bluuebunny/crossref_metadata_2025 huggingface dataset:
HAPTEN-SPECIFIC IgE ANTIBODY RESPONSES IN MICE
Authors: Hamaoka Toshiyuki , Katz David H. , Benacerraf Baruj
Published: June 2004

The present studies have established conditions for the demonstration of cooperative interactions between specific T and B lymphocyte populations in the development of IgE antibody responses in vivo in mice. This has been accomplished by utilizing a system which permits the successful adoptive transfer to irradiated recipients of DNP-specific secondary IgE responses with spleen cells from suitably primed syngeneic donor mice. Thus, adoptively transferred DNP-KLH or DNP-ASC-primed spleen cells produced high levels of anti-DNP antibodies of both IgE and IgG antibody classes in response to challenge with the appropriate homologous priming conjugate but failed to develop more than meager responses to the reciprocal heterologous conjugate. However, when spleen cells from donors primed to the second carrier were concomitantly transferred with hapten-primed lymphocytes, secondary IgE ant-DNP responses were consistently obtained upon challenge with the heterologous conjugate. Moreover, we have been able to elicit augmented primary IgE anti-DNP antibody responses to either DNP-ASC or DNP-KLH after adoptive transfer of spleen cells from donors primed only to the carrier, ASC or KLH, respectively.

This adoptive transfer system has enabled us to provide direct proof for the participation of θ-bearing T lymphocytes in antibody responses of the IgE class. Thus, the capacity of ASC-primed spleen cells to effectively cooperate with the DNP-KLH-primed lymphocytes in the adoptive secondary response to DNP-ASC could be abolished by in vitro treatment of such cells with anti-θ serum plus complement. This was true not only for the anti-DNP response of the IgG antibody class, but for the IgE antibody class as well.

These studies have, furthermore, demonstrated the capacity to stimulate secondary anti-DNP antibody production in vivo by the concomitant administration of the DNP and relevant carrier determinants on separate molecules. This was more readily seen in the IgE than in the IgG antibody class. Thus, DNP-ASC-primed cells developed significant IgE, but more variable IgG, anti-DNP responses upon challenge with DNP-KLH plus unconjugated ASC. Antibody responses of both classes elicited in this manner were appreciably improved by the transfer of additional carrier (ASC)-primed cells. These and other results presented herein suggest that IgE B lymphocyte precursors may be inherently more sensitive than IgG B cells to at least certain of the functions of T lymphocytes concerned with regulatory mechanisms involved in antibody production.
CHARACTERIZATION OF SPLENIC LYMPHOID CELLS IN FETAL AND NEWBORN MICE
Authors: Spear Patricia G. , Wang Ai-Lan , Rutishauser Urs , Edelman Gerald M.
Published: June 2004

In order to clarify the cellular events that precede the onset of immunological competence in the mouse, we have characterized and quantitated the lymphoid cells of the spleen as a function of age. Our results show that T cells and B cells both appeared in the spleens of Swiss-L mice as early as the 15th-16th day of gestation. Antigen-binding cells specific for each of three different antigens were also first detected during this same 24 h interval. The B cells and three varieties of antigen-binding cells increased in number rapidly and in parallel until about 1 wk after birth. The T cells, which were more numerous than B cells at first, increased in number somewhat more slowly.

Coincident with the onset of response to antigen, there was a further increase in B cell numbers and a decrease in the T cell to B cell ratio. The capacity to respond to antigen by cellular proliferation and synthesis of antibody did not arise until about 2 wk after birth although there were no quantitative changes in the total numbers of T cells, B cells, and antigen-binding cells between 1 and 2 wk of age. Some qualitative change, such as the functional maturation of an antigen-reactive cell, may be required during this interval for the onset of this immunological response. Although the numbers of antigen-binding cells present in fetuses and young animals were smaller than in adults, we have as yet been unable to detect any restriction in the variety of specificities that can be expressed in fetuses, either in the kinds of antigens bound or in the range of avidities with which a single antigen is bound.
TRANSFORMATION OF A HISTOCOMPATIBILITY IMMUNOGEN INTO A TOLEROGEN
Authors: Nimelstein Stephen H. , Hotti Allan R. , Holman Halsted R.
Published: June 2004

H-2 antigens on spleen cell membranes absorb antibody to H-2 antigens and induce both humoral and cellular responses. Liver cell membrane H-2 antigens by contrast also absorb antibody but do not influence cellular response and are tolerogenic for the humoral response. This paper demonstrates that syngeneic liver cells contain a substance which can transform the properties of allogeneic spleen cell membranes into those of allogeneic liver cell membranes, i.e., transform a humoral immunogen into a humoral tolerogen. The process appears to be accompanied by cleavage of an antigen component from the spleen membrane and hence to result in a structural change in the H-2 antigen.
SELF-REGULATION OF GROWTH IN THREE DIMENSIONS
Authors: Folkman Judah , Hochberg Mark
Published: June 2004

Multi-cell spheroids were grown in soft agar. When each spheroid was cultured in a large volume of medium, frequently renewed, all spheroids eventually reached a dormant phase at a diameter of approximately 3–4 mm and a population of approximately 106 cells. In the dormant spheroid, newly generated cells at the periphery balanced those lost by necrosis in the center. We propose that this dormant phase is due to a gradual reduction in the ratio of surface area to volume: a size is achieved beyond which there is insufficient surface area for the spheroid to eliminate catabolites and absorb nutrients. Thus, in the face of unlimited space and of new medium, three-dimensional cell populations become self-regulating. This phenomenon contrasts with standard tissue culture in which cell populations, living on a flat plane in two dimensions, will not stop growing in the face of unlimited space and new medium because the ratio of surface area to volume remains constant.

These experiments provide a mechanism for our observations in vivo: before vascularization, solid tumors live by simple diffusion as three-dimensional spheroids or ellipsoids. They become dormant at a diameter of only a few millimeters; once vascularized, they are released from this dormant phase and begin exponential growth. Thus, tumor dormancy resulting from absence of angiogenesis in vivo, may operate by the same mechanism responsible for dormancy of spheroids in vitro.
STUDIES IN PORPHYRIA
Authors: Bradlow H. Leon , Gillette Peter N. , Gallagher T. F. , Kappas Attallah
Published: June 2004

Patients with the genetic liver disease, acute intermittent porphyria (AIP), have a defect in the reductive transformation of steroid hormones that is manifest by the disproportionate generation of 5ß-steroid metabolites from precursor hormones. 5ß-steroid metabolites were earlier shown to be potent inducers experimentally of δ-aminolevulinate synthetase (ALAS), the mitochondrial enzyme that is rate-limiting in porphyrin synthesis, and that is found at high levels of activity in the livers of AIP patients.

In this report, the basis for the defective steroid metabolism in AIP has been shown, through studies with the 14C-labeled adrenal hormone 11ß-hydroxy-Δ4-androstenedione, to reside in a substantial deficiency of hepatic steroid Δ4-5α-reductase activity. This enzymic deficiency was found in all seven AIP patients studied, and ranged from 34% to as much as 70% below the mean enzyme activity characterizing normal subjects.

The functional consequence of the low levels of 5α-reductase activity in AIP is to divert the reductive transformation of certain natural hormones from the 5α- to the 5ß-pathway; the latter is the metabolic route through which endogenous steroids having the potential for inducing hepatic ALAS are generated. It is not presently known whether the 5α-reductase deficiency in AIP is acquired in some fashion or whether it has partial genetic determinants. It seems probable, however, that this enzymatic abnormality, coupled with the dramatic increase in hormone synthesis that occurs at puberty, may be of major importance in determining clinical expression of the latent gene defect for AIP in many individuals. The 5α-reductases for steroid hormones are known to be localized in the endoplasmic reticulum of hepatic cells and the present findings in AIP thus represent the first demonstration that an enzymic component of these membranous structures is functionally abnormal in this hereditary liver disease.
GENETIC CONTROL OF IMMUNE RESPONSES IN VITRO
Authors: Kapp Judith A. , Pierce Carl W. , Benacerraf Baruj
Published: June 2004

In vivo, the antibody response in mice to the random terpolymer L-glutamic acid50-L-alanine30-L-tyrosine10 (GAT) is controlled by a histocompatibility-linked immune response gene(s). We have studied antibody responses by spleen cells from responder and nonresponder mice to GAT and GAT complexed to methylated bovine serum albumin (GAT-MBSA) in vitro. Cells producing antibodies specific for GAT were enumerated in a modified Jerne plaque assay using GAT coupled to sheep erythrocytes as indicator cells. Soluble GAT stimulated development of IgG GAT-specific plaque-forming cell (PFC) responses in cultures of spleen cells from responder mice, C57Bl/6 (H-2b), F1 (C57 x SJL) (H-2b/s), and A/J (H-2a). Soluble GAT did not stimulate development of GAT-specific PFC responses in cultures of spleen cells from nonresponder mice, SJL (H-2s), B10.S (H-2s), and A.SW (H-2s). GAT-MBSA stimulated development of IgG GAT-specific PFC responses in cultures of spleen cells from both responder and nonresponder strains of mice. These data correlate precisely with data obtained by measuring the in vivo responses of responder and nonresponder strains of mice to GAT and GAT-MBSA by serological techniques. Therefore, this in vitro system can effectively be used as a model to study the cellular events regulated by histocompatibility-linked immune response genes.
FUNCTION OF MACROPHAGES IN ANTIGEN RECOGNITION BY GUINEA PIG T LYMPHOCYTES
Authors: Rosenthal Alan S. , Shevach Ethan M.
Published: June 2004

Antigen activation of DNA synthesis in immune thymus-derived lymphocytes of guinea pigs requires the cooperation of macrophages and lymphocytes. We have investigated the role of histocompatibility determinants in this macrophage-lymphocyte interaction using cells from inbred strain 2 and 13 guinea pigs. The data demonstrate that efficient presentation of macrophage-associated antigen to the lymphocyte requires identity between macrophage and lymphocyte at some portion of the major histocompatibility complex. The failure of allogeneic macrophages to effectively initiate immune lymphocyte proliferation was not the result of the presence of an inhibitor of blastogenesis released in mixtures of allogeneic cells, peculiarities of the antigen or lymphoid cells employed, nor differing kinetics of activation by allogeneic macrophages. In addition, data were presented that demonstrated that alloantisera inhibit lymphocyte DNA synthesis by functional interference with macrophage-lymphocyte interaction.
A MOUSE B-CELL ALLOANTIGEN DETERMINED BY GENE(S) LINKED TO THE MAJOR HISTOCOMPATIBILITY COMPLEX
Authors: Sachs David H. , Cone James L.
Published: June 2004

Antibodies cytotoxic for only a subpopulation of C57Bl/10 lymph node and spleen cells were detected when rat antiserum against B10.D2 was exhaustively absorbed with B10.A lymphocytes. Antibodies of similar specificity were also detected in B10.A anti-B10.D2 and in B10.A anti-C57Bl/10 alloantisera. Reactions with recombinant strains of mice indicate that the cell-surface antigen(s) responsible for this specificity is determined by gene(s) in or to the left of the Ir-1 region of the major histocompatibility complex. A variety of criteria implicate B cells as the subpopulation of lymphocytes bearing this antigen. In view of these data and the recent report by others of a T-cell alloantigen determined by gene(s) in the major histocompatibility complex, it seems possible that there may be a variety of H-2-linked alloantigens expressed preferentially on subclasses of lymphocytes.
MECHANISM OF ACTIVATION OF THE BONE MARROW-DERIVED LYMPHOCYTE
Authors: Schrader John W.
Published: June 2004

Peritoneal exudate cells from nu/nu mice stimulated with proteose peptone broth, but in general not from unstimulated mice, permitted cultures of spleen cells from congenitally athymic (nu/nu) mice to respond to the thymus-dependent antigen fowl gamma globulin (FγG). Supernatants of cultures of peritoneal cells were also effective, the activity being sensitive to trypsin. Since nu/nu mice were effective sources of the peritoneal cells it would not seem obligatory for the thymus-derived (T) cell to be involved in the triggering of the bone marrow-derived (B) cell by a thymus-dependent antigen FγG. It is proposed that the B cell is triggered at the macrophage surface where it encounters two signals (a) the antigen and (b) a protein secreted by the activated macrophage. In vivo the T cell may have a role in B-cell triggering, either in activating the macrophage or in aiding in presentation of antigen on the macrophage surface. Thymus-independent antigens are proposed to induce an IgM response because they are able to provide "signal two" either by direct interaction with the B cell or via irritation or activation of the macrophage. The stimulatory effect of T cells activated by an allogeneic interaction was used as a model of one influence of the T cell on the development of an antibody response. The presence in cultures of nu/nu spleen of an allogeneic interaction had no effect on the inability of these cells to respond to FγG. However when a source of the postulated second signal such as the supernatant of a macrophage culture was present, an allogeneic interaction had a powerful amplifying effect on the anti-FγG response. In contrast the response of nu/nu spleen cultures to heterologous erythrocytes was greatly enhanced by the presence of an allogeneic interaction. It is suggested that since there was a definite basal response to the heterologous erythrocytes added alone, the enhancement represented not an activation of more B cells but rather an amplification of this basal response. Thus the anti-FγG response in cultures of nu/nu spleen differentiates between factors such as those released by activated macrophages that are involved in B-cell triggering and factors released by activated T cells that amplify the numbers of antibody-forming cells resulting from a B cell already triggered.
THYMUS-DEPENDENT LYMPHOCYTES IN TISSUE SECTIONS OF REJECTING RAT RENAL ALLOGRAFTS
Authors: Balch C. M. , Wilson C. B. , Lee S. , Feldman J. D.
Published: June 2004

Lewis kidneys were grafted into BN recipients and examined at daily intervals up to 6 days after grafting with immunofluorescent reagents. A horse antiserum specific for T lymphocytes revealed an increasing number of T lymphocytes in the cellular infiltrates of rejecting allografts. These were detectable 1 day after grafting, reached a maximum 3 days later, and were relatively diminished at 6 days. In control isografts and nonimmunological inflammations of kidney, a small number of dispersed T lymphocytes was seen. A rabbit antirat thymocyte antiserum, given to allografted BN rats, prolonged survival of the grafts and decreased the cellular infiltrate and the number of T lymphocytes in the infiltrates. We conclude that in graft rejection there is a flow of T lymphocytes into areas of tissue damage and these T lymphocytes are immunologically reactive to graft antigens.
IMMUNOCHEMICAL ANALYSIS OF THE IDIOTYPES OF MOUSE MYELOMA PROTEINS WITH SPECIFICITY FOR LEVAN OR DEXTRAN
Authors: Weigert Martin , Raschke William C. , Carson Dennis , Cohn Melvin
Published: June 2004

This paper deals solely with idiotypic determinants, the configurations of which are modified when the antibody bearing them interacts with its ligand. This phenomenon is measured as an inhibition of the reaction between anti-idiotype and idiotype.

Two points are made: (a) The assay for ligand-modifiable determinants can be used to determine the "size" of the combining site. This is illustrated here with the anti-α(1 → 6) dextran mouse myeloma immunoglobulin W3129. Whether the interaction between a homologous series of α(1 → 6) oligosaccharide ligands and the combining site of W3129 is measured by inhibition of precipitation with α(1 → 6) dextran (4) or of binding of W3129 to anti-W3129 idiotype, the finding is the same. The order of inhibition is isomaltohexaose = isomaltopentaose >> isomaltotetraose > isomaltotriose >>> isomaltose. The combining site is optimally complementary to isomaltopentaose.

(b) Cross-idiotypic specificity is closely correlated with cross-combining specificity; the converse is not true. This is illustrated here with three groups of mouse myeloma immunoglobulin, each specific for α(1 → 3) dextran, α(1 →6) dextran, ß(2 → 1) or ß(2 → 6) levan. If a given anti-idiotypic serum cross-reacted with several myeloma proteins, they always had similar combining specificity. Thus the three proteins, J558, MOPC 104E, and UPC 102, which cross-react with anti-J558 have combining specificity for α(1 → 3) dextran; cross-reacting W3082, UPC 61, and Y5476 have specificity for levan; and cross-reacting W3129 and W3434 have specificity for α(1 → 6) dextran. This extends previous studies with proteins specific for phosphorylcholine (7) or γ-globulin (8). As expected, the converse is not true, for proteins may have combining specificity for α(1 → 6) dextran e.g. QUPC 52, or levan e.g. J606, UPC 10 and yet not carry the above-mentioned reference idiotypes.

The correlation between cross-idiotypic and combining specificity breaks down when idiotypic determinants which are not modifiable by ligand are studied. The implications of this are pointed out since most investigations deal with ligand-nonmodifiable determinants.
MIXED LYMPHOCYTE REACTIVITY AGAINST NORMAL CELLS BY SPLENIC LYMPHOCYTES FROM TUMOR-BEARING MICE
Authors: Devlin R. G. , McCurdy J. D. , Baronowsky P. E.
Published: June 2004

A possible consequence of an antilymphocytic autoimmune process would be serious impairment of an animal's ability to destroy tumor cells. One measure of autoimmune reactivity of this type would be the demonstration of cellular immune responsiveness by cells from tumor-bearing mice against syngeneic normal cells. These experiments demonstrate that spleen cells from mice bearing a lymphocytic leukemia of identical histocompatability type as the host mounted a vigorous immune response against normal syngeneic cells in a mixed lymphocyte reaction (MLR). Moreover, ascitic cells from leukemic mice responded significantly to normal syngeneic spleen cells in MLR's. The former reactions are usually much more vigorous than the responses of normal to malignant cells. These results are discussed in terms of the relationship between autoimmunity and neoplasia. Alternative explanations necessitated by the dangers involved in the interpretation of the immunology of transplantable tumors are considered.
MECHANISM OF THYMUS-INDEPENDENT IMMUNOCYTE TRIGGERING
Authors: Coutinho Antonio , Gronowicz Eva , Bullock Wesley W. , Möller Göran
Published: June 2004

The present experiments were performed in order to analyze the mechanism by which thymus-independent antigens (nonspecific B-cell mitogens) can induce specific immune responses to antigenic determinants present on the same molecule. The hapten NNP was coupled to the B-cell mitogen, lipopolysaccharide (LPS). The conjugate retained full mitogenic activity and bound specifically to NNP-reactive cells. NNP-LPS activated polyclonal as well as specific anti-NNP antibody synthesis, but the optimal concentrations for induction of specific anti-NNP cells were several orders of magnitude lower than the concentrations required for polyclonal activation. These low concentrations failed to activate nonspecific cells, but they induced specific thymus-independent responses of high-avidity NNP-specific cells with the typical kinetics of antigenic responses in vitro. Furthermore, hapten-specific cells were paralyzed by NNP-LPS concentrations that were optimal for induction of polyclonal activation. Specific activation and paralysis could be abolished by free hapten indicating that selective binding of NNP-LPS to hapten-specific cells was responsible for the specificity of the response. However, the triggering signal lacked specificity, since high-avidity specific anti-NNP cells could still be activated by stimulating concentrations of NNP-LPS in the presence of free hapten, even though the Ig receptor combining sites were presumably occupied by NNP.

The findings show that B cells with specific Ig receptors for the antigenic determinants on mitogen molecules preferentially bind these molecules and become activated at concentrations still unsufficient to trigger other B cells that lack specific receptors. It is suggested that activation for primary IgM responses in B cells is the result of "one nonspecific signal." This nonspecific signal is provided by the mitogenic properties of some antigens (highly thymus independent or, alternatively, by nonspecific T-cell factors (for highly T cell-dependent antigens), or both, and the surface structures responsible for triggering are not the Ig receptors. The specific Ig receptors only act as passive focusing devices for nonspecific stimuli, entitling the cell to be selectively activated, even though both the signal and the receptors for the triggering are nonspecific.
PERSISTENCE OF HAPTEN-ANTIBODY COMPLEXES IN THE CIRCULATION OF IMMUNIZED ANIMALS AFTER A SINGLE INTRAVENOUS INJECTION OF HAPTEN
Authors: Schmidt Donald H. , Kaufman Bette M. , Butler Vincent P.
Published: June 2004

To study the fate of a low molecular weight antigen (hapten) in the circulation of animals whose sera contain antibodies specific for that low molecular weight antigen, a single injection of digoxin-3H (0.4 mg/kg) was administered intravenously to 18 rabbits. Thirteen animals (nine nonimmunized and four immunized with bovine serum albumin) served as control animals. In five rabbits which had been immunized with a digoxin-bovine serum albumin conjugate and whose sera contained digoxin-specific antibodies, the mean 12-h serum digoxin concentration was 8,300 ng/ml (control: 92 ng/ml) and the mean serum concentration 12 mo after the single injection of digoxin-3H was 85 ng/ml. In digoxin-immunized rabbits, less than 10% of the digoxin-3H was excreted in the first 10 days (control: 77% recovered in urine and feces) and the mean biological half-life of digoxin, as calculated from serum digoxin-3H disappearance curves, was 72 days (control: 3.4 days). In sera of digoxin-immunized rabbits, more than 90% of the circulating digoxin-3H was immunoglobulin bound, as determined by the double-antibody and dextran-coated charcoal methods.

The serum disappearance rate of 125I-antidigoxin antibodies was similar in nonimmunized and in immunized animals and in the presence or absence of digoxin.

It is concluded that the biological half-life of a hapten may be markedly prolonged when the hapten is bound to specific antibody. The persistence of antibody-hapten complexes in the circulation suggests that these complexes may not be deposited in tissues and raises the possibility that low molecular weight determinants may be capable of preventing or reversing the deposition of immune complexes, containing macromolecular antigens, in the tissues of experimental animals and man.
LIGAND-INDUCED MOVEMENT OF LYMPHOCYTE SURFACE MACROMOLECULES
Authors: Unanue Emil R. , Ault Kenneth A. , Karnovsky Morris J.
Published: June 2004

Microscopic examination of spleen lymphocytes discloses a small number moving at random at a given time. The majority of lymphocytes with this spontaneous movement are thymic derived. Addition of anti-Ig antibodies stimulates random movement of B lymphocytes. This movement depends upon a bivalent antibody and a metabolically active cell. The movement is inhibited by DFP, suggesting the involvement of a serine esterase. Also the anti-Ig stimulated movement of the lymphocyte is inhibited by cytochalasin B or by not allowing the cells to settle onto a surface. Lymphocytes treated with DFP or cytochalasin B, or untreated lymphocytes in suspension, capped the anti-Ig-Ig complexes. Hence, one can dissociate the surface capping of anti-Ig-Ig complexes from cell movement. We postulate that capping may result from superficial movements of the surface and/or from membrane flow, both of which are not related to actual translation of the cell on a surface. Four effects have now been observed following combination of a ligand with the antigen receptor on the B lymphocytes: redistribution on the surface of the complexes; pinocytosis and catabolism; shedding into the extracellular environment; and stimulation of translational movement.
ELECTROPHORETIC MOBILITY AND AGGLUTINABILITY OF RED BLOOD CELLS: A "NEW" POLYMORPHISM IN MICE
Authors: Rubinstein Pablo , Liu Ngukoy , Streun Edwin W. , Decary Francine
Published: June 2004

A quantitative method has been developed to determine agglutinability of mouse red blood cells. Tests with different inbred strains of mice revealed only two phenotypes. The same inbred strains were tested with the cytopherometer to determine the electrophoretic mobility of the corresponding red cells. Again, two phenotypes were uncovered, and faster mobility was found in the red cells that had higher agglutinability. The genetic control of this character is autosomal and codominant, and segregates independently of H-2 and coat color.
PROPERDIN FACTOR D: CHARACTERIZATION OF ITS ACTIVE SITE AND ISOLATION OF THE PRECURSOR FORM
Authors: Fearon Douglas T. , Austen K. Frank , Ruddy Shaun
Published: June 2004

The activity of properdin factor D was measured by the generation of the hemolytically active cellular intermediate, EAC43B(D), bearing the C3b-dependent alternate pathway C3 convertase. Treatment of factor D with DFP prevented formation of EAC43B(D); thus, a serine esterase is essential for the generation of the alternate pathway C3 convertase, a situation analogous to the role of C1 in the formation of the classical C3 convertase, C42. The definition of factor D as a serine esterase prompted a search for its proenzyme form, and resulted in the chromatographic isolation from plasma of a single peak of trypsin-inducible factor D activity, distinct from activated factor D. Analytical gel filtration indicated an apparent mol wt of 25,000. This protein from which trypsin elaborated factor D activity, as assessed by the formation of EAC43B(D), the generation of the CoVF-dependent C3 convertase, and the cleavage of factor B in the presence of C3b, was designated "precursor factor D." The DFP resistance of precursor factor D, and the susceptibility of its trypsin-activated form to inactivation by DFP is analogous to the behavior of other plasma serine esterases, including C1.
SPECIFIC HETEROLOGOUS ENHANCEMENT OF IMMUNE RESPONSES
Authors: Rubin Arnold S. , MacDonald A. Bruce , Coons Albert H.
Published: June 2004

The mixing of two histoincompatible human lymphocyte cell lines generated the release of a soluble factor which was capable of nonspecifically enhancing the in vitro immune response of normal mouse spleen cells against sheep erythrocytes. When active supernates were subjected to exclusion chromatography on Sephadex G-150 and BioGel P-150, the active principle eluted with molecules of approximately 35,000 mol wt. Column aliquots from similarly treated supernates from independent cultures of each lymphoid cell line were inactive. The human enhancing factor was concentrated and purified by ammonium sulfate fractionation, followed by Sephadex gel filtration and polyacrylamide gel electrophoresis.
CHARACTERIZATION OF A HUMAN ACUTE PHASE PROTEIN FOUND IN ASSOCIATION WITH RUBELLA VIRUS INFECTION
Authors: Cappel Roger , Schluederberg Ann , Gifford Robert H. , Horstmann Dorothy M.
Published: June 2004

A precipitating antigen, rho, was first detected in the blood of persons with rubella and in rubella virus-infected cell culture fluids (1). Partially purified antigens from both sources were examined and shown to have similar properties, although antigen from serum sedimented more heterogeneously, with estimated coefficients from 15 to 21 S, while that from culture fluids sedimented in the 11–14 S region. In each case, antigen was located in the ß-1 zone after electrophoresis in agarose, and at a density of 1.305 g/ml after centrifugation in CsCl. Stability characteristics were typical of protein antigens.

Immunofluorescent microscopy revealed that rubella virus induced the appearance of rho antigen scattered throughout the cytoplasm of infected cells. When cells containing antigen were exposed for 24 h to 5 µg/ml actinomycin D rho was no longer detectable, indicating the probable cellular origin of the antigen. Also, titers in medium of infected cultures showed a reduction after actinomycin treatment, but levels of the virus-specified antigen, iota, were relatively unaffected.

Rho appears to be a protein common to man and many animals. In vitro, it was induced by rubella virus and by adenovirus. In vivo, rho titers were shown to be elevated after rubella virus infection and, to a lesser extent, after infection with certain other viruses. High titers were also demonstrated in women late in pregnancy and in patients with rheumatoid arthritis. In man and the chimpanzee, the appearance and decline of rho in the blood after rubella virus infection were temporally similar to the patterns of CRP, although rho seemed to be a more sensitive indicator of infection.

The data presented indicate that rho is a newly recognized acute phase protein inducible by certain virus infections and by other unidentified stimuli present prominently in pregnancy and rheumatoid arthritis.
ALLOANTISERUM-INDUCED INHIBITION OF IMMUNE RESPONSE GENE PRODUCT FUNCTION
Authors: Shevach Ethan M. , Paul William E. , Green Ira
Published: June 2004

It has been previously shown that alloantisera prepared by reciprocal immunization of strain 2 and strain 13 guinea pigs specifically block the activation of T lymphocytes from immune guinea pigs by antigens, the response to which is controlled by Ir genes. In this report we have examined the effect of absorption of the 13 anti-2 serum with different populations of lymphoid cells. It is unlikely that the inhibitory activity of the anti-2 serum on the proliferation of (2 x 13)F1 lymphocytes to a DNP derivative of a copolymer of L-glutamic and L-lysine (DNP-GL) is due to the presence of antibodies specific for the unique antigenic determinants (idiotypes) of clonally distributed T-lymphocyte receptors. Thus, cells obtained from a normal animal and a DNP-GL immune animal were equivalent in their absorptive capacity. Populations of T lymphocytes were ineffective in absorbing either the cytotoxic or inhibitory activity of the anti-2 serum, while L2C leukemia cells, a malignant B-cell population, were most efficient in absorbing both activities. Thus, the antigen(s) against which the cytotoxic and inhibitory activities are directed are present to a greater extent on B lymphocytes than on T lymphocytes. However, these results do not allow us to definitively determine whether the inhibitory activity of the alloantisera is due to antibodies specific for Ir gene products or antibodies specific for linked antigens in the MHC.

We also examined the effect of a number of anti-immunoglobulin reagents which had specificity for the heavy and/or light chains of guinea pig immunoglobulin on the in vitro lymphocyte proliferative response to antigen. Under conditions in which we were able to completely and specifically suppress the response of (2 x 13)F1 lymphocytes to DNP-GL with anti-2 serum, the anti-immunoglobulin reagents were devoid of inhibitory effect on the response of these same F1 cells to DNP-GL, a copolymer of L-glutamic and L-tyrosine (GT), or purified protein derivative of tuberculin (PPD). These results strongly suggest that conventional serum-type immunoglobulin is not important in antigen recognition by the T cells involved in the DNA synthetic response.
STUDIES OF IDIOTYPIC ANTIBODIES
Authors: Rodkey L. Scott
Published: June 2004

Rabbits were immunized with a hapten-protein conjugate and sera were collected for 189 days. The antihapten antibodies were purified by affinity chromatography, then the same animal that synthesized the antibody was reinjected with polymerized F(ab')2 fragments of antihapten antibodies. Sera were collected after autoimmunization and tested by an indirect radioimmunoassay technique for reaction with [125I]F(ab')2 fragments of the original antihapten antibody. Results showed that each individual responded to its own F(ab')2 and the antisera were specific for antihapten antibodies of that individual. Quantitative allotype assays established the immunoglobulin nature of the labeled test antigen. Inhibition assays showed that the reaction was specifically inhibitable with hapten. The relationship of this system with other idiotypic systems and the possible autoimmune implications of autoantiidiotypic antibodies are discussed.
GRAFT-VS.-HOST REACTIONS IN RECIPROCAL HYBRID MICE
Authors: Morse Herbert C. , Harrison Michael R. , Asofsky Richard
Published: June 2004

Spleen cells from BALB/c and C57BL/6 mice were tested for their reactivity against reciprocal hybrid tissues ((BALB/c x C57BL/6) F1 and (C57BL/6 x BALB/c) F1) in three assay systems: the mixed lymphocyte reaction (MLR); the Simonsen spleen-weight graft-vs.-host (GVH) assay; and a GVH mortality assay. It was shown that both F1's serve as equally effective stimulators of parental cells in the MLR. In the spleen-weight assay, BALB/c and C57BL/6 cells were equally active in a given host, but greater splenomegaly was observed in (BALB/c x C57BL/6) F1 hosts regardless of the donor strain. By contrast, BALB/c cells were much less lethal than C57BL/6 cells in (BALB/c x C57BL/6) F1 hosts than in (C57BL/6 x BALB/c) F1 hosts, and to a lesser degree C57BL/6 cells were less lethal than BALB/c cells in (C57BL/6 x BALB/c) F1 hosts. The possibility that modifying substances may differentially alter reactivity of parental lymphocytes and that considerations other than genotype determine the outcome of a GVH reaction are discussed in detail.
AUTORADIOGRAPHIC STUDIES ON THE PROLIFERATION OF ANTIBODY-PRODUCING CELLS IN VITRO
Authors: Hünig T. , Schimpl A. , Wecker E.
Published: June 2004

A rapid and reliable autoradiographic technique for plaque-forming cells (PFCs) using 14C rather than tritiated thymidine is described. Its application to PFCs developing in vitro shows that (a) practically all PFCs derive from precursors dividing steadily during the culture period, (b) PFC precursors divide in the absence of T-cell helper function, and (c) at least some PFCs may continue to divide.
INHIBITION OF A T-CELL-DEPENDENT IMMUNE RESPONSE IN VITRO BY THYMOMA CELL IMMUNOGLOBULIN
Authors: Stocker John W. , Marchalonis John J. , Harris Alan W.
Published: June 2004

Concentrated medium obtained from cultures of a continuous thymus-derived mouse lymphoma cell line (WEHI-22.1) was found to inhibit a T-cell-dependent (antidonkey red blood cell), but not a T-cell-independent (anti-DNP) immune response in vitro. Passage of such a concentrate through an anti-mouse Ig immunoadsorbent column removed its inhibitory activity. It is suggested that the tumor cell Ig can compete with specific normal T-cell Ig in its collaborative function in immune responses. A similar mechanism may account for anergy associated with some human lymphoid neoplasms.
THE K-CHAINS OF THE IMMUNOGLOBULIN FROM A CONTINUOUS CULTURE OF HUMAN LYMPHOCYTES (DAUDI) HAVE AN UNUSUAL MOLECULAR SIZE
Authors: Kennel Stephen J.
Published: June 2004

Membrane-associated immunoglobulin (M-Ig) was isolated from Daudi cells which had been radioiodinated using lactoperoxidase. This M-Ig was found to have a molecular size on SDS-PAGE of 330,000 daltons. The component protein chains (µ and K) have molecular sizes of 75,000 and 33,000 daltons, respectively. Further studies showed that [3H]galactose can be incorporated into K-chains, but that extra protein and not carbohydrate was responsible for the increased molecular size of this molecule.
SECRETION OF PLASMINOGEN ACTIVATOR BY STIMULATED MACROPHAGES
Authors: Unkeless Jay C. , Gordon Saimon , Reich E.
Published: June 2004

Cultured thioglycollate-stimulated peritoneal macrophages synthesize, accumulate, and continuously release high levels of plasminogen activators for at least 4 days whereas cultures of unstimulated macrophages do not; the higher specific catalytic activity of released vs. cell-associated enzyme suggests that the plasminogen activators are actively secreted. The major macrophage plasminogen activator is a serine protease of mol wt 48,000, and thus resembles the comparable enzyme released by virally transformed fibroblasts. Macrophages release a second plasminogen activator of mol wt 28,000 that is also a serine enzyme.

The secretion products released by stimulated and unstimulated macrophages have been compared by SDS-polyacrylamide gel electrophoresis after chemical labeling with 3H-DFP or biosynthetic labeling with 14C-amino acids. These procedures show that some proteins are formed in both cultures, whereas others are uniquely secreted by each type of macrophage. The serine enzymes released by the two kinds of macrophages differ in specificity and electrophoretic mobility.
CELL SURFACE IMMUNOGLOBULIN
Authors: Vitetta Ellen S. , Grundke-Iqbal Inge , Holmes Kathryn V. , Uhr Jonathan W.
Published: June 2004

Lymphoid cells from the spleen, lymph nodes, and thoracic duct of axenic and control mice were incubated with [3H]tyrosine and synthesis and secretion of protein and Ig studied. It was found that only IgM was synthesized by cells from axenic mice whereas cells from control mice also synthesized IgG. Splenocytes from both axenic and control mice had 8S IgM on their surface. Radiolabeled splenocytes from axenic mice were incubated to determine the kinetics of release of 125I-labeled cell surface IgM and [3H]tyrosine-labeled IgM. Cell surface IgM was shed as 8S with an initial half-life of release of 5–8 h whereas [3H]tyrosine-labeled Ig was secreted as 19S with an initial half-life of 2–3 h. These findings suggest that two independent pathways are involved. It is suggested that small lymphocytes shed 8S IgM and plasma cells secrete 19S IgM.

It was observed that lymphoid cells from axenic mice synthesize a higher proportion of IgM relative to total protein. Electron microscopic examination of splenocytes from such mice revealed a markedly higher proportion of plasma cells and a paucity of lymphoblasts compared to controls. It was suggested, therefore, that axenic mice lack a population of stimulated T cells which can induce a switch from IgM to IgG synthesis and which is capable of suppressing IgM synthesis. Lymphoid cells from axenic mice synthesize and secrete less protein that coprecipitates with antigen-antibody complexes.
EFFECT OF B-CELL MITOGENS ON LYMPHOCYTE SUBPOPULATIONS POSSESSING C'3 AND Fc RECEPTORS
Authors: Möller Göran
Published: June 2004

By the use of a rosette method allowing the detection at the cellular level of lymphocytes simultaneously binding Fc- and C'3-sensitized red cells it was found that about 70% of the rosette-forming cells from spleens of nude and normal mice possessed receptors for both Fc and C'3, whereas 30% only had Fc receptors. Very few, if any, lymphocytes possessed only C'3 receptors.

The B-cell mitogens, purified-protein derivative of tuberculin (PPD), lipopolysaccharide from Escherichia coli (LPS), and pneumococcal polysaccharide type SIII (SIII), induced marked changes of these receptor-bearing lymphocytes. PPD caused a rapid loss of cells capable of binding C'3 and a concomitant increase of only Fc-binding cells, which was detected after only 24 h. LPS and SIII induced analogous changes, but they were not detected until 48 h and were not complete until after 72 h. It is suggested that immature lymphocytes possess both Fc and C'3 receptors and lose the latter receptor upon differentiation induced by B-cell mitogens PPD. and LPS would affect different populations of B cells, PPD-activating cells that have already reached a higher differentiation stage, whereas LPS and SIII would activate more immature B cells.
GENETICS OF A NEW IgVH (T15 IDIOTYPE) MARKER IN THE MOUSE REGULATING NATURAL ANTIBODY TO PHOSPHORYLCHOLINE
Authors: Lieberman R. , Potter M. , Mushinski E. B. , Humphrey W. , Rudikoff S.
Published: June 2004

The idiotype present on the Fab of a phosphorylcholine-binding IgA myeloma protein TEPC 15 (T15) of BALB/c origin was found in normal serum of BALB/c mice. Molecules carrying the T15 idiotype in normal serum could be adsorbed with Sepharose phosphorylcholine beads and R36A pneumococci. The T15 idiotype is absent in germ-free BALB/c but appears when the mice are conventionalized. A survey of normal sera of inbred strains for the T15 idiotype showed it to be present in BALB/c, 129, C57L, C58, and ST and absent or in low levels in CBA, C3H, C57BL/6, C57BL/Ka, C57BL/10, SJL, B10.D2, DBA/2, RIII, A, AL, AKR, NZB, and NH inbred strains of mice. The T15 idiotype is associated with some but not all strains carrying the IgCH allotypes found in BALB/c. Linkage of genes controlling the T15 idiotype in normal serum to the IgCH locus of BALB/c was demonstrated in F2 progeny of a BALB/c and C57BL cross, Bailey's recombinant inbred strains, C x BD, C x BE, C x BG, C x BH, C x BI, C x BJ, C x BK, and CB20 congenic strains. Among these strains, only those possessing the IgCH locus of BALB/c including the F2 progeny consisting of BALB/c homozygotes and BALB/c/C57BL heterozygotes and C x BG and C x BJ recombinants showed the T15 idiotype.
IMMUNOHISTOCHEMICAL DEMONSTRATION OF IgG IN REED-STERNBERG AND OTHER CELLS IN HODGKIN'S DISEASE
Authors: Garvin A. J. , Spicer S. S. , Parmley R. T. , Munster A. M.
Published: June 2004

Spleens and lymph nodes fixed lightly for optimal immunocytochemistry or processed routinely for surgical diagnosis disclosed strong selective immunostaining for IgG in numerous immunocytes in tumor-free areas. Areas involved by Hodgkin's disease revealed, in addition, strong immunostaining for IgG but not IgM in Reed-Sternberg cells and faint to strong staining in Hodgkin cells as well. Ultrastructurally the Reed-Sternberg and Hodgkin cells displayed abundant polyribosomes and sparse granular reticulum and appeared to form unexportable IgG on unbound ribosomes.
UPTAKE AND REDUCTION OF OXIDIZED AND REDUCED ASCORBATE BY HUMAN LEUKOCYTES
Authors: Bigley Robert H. , Stankova Libuse
Published: June 2004

Incubation of human leukocytes with dehydroascorbate (DHA) results in an increase in their reduced ascorbate (AA) content and hexose monophosphate shunt (HMS) activity, independent of oxygen supply. Incubation with AA induces these changes only in the presence of oxygen. The increase in HMS activity observed as cell AA increases by 1 µmol is the same during incubation with either DHA or AA.

We propose that human leukocytes take up ascorbate as DHA (AA after oxidation to DHA) and reduce it promptly to AA, and that HMS stimulation upon incubation with either AA or DHA is a result of DHA reduction.
GENETIC CONTROL OF THE IMMUNE RESPONSE TO STAPHYLOCOCCAL NUCLEASE
Authors: Lozner Eugene C. , Sachs David H. , Shearer Gene M.
Published: June 2004

A number of inbred and congenic resistant strains of mice were immunized with staphylococcal nuclease (Nase). Antibody responses were measured in the sera of the animals by a sensitive method involving inhibition of enzymatic hydrolysis of DNA, High responder strains included A/J, DBA/2, BALB/c, AKR/J, C57BR, and SJL/J. DBA/1 and C57BL/6 mice were low responders. The strain distribution of anti-Nase response potential was compatible with the relevant immune response gene(s) being linked to the murine major histocompatibility complex. Linkage of this response to H-2 was demonstrated by the findings that: (a) the congenic C3H/HeJ and C3H.SW mice were respectively high and low responders; (b) the congenic lines B10.A and B10.D2 were high responders, whereas the C57BL/10 strain was a poor responder; and (c) anti-Nase response potential of F2 progeny from DBA/1 x SJL/J matings correlated with their H-2 type. Three B10.A recombinant lines were used to map this Ir gene within H-2. B10.A(4R) was a high responder to Nase, whereas B10.A(2R) and B10.A(5R) were both low responders. We wish to propose the name Ir-Nase for the gene(s) controlling antibody responsiveness to this immunogen. Our data indicate that Ir-Nase is located within the same chromosomal segment of the H-2 complex as is Ir-IgG.
IN VITRO SYNTHESIS AND SECRETION OF LYSOZYME BY MONONUCLEAR PHAGOCYTES
Authors: Gordon Salmon , Todd Jean , Cohn Z. A.
Published: June 2004

Pure cultures of three types of mononuclear phagocytes—mouse peritoneal macrophages, unstimulated or after thioglycollate stimulation, and human monocytes—synthesize and secrete large amounts of lysozyme in vitro. The macrophage lysozyme is indistinguishable from authentic lysozyme in its ability to lyse M. lysodeikticus, inhibition by specific antisera, a similar size of 14,000 and cationic charge. Lysozyme secretion in culture is characterized by a large net increase in total lysozyme, 4–20-fold in 3 h, 75–95% of which is in the medium, and its continued extracellular accumulation over at least 2 wk in culture. Lysozyme is the major 14C-labeled protein secreted into the medium by both unstimulated and thioglycollate-stimulated macrophages and the 0.75–1 µg produced per 1 x 106 cells/day represents 0.5–2.5% of the total cell protein.

Lysozyme is a cell-specific marker for mononuclear phagocytes and the PMN, which contains preformed enzyme, since it is absent in lymphoid cells and a variety of fibroblast and epithelioid cell lines. Lysozyme production is also a useful measure of mononuclear phagocyte cell number.

The rate of lysozyme production and secretion is remarkably constant for all cell types under a variety of culture conditions. Production by the mouse macrophage increases threefold on the 2nd day in culture and then remains linear with time. Production is optimal at a relatively low serum concentration, but can be maintained, in the absence of serum, in lactalbumin hydrolysate or, at a reduced level in basal media.

The production and secretion of lysozyme are independent of the production of macrophage acid hydrolases. Net increase and secretion of lysozyme occur under conditions where acid hydrolases like N-acetyl ß-glucosaminidase, ß-glucuronidase, ß-galactosidase, and cathepsin D are neither accumulated nor secreted. Massive phagocytosis of latex particles has no effect on lysozyme production and secretion.

Lysozyme production can be rapidly inhibited by treatment with cycloheximide (0.4 µg/ml) whereas inhibition of its production by colchicine (10–6 M) occurs only after a lag period of more than 8 h, and is probably due to a secondary effect.

These results show that mouse macrophages provide a simple in vitro system to measure lysozyme secretion and its control. These studies also indicate the possible importance of mononuclear phagocytes in the secretion of a variety of biologically active products and in the modification of their environment.