ANTIBODY RESPONSE IN VITRO
Authors: Ambrose Charles T.
Published: June 2004

Two opposite effects of actinomycin D on antibody synthesis have been studied in organ cultures of rabbit lymph node fragments. These cultures were prepared from previously primed rabbits and stimulated with antigen(s) on day 0 to yield a secondary response, whose inductive phase extended to about day 9 and whose productive phase may last for several months in the serum-free medium described here.

Concentrations of actinomycin D above 0.01 µM (0.012 µg/ml) produce inhibition of antibody synthesis during both phases of the response. However, antibody synthesis is about 10 times more sensitive to inhibition by this drug when it is added during the inductive phase than during the productive phase. During the latter phase, synthesis is more rapidly terminated as the drug level approaches 10 /µM (12.5 µg/ml). At this level the 50% synthesis time is about 2.8 hr, which is identical with that found when 5–10 µM puromycin is added to the medium of parallel cultures.

Transient enhancement of antibody synthesis is frequently produced by a brief exposure to low levels of actinomycin D (generally less than 0.01 µM). Enhancement appears in precise temporal association with actinomycin pulses added for 2 days or less only between days 6 and 16. This apparent enhancement of antibody synthesis resembles the increased enzyme synthesis described by Garren et al. (6) and led to a search for an antibody-inhibitory material (AIM) whose synthesis might be stopped preferentially by low levels of the drug.

Stimulated lymph node cultures produce between days 6 and 15 a nondialyzable material which inhibits antibody synthesis during the productive phase of heterologous antigen-antibody culture systems. Just as enhancement with low levels of actinomycin D appears within 2 hr after the drug has been added to cultures, so inhibition occurs within 4 hr of adding AIM to cultures during their productive phase. These observations suggest that AIM is analogous to the translational "repressor" postulated by Garren et al. (6). AIM has relevance in two areas of immunology: (a) it may be the explanation for many examples of antigenic competition, and (b) it may represent a normal control mechanism for the productive phase.
ANTIGEN RECOGNITION: IN VITRO STUDIES ON THE SPECIFICITY OF THE CELLULAR IMMUNE RESPONSE
Authors: Schlossman Stuart F. , Herman Judith , Yaron Arieh
Published: June 2004

Studies of the immunochemical specificity of antigen-induced thymidine-2-14C incorporation in lymph node cells obtained from animals immunized to a series of closely related α-DNP-oligolysines, ϵ-DNP-oligolysines, and oligolysines have shown that the sensitized cell exhibits an extraordinary degree of specificity for antigen. The sensitized cell is maximally stimulated by the homologous immunizing antigen and can discriminate among compounds which differ from one another only in the position of a dinitrophenyl group or D-lysine residue on an identical oligolysine backbone. These studies support the view that the immunogen is not degraded prior to the induction of the immune response, and that the majority of cells produced as a consequence of immunization have stereospecific antigen receptors for the DNP-oligolysine used to induce the response; a smaller and more variably sized population of cells is produced with receptors specific for the oligolysine portion of the immunizing antigen. When specifically sensitized lymph node cell cultures are stimulated in vitro by heterologous DNP-oligolysines, the oligolysine- and not the DNP-oligolysine-sensitive population of cells appears to play a crucial role in the specificity of such cross-reactions. It is concluded from these studies that the antigen receptor on the sensitized lymph node cell differs in both kind and degree from conventional antibody. The chemical nature of the receptor and the means by which this receptor reacts with antigen to initiate the biosynthetic or proliferative cellular immune response still remain undefined.
HEAT LABILE OPSONINS TO PNEUMOCOCCUS
Authors: Smith Mary Ruth , Wood W. Barry
Published: June 2004

Heat labile opsonins (HLO) in normal rat serum to both encapsulated and unencapsulated pneumococci (a) have the same heat lability as complement (C); (b) are active at 37°C but not at 0°C; (c) are inactivated proportionately to hemolytic C by the addition of immune aggregates to the serum; (d) are adsorbed from serum nonspecifically by bacteria at 37°C but not at 0°C; (e) are Ca++- and/or Mg++-dependent in their action; and (f) are inactivated by zymosan and a purified cobra venom factor, and in the case of encapsulated pneumococci, at least, by NH4OH. Like other opsonins, HLO to pneumococci act primarily on the bacteria rather than on the phagocytes. Their combined properties indicate that they involve multiple components of the hemolytic C system.

Since HLO are immunologically polyspecific, they presumably play a broad protective role in the early (preantibody) phase of acute bacterial infections.
STUDIES ON THE MIGRATORY BEHAVIOR OF MELANOCYTES IN GUINEA PIG SKIN
Authors: Billingham R. E. , Silvers Willys K.
Published: June 2004

Pigment spread is the natural or experimentally procured (through grafting) progressive encroachment of pigmentation from black or red skin areas into juxtaposed white skin areas, or from black skin areas into red skin areas in spotted guinea pigs and other mammals. So far as spread from black into white skin is concerned, it had previously been shown that migration of epidermal melanocytes into skin lacking homologues of these cells was responsible. However, since red skin already has its own complement of phenotypically "red" melanocytes, the intriguing possibility remained that when black pigment encroaches upon red, rather than melanocyte migration being responsible, phaeomelanin (red)-producing melanocytes are transformed into eumelanin (black)-producing cells by some kind of serially transmissible factor derived from contiguous eumelanotic melanocytes.

By utilizing two isogenic strains (Nos. 2 and 13) of spotted guinea pigs and their F1 hybrids, the mechanism underlying the spread of pigment from black into red skin has been analyzed, employing cellular transplantation antigens as melanocyte "markers." The findings demonstrate unequivocally that a physical migration of pigment cells is responsible.

By comparing the extents of pigment spread from black ear skin grafts, or from epidermal cell suspensions prepared therefrom, from parental strain or from F1 hybrid donors in white host skin areas of F1 hybrid guinea pigs, it has been possible to evaluate the influence of the intimate contact of melanocytes with alien transplantation antigens on their survival and migratory behavior. No evidence was forthcoming that pigment spread takes place less readily when the cells responsible are confronted by epidermal cells bearing foreign antigens than when they are confronted by cells of their own antigenic constitution. These findings are contrary to expectation if the phenomena of allogeneic inhibition or contact-induced cytotoxicity apply to normal cells in in vivo situations.
STUDIES ON THE PATHOGENESIS OF FEVER
Authors: Hahn Helmut H. , Cheuk S. Fai , Moore Douglas M. , Wood W. Barry
Published: June 2004

Evidence has been presented that the release of active endogenous pyrogen from rabbit exudate granulocytes incubated in isotonic NaCl is a relatively prompt energy-dependent process that is preceded by a rise in intracellular pyrogen, and involves a rise in total intracellular cations and an increased permeability of the cell membranes, but does not require the synthesis of new proteins.
INDUCTION OF RABBIT ANTIBODY WITH MOLECULAR UNIFORMITY AFTER IMMUNIZATION WITH GROUP C STREPTOCOCCI
Authors: Eichmann Klaus , Lackland Henry , Hood Leroy , Krause Richard M.
Published: June 2004

Antibodies with uniform properties may occur in rabbits after immunization with Group C streptococci. These precipitating antibodies possess specificity for the group-specific carbohydrate. Not uncommonly, their concentration is between 20 and 40 mg/ml of antiserum. Evidence for molecular uniformity in the case of one of these antibodies, described in detail here, includes: individual antigenic specificity; monodisperse distribution of the light chains by alkaline urea polyacrylamide disc electrophoresis; and a single amino acid in each of the first three N-terminal positions of the light chains. When the amino acid sequence of rabbit antibody b+ light chains (κ type) are aligned against their human κ counterparts, a definite homology is observed between the N-terminus of the human and the rabbit variable region.
SPECIFIC ANTIBODY WITHIN LYMPHOID GERMINAL CENTER CELLS OF MICE AFTER PRIMARY IMMUNIZATION WITH HORSERADISH PEROXIDASE: A LIGHT AND ELECTRON MICROSCOPIC STUDY
Authors: Sordat Bernard , Sordat Martine , Hess Max W. , Stoner Richard D. , Cottier Hans
Published: June 2004

The appearance in mice of specific antibody within newly formed germinal centers in lymph nodes was demonstrated by light and electron microscopy after regional primary antigenic stimulation with horseradish peroxidase (HRP).

Lymphoid germinal center cells containing anti-HRP antibody in the perinuclear space and in the cytoplasm were detected from 17 to 26 days after antigenic stimulation. Extracellular anti-HRP antibody within germinal centers, localized between dendritic reticular cells and lymphoid elements, could not be found before the appearance of intracellular antibody. These findings strongly suggest antibody formation by lymphoid germinal center cells. Both antigen and corresponding antibody persisted in intercellular spaces up to 35 days after primary stimulation.

The concomitant presence in a given lymph node of germinal centers which are positive or negative with regard to specific antibody provide evidence in favor of monospecificity of individual centers. The mechanisms of antigen-trapping within germinal centers are discussed in the light of the present observations.
REACTIVE LYSIS: THE COMPLEMENT-MEDIATED LYSIS OF UNSENSITIZED CELLS
Authors: Lachmann P. J. , Thompson R. A.
Published: June 2004

It has been shown that the "activated reactor" that is produced in certain human sera by complement activation is a stable complex of the fifth and sixth component of complement (C56). On interaction with C7, the indicator factor, a complex C567 is formed which for a short time (half-life less than 1 min) has an activated binding site and can attach itself to normal red cell membranes, conferring on them the hemolytic properties of the "heat stable" complement intermediate EC 1 ∼ 7, the capacity to be lysed by C8 and C9. These cells have neither antibody nor the complement components up to C3 bound on them.

The binding site—activated C567c—can similarly bind to other hydrophobic surfaces, including agarose gel where it forms a "stainable line".

If the complex is not bound to a surface, the binding site decays and the resulting complex will no longer give rise to lysis. However it will still inactivate C8 and C9 in solution. The sera that can generate activated reactor apparently do so because they have an excess of C5 and C6, compared to their content of C7.

The phenomenon of reactive lysis thus represents complement-mediated lysis of unsensitized cells initiated at the C5 stage by a stable complex (C56) which was generated by complement activation at a distance.

The immunochemistry of the phenomenon is described and some of its implications discussed.
EVIDENCE FOR TRANSFORMATION OF SPLEEN CELLS ONE DAY AFTER INFECTION OF MICE WITH FRIEND LEUKEMIA VIRUS
Authors: Rossi Giovanni B. , Cudkowicz Gustavo , Friend Charlotte
Published: June 2004

Proliferation and erythroid differentiation of transplanted DBA/2 marrow cells and Friend virus-induced leukemic cells were assessed in syngeneic, allogeneic (H-2 compatible), and (BALB/c x DBA/2)F1 hybrid mice (CDF1). Measurements were made 5 days after transplantation of donor cells into nonirradiated or X-irradiated mice by the spleen colony or the 125IUdR-59Fe uptake methods. Growth of DBA/2J (Jackson subline) marrow grafts was poor in irradiated CDF1J hybrids as compared with growth in syngeneic and allogeneic hosts. The DBA/2J transplants proliferated, however, without impairment in irradiated CDF1 hybrids which were the progeny of DBA/2 male parents of other sublines, e.g. DBA/2Ha, DBA/2Cr, and DBA/2Cum. In contrast, tissue-cultured Friend leukemic cells of DBA/2J origin grew deficiently in all CDF1 hybrids tested, regardless of irradiation and of the DBA/2 parent's subline. The growth pattern of transplanted DBA/2J cells was a manifestation of hybrid resistance. The results with DBA/2J and other DBA/2 subline grafts suggested that hybrid histocompatibility alleles were expressed to a greater extent in leukemic than in normal marrow cells, for the former were consistently recognized as "nonself" by CDF1 mice, but not the latter cells.

The property of deficient growth in irradiated CDF1Ha hybrids was acquired by DBA/2J hemopoietic cells within 6 hr from infection in vivo with Friend leukemia virus, and persisted during the following 8 days. It was ascribed to enhanced expression of hybrid histocompatibility gene(s) (Hh) induced by the virus. Autonomous growth potential of hemopoietic cells, manifested by proliferation in nonirradiated recipients, was first detected 24 hr from infection, and likewise persisted at the later intervals. At the same time, the infected cells grew deficiently also in nonirradiated CDF1Ha mice. The two irreversible cellular changes were regarded as the earliest signals of virus-induced transformation.
THE FIXATION OF COMPLEMENT AND THE ACTIVATED FIRST COMPONENT (C1) OF COMPLEMENT BY COMPLEXES FORMED BETWEEN ANTIBODY AND DIVALENT HAPTEN
Authors: Hyslop N. E. , Dourmashkin R. R. , Green N. M. , Porter R. R.
Published: June 2004

Hapten-antibody complexes prepared at equivalence with the bivalent hapten bis-DNP-octamethylene-diamine and purified rabbit anti-DNP antibody were fractionated by Sepharose gel-filtration and the fractions examined by electron microscopy. Individual fractions were tested for whole-complement fixation and C1 fixation. Dimer forms did not show this type of biological activity, while fractions containing tetramers and larger polymers exhibited both C and C1 fixation, which could be inhibited by prior exposure of the complexes to the univalent hapten epsilon-DNP-caproic acid. The dose-response result indicated that the C-fixation observed was not due to interpolymeric cooperative effects.

It was concluded that in the generation of biological activity by soluble antigen-antibody complexes made with complement-fixing antibody, quaternary structural changes following specific combination with antigen may be as important as any tertiary structural alterations that occur in the individual immunoglobulin molecule.
INTERACTIONS OF THE COMPLEMENT SYSTEM WITH ENDOTOXIC LIPOPOLYSACCHARIDES IN IMMUNOGLOBULIN-DEFICIENT SERA
Authors: Gewurz H. , Pickering R. J. , Snyderman R. , Lichtenstein L. M. , Good R. A. , Mergenhagen S. E.
Published: June 2004

Bacterial lipopolysaccharides (LPS) derived from a variety of organisms effectively induced C consumption in humans, bovines, and porcines with developmental agammaglobulinemia; birds with experimental agammaglobulinemia; and humans with agammaglobulinemia syndromes. This interaction proceeded even in precolostral piglet sera which contained less than 2.5 x 10–6 mg/ml gamma globulin, and led to generation of neutrophil chemotactic factor and anaphylatoxin in these sera. Hence, the LPS-C interaction can proceed in sera markedly deficient in immunoglobulin.

The question of whether immunoglobulins can be bypassed in the LPS-C interaction, or whether they are regularly utilized in a way so efficient that their participation is masked, was considered.
THE ENHANCING EFFECT OF BONE MARROW CELLS ON THE PRIMARY IMMUNE RESPONSE OF THE ISOLATED PERFUSED SPLEEN
Authors: Atkins Robert C. , Robinson William A. , Eiseman Ben
Published: June 2004

The addition of bone marrow cells or peripheral lymphocytes to the isolated pig spleen markedly enhanced the primary antibody response after 3-day perfusion and antigenic challenge in vitro. The splenic preparation without added cells or with the addition of marrow cells to an irradiated spleen gave a limited response. Contributory evidence is provided that at least two distinct cell types are needed for antibody production. For optimal antibody response by an isolated perfused spleen, marrow cells or peripheral lymphocytes should be added to the system.
MACROPHAGE-MELANOCYTE HETEROKARYONS
Authors: Gordon Saimon , Cohn Zanvil
Published: June 2004

High yields of mouse macrophage-melanocyte heterokaryons and macrophage-macrophage homokaryons were obtained through the virus-induced fusion of cells spread on a glass surface. After fusion there was a striking reorganization of cellular architecture by means of a colcemid-sensitive process. Heterokaryons were isolated through the use of differential trypsinization and many underwent division to form melanocyte-like hybrids. The selective uptake of dextran sulfate by macrophages served as a useful cytoplasmic marker in identifying hybrids.

Many characteristic macrophage properties were altered in the heterokaryons. Within an hour of fusion macrophage nuclei became swollen, nucleoli were more prominent, and increased nuclear RNA synthesis occurred. 3 hr after fusion, a wave of DNA synthesis took place in the previously dormant macrophage nuclei.

The fate of typical macrophage markers was examined in both heterokaryons and homokaryons. Macrophage homokaryons continued to exhibit active phagocytosis of sensitized erythrocytes, whereas this capacity was lost irreversibly in heterokaryons. The loss of phagocytic activity of heterokaryons occurred at an exponential rate and was accelerated by high concentrations of calf serum.

Another macrophage surface marker, a divalent cation-dependent adenosine triphosphatase (ATPase), could be demonstrated histochemically on heterokaryons. Shortly after fusion, it was present in discrete regions, but it became more diffuse and disappeared within a day.

Acid phosphatase-positive secondary lysosomes and retractile lipid droplets disappeared from heterokaryons but continued to accumulate in macrophage homokaryons.

These observations indicate that typical macrophage properties cease to be expressed in heterokaryons, and melanocyte functions presumably predominate in heterokaryons and hybrids.
MECHANISM OF EOSINOPHILIA
Authors: Basten Antony , Boyer Markley H. , Beeson Paul B.
Published: June 2004

The phenomenon of eosinophilia was studied in rats using inoculation with Trichinella larvae as the experimental stimulus. Comparisons were made between the eosinophil response accompanying active infestation via the gastrointestinal tract and that resulting from parenteral inoculation of larvae or their products.

A vigorous eosinophilia could be provoked by a single intravenous injection of intact parasites. In this circumstance the larvae lodged in the lungs causing an acute inflammatory reaction which led to their disintegration within 24 hr. Intraaortic injection also produced a significant response, whereas inoculation of the same number of parasites by the intramuscular, intraperitoneal, or subcutaneous routes did not cause eosinophilia. Eosinophilia likewise failed to develop if parasites were homogenized before intravenous injection, so that they were not arrested in the lungs.

Antibody levels, as measured by a hemagglutination technique, using whole larval extract as antigen, did not correlate closely with the eosinophil response.

The findings are interpreted as suggesting that increased eosinophil production is induced under some circumstances as a consequence of interaction between intact parasites and certain host cells in blood and tissue. No evidence was found for the existence of a specific constituent of the parasite capable of stimulating eosinophil production.

Attention is directed to features of eosinophilia which fit with the concept that this phenomenon belongs in the category of immunologic reactions.
STUDIES ON THE PATHOGENESIS OF FEVER
Authors: Cheuk S. Fai , Hahn Helmut H. , Moore Douglas M. , Krause Donald N. , Tomasulo Peter A. , Wood W. Barry
Published: June 2004

Suppression of the pyrogen-producing capacity of exudate granulocytes results from incubation of the cells in plasma, serum, or Ringer's solution. When transferred in this state and incubated in isotonic NaCl, the cells release much less pyrogen than untreated exudate cells. The suppressive effect is reversible and appears to involve the cellular uptake of calcium ions.

In contrast, regeneration of pyrogen-producing capacity in depleted exudate cells occurs only when the cells are incubated in serum. The process resembles activation and requires the cellular synthesis of protein.
A PLAQUE ASSAY FOR ENUMERATING ANTIGEN-SENSITIVE CELLS IN DELAYED-TYPE HYPERSENSITIVITY
Authors: Bloom Barry R. , Jimenez Luis , Marcus Philip I.
Published: June 2004

A general method is described for enumerating antigen-sensitive lymphocytes obtained from individuals having delayed hypersensitivity, in this case from highly tuberculin-sensitive guinea pigs. The method is based on the observation that resting lymphocytes are generally unable to support replication of RNA viruses, whereas antigen-"activated" lymphocytes can.

Lymph node lymphocytes from individual animals were cultured in the presence or absence of tuberculin purified protein derivatives (PPD). After various periods of time, the cells were infected either with Newcastle disease virus or vesicular stomatitis virus, and plated in agar over a monolayer of cells susceptible to the virus. Wherever a lymphocyte yielded infectious virus, a discrete plaque in the monolayer could be seen. The increase in plaques of the antigen-stimulated cells over the background of the control sample was taken as the number of antigen-sensitive cells in the population. When lymphocytes from normal guinea pigs or from guinea pigs immunized to produce only circulating antibody to PPD were similarly tested, no increase in plaque-forming units (PFU) was observed.

The average increase in PFU due to antigenic stimulation varied from 1 per 1000 lymphocytes at 24 hr to 16 per 1000 lymphocytes at 96 hr. By employing inhibitors of mitosis (colchicine, vinblastine, and thymidine) it was evident that the increase in PFU at least up to 48 hr was primarily due to initial antigen-reactive cells and not their progeny.
SYNERGISTIC INTERACTION OF MACROPHAGES AND LYMPHOCYTES IN ANTIGEN-INDUCED TRANSFORMATION OF LYMPHOCYTES
Authors: Seeger Robert C. , Oppenheim Joost J.
Published: June 2004

The role of macrophages and lymphocytes in antigen-induced transformation of lymphocytes has been investigated. Lymphocytes and macrophages were obtained from inbred strain 13 guinea pigs which were either unimmunized or immunized with complete Freund's adjuvant (CFA) or tetanus toxoid in CFA. The transformation response to PPD or tetanus toxoid was assayed by tritiated thymidine incorporation.

Addition of macrophages to immune lymphocytes significantly increased their response to purified protein derivative (PPD) or tetanus toxoid. This was observed if the macrophages were (a) "immune" or "nonimmune", (b) unirradiated or irradiated (3000 R), (c) 99% pure, and (d) peritoneal or alveolar in origin. Neither immune nor nonimmune macrophages were able to induce nonimmune lymphocytes to respond to PPD or tetanus toxoid. When macrophages were incubated with PPD or tetanus toxoid and then washed, they stimulated immune lymphocytes to transform. An incubation time of ½ hr was adequate, however, 2–4: hr was optimal.

These studies indicate (a) that antigen-induced transformation of lymphocytes is greatly enhanced by macrophages; (b) that macrophage-antigen interaction can antecede lymphocyte-antigen interaction and results in macrophages which are able to stimulate lymphocyte transformation; and (c) that the immunological memory requisite to elicit specific transformation responses is a property of the lymphocyte and not the macrophage.
SOME PHYSICAL AND RADIOBIOLOGICAL PROPERTIES OF IMMUNOLOGICALLY REACTIVE MOUSE SPLEEN CELLS
Authors: Osoba David
Published: June 2004

Three classes of immunologically reactive cells, differing only slightly in size from each other, are required for the production of hemolysin-forming cells in culture. The three classes of cells can be detected in the normal mouse spleen by the combined use of rosette formation, velocity sedimentation, and irradiation.

One class of cells (peak sedimentation velocity, 3.2 mm per hr) forms rosettes. The capacity of these cells to participate in immune responses to foreign erythrocytes is inhibited by relatively low doses of irradiation. These cells may be the immediate precursors of hemolysin-forming cells.

A second class of cells (peak sedimentation velocity, 3.6 mm per hr) facilitates the production of hemolysin-forming cells by small numbers of normal spleen cells. Their facilitative activity is resistant to a relatively large dose of radiation. They do not form rosettes.

The requirement of a third class of cells was deduced from the results of mixing experiments. Neither rosette-forming cells nor spleen cells largely depleted of rosette-forming cells could give rise to hemolysin-forming cells when cultured either alone or in the presence of large numbers of heavily irradiated cells. However, when rosette-forming cells, cells depleted of rosette-forming cells, and heavily irradiated cells were mixed together, hemolysin-forming cells were produced. The peak responses were found in fractions sedimenting at 4 mm per hr. Thus, it is suggested that these fractions contain a third class of cells. This class of cells does not form rosettes, but its function is inhibited by relatively low doses of radiation.
PHYLOGENY OF IMMUNOGLOBULIN STRUCTURE AND FUNCTION
Authors: Clem L. W. , Small P. A.
Published: June 2004

The giant grouper, Epinephelus itaira, was shown to synthesize 16 and 6.4S antibodies specific for the dinitrophenyl determinant (DNP). Sera obtained at various intervals between 1 month and 2 yr after initial immunization contained both species of antibody; no temporal synthesis was evident.

Equilibrium dialysis studies employing ϵ-dinitrophenyl-amino caproic acid were conducted with purified grouper antibodies specific for DNP. The 16S antibody preparations obtained at 1 and 2 months of immunization showed heterogeneity of hapten binding indicative of two populations of combining sites. One-half of these sites (an average of four sites per 16S molecule) exhibited an average intrinsic association constant (Ko) of ∼106M–1; the Ko of the remaining four-sites was ∼104M–1. Thus, the valence of the grouper 16S antibody molecule appears to be eight although the distribution of the high and low Ko sites is unknown, i.e., are they each on the same or on different molecules? The 16S antibody preparations obtained after more prolonged immunization exhibited increasingly lower Ko values; the so-called low Ko sites were no longer detectable. These findings are in contrast to reports of rabbit IgG antibodies showing an increase in Ko with increased time.

The 6.4S antibody preparations obtained from the 1 and 2 month antisera had Ko values of ∼106M–1 and a valence of one. These antibodies would not precipitate with antigen. The 6.4S antibody preparations obtained at later times showed decreasing Ko values comparable to those of the 16S antibodies from the same bleedings.

Studies on the thermodynamic parameters of the hapten-antibody interaction showed the grouper 16 and 6.4S antibodies to be similar to each other. These data also showed that the enthalpy and entropy changes of grouper antibody-hapten reactions resemble those reported for rabbit IgG antibodies to this hapten. It is thus suggested that, although considerable evolution of immunoglobulin classes has occurred between fish and rabbits, the antibody combining site may have remained relatively unchanged during a large part of evolutionary time.
STUDIES ON THYMUS FUNCTION
Authors: Stutman Osias , Yunis Edmond J. , Good Robert A.
Published: June 2004

Significant immunological restoration of 45-day old, neonatally thymectomized C3Hf mice was obtained by the cooperation of syngeneic newborn or embryonic hemopoietic liver cells with thymic function. Thymic function or cells alone are almost ineffective or restore approximately 10% of the animals. Newborn liver cells are effective in association with thymus grafts or humoral thymic function (thymoma grafts and thymus or thymomas in diffusion chambers). Embryonic liver cells are ineffective, even in large numbers, when associated with humoral thymic function. On the other hand, embryonic liver cells are effective in the cooperative effect only in association with viable thymus grafts, preferably syngeneic, whether the grafts were placed subcutaneously, intraperitoneally, or under the kidney capsule. Dispersed viable thymic cells are ineffective in association with embryonic liver cells. Cells capable of cooperating with humoral thymic function start to appear to embryonic liver by day 19–21 of gestation and are detectable until day 5–6 postbirth. Embryonic hemopoietic liver cells from 12 to 18 days of gestation contain cells capable of cooperation only with viable free thymus grafts and not with humoral thymic function.

A prethymic cell population of partially differentiated cells of hemopoietic origin, insensitive to humoral activity of the thymus but requiring thymic stroma and traffic through the thymus is postulated to explain our results. This population of prethymic cells can become postthymic through this process and eventually develop into competent cells. Postthymic cells are characterized by their sensitivity to humoral activity of the thymus and by their wide distribution in the lymphohemopoietic tissues of newborn and young adult mice.
STUDIES ON THE LEUKOCYTOSIS AND LYMPHOCYTOSIS INDUCED BY BORDETELLA PERTUSSIS
Authors: Morse Stephen I. , Barron Bruce A.
Published: June 2004

Peripheral blood lymphocytes were isolated from normal mice and mice undergoing pertussis-induced lymphocytosis. After labeling in vitro with tritiated uridine the cells were transfused into normal or pertussis-treated mice. It was found that the lymphocytes from pertussis-treated mice entered the lymph nodes of both normal mice and pertussis-treated mice to a significantly lesser extent than did normal lymphocytes which had been transfused into either class of recipient. In addition, an interdependence of changes in the various body compartments examined was found when normal lymphocytes were injected into either type of recipient. However, when pertussis lymphocytes were injected into normal mice there was no interrelationship between the changes in the node with those in the blood, liver, lung, or spleen. In the case of pertussis lymphocytes transfused into pertussis-treated mice no interrelationship between any two compartments was observed. It was concluded that in pertussis-treated mice there is an inhibition of lymphocyte emigration which is primarily the consequence of an effect on the cell.
COMPLEMENT FIXATION BY A TWO-COMPONENT ANTIBODY SYSTEM: IMMUNOGLOBULIN G AND IMMUNOGLOBULIN M ANTI-GLOBULIN (RHEUMATOID FACTOR)
Authors: Schmid Frank R. , Roitt Ivan M. , Rocha Maria J.
Published: June 2004

Complement-mediated lysis of sheep erythrocytes coated with optimal concentrations of rabbit IgG hemolysin was inhibited by euglobulin fractions from the sera of patients with seropositive rheumatoid arthritis. That this was due to direct interaction with the IgG coat on the red cell rather than a nonspecific reaction with complement in the fluid phase was confirmed by controls using cells coated with IgM hemolysin. The inhibitory activity was recovered in purified IgM rheumatoid factor preparations and could be absorbed out with insoluble aggregated human IgG. The inhibitory potency of the rheumatoid factors correlated well with their sheep cell agglutination titers. Inhibition was not the result of physical aggregation of the erythrocytes by rheumatoid factor. Kinetic studies were consistent with the view that rheumatoid factor displaces C1q from its binding to IgG. Paradoxically, at suboptimal sensitizing concentrations of IgG hemolysin, rheumatoid factor enhances the fixation of complement. These results can be interpreted on the basis of the blockage of complement fixation by IgG and its replacement by a relatively weak direct fixation by the IgM rheumatoid factor. Thus, the interaction of RF with IgG generates only a limited ability to fix complement which, when contrasted with the fixation at suboptimal concentrations of IgG hemolysin alone, appears as net enhancement; when this is contrasted with fixation occurring with optimal concentrations of IgG, it appears as net inhibition.
MORPHOLOGY AND PEROXIDASE CYTOCHEMISTRY OF MOUSE PROMONOCYTES, MONOCYTES, AND MACROPHAGES
Authors: van Furth Ralph , Hirsch James G. , Fedorko Martha E.
Published: June 2004

Mouse promonocytes have been identified and studied in cultures of bone marrow cells. These cells have a diameter of 14–20 µ, and in stained preparations reveal a large, indented or folded nucleus, and basophilic, finely granular cytoplasm. The living promonocyte viewed by phase contrast shows additional features: nucleoli, small dense bodies, and vesicles in the cytoplasm adjacent to the nuclear hilus, and slight membrane ruffling. Prominent ultrastructural components of promonocytes include a well developed Golgi apparatus, small numbers of centrosomal granules and vacuoles, extensive ribosomal aggregates, and finger-like projections of the cell surface.

Promonocytes engage in pinocytosis and phagocytosis, but they are less active in these functions than are peripheral blood monocytes of peritoneal macrophages.

Promonocytes are positive for peroxidase, the reaction product being localized to granules most of which are centrally situated in the cell. Monocytes in blood or in inflammatory peritoneal exudates display much smaller numbers of peroxidase-positive granules, and various types of mature mouse macrophages are peroxidase negative.