bluuebunny/crossref_metadata_2025 huggingface dataset:
LOCALIZATION OF FREE AND BOUND SECRETORY COMPONENT IN HUMAN INTESTINAL EPITHELIAL CELLS
Authors: Poger Marshall E. , Lamm Michael E.
Published: June 2004

Antibody reagents were made specific for each of the two forms of human SC, FSC and BSC, which is an integral part of the sIgA molecule. With the fluorescent antibody method the cytological and histological localization of FSC, BSC, and α-chains has been studied in various human mucous membranes. SC was present in columnar epithelial cells of the intestines and in the cells of serous acini of bronchial and salivary glands. In contrast, SC was not found in intestinal goblet cells or cells of mucous acini of bronchial and salivary glands. In the columnar epithelial cells of the small and large bowel, FSC was present most prominently in the Golgi zone, and much less prominently in the apical cytoplasm. On the other hand, BSC and α-chains were located only in the apical cytoplasm in an overlapping manner. The results favor a model in which sIgA is assembled inside epithelial cells from SC, which was synthesized in the same cell, and IgA, which entered the epithelial cell after synthesis in and secretion by a plasma cell.
A DEPRESSION OF CELL-MEDIATED IMMUNITY TO MEASLES ANTIGEN IN PATIENTS WITH SYSTEMIC LUPUS ERYTHEMATOSUS
Authors: Utermohlen Virginia , Winfield John B. , Zabriskie John B. , Kunkel Henry G.
Published: June 2004

Using the direct migration inhibition test, response to measles antigen in patients with systemic lupus erythematosus (SLE) was found to be decreased when compared with that of normal subjects. No alteration was observed in similar experiments using parainfluenza type 1 and rubella antigens. The specific decrease in measles antigen effect showed no obvious correlation with activity of SLE or with the presence of lymphocytotoxic antibodies. Whether the specificity of the decrease in reactivity is due to some particular relationship between the measles virus or antigen and SLE, or to the possibility that measles reactivity is a more sensitive indicator of a generalized defect of cell-mediated immunity, remains unclear.
THE EFFECT OF 2-MERCAPTOETHANOL ON MURINE MIXED LYMPHOCYTE CULTURES
Authors: Bevan Michael J. , Epstein Ruth , Cohn Melvin
Published: June 2004

Mouse spleen cells which have been depleted of adherent cells do not respond to allogeneic lymphocytes in vitro. Their cytotoxic response can be restored by inclusion of mercaptoethanol in the medium. Mercaptoethanol is shown to have a stimulatory effect also on the response of normal (unseparated) spleen cells to alloantigens. The enhancement of the DNA-synthetic and cytotoxic response is similar, varying from 3.5–15-fold. Cytotoxic cells also appear in unmixed lymphocyte cultures in the presence of mercaptoethanol and fetal calf serum. The specificity of these background cytotoxic cells is not known.
CYTOTOXIC ACTIVITY OF MOUSE MACROPHAGES STUDIED BY VARIOUS INHIBITORS
Authors: Melsom Haakon
Published: June 2004

The influence of various inhibitors on the cytolytic potential of mouse macrophages against syngeneic erythrocytes has been investigated in vitro by isotope techniques. Intact macrophage membrane and cell metabolism was essential for full cytotoxic activity. The process was completely blocked by anaerobiosis and cold. ATP from both mitochondrial respiration and glycolysis seems to be the high energy intermediate which is utilized during the cytotoxic activity of macrophages leading to target cell lysis. The process did not depend on concomitant DNA transcription, translation, or protein synthesis.
EPSTEIN-BARR VIRUS-NEGATIVE HUMAN MALIGNANT T-CELL LINES
Authors: Kaplan Joseph , Shope Thomas C. , Peterson Ward D.
Published: June 2004

Two lymphoblastoid lines, CCRF-CEM and HSB-2, with properties of malignant cells, derived from children with leukemia secondary to lymphosarcoma, have T-cell properties and lack Epstein-Barr virus antigens.
EVENTS AFTER THE BINDING OF ANTIGEN TO LYMPHOCYTES
Authors: Ault Kenneth A. , Unanue Emil R.
Published: June 2004

The behavior of the immunoglobulin antigen receptor on lymphocytes was studied using both fluorescent antiimmunoglobulin antibody to detect B cells and autoradiography with radiolabeled antigens to detect antigen-binding cells. It was shown that after binding of antiimmunoglobulin antibody to the lymphocyte there was a rapid loss of surface immunoglobulin and then a progressive reappearance over 18 h. This could be quantitated using an inhibition assay for surface immunoglobulin. Similarly, after binding various dinitrophenyl-conjugated proteins or keyhole limpet hemocyanin to their specific antigen-binding cells, there was a loss of the antigen receptor from the surface and then a progressive reappearance of the receptor. The reappearance of surface immunoglobulin and of the antigen receptor proceeded at about the same rate. Repeated exposure to antibody or prolonged exposure to antigen did not diminish the capacity of the lymphocyte to re-express its receptor. These events, which follow the interaction of antigen and its receptor, are of possible importance in understanding the mechanism of triggering of the immune response and of tolerance.
CELLULAR REQUIREMENTS FOR THE PRIMARY IN VITRO ANTIBODY RESPONSE TO DNP-FICOLL
Authors: Mosier D. E. , Johnson B. M. , Paul W. E. , McMaster P. R. B.
Published: June 2004

The cellular requirements for the primary in vitro IgM and IgG response to dinitrophenyl-substituted Ficoll were examined. Neither thymus-derived lymphocytes nor macrophage-rich splenic adherent cells were required for anti-DNP antibody synthesis. DNP-Ficoll is therefore tentatively classified as a "thymic-independent" antigen.
CLONAL MEMORY
Authors: McMichael A. J. , Williamson A. R.
Published: June 2004

A single clone of B cells producing anti-DNP antibody recognizable by the isoelectric-focusing spectrum has been used, in a double transfer system, to study clonal memory. Trasnsferable B memory develops between 4 and 7 days after the first transfer with antigen. B-memory cells thus proliferate before or concomitantly with antibody-forming cells.
IMMUNOLOGICAL TOLERANCE IN BONE MARROW-DERIVED LYMPHOCYTES
Authors: Hamaoka Toshiyuki , Katz David H.
Published: June 2004

The studies presented here have focused on the important question of reversibility of inactivation of DNP-specific B lymphocytes induced by the DNP derivative of the copolymer of D-glutamic acid and D-lysine (D-GL). In so doing, we have analyzed the capacity of a strong T-cell stimulus, such as that provided by the allogeneic effect, and of gentle enzymatic treatment with trypsin to alter, prevent, or reverse the tolerance induced by DNP-D-GL. Under experimental conditions in which DNP-specific B lymphocytes were exposed first to the tolerogenic molecule, and rendered markedly unresponsive by such exposure either in vitro or in vivo, subsequent exposure to an allogeneic effect failed to appreciably reverse or alter the tolerant state. This contrasts directly with the capacity of DNP-D-GL to serve as a stimulus for DNP-specific B lymphocytes when the critical moment of specific binding occurs subsequent to the development of an allogeneic effect. In another series of experiments, the effects of enzymatic treatment with trypsin on the tolerant B-cell population were found to vary depending on the stage of tolerance at which such treatment was performed. Thus, when exposure of cells to DNP-D-GL for a relatively short time in vitro is carried out at low temperature (4°C), the development of tolerance can be interceded by immediate trypsinization. In contrast, cells exposed to DNP-D-GL for longer periods of time and/or at 37°C were not reversed to responsiveness by trypsinization. These data were interpreted to indicate that: (a) the effect(s) of trypsin in reversing (or preventing) tolerance at the cellular level does not depend necessarily on the susceptibility of the tolerogenic moiety to the action of the enzyme, and (b) the generation of the tolerance-inducing signal involves metabolic cellular processes that can be delayed somewhat by low temperature leaving such cells relatively more susceptible to intercedent manipulations such as trypsinization. Taken collectively, therefore, the evidence obtained in these studies reinforces the concept of central tolerance in B cells induced by DNP-D-GL as reflecting sub- or intracellular inactivating events.
EVIDENCE OF SUPPRESSOR CELL ACTIVITY IN SPLEENS OF MICE BEARING PRIMARY TUMORS INDUCED BY MOLONEY SARCOMA VIRUS
Authors: Kirchner H. , Chused T. M. , Herberman R. B. , Holden H. T. , Lavrin D. H.
Published: June 2004

Spleens from Moloney sarcoma virus (MSV) tumor-bearing C57BL/6N mice contained four times the normal number of mononuclear cells and displayed a markedly elevated "spontaneous" (mitogen-independent) DNA synthesis on a per cell basis. The number of macrophages were increased three-fold while there was a slight reduction in the percentage of T lymphocytes. The phytohemagglutinin (PHA) response on a per cell basis of spleens from tumor-bearing mice was decreased about 90% when compared with normal control mice. The primary in vitro immune response to sheep red blood cells was also suppressed to levels of less than 10% of normals.

The PHA response could be restored by purification of MSV spleen cells by rayon adherence columns and by removal of phagocytic cells by an iron/magnet technique. The activity of suppressor cells in MSV spleens was demonstrated in mixtures with syngeneic normal spleen cells where a marked impairment of the PHA response was observed. Spleen cells from tumor-free nude mice and normal spleen cells treated by anti-θ serum plus guinea pig complement (C'), both totally unreactive to PHA, had no such effect. The inhibitor cell in MSV spleens was shown to be insensitive to inactivation by anti-θ plus C', but could be removed by the adherence columns and the iron/magnet technique. These data suggest that this suppressor cell is a cell of the monocyte/macrophage series. Suggestive evidence was also presented that the suppressor cells belong to a proliferating population in MSV spleens. Similar suppressor cells have been previously demonstrated in spleens of mice during a variety of immune responses. Our data show, that a tumor, although stimulating the immune system, nevertheless may be suppressive on certain immune functions through the activation of suppressor cells.
SYNERGY AMONG RESPONDING LYMPHOID CELLS IN THE ONE-WAY MIXED LYMPHOCYTE REACTION
Authors: Tittor Wolfgang , Gerbase-Delima Maria , Walford Roy L.
Published: June 2004

The present studies have shown that two subpopulations of thymus-dependent lymphocytes may act synergistically in the mixed lymphocyte reaction (MLR) in the mouse. One subpopulation was well represented in the young adult thymus and the other in lymph nodes. For optimum synergy, both populations must be allogeneic to the stimulator cells. Pretreatment of either population with mitomycin-C abolished synergy. Anti-θ serum abolished both MLR responding and synergizing activities of lymphoid cells. The two thymus-dependent subpopulations were both present in the spleen, and displayed different migratory patterns when injected into irradiated mice: one population went to spleens of the irradiated mice, the other to lymph nodes. The effects of anti-thymocyte serum on the MLR and upon synergy were assessed. While minor differences exist and are herein described, our overall results strongly suggest that in our experiments with synergy in MLR, we may be dealing with the same T1- and T2-cell subpopulations described by Cantor and Asofsky and coworkers (1, 2, 4, 5, 14) as displaying synergy in the graft-vs.-host reaction.
GENETIC CONTROL OF THE ANTIBODY RESPONSE TO TYPE III PNEUMOCOCCAL POLYSACCHARIDE IN MICE
Authors: Amsbaugh Diana F. , Hansen Carl T. , Prescott Benjamin , Stashak Philip W. , Asofsky Richard , Baker Phillip J.
Published: June 2004

Serum IgM immunoglobulin levels and antibody responses to an optimally immunogenic dose of Type III pneumococcal polysaccharide (SSS-III) were assessed for F1, F2, and backcross progeny derived from crosses between high responding BALB/cAnN (B) and low responding CBA/HN (C) mice. The results obtained confirmed our original hypothesis, namely, that a major component, present on the X chromosome, governs the ability to respond to SSS-III in a decisive manner. Although all low responding C mice had low IgM levels, both intermediate and high responders had high IgM levels of the same magnitude. Treatment with bacterial lipopolysaccharides (LPS) resulted in a significant increase in the IgM levels of low responding C mice. While the IgM levels attained were similar to those of high responding B mice, not given LPS, no antibody specific for LPS appeared to be produced. These findings suggest that C mice are unable to make an IgM antibody response to SSS-III and other polysaccharide antigens, despite the fact that they possess the capacity to synthesize normal amounts of IgM immunoglobulin.
AUTOGENOUS IMMUNITY TO ENDOGENOUS RNA TUMOR VIRUS
Authors: Ihle J. N. , Hanna M. G. , Roberson L. E. , Kenney F. T.
Published: June 2004

The viral antigenic determinants recognized in an autogenous immune response in mice against their endogenous C-type virus have been identified by SDS-polyacrylamide gel electrophoresis of immune precipitates between various sera and H3-labeled intact or disrupted AKR leukemia virus. Normal B6C3F1 [(C57BL/6 x C3H/Anf)F1] serum reacts with viral envelope antigens having mol wt of approximately 68,000, 43,000, and 17,000. In addition, minor reactions with viral antigens having mol wts of approximately 19,000 and 15,000 are demonstrable. The 68,000 and 43,000 mol wt antigens can be labeled with [3H]glucosamine and may correspond to the major viral envelope antigens M2 and M1, respectively.

The antigens recognized by autogenous immune sera do not differ with respect to age of the animal, nor are they significantly different in sera from various strains of mice (BALB/c, C57BL/6, and C3H/Anf). These results suggest that the age-asociated and strain variations in the autogenous immune response, as determined by radioimmune precipitation assays against intact virus, are due to quantitative and qualitative alterations of antibody levels against common antigens.
CELL SURFACE IMMUNOGLOBULIN
Authors: Vitetta Ellen S. , Uhr Jonathan W.
Published: June 2004

A new method for the detection of cell surface immunoglobulin labeled with isotopic precursors is described. The method consists of the aggregation of surface Ig on cells with specific antibody (heterologous) and the subsequent removal of antigen-antibody complexes by the combination of high speed centrifugation and immunoprecipitation of remaining soluble complexes using antibody to the heterologous Ig. Using this method, the kinetics of appearance of cell surface Ig and its turnover were studied in murine splenocytes. The results suggest that cell surface Ig is synthesized and transported in the same manner as secretory Ig rather than being synthesized on the plasma membrane. The turnover of intracellular and cell surface Ig in lymphocytes is slow. In contrast, intracellular Ig in plasma cells is rapidly secreted and usually without a cell surface phase. Cell surface Ig was shown to be radiolabeled with [3H]glucosamine, -galactose, and -fucose. The proportion of cell surface to intracellular (nonsurface) Ig labeled with these precursors suggests the same sequence of addition of sugars to Ig destined to be on the surface of lymphocytes as with Ig which will be secreted by plasma cells. Results with this new method also confirm earlier conclusions based on experiments using cell surface iodination: 8S IgM is the predominant Ig on the surface of murine splenocytes and the molecule appears to be attached by its µ-chains.
EXPERIMENTAL IMMUNE COMPLEX DISEASE OF THE LUNG
Authors: Brentjens Jan R. , O'Connell David W. , Pawlowski Irene B. , Hsu Konrad C. , Andres Giuseppe A.
Published: June 2004

Membranous and/or proliferative pneumonitis, similar in certain features to human interstitial pneumonitis, developed in rabbits making hyperactive antibody response to daily injections of bovine serum albumin (BSA) administered in multiple large doses sufficient to maintain the state of relative antigen-antibody equivalence. The pulmonary lesions were associated with deposition in alveolar capillary walls and interstitium of antigen, host globulin and complement, presumably in immune complexes. In some rabbits chronic interstitial pneumonitis, characterized by thickening of alveolar capillary walls, interstitial fibrosis and deposition of fibrinogen, was observed.

The production of immune complex pneumonitis seems to depend on the degree of the antibody response because rabbits developing chronic serum sickness with low doses of BSA, rabbits with acute serum sickness as well as nonresponders showed no pulmonary alterations. This observation is comparable to that described by Dixon in his studies on experimental immune complex glomerulonephritis. It is conceivable that the pulmonary pathology shown here is produced by formation of larger amounts of complexes which may persist longer at critical levels in the circulation than in rabbits immunized with a single daily injection of BSA.

In conclusion this study suggests: first, that experimental chronic serum sickness can be used as a model, not only for glomerulonephritis, but also for experimental systemic disease, comparable to human systemic diseases produced by circulating antigen-antibody complexes; and second, that the pathogenesis proposed here offers an alternative to using antilung basement membrane pneumonitis for the experimental approach to the study of human lung immunopathology.
SCANNING ELECTRON MICROSCOPY OF HUMAN LYMPHOCYTE-SHEEP ERYTHROCYTE ROSETTES
Authors: Polliack A. , Fu Shu Man , Douglas S. D. , Bentwich Z. , Lampen N. , De Harven E.
Published: June 2004

Human lymphocytes of known B or T derivation were examined by scanning electron microscopy (SEM) before and after rosetting with SRBC. After collection of the cells onto silver membranes the samples were prepared for SEM by the critical point drying method. Sheep RBC frequently underwent sphero-echinocyte transformation and multiple projections extended from their surfaces. This was readily noticeable after storage of SRBC in the cold and washing in Hanks, but more prominent after rosetting. These erythrocyte surface alterations were less apparent when freshly withdrawn cells were used. Spontaneous sheep erythrocyte rosettes (E-R), a marker for human T lymphocytes, were prepared with normal peripheral blood lymphocytes (PBL), thymic cells, and cultured T cells. EAC-rosettes (EAC-R), used to identify B lymphocytes with complement receptors, were prepared with normal PBL and cultured B cells. The majority of rosetting T lymphocytes had generally smooth surfaces while about 20% had an intermediate number of microvilli and 15% were more villous and indistinguishable from villous B cells. Studies of rosetting thymocytes and cultured T cells however indicated that the surface of some T cells alters on rosetting, becoming more villous and thus account for the higher numbers of villous T cells seen in E-rosettes. Point to point contact sites between SRBC and T lymphocytes were more frequent than broad zones of attachment. The majority of rosetting B lymphocytes had multiple microvilli, about 25% had a moderate number of microvilli and less than 10% had smooth surfaces similar to those of most T cells. Areas of contact between EAC and B lymphocytes were frequently broad zones of attachment.

The study confirms that in many cases B and T lymphocytes can be distinguished by their surface architecture as seen under the SEM; however, about 20% of rosetting B and T cells have similar surfaces with intermediate numbers of surface microvilli and cannot be distinguished by SEM without parallel immunologic identification.
GENETIC CONTROL OF IMMUNE RESPONSES IN VITRO
Authors: Kapp Judith A. , Pierce Carl W. , Benacerraf Baruj
Published: June 2004

Although nonresponder, H-2s and H-2q, mice fail to develop GAT-specific PFC responses to GAT, they do develop GAT-specific PFC responses when stimulated by GAT complexed to an immunogenic carrier such as methylated bovine serum albumin. The studies described in this paper show that injection of nonresponder mice with GAT specifically decreases their ability to develop anti-GAT PFC responses to a subsequent challenge with GAT-MBSA. Addition of GAT to cultures of spleen cells from nonresponder mice also prevents development of the GAT-specific PFC responses stimulated by GAT-MBSA. Thus, interaction of nonresponder spleen cells with GAT leads to the induction of unresponsiveness in vivo and in vitro.

Various parameters of the tolerance induction have been investigated and described. A comparison of the effects of GAT on B cells indicates that nonresponder B cells are more readily rendered unresponsive by soluble GAT than are responder B cells. The significance of these data for our understanding of Ir gene regulation of the immune response is discussed.
HUMAN MIXED LEUKOCYTE CULTURE: IDENTIFICATION OF THE PROLIFERATING LYMPHOCYTE SUBPOPULATION BY SEX CHROMOSOME MARKERS
Authors: Lohrmann Hans-Peter , Whang-Peng Jacqueline
Published: June 2004

The nature of the cells which proliferate in response to allogeneic histoincompatible lymphocytes in human mixed leukocyte cultures (MLC) was investigated. Direct information on the differentiation of proliferating cells was obtained in an in vitro chimeric system with T- and B-cell sex chromosome markers. Highly purified T and B cells were prepared from the peripheral blood of three pairs of HL-A genotype identical, MLC-negative siblings of opposite sex by E-rosette sedimentation techniques. Recombined T and B cells of the siblings were stimulated with allogeneic cells, and karyotype analysis of proliferating cells performed. 96% of the proliferating cells were found to carry the sex chromosome marker of the respective T-cell population. These results indicate that proliferation in human MLCs is a feature of T cells.
CELL SURFACE ANTIGENS OF A MOUSE TESTICULAR TERATOMA
Authors: Gooding Linda R. , Edidin Michael
Published: June 2004

Rabbit antisera to a mouse testicular teratoma, absorbed with normal mouse tissues, react by immunofluorescence with plasma membrane antigens of a variety of transplantable mouse tumor cells and transformed fibroblast cell lines including Clone 1D, SV-40-3T3, and 3T12. Trypsin treatment of cells of "normal" lines, 3T3 and FR-SV-3T3, uncovers reactivity on these as well. Early passage mouse embryo fibroblast cell cultures do not react even after trypsinization. By cross-absorbtion studies, the anti-teratoma serum appears to react with an antigen common to most tumor cells investigated thus far. When this antigen on Clone 1D cells is "capped," H-2 antigens collect with the teratoma antigens in the cap indicating a physical association between the molecules. Molecules specified by both the H-2D and H-2K regions are bound to the teratoma antigens in the Clone 1D plasma membrane. This antigen is also found in soluble tumor cell fractions where it is believed to be free of H-2. A second cell surface antigen defined by anti-teratoma serum is expressed only by hepatoma and teratoma itself. This second antigen is apparently a secretory product of teratoma cells. A third surface antigen defined by anti-teratoma serum appears to be specific for the teratoma.
ANTIGEN-SPECIFIC THYMUS CELL FACTORS IN THE GENETIC CONTROL OF THE IMMUNE RESPONSE TO POLY-(TYROSYL, GLUTAMYL)-POLY-D, L-ALANYL--POLY-LYSYL
Authors: Taussig M. J. , Mozes Edna , Isac Ronit
Published: June 2004

The genetic control of the antibody response to a synthetic polypeptide antigen designated poly-L(Tyr, Glu)-poly-D,L-Ala--poly-L-Lys [(T, G)-A--L] has been studied in congenic high responder C3H.SW (H-2b) and low responder C3H/HeJ (H-2k) strains of mice. This response is controlled by the Ir-1 gene and is H-2 linked. The method employed was to study the ability of specifically primed or "educated" T cells of each strain to produce cooperative factors for (T, G)-A--L in vitro. Such factors have been shown to be capable of replacing the requirement for T cells in the thymus-dependent antibody response to (T, G)-A--L in vivo. The T-cell factors produced were tested for their ability to cooperate with B cells of either high or low responder origin by transfer together with bone marrow cells and (T, G)-A--L into heavily irradiated, syngeneic (for bone marrow donor) recipients. Direct anti-(T, G)-A--L plaque-forming cells were measured later in the spleens of the recipients. The results showed that (a) educated T cells of both high and low responder origin produced active cooperative factors to (T, G)-A--L, and no differences between the strains in respect to production of T-cell factors could be demonstrated; and (b) such factors, whether of high or low responder origin, cooperated efficiently with B cells of high responder origin only, and hardly at all with B cells of low responder origin. The conclusion was drawn that the cellular difference between the two strains lies in the responsiveness of their B cells to specific signals or stimuli received from T cells. As far as could be discerned by the methods used, no T-cell defect existed in low responder mice and the expression of the controlling Ir-1 gene was solely at the level of the B cells in this case.
RESTRICTION OF GENE EXPRESSION IN B LYMPHOCYTES AND THEIR PROGENY
Authors: Jones Patricia P. , Craig Susan W. , Cebra John J. , Herzenberg Leonard A.
Published: June 2004

To determine whether or not B lymphocytes are committed to the synthesis of a single immunoglobulin heavy chain isotype during their differentiation into plasma cells, rabbit lymph node and Peyer's patch cells were separated into populations with and without membrane IgM, using a fluorescence-activated cell sorter (FACS). The potential of the µ-bearing (µ+) and non-µ-bearing (µ-) cells to give rise to plasma cells both in vivo after transfer into irradiated recipients and in vitro in the presence of pokeweed mitogen was assessed by immunofluorescence techniques, and the relative proportions of the cytoplasmic Ig-stained cells (CSC) synthesizing each class of heavy chains were determined.

Most of the CSC arising in vitro from µ-bearing lymph node and Peyer's patch cells contained IgM; all IgM CSC appeared to be derived from µ+ cells. Peyer's patch lymphocytes, however, did not generate IgM CSC after cell transfer and thus may be functionally different from lymph node µ+ cells. It was found also that nearly all of the many IgA CSC generated by Peyer's patch lymphocytes either in culture or after transfer were derived from µ- cells. Further fractionation of these µ- cells with the FACS after they had been membrane stained with anti-b locus allotype reagents revealed that the precursors of IgA CSC belong to a minor population of cells which do have b locus light chain determinants on their membranes, although they do not have detectable µ-chains. These cells are not found in lymph nodes.

Although the majority of Peyer's patch and lymph node cells were found to be precommitted to the synthesis of a single heavy chain isotype, a small proportion of cells may not be similarly restricted. Some of the CSC with membrane IgM were found to contain cytoplasmic IgA or IgG. In addition, µ+ populations did give rise to low numbers of IgA and IgG CSC. The implications of these results, obtained under experimental conditions, on the normal differentiation of B lymphocytes in situ are discussed.
STUDIES OF THE HUMAN LYMPHOCYTE RECEPTOR FOR HEAT-AGGREGATED OR ANTIGEN-COMPLEXED IMMUNOGLOBULIN
Authors: Dickler Howard B.
Published: June 2004

The lymphocyte receptor for complexed immunoglobulin was shown not to bind heat-aggregated human serum albumin, bovine serum albumin, transferrin, F(ab')2, reduced and alkylated Ig, and mildly oxidized Ig, which indicated that the receptor is specific for a site dependent on disulfide bond(s) on the Fc portion of complexed Ig. Inhibition experiments provided evidence that the same receptor binds both heat-aggregated Ig and antigen-antibody complexes.

Lymphocytes treated with pronase were no longer able to bind Ig complexes, which suggested that the receptor is a protein or glycoprotein. Additional evidence was obtained that lymphocyte surface Ig and the receptor for complexed Ig are distinct since the former could be capped without affecting the distribution of the latter, and surface Ig was not detectable after trypsinization of lymphocytes, whereas the binding of Ig complexes was unaffected by such treatment. Incubation of lymphocytes which had bound Ig complexes in tissue culture medium at 37°C revealed that the complexes remained on the surface membrane for several hours, and that only a minority of lymphocytes binding complexes showed cap formation.

Lymphocytes which had heat-aggregated IgG specifically bound to their receptors for complexed Ig were markedly inhibited in their ability to mediate antibody-dependent cytotoxicity, thus providing strong evidence for the necessity of the receptor in this immune activity. Titration of this inhibition with varying amounts of complexes revealed distinct plateaus in the dose-response curve. This suggested that there may be more than one kind of receptor and/or different populations of lymphocytes which bear the receptor.
HAPTEN-SANDWICH LABELING
Authors: Wofsy L. , Baker P. C. , Thompson K. , Goodman J. , Kimura J. , Henry C.
Published: June 2004

A hapten-sandwich procedure has been developed for specific labeling of cell surface antigens for fluorescence or electron microscopy. Haptens are azo-coupled to immunoglobulins specific for a cell surface antigen; the hapten-modified cell-bound antibodies can then be visualized by adding fluorescent antihapten antibody, or by adding antihapten antibody followed by hapten-modified markers for electron microscopy. Virus or high molecular weight protein markers are lightly cross-linked before conjugation with hapten to prevent their disruption. Such stable hapten-modified markers, and the accessibility of many different purified anti-azophenyl-hapten antibodies, make it feasible to distinguish more than one membrane antigen in a given labeling experiment. When mouse lymphoid cell populations are labeled with separate markers for Ig and for thymus-associated antigens, many cells exhibit the Ig marker exclusively or the thymic marker predominantly, and some cells are completely free of label.
QUANTITATIVE STUDIES ON TUMOR ENHANCEMENT IN MICE
Authors: Rubinstein Pablo , Decary Francine , Streun Edwin W.
Published: June 2004

The concentration of specific alloantibody in purified mouse immunoglobulin preparations was determined. When passively transferred in adequate doses, IgM, IgG1, and IgG2 antibodies all induced tumor enhancement in allogeneic hosts. IgM and IgG2 antibodies in high concentration led to inhibition of tumor growth. IgM and either IgG1 or IgG2 had additive effects on tumor enhancement. IgG1, but not IgG2, suppressed the inhibitory effect of IgM in high concentration.
SPONTANEOUS RELEASE OF T-CELL RECEPTORS FOR ALLOANTIGENS
Authors: Ramseier Hansruedy
Published: June 2004

In vitro cultivation of murine spleen cells resulted in a spontaneous release of receptors for alloantigens. This was revealed by the capacity of cell-free culture supernates to recognize alloantigens as measured in the PAR assay. Qualitatively, recognition responses obtained with these supernates reproduced faithfully those found with the corresponding cells.

Large amounts of receptors were released by untreated spleen cells and by spleen cells treated with a rabbit antimouse immunoglobulin serum and complement, smaller amounts were released by bone marrow cells, and native thymus cells released none. Spleen cells from nude mice and spleen cells from normal mice treated with anti-θ serum and complement showed no release of receptors. It was concluded that receptors active in the PAR test were of T-cell origin.

Release of T-cell receptors was found to be discontinuous and proceeded in waves. The amount of released receptors depended on the number of cells cultivated. Release occurred at 37°C but not at 4°C. Interaction with antigen, however, was temperature-independent. In contrast to T-cell receptors, a release of H-2 antigens could not be detected with the culture conditions employed.
THE STRANGE CROSS-REACTION OF MENADIONE (VITAMIN K3) AND 2,4-DINITROPHENYL LIGANDS WITH A MYELOMA PROTEIN AND SOME CONVENTIONAL ANTIBODIES
Authors: Michaelides Maria C. , Eisen Herman N.
Published: June 2004

To explore the possibility that the affinity of some myeloma proteins for 2,4-dinitrophenyl (DNP) ligands is the consequence of a "strange" (i.e., unexpected) cross-reaction for more natural ligands, a variety of substances (primarily derivatives of purines, pyrimidines, naphthaquinone) were tested for ability to block the binding of [3H]-ϵ-DNP-L-lysine by protein 315, an IgA mouse myeloma protein with high affinity for DNP ligands. The most impressive inhibiting activity was observed with 2-methyl-1,4-napthaquinone (menadione, vitamin K3). The affinity (intrinsic association constant) of protein 315 for menadione was 5 x 105 L/M (at 4°C). Because the same affinity was measured in direct-binding assays (e.g., equilibrium dialysis) and in an indirect one based on the assumption of competitive binding with DNP-lysine, it is likely that menadione and DNP bind at overlapping sites in the protein's combining region. This conclusion is supported by molecular models which reveal some common structural features in these ligands. Hence it is not surprising that antinitrophenyl antibody preparations, raised by conventional immunization procedures (anti-2,4-DNP; anti-2,6-DNP; anti-2,4,6-TNP) also bind menadione with considerable affinity. As with DNP ligands, when menadione binds to protein 315 or to conventional antinitrophenyl antibodies, some of the protein's tryptophan fluorescence is quenched, there is a change in the ligand's absorption spectrum (hypochromia and/or red shift), and the binding is temperature-dependent (exothermal).
GENERATION OF CYTOTOXIC T LYMPHOCYTES IN VITRO
Authors: Cerottini Jean-Charles , Engers Howard D. , MacDonald H. Robson , Brunner K. Theodor
Published: June 2004

Mouse cytotoxic T lymphocytes (CTL) were generated in mixed leukocyte cultures (MLC) using spleen cells as responding cells and irradiated allogeneic spleen cells as stimulating cells. Cytotoxicity was assessed by a quantitative 51Cr assay system and the relative frequency of CTL in individual cell populations was estimated from dose-response curves. Inclusion of 2-mercaptoethanol in the MLC medium resulted in a 20–40-fold increase in the relative number of CTL generated at the peak of the response. Under these culture conditions, cell-mediated cytotoxic activity was detectable in MLC populations as early as 48 h after the onset of the cultures.

When spleen cells from mice immunized with allogeneic tumor cells 2–4 mo previously were cultured with irradiated spleen cells of the same alloantigenic specificity (MLC-Imm), it was found that the cell-mediated cytotoxic response was detectable earlier and reached higher levels than that observed in a primary MLC. At the peak of the response, MLC-Imm populations were observed to lyse up to 50% of the target cells within 3 h at a lymphocyte: target cell ratio of 0.3:1. Immunological and physical characterization of the effector cells generated in MLC-Imm indicated that they were medium to large-sized T lymphocytes. Altogether, these studies suggested the existence of an anamnestic cell-mediated cytotoxic response in MLC-Imm.
GENERATION OF CYTOTOXIC T LYMPHOCYTES IN VITRO
Authors: MacDonald H. Robson , Engers Howard D. , Cerottini Jean-Charles , Brunner K. Theodor
Published: June 2004

Mouse cytotoxic T lymphocytes (CTL) were generated in unidirectional mixed leukocyte cultures (MLC) using normal C57BL/6 spleen cells as responding cells and irradiated DBA/2 spleen cells as stimulating cells. Cytotoxicity was assayed on 51Cr-labeled P-815 (DBA/2) target cells, and the relative frequency of CTL in individual cell populations was estimated from dose-response curves. Upon inclusion of 2-mercaptoethanol in the culture medium, it was found that significant CTL activity could be detected for as long as 3 wk in primary MLC. Reexposure of MLC cells to the original stimulating alloantigens after 14–41 days in culture resulted in significant cell proliferation and rapid regeneration of high levels of immunologically specific cytotoxicity. CTL activity in these secondary cultures increased dramatically within the first 24 h and reached higher peak levels than those found at the peak of the primary response. Furthermore, proliferation and reappearance of CTL activity could be demonstrated following each of as many as four sequential alloantigenic stimulations of the same initial cell population at 20-day intervals. Interestingly, cells recovered from MLC at the peak of the primary response on day 4 were insensitive to further allogeneic stimulation. Taken together, these results are consistent with the hypothesis that CTL differentiate in MLC to become long-lived memory cells which gradually lose their cytotoxic activity. Upon reexposure to specific alloantigen, such memory CTL rapidly regain their functional activity and proliferate to generate an expanded CTL population.
CELLULAR IMMUNITY TO HERPES SIMPLEX VIRUS MEDIATED BY INTERFERON
Authors: Lodmell Donald L. , Notkins Abner Louis
Published: June 2004

Rabbit kidney cell monolayers infected with herpes simplex virus (HSV) were incubated with leukocytes from rabbits immunized with complete Freund's adjuvant. When the leukocytes were exposed to tuberculin purified protein derivative (PPD), viral replication and plaque formation were markedly inhibited. Similarly, when leukocytes from animals immunized with HSV were exposed to UV-inactivated HSV, viral replication was markedly inhibited. Exposure of leukocytes from unimmunized animals or animals immunized with incomplete Freund's adjuvant to UV-inactivated virus or PPD produced relatively little inhibition of viral replication. Examination of supernatant fluids from stimulated cultures revealed a soluble mediator that had the properties of interferon. Interferon production was detected within several hours after exposure of sensitized leukocytes to antigen. Supernatant fluids from as few as one sensitized leukocyte per 200 rabbit kidney cells inhibited HSV replication by over 90%. These findings support the concept that the cellular immune response to HSV consists of two phases: an immunologically specific antigen recognition phase, and a nonspecific effector phase that stops HSV spread by generating interferon.
GENETIC ASPECTS OF CANINE MIXED LEUKOCYTE CULTURES
Authors: van den Tweel J. G. , Vriesendorp H. M. , Termijtelen A. , Westbroek D. L. , Bach M. L. , van Rood J. J.
Published: June 2004

Recent data stress the importance of matching donor and recipient of an organ graft for both the serologically defined (SD) and lymphocyte-defined (LD) determinants. To allow experimental evaluation of the effect of these SD and LD structures in a noninbred experimental animal, mixed leukocyte culture tests were performed between SD identical and nonidentical dogs to clarify the LD system in these animals.

The results of these experiments can be summarized as follows: (a) In the dog there is a LD locus distinct from the known SD loci, which in all probability is localized outside the first (SD-1) series locus on the chromosome. (b) The crossing-over frequency between the SD and LD loci on the chromosome is low. (c) Studies in SD identical unrelated dogs and random unrelated dogs show an apparent high linkage disequilibrium between SD and LD loci. (d) The LD system in dogs is polymorphic.
EVIDENCE FOR A CYTOLYTIC FACTOR RELEASED BY MACROPHAGES
Authors: Melsom Haakon , Kearny Gilla , Gruca Stanislav , Seljelid Rolf
Published: June 2004

Mouse peritoneal macrophages cultivated in vitro acquire a strong extracellular cytotoxic activity towards isotope labeled syngeneic erythrocytes as demonstrated by isotope release to the medium. This lytic process is mediated by an extremely labile macrophage cytolytic factor (MCF) which is not detected under ordinary tissue culture conditions with serum present in the medium. By the use of serum-free medium containing low doses of 2-mercaptoethanol MCF is stabilized and found to be an easily dialysable, low molecular substance which resists heating at 60°C for 30 min.
BLOCKAGE OF AMYLOID INDUCTION BY COLCHICINE IN AN ANIMAL MODEL
Authors: Shirahama Tsuranobu , Cohen Alan S.
Published: June 2004

Colchicine was found to have a strong inhibitory effect on amyloid induction in an animal model. When CBA/J mice were treated with colchicine concurrently with the amyloid induction regimen, the incidence of amyloidosis was, depending upon the dosage of colchicine, significantly decreased (0.005–0.010 mg colchicine per day) or completely blocked (more than 0.015 mg colchicine per day). The colchicine treatment was effective not only when colchicine was given for the entire course of the amyloid induction regimen but also when it was given only in the late pre-amyloid or the amyloid phase of the regimen or to the recipients after the transfer of amyloid. The data suggest the colchicine is effective in blocking amyloidogenesis at its final stage(s), while it may not affect significantly amyloid already deposited in the tissue.
ANTIGENIC MODULATION IN VITRO
Authors: Stackpole Christopher W. , Jacobson Janet B. , Lardis Michael P.
Published: June 2004

The modulation or loss of thymus-leukemia (TL) antigenicity from the surfaces of mouse RADA1 leukemia cells and normal thymocytes during incubation with TL antibody in vitro at 37°C was investigated by cytotoxicity, immunofluorescence, and immunoelectron microscopy. The fate of bivalent and monovalent antibody during modulation was visualized by fluorescence microscopy. Considerable antibody remained bound to the cell surface after modulation, bivalent antibody being displaced topographically into "patches" and "caps" while monovalent antibody was only slightly aggregated on the cell surface. Some antibody was internalized, presumably by pinocytosis, and was sequestered into the Golgi region of the cell. Capping usually occurred over the pole of the cell opposite from the Golgi region, which may explain the lack of extensive pinocytosis of modulating bivalent antibody. Since modulation with monovalent antibody occurs without patch or cap formation, gross topographical redistribution of TL antigen-antibody complexes is not required for modulation, although more subtle displacement of these complexes may be involved.

Modulation was demonstrable by cytotoxicity with guinea pig C' but not with absorbed rabbit C', indicating that modulated TL antigens remain bound to the cell surface. A heat-labile factor in TL antiserum and in mouse serum in general is responsible for "blocking" the cytolytic interaction of guinea pig C' with modulated TL antigen-antibody complexes.
COMPARATIVE ANALYSIS OF ANTIGEN-BINDING T CELLS IN GENETIC HIGH AND LOW RESPONDER MICE
Authors: Hämmerling Günter J. , McDevitt Hugh O.
Published: June 2004

[125I](T,G)-A--L-binding T cells have been studied in mice whose ability to mount an immune response to (T,G)-A--L is under control of the H-2-linked Ir-1A gene. Nonimmunized high and low responder mice have approximately the same frequency of T-ABC. Following immunization, T-ABC proliferated only in high responders, but not in low responders, indicating expression of Ir-1A in T cells. When, for comparison, [125I]arsanyl-mouse serum albumin binding B and T cells were investigated in mice whose antibody response to the hapten arsanyl is controlled by an allotype-linked Ir gene, it was found that following immunization the number of B-ABC increased only in high responders. In contrast, T-ABC proliferated to the same extent in both high and low responders, suggesting exclusive expression of the allotype-linked Ir gene in the B-cell line. Preliminary studies indicate that anti-Ia sera inhibit neither B-ABC nor T-ABC.
ONTOGENY OF B-LYMPHOCYTE FUNCTION
Authors: Goidl Edmond A. , Siskind Gregory W.
Published: June 2004

The ontogeny of the ability of B lymphocytes to produce an antihapten response which is heterogeneous with respect to affinity for the antigenic determinant was studied in a cell transfer system. The heterogeneity of affinity of the immune response of lethally irradiated mice reconstituted with syngeneic, adult thymus cells and fetal or neonatal tissues as a source of B lymphocytes was studied. It was found that B cells from 17 day fetal liver or neonatal liver are highly restricted with respect to heterogeneity of affinity as compared with adult spleen or bone marrow. The B-cell population achieves an adult character with respect to heterogeneity of affinity by 2 wk of age. The peripheral lymphoid tissues (spleen) appear to mature in this respect more rapidly than do central lymphoid tissues (bone marrow). Spleens from 10-day old donors behave in an adult, heterogeneous manner while bone marrow from the same donors exhibit a marked restriction in heterogeneity of affinity. Germfree mice produce an immune response which is indistinguishable from conventionally reared adult animals with respect to heterogeneity of affinity.

The earlier appearance of the ability to transfer a heterogeneous immune response in spleen as compared with bone marrow suggests that the increasing heterogeneity of the B-lymphocyte population which occurs between birth and 2 wk of age is the result of a differentiation event and not of a somatic mutation or recombination event.
I-REGION GENES ARE EXPRESSED ON T AND B LYMPHOCYTES
Authors: Lonai Peter , McDevitt Hugh O.
Published: June 2004

Unidirectional mixed lymphocyte reactions (MLR) were performed between mouse strains differing for various segments within the H-2 complex. Thymocytes and purified lymph node T cells and B cells were used as stimulator cells. In three of five combinations studied, differing only within the I region, both T and B cells stimulated in the MLR. This suggests that the region codes for both T- and B-cell surface structures. However, if the difference was restricted to one I subregion (I-C), only T cells stimulated. This finding suggests that some of the I-region genes may be expressed either in T or in B cells.
FOURTH COMPONENT OF HUMAN COMPLEMENT: DESCRIPTION OF A THREE POLYPEPTIDE CHAIN STRUCTURE
Authors: Schreiber Robert D. , Müller-Eberhard Hans J.
Published: June 2004

The fourth component of human complement (C4) was shown to be composed of three distinct polypeptide chains linked by disulfide bonds and noncovalent forces. The sum of the molecular weights of the chains equalled that of the intact molecule. The mol wt of the α-, ß-, and γ-chains were respectively, 93,000, 78,000, and 33,000 daltons. Action of C1s on C4 affected only the α-chain, reducing its mol wt to 87,000 daltons. The size of the activation peptide. C4a, is therefore estimated to be 6,000 and that of the major fragment C4b, 198,000 daltons. Periodic acid-Schiff-stained SDS polyacrylamide gels of reduced C4 revealed carbohydrate to be associated with all three chains. A modification of the original method of isolation of C4 is presented.
ROLE OF H-2 LYMPHOCYTE-DEFINED AND SEROLOGICALLY-DEFINED COMPONENTS IN THE GENERATION OF CYTOTOXIC LYMPHOCYTES
Authors: Alter Barbara J. , Bach Fritz H.
Published: June 2004

The cell-mediated lympholytic capability of mouse spleen cells stimulated in mixed lymphocyte culture is determined by lymphocyte-defined (LD) and serologically-defined (SD) antigenic differences present during sensitization. Cells which are activated by LD differences alone are markedly less effective in causing lysis of target cells. This lack of cytotoxicity is shown to be, at least in part, due to the inability of LD differences to allow the efficient generation of cytotoxic lymphocytes. SD antigens not only serve as good targets for CML but are also shown to be important for the generation of cytotoxic lymphocytes during the mixed lymphocyte culture.