bluuebunny/crossref_metadata_2025 huggingface dataset:
Preparation of a Rabbit Anti-Guinea Pig Basophil Serum: in Vitro and in Vivo Characterization
Authors: Galli Stephen J. , Colvin Robert B. , Verderber Evie , Galli Anne S. , Monahan Rita , Dvorak Ann M. , Dvorak Harold F.
Published: December 2022

A high titer anti-basophil serum was prepared by immunizing rabbits with partially purified guinea pig peripheral blood basophils. Successive absorptions with normal guinea pig serum, red blood cells, polymorphonuclear leukocytes, and peritoneal-exudate macrophages and lymphocytes rendered the anti-basophil serum (ABS) reactive with basophils but no other leukocytes as determined by immunofluorescence, immunoferritin electron microscopy, and cytotoxicity assay. The basophil antigen(s) was expressed on the cell surface in close proximity to the plasma membrane and could be capped. ABS induced complement (C)-independent basophil degranulation and histamine release, but C was required for substantial basophil cytotoxicity and maximal degranulation and histamine release. ABS may recognize a basophil antigen(s) shared with tissue mast cells, because intradermal administration of small amounts of ABS produced local mast cell degranulation, reduction in skin histamine content, and an increase in vascular permeability that could be partially blocked by promethazine.

ABS was tested in vivo in guinea pigs sensitized both with ovalbumin for cutaneous basophil hypersensitivity (CBH) and with mycobacteria for classic delayed hypersensitivity (DH). Administration of 0.25 to 1.0 ml of ABS i.v. reduced circulating basophils to 30% control values (p < 0.05) at 2 hr and reduced the number of bone marrow basophils to 7% control values (p < 0.001) at 24 hr. Other marrow and circulating leukocytes were not significantly affected. ABS also markedly reduced the number of basophils infiltrating 24-hr CBH reactions as determined by histology and histamine content. Other leukocytes were not altered nor was the cellular composition of DH or Con A-induced reactions affected. Systemic administration of ABS had no appreciable effect on subepidermal mast cells: the number and morphology of subepidermal mast cells, and the histamine content of normal skin were the same in animals that had received either i.v. ABS or NRS 24 hr before sacrifice. The CBH reactions in ABS-treated animals were more intensely erythematous than those in NRS-treated controls (p = 0.015). ABS may prove useful in analyzing the basophil's role in inflammatory reactions.
Opposing Reactivities of Subpopulations of T Lymphocytes Toward Syngeneic Tumor Cells: Separation of Early Thymocytes
Authors: Small Myra
Published: December 2022

The present communication is a continuation of earlier studies which indicated that interaction between syngeneic tumors and those lymphocytes in the early stages of thymic processing can result in enhanced tumor growth in vivo. The thymocytes involved in this tumor enhancement were found previously in the rapidly dividing subpopulation of subcapsular cortical thymocytes, both in the untreated thymus and in the thymus undergoing repopulation after cortisone depletion. In the present experiments we have isolated this small subpopulation of early thymocytes. After cortisone injection such cells could be separated from the medullary cortisone-resistant thymocytes since the latter cells exhibit a high level of surface H-2 antigens and were thus lysed preferentially by anti-H-2 serum and complement. The repopulating subcapsular early thymocytes, which were resistant to this treatment, were incapable of responding to PHA while their basal proliferation rate was undiminished, and the majority of the cells were found to be dividing. When such low H-2 early thymocytes were injected together with three different tumors into syngeneic mice their tumor-enhancing activity was evident. It is clear that such early thymocytes are not devoid of biologic reactivity and their release from the thymus could have decisive results.
Separate Pathways of C Activation by Measles Virus Cytotoxic Antibodies: Subclass Analysis and Capacity of F(ab) Molecules to Activate C via the Alternative Pathway
Authors: Ehrnst Anneka
Published: December 2022

Commercially available IgG was fractionated into different subclasses a) by partial papain-digestion and exclusion chromatography, separating intact IgG2 and IgG4 from F(ab) and Fc of IgG1 and IgG3 and b) by passage through a staphylococcus protein A-coated Sepharose column to yield IgG3 separate from IgG1, IgG2, and IgG4. Measles antibody titers were analyzed by hemagglutination-inhibition (HI), hemolysis-inhibition (HLI), and by cytotoxicity assays. Diluted ordinary rabbit complement (C) was employed to exclude activation via the alternative pathway in studies of the classical pathway. C-4 deficient guinea pig C and rabbit C in ethyleneglycoltetraacetic acid (EGTA), and as a control, ethylenediaminetetraacetic acid (EDTA), were employed for studies of the alternative pathway. The discrepancy in that HI but not hemolysin antibodies activate C by the alternative pathway but both may activate C by the classical way was then further investigated.

Measles virus HI and HLI antibody activity per milligram protein was highest in fractions containing IgG1. The titers in the IgG3 and IgG2 ± 4 materials were low and might result from contamination by IgG1. This demonstrated a segregation of measles virus HI and HLI antibodies into IgG1 in normal individuals. Cytotoxicity by both pathways of complement activation was most pronounced in the presence of IgG1. Since both HI and hemolysin antibodies were mainly of the IgG1 subclass, their separate pathways of C activation could not depend on subclass differences. F(ab) fragments of human and rabbit IgG and human F(ab′) molecules, carrying measles virus HI activity, were as efficient in cytotoxicity tests via the alternative pathway as intact IgG with a corresponding HI titer. It is suggested that characteristics of the surface antigens might determine the pathway of C activation.
The Activation of Mononuclear Phagocytes: Fact, Fancy, and Future
Authors: Cohn Zanvil A.
Published: December 2022

Activation revisited. Since the time of Metchinkoff (1) and his contemporaries it has been clear that tissue macrophages exist in various states of activity. One reads about the resting-wandering cells of the connective tissue, arising from the mesenchyme and stimulated by the products of local injury. Such observations performed with the light microscope, vital dyes, and keen insight have in the past two decades been expanded with our increasing knowledge of the mononuclear phagocyte system (2).

In the recent past the term “activated macrophage” has largely been relegated to the bactericidal process and to the role of these cells in cell-mediated immunity. Unraveled largely through the imaginative in vivo studies of Mackaness and North and their colleagues (3, 4), the two cell nature of the process, the effector role of the macrophage, and the unique sensitizing qualities of viable organisms were elucidated. Yet, it is apparent when one scans the varied roles of tissue macrophages in the physiology and pathology of mammals that this is a much restricted definition.
Sodium Polyanethole Sulfonate: A New Macrophage-Dependent Polymyxin-Inhibitable, Polyclonal B Cell Activator
Authors: Smith C. I. Edvard , Hammarström Lennart
Published: December 2022

The anionic detergent sodium polyanethole sulfonate was found to activate mouse B lymphocytes to increased cell division and formation of plaque-forming cells, possibly by an indirect mechanism. An increased cell division was also obtained in guinea pig and in rabbit lymphoid cell cultures. A strong similarity was found when comparing the dextran sulfate- and the sodium polyanethole sulfonate-sensitive cell population. Thus, the responding mouse cells resided in spleen and lymph node and also in fetal liver and bone marrow. The sensitive lymphocytes were dependent on the presence of macrophages or macrophage-replacing factors, since depletion of phagocytic cells resulted in loss of response. Addition of polymyxin B inhibited the stimulation, possibly through a direct interaction with the polyanion. In potency sodium polyanethole sulfonate was found to surpass dextran sulfate, making this benzene sulfonate a useful tool when studying lymphocyte responses.
Possible Role of Macrophage Glycolipids as Receptors for Migration Inhibitory Factor (MIF)
Authors: Higgins Terry Jay , Sabatino Ann P. , Remold Heinz G. , David John R.
Published: December 2022

Guinea pig peritoneal exudate cells incubated with water soluble glycolipids obtained from macrophages show an enhanced response to migration inhibitory factor. Incorporation of these glycolipids into liposomes greatly facilitates their interaction with indicator cells. Enhancement of peritoneal exudate cell responsiveness to migration inhibitory factor was specific for glycolipids from guinea pig macrophages. Glycolipids extracted from guinea pig brain and polymorphonuclear leukocytes as well as several bovine and porcine glycolipids had no effect. Specificity of enhancement was not due merely to a preferential association of macrophage glycolipids with indicator cells. The possible role of macrophage glycolipids as receptors for MIF is discussed.
Immunologic Block against Antigen Absorption from Isolated Perfused Rabbit Lungs
Authors: Braley Janet F. , Dawson Christopher A. , Moore Vernon L.
Published: December 2022

In previous studies with isolated perfused rabbit lungs, we observed that inhaled human serum albumin (HSA) or ovalbumin (OA) entered the pulmonary circulation antigenically intact. The inhaled proteins were also metabolized in the lung. Immunization reduced the amount of intact protein and increased the amount of metabolites absorbed.

In the present study, we have begun to characterize the immune mechanisms responsible for reduced antigen absorption. A humoral immune mechanism appeared to be involved because the phenomenon could be passively transferred to normal animals by administering immune serum either 18 hr or immediately before antigen inhalation. The reduction was also observed when lungs from immunized rabbits were perfused with normal rabbit blood, indicating that antibodies in both lung and blood may be involved. Experiments, in which lungs from immunized rabbits were simultaneously insufflated with the immunizing antigen and with a nonspecific protein, demonstrated that the block against antigen absorption was specific for the immunizing antigen and was not due to some antigen-induced nonspecific changes in lung physiology.
Lymphocyte-Mediated Activation of Fibroblast Proliferation and Collagen Production
Authors: Wahl S. M. , Wahl L. M. , McCarthy J. B.
Published: December 2022

Among the many activities of antigen-triggered lymphocytes may be the control of fibroblast function. Thymus-dependent lymphocytes challenged with the specific antigen, dinitrophenylated ovalbumin or tetanus toxoid produced a nondialyzable factor(s) capable of causing dermal fibroblasts to undergo DNA synthesis. These fibroblasts, which exhibit basal proliferative levels in the absence of serum, responded to the lymphocyte factor with maximal thymidine incorporation at 48 to 72 hr. In addition, these activated fibroblasts significantly increased their production of protein of both the collagenous and noncollagenous types. This increase in protein synthesis preceded maximal proliferation. Thus, the fibroplasia consisting of increased numbers of fibroblasts and of increased collagen deposition associated with chronic inflammatory diseases may be the direct consequence of a specific antigenic challenge.
Sequential Precipitation of Mouse Thymocyte Extracts with Anti-Lyt-2 and Anti-Lyt-3 Sera
Authors: Durda Paul J. , Gottlieb Paul D.
Published: December 2022

Anti-Lyt-2.1 and anti-Lyt-3.1 sera were employed for sequential precipitation of NP-40 extracts of 125I-labeled C57BL/6-Lyt-2a, Lyt-3a thymocytes (Lyt-2.1, Lyt-3.1) to determine whether these alloantigenic determinants are present on the same or different molecular species. Treatment of extracts with anti-Lyt-3.1 serum and SaCI completely precipitated both Lyt-3.1 and Lyt-2.1-specific components, whereas treatment with anti-Lyt-2.1 serum reduced by approximately 37% the quantity of labeled species subsequently precipitable by anti-Lyt-3.1 serum. When 125I-labeled thymocytes were subjected to mild trypsinization before NP-40 extraction, the quantity of radioactive components precipitated by anti-Lyt-2.1 serum was essentially unchanged, but that of anti-Lyt-3.1-precipitable components was greatly reduced. Moreover, sequential precipitation of extracts of trypsinized thymocytes with anti-Lyt-2.1 and anti-Lyt-3.1 sera demonstrated that these molecular species were precipitated independently. Thus 1) Lyt-2.1 and Lyt-3.1 antigenic determinants appear to reside on different molecular species; 2) some Lyt-2.1- and Lyt-3.1-positive molecules appear to be complexed with each other in the NP-40 extract; and 3) this association of Lyt-2.1- and Lyt-3.1-positive species was dependent upon components that were labile to trypsinization of intact thymocytes.
Syngeneic Mixed Lymphocyte Reaction in Mice: Strain Distribution, Kinetics, Participating Cells, and Absence in NZB Mice
Authors: Bruce Smith J. , Pasternak Richard D.
Published: December 2022

Co-culture of mouse spleen nonadherent (T-enriched) cells with mitomycin C-treated unfractionated syngeneic spleen cells resulted in increased DNA synthesis in the responding T cells. The kinetics of this syngeneic mixed lymphocyte reaction (SMLR) showed that peak DNA synthesis occurred on day 5 of culture compared to day 4 for conventional mixed lymphocyte reaction (MLR). Anti-T cell antiserum plus complement treatment of the responding cell population abolished the reaction, and similar treatment of the stimulator population enhanced SMLR. These studies indicate that SMLR represents the response of T cells to non-T cells. Studies on the generation of cytotoxic T lymphocytes (CTL) in parallel cultures of T cells activated by syngeneic or allogeneic spleen cells showed no cytotoxicity of SMLR-activated cells for either PHA- or LPS-induced blasts but did show a good CTL response of allo-activated cells to both targets. Studies on the strain distribution of SMLR revealed that NZB mice manifested poor or no stimulation in SMLR whereas all other strains tested exhibited strong SMLR. This defect in NZB mice may be pathogenetically related to the autoimmune disease that develops in these mice.
Nonspecific Activation of Murine Lymphocytes
Authors: Goodman Michael G. , Fidler John M. , Weigle William O.
Published: December 2022

The lymphocyte subpopulations that are activated by 2-ME, LPS, poly IC, and PPD were studied in terms of their maturational characteristics. Attempts to stimulate hepatic and splenic lymphoid cells from mice of different ages with these mitogens demonstrated a well ordered sequence for the emergence of mitogen responsiveness in C3H mice: reactivity to LPS and Poly IC was observed early in maturation and was followed by that to PPD, and finally by the development of responsiveness to 2-ME. The same sequence appeared when the mitogen responsiveness of lethally irradiated, fetal liver-reconstituted syngeneic adult recipients was examined. The mitogenic action of 2-ME was dissociated from its ability to enhance lymphocyte reactivity to other mitogens in mice too young to respond to 2-ME as a mitogen. Experiments in which additivity of responses was assayed by adding mitogens to culture singly or conjointly indicated that LPS and Poly IC activate nearly identical B lymphocyte subpopulations, whereas PPD stimulates a subset of cells distinct from that which is responsive to the former two mitogens. The mitogen responsiveness of CBA/N mice, relative to normal CBA/WEHI mice, was shown to decrease as a function of the maturity of the subpopulation of lymphocytes activated. The CBA/N mouse was shown to be unresponsive to stimulation by 2-ME.
Enhancement of T Cell Cytotoxic Responses by Purified Human Fibroblast Interferon
Authors: Zarling Joyce M. , Sosman Jeffrey , Eskra Linda , Borden Ernest C. , Horoszewicz Julius S. , Carter William A.
Published: December 2022

Purified polyribonucleotide-induced human fibroblast interferon (HFIF) was tested for its effects on proliferative and cytotoxic human T cell responses to alloantigens. The addition of HFIF (100 to 400 IFU/ml) to mixed leukocyte cultures decreased alloantigen-induced lymphocyte proliferative responses as determined by both recovery of responding cells and by 3H-thymidine incorporation into responding cells. However, HFIF, but not the mock interferon preparation, increased the cytotoxic response of T cells to allogeneic cells by 4- to 5-fold when expressed in terms of lytic units. Although fibroblast and leukocyte interferons have different physicochemical and biologic properties, the results reported here are in concert with previous findings concerning the effects of virus-induced leukocyte interferon on human T cell functions.
Opinion
Authors: Frangione Blas , Franklin Edward C.
Published: December 2022

In 1965, the “two gene-one polypeptide” theory for the synthesis of antibody chains was first proposed (1). Since then, amino acid sequence studies and genetic analyses of the variable (V)2 and constant (C) regions of light (L) and heavy (H) chains have been consistent with this hypothesis (2). A direct demonstration of the existence of separate genetic units was achieved in 1977 (3), when it was shown that the DNA segments (exon DNA) coding for V and C regions of mouse L chains (λ type) are separated from one another by a large DNA segment (intron or intervening sequence or IVS) that is missing from the mRNA that serves as the template for the L chain. In addition, in DNA derived from embryonic cells, the V gene is shorter than expected (4, 5), ending at the codon specifying position 98 instead of 112 commonly accepted at the VC joining region.
Tryptic Degradation of the CH1 and VL Regions of IgD and IgE
Authors: Kocher Hans P. , Spiegelberg Hans L.
Published: December 2022

Fab fragments of all human Ig classes were digested with trypsin and analyzed by SDS-PAGE for degradation products. Fab fragments of all four IgG subclasses, IgM and IgA1 were resistant whereas IgD and IgE Fab fragments were degraded into smaller fragments. IgE Fab fragments had already become degraded during the papain digestion used to produce them. The degradations of IgD and IgE Fab fragments were independent of κ and λ light chain types. After prolonged tryptic digestion of a λ IgD Fab fragment, three major fragments remained, which were isolated and characterized by partial amino acid sequence analysis. The fragments consisted of 1) λ-chains, 2) the constant region of the λ-chains minus the first three amino acids, and 3) the variable region of the δ-chain including the first four amino acids of the Cδ1 region. In contrast to the Fab fragments, IgD Fc fragments and isolated λ-chains were not degraded into small fragments. The data indicate that the first constant regions of IgD and IgE, but not IgG, IgM, and IgA, are highly susceptible to enzymatic degradation and that association of the light chains with δ- and ε-chains renders the VL regions susceptible to enzymatic attack. Since it is unlikely that the CH1 regions of δ- and ε-chains change the tertiary structure of the VL regions, it is suggested that monomeric light chains are released after digestion of the CH1 regions and that the VL region of these monomers is degraded.
Cell-Mediated Immune Response to Syngeneic UV-Induced Tumors
Authors: Woodward Jerold G. , Fernandez Paul A. , Daynes Raymond A.
Published: December 2022

An Ia-positive accessory cell with macrophage characteristics was demonstrated to be required for the in vitro proliferation and differentiation of lymph node T lymphocytes from tumor-immunized mice into cytotoxic T lymphocytes (CTL). An experimental system is described that provides for the measurement of both proliferation and CTL capacity in in vitro cultures of draining lymph node (DLN) cells from tumor-immunized mice. DLN cells obtained from the popliteal lymph nodes of mice immunized in the hind footpads with a syngeneic UV-induced fibrosarcoma proliferate and differentiate into highly effective CTL (as assessed in a 6-hr chromium release assay) when placed in culture without the addition of tumor antigen. Both proliferation and CTL generation by DLN cells in vitro were shown to result from the interaction of at least two physically separable cell populations: a T cell population responsible for the proliferation and expression of CTL activity, and an accessory cell population required for the expression of the T cell functions. The accessory cell was shown to be resistant to γ-radiation, adherent to nylon and glass wool columns, sensitive to treatment with anti-Ia antisera plus complement (C), and resistant to treatment with anti Thy-1 antisera plus C. Antigen presentation did not appear to be required for effective cooperation between lymphocytes from tumor-immunized mice and accessory cells, although antigen presentation did occur when unprimed lymphocytes were mixed with accessory cells present in the DLN of tumor-immunized mice. The regulation of CTL development and the role of the Ia-positive accessory cell in anti-tumor immunity are discussed.
In Vitro Differentiation of B Lymphocytes from Pre-B Cells
Authors: Lau Catherine Y. , Melchers Fritz , Miller Richard G. , Phillips Robert A.
Published: December 2022

Adult bone marrow contains both B lymphocytes and their immediate precursors, pre-B cells. These two cells differ in size and can be separated by velocity sedimentation; B cells are enriched in the subpopulation of cells sedimenting at between 2.0 and 3.5 mm/hr and pre-B cells in the subpopulation between 5.0 and 7.0 mm/hr. Incubation of pre-B cells in vitro for 4 or 5 days leads to their differentiation into functional B lymphocytes. The transition from pre-B to B appears to occur in two steps. The first step gives mitogen responsive B cells with an intermediate sedimentation velocity and the second step produces typical small, slowly sedimenting B cells. pre-B cells can be quantified by using a limiting dilution assay and occur at a frequency of 1/60 in the subpopulation of rapidly sedimenting bone marrow cells.
Polyclonal Immunoglobulin Secretion by Human B Lymphocytes Exposed to Epstein-Barr Virus in Vitro
Authors: Kirchner Holger , Tosato Giovanna , Michael Blaese R. , Broder Samuel , Magrath Ian T.
Published: December 2022

Epstein-Barr virus (EBV)-induced activation of human peripheral blood lymphocytes was studied by the use of a reverse hemolytic plaque assay (RHPA) for the detection of immunoglobulin-producing cells. The results were compared with the effects of pokeweed mitogen (PWM) on the same cell population. Both agents caused the development of immunoglobulin-producing cells in cultures of unseparated mononuclear cells. However, B cell populations sufficiently depleted of T cells by a variety of techniques to be unresponsive to PWM showed a marked response to EBV. The reactivity of B cells to PWM could be restored by irradiated T cells, whereas there was no effect of irradiated T cells on reactivity to EBV. These data suggest that the response to EBV in contrast to the PWM response is T cell independent. Lymphocytes secreting each class of immunoglobulin (IgG, IgA, and IgM) were found in EBV-stimulated cultures of both unseparated mononuclear cells and T cell-depleted cultures, demonstrating that the response in each immunoglobulin class is also T cell independent in this system. When unseparated cell populations and B cell populations cultured at the same cell concentration were compared, the latter showed a 2- to 5-fold increased reactivity to EBV. This difference appeared to be caused primarily by an enrichment of B cells as was suggested by experiments in which the two cell populations were compared at different cell concentrations.
T100: A New Murine Cell Surface Glycoprotein Detected by Anti-Lyt-2.1 Serum
Authors: Durda Paul J. , Boos Stephen C. , Gottlieb Paul D.
Published: December 2022

A cell surface glycoprotein (designated T100) of apparent m.w. 100,000 by SDS-PAGE under reducing and non-reducing conditions was precipitated from NP-40 extracts of surface radiolabeled thymocytes from a variety of inbred strains of mice by the standard noncongenic Lyt-2.1-typing serum. The inbred strain distribution, trypsin sensitivity on intact cells, and apparent m.w. of T100 suggest that it is different from Lyt-2.1. Inheritance and expression of T100 suggest that it is determined by an allele at a single genetic locus, and testing of CXB recombinant inbred strains and B6.C minor histocompatibility congenic strains suggest that this locus is linked to H-25. Antiserum absorption experiments, two-stage cytotoxicity assays, and results of immunoprecipitations performed after prebinding antibody to radio-labeled thymocytes suggest that some T100 is accessible to antibody on the intact cell surface. However, for unknown reasons the number of cells required to absorb anti-T100 precipitating activity from antiserum was much higher than for removal of anti-Lyt-2.1 activity. A molecule with properties of T100 was also detected on lymph node cells and on the AKTB-1 lymphoma.
Immune Responses During Human Schistosomiasis Mansoni
Authors: Colley Daniel G. , Todd Charles W. , Lewis Fred A. , Goodgame Richard W.
Published: December 2022

Simultaneous in vitro exposure of human peripheral blood mononuclear cells to phytohemagglutinin-P (PHA) and either soluble schistosomal egg antigenic preparation (SEA) or soluble cercarial antigenic preparation (CAP) obtained from Schistosoma mansoni resulted in decreased responsiveness as compared to exposure to PHA alone. The addition of a soluble adult worm antigenic preparation (SWAP) did not predictably alter PHA responses in this system. The suppression due to in vitro exposure to either SEA or CAP was expressed whether the lymphocyte donors were S. mansoni patients (early infection, chronic, or treated), or uninfected subjects. The degree of suppression was related to the concentration of SEA used, and the timing of exposure. Preexposure to SEA for 3 days before the addition of PHA resulted in more potent suppression. However, a delay in the time of the addition of SEA of 6 and 24 hr after PHA exposure decreased and eliminated, respectively, its suppressive capacity. SEA and CAP were not directly toxic to responding cells, and appeared to exert their nonspecific suppressive influences through T lymphocyte-related mechanisms. It was observed that although these suppressive events could be induced and observed in vitro, the responsiveness of S. mansoni patient lymphocytes to PHA was equal with that of uninfected controls.
Reduced Responsiveness of Immature B Cells to the B Cell Mitogen, Lipoprotein
Authors: Phillips Robert A. , Melchers Fritz
Published: December 2022

When pre-B cells are isolated from bone marrow and incubated in vitro for several days, they spontaneously differentiate into LPS-reactive B cells. These newly produced B cells do not respond to another B cell mitogen, lipoprotein (Lp). The lack of Lp reactivity of newly produced B cells represents a qualitative difference between these cells and B cells in peripheral tissues that respond equally well to both LPS and Lp. Thus, responsiveness to different mitogens provides unique markers to identify B cells at early stages of development.
Hapten-Specific IgE Antibody Responses in Mice
Authors: Chiorazzi Nicholas , Tung Amar S. , Katz David H.
Published: December 2022

These studies were conducted to investigate the effects of various adjuvants on antibody responses of the IgE and IgG classes in mouse strains of the high IgE response phenotype. Administration of either complete or incomplete Freund's adjuvant or aluminum hydroxide gel (alum) at appropriate times was found to generate a nonspecific suppression of antibody responses of both the IgE and IgG classes. Such “adjuvant-generated suppression” could be induced both in intact animals and could be passively transferred with cells from adjuvant-pretreated donors. It was shown that T cells were essential for the suppression, since pretreatment of spleen cells from adjuvant-inoculated mice with anti-ϑ serum plus complement abolished the suppressive effects of such transferred cells. Moreover, the generation of these suppressor T cells could be inhibited by pre-exposing donor animals to low doses of x-irradiation at an appropriate time before adjuvant administration. On the other hand, once suppressor cells have been induced by inoculation with complete Freund's adjuvant, x-irradiation had little effect in diminishing the suppressive activity of such cells. The relevance of adjuvant administration to induce suppressor mechanisms for both IgE and IgG antibody responses to regulatory control mechanisms of the immune response is discussed.
Studies of the Mechanisms for the Induction of in Vivo Tumor Immunity
Authors: Ting Chou-Chik , Rodrigues Dennis , Igarashi Tsuyoshi
Published: December 2022

In studying the cell-mediated immunity to FBL-3, a Friend virus-induced leukemia syngeneic to C57BL/6 mice, it was found that the levels of cytotoxic response induced by the in vivo adoptive transfer experiments were always considerably higher than those induced by the in vitro mixed lymphocyte tumor cell culture (MLTC). This was especially true in using lymph node cells as responders. Further investigation showed that this difference was not due to the generation of suppressor cells in vitro or to the lack of precursor killer cells in lymph nodes. During in vivo sensitization, it appeared that helper cells were recruited by T cells to augment the cell-mediated cytotoxic response. In the in vivo experiments, pretreatment of the recipient hosts with antimacrophage agents (silica or carrageenan) caused significant reduction of cytotoxic response induced in the adoptive transfer experiments; the effect was more pronounced on the adoptive transfer of lymph node cells than spleen cells, indicating that macrophages are needed for generating an efficient in vivo T cell-mediated immunity. In the in vitro experiments, it was found that the peritoneal cells or spleen cells from normal x-irradiated syngeneic mice could also augment the MLTC reactions, thus, these helper cells were presumably macrophages. The lack of sufficient supply of exogenous macrophages could be one of the major cause of the low level of cytotoxic response generated by in vitro sensitization. This is especially true for the lymph node cells that have more stringent dependency on the presence of excess amounts of macrophages. These results indicate that recruitment of macrophage-like helper cells may play an important role in the induction of an efficient in vivo tumor immunity.
Onco-Neural Antigen: A New Neural Differentiation Antigen Expressed by Neuroblastoma, Oat Cell Carcinoma, Wilms' Tumor, and Sarcoma Cells
Authors: Seeger Robert C. , Zeltzer Paul M. , Rayner Sylvia A.
Published: December 2022

This investigation defined a new neural differentiation antigen that is expressed by a number of tumors derived from neuroectoderm and surprisingly by some derived from mesoderm. Heteroantisera were prepared by immunizing rabbits with cultured human neuroblastoma cells followed by extensive absorption with nine different types of normal non-neural tissues and cells; sera were evaluated with cultured cells and radioiodinated staphylococcal protein A to detect binding of antibodies to cell surface antigens (CSA). One antiserum that was raised against LA-N-1 neuroblastoma cells was analyzed in detail. It reacted well with seven neuroblastoma cell lines including LA-N-1; it also reacted, but to a lesser degree, with medulloblastoma, rhabdomyosarcoma, and leiomyosarcoma cell lines. Seventeen additional cell lines and noncultured acute leukemia cells were minimally reactive or nonreactive with the serum. Absorption of the serum with homogenates of adult brain or adrenal reduced its binding to neuroblastoma cells more than 80%. Greater than 65% of the reactivity with LA-N-1 neuroblastoma cells also was absorbed by homogenates of 10 of 10 neuroblastomas, seven of seven Wilms' tumors, five of eight sarcomas, two of three oat cell carcinomas, and one of six melanomas. Eight other types of malignancies did not remove significant activity compared to control non-neural normal tissue absorbents. Sequential absorption with adult brain or adrenal followed by the positive tumors demonstrated that both normal and tumor cells expressed the same antigen. We term the neural differentiation antigen(s) that is expressed by tumors derived from neuroectoderm and mesoderm an onco-neural antigen (ONA).
In Situ Lymphoid Cells of Mouse Mammary Tumors
Authors: Ruppert Barbara , Blazar Beverly , Medina Daniel , Heppner Gloria
Published: December 2022

Lymphoid cells isolated from several types of mouse mammary tumors are capable of stimulating tumor cell growth or survival in MCT assays. Lymph node and spleen cells of mice bearing such a tumor are specifically cytotoxic to the tumor cells. Surgical removal of the tumor is followed in 4 to 7 days by the appearance of stimulatory capacity in spleens and lymph nodes. By day 10, cytotoxic cells specific for the sensitizing tumor are again detected. These reach a peak on day 13. By day 17 no reactivity is detectable. The functional distribution of tumor-reactive lymphoid cells is different between tumor masses and peripheral lymphoid organs.
In Vitro Studies on Lymphocyte Differentiation
Authors: Schrader John W. , Goldschneider Irving , Bollum Fred J. , Schrader Sabariah
Published: December 2022

The enzyme TdT is the earliest known marker of lymphocytic differentiation in rodents. Cells containing this enzyme were demonstrated in suspension cultures of mouse bone marrow cells that had been maintained in vitro for periods of 7 to 45 days. The cells were detected by immunofluorescence using purified antibodies to homogeneous TdT. Between 0.04 and 2.0% of cultured bone marrow cells from a variety of mouse strains were positive. More than 40% of the TdT-positive cells incorporated 3H-thymidine during a 20-min pulse. Surface Ig and Thy-1 antigens were not detected on the TdT-positive cells. The prevalence of TdT-positive cells was decreased 10-fold in cultures that had been treated with 10-6 M hydrocortisone 24 hr before harvesting. The results indicate that lymphoid progenitor cells can be generated in vitro.
Variations in Expression of Xenotropic Murine Leukemia Virus Genomes in Lymphoid Tissues of NZB Mice
Authors: Morse Herbert C. , Chused Thomas M. , Sharrow Susan O. , Hartley Janet W.
Published: December 2022

Lymphocytes from Thy, Sp, LN, and BM of NZB mice were tested for expression of X-MuLV genomes as cell surface gp70 (XenCSA) or infectious virus. The results demonstrate major dissociations for these parameters of X-MuLV expression in different lymphoid compartments and suggest that factors involved in T lymphocyte differentiation modify the levels of expression in these two modes.
Binding Studies with Antibodies Having Phosphorylcholine Specificity and Fragments Derived from Their Homologous Streptococcus Pneumoniae Type 27 Capsular Polysaccharide
Authors: Bennett Larry G. , Glaudemans Cornelis P. J.
Published: December 2022

The capsular polysaccharide from Streptococcus pneumoniae type 27 (S-27) has been fragmented into oligosaccharides by controlled degradations. Binding of these derived small ligands with heterogeneous rabbit anti-S-27 antibody of selected phosphorylcholine affinity indicates that a major immunodeterminant of S-27 is the phosphorylcholine moiety, but that these anti-P.C. antibodies are also capable of binding to the polysaccharide backbone from which the P.C. has been removed.
Purine Nucleoside Phosphorylase Deficiency: A Molecular Model for Selective Loss of T Cell Function
Authors: Ochs Ute H. , Chen Shi-Han , Ochs Hans D. , Osborne William R. A. , Scott C. Ronald
Published: December 2022

Absence of purine nucleoside phosphorylase (NP) is associated with severe T cell immune deficiency and normal B cell function. Patients with this enzyme defect accumulate inosine, guanosine, and their respective deoxycompounds, all of which are substrates for NP. We have evaluated the effect of these four NP substrates on PHA-stimulated lymphocytes and lymphoblastoid cell lines with B and T cell characteristics. Inosine and deoxyinosine had little inhibitory effect on human lymphocytes, whereas guanosine and deoxyguanosine inhibited DNA and protein synthesis in both PHA-stimulated human lymphocytes and hyman lymphoblastoid cells. In all experiments, deoxyguanosine was more toxic than guanosine.

Human lymphoblastoid cells with T cell characteristics (T-LCL) were found to be particularly sensitive to the presence of deoxyguanosine. At low µM concentrations 3H-thymidine and 3H-leucine incorporation into the T-LCL was markedly decreased. At a concentration of 10 µM, no cell growth occurred and 50% of the cells were killed. Partial reversal of the toxic effect of deoxyguanosine on the T-LCL could be achieved with simultaneous addition of deoxycytidine. Human lymphoblastoid cells with B cell characteristics (B-LCL) were not inhibited at these low deoxyguanosine levels but cell growth and incorporation of 3H-thymidine and 3H-leucine were moderately decreased at higher deoxyguanosine concentrations. In a B-LCL established from a patient with NP deficiency, neither DNA nor protein synthesis was inhibited by guanosine or deoxyguanosine at concentrations that affected other B cell lines.

The extreme sensitivity of the T lymphoblasts and relative resistance of B lymphoblasts to the toxicity of deoxyguanosine or one of its metabolic products may explain the selective loss of T cell function in patients with NP deficiency.
Genetic Control of the IgG2a Response to Sheep Erythrocytes in Mice: Isotype- and Antigen-Specific T Cell-Mediated Suppression in Low Responders
Authors: Seman Michel , Zilberfarb Vladimir
Published: December 2022

The IgG2a response to sheep erythrocytes is examined in different congenic strains of mice. B10, B6, and C57BL/Ks animals produce a low level of IgG2a antibodies to SRBC during the primary response in vivo. They remain low responders after secondary challenge in vitro. Total spleen cells or nylon-purified T cells from these low responders inhibit the IgG2a response of H-2 compatible-responding mice in a mixed culture system. This suppression is mediated by Thy-1+, Ly-1-, Ly-2+, and I-J+ T cells only present in the spleen of low responding animals. These suppressor T cells appear to be IgG2a-and SRBC-specific. Function of non-H-2-linked genes as regulators of suppressor T cells differentiation is discussed.
Natural Killing of Measles-Infected Cells by Human Lymphocytes
Authors: Ault Kenneth A. , Weiner Howard L.
Published: December 2022

HeLa cells persistently infected with measles virus (M-HeLa) show markedly increased susceptibility to natural killing (NK) by normal human blood lymphocytes compared to uninfected HeLa cells. The killing is mediated by the same population of non-B, non-T lymphocytes with avid Fc receptors for IgG that mediate other forms of NK and antibody-dependent cellular cytotoxicity (ADCC). There is no evidence that this NK is mediated by antibody; specifically, the killing regenerates after being abolished by extensive treatment of the effector cells with trypsin or pronase, and it cannot be inhibited by large amounts of F(ab′)2 fragments of anti-measles antibody or by a large excess of normal γ-globulins. In addition, NK of M-HeLa is strongly correlated (r = 0.79, p < 0.001) with NK of an unrelated cell line (K-562) that does not contain measles virus and is a standard target for NK. The strong correlation between NK of M-HeLa and K-562 is confirmed by cold-target inhibition studies showing cross-competition between the two cell lines. Killing cannot be enhanced by addition of supernatants from lymphocytes cultured with M-HeLa to fresh lymphocytes cultured with M-HeLa, HeLa, K-562, or other cell lines, making it unlikely that interferon or other soluble factors have a role in this system. These studies suggest that persistent infection of HeLa cells with measles virus results in the modification of the HeLa cell membrane and that this modification is similar to some structure on K-562 that is important for NK. In addition, these studies raise the possibility that NK could play a role in host defenses against measles virus infection.
Serologically Detectable Human Melanoma-Associated Antigens Are Not Genetically Linked to HLA-A and B Antigens
Authors: Curry Russell A. , Quaranta Vito , Pellegrino Michele A. , Ferrone Soldano
Published: December 2022

Serologic, structural and immunogenic data accumulated in animal model systems suggest that some tumor-specific antigens are newly expressed or modified histocompatibility antigens (see for review 1). This evidence raises the possibility that in man some tumor-associated antigens may derive from histocompatibility (HLA) antigens. We tested this hypothesis in the human melanoma system by investigating whether human chromosome 6, which carries the major histocompatibility complex region (2, 3), controls the expression of serologically detectable melanoma-associated antigens (sd-MAA). An absence of genetic linkage between sd-MAA and HLA antigens would cast serious doubts on the proposition that sd-MAA are modified HLA antigens. In our studies, we prepared hybrid clones by fusing cultured human melanoma cells with murine fibroblasts. We tested these hybrids for expression of sd-MAA and HLA-A and B antigens after a number of passages in vitro, during which a preferential loss of human chromosomes resulted in cells with a few human chromosomes superimposed on essentially complete murine genomes.
Characterization of Human Breast Milk Macrophages Cytostatic for Human Cell Lines
Authors: Balkwill Frances R. , Hogg Nancy
Published: December 2022

Approximately 40% of the cells in human early lactation milk possessed the characteristics of macrophages, being adherent, phagocytic, α napthyl acetate esterase positive, and possessing C3b and Fc receptors. These cells were also cytostatic for MDA 157, a cell line derived from a pleural effusion of human breast cancer, and HEL 23, a strain of human fetal lung fibroblasts. Investigations into the cytostatic phenomenon indicated that the cytostasis could not be mediated by macrophage-conditioned medium and that very close contact between breast milk macrophages and target cells was required. Cytostasis was not fully effective until 18 hr after the initial interaction between macrophages and target cells.
Augementation of Antigen-Stimulated Histamine Release from Human Basophils by Serum-Treated Zymosan Particles
Authors: Thomas Larry L. , Lichtenstein Lawrence M.
Published: December 2022

Although antigen-stimulated histamine release from human basophils is well characterized, the response of basophils to other stimuli is less well studied. Complement-coated particles are prominent among the stimuli generated during inflammatory reactions; accordingly, the effect of serum-treated zymosan particles (Zx) on histamine release from human basophils was examined. Although Zx alone failed to stimulate histamine release from basophil-containing mononuclear cells of allergic and nonallergic individuals. Zx consistently augmented antigen or anti-IgE stimulated release. The percent enhancement ranged from approximately 20 to 100%, depending on the level of antigen-stimulated release, and occurred in a concentration-dependent fashion. The kinetics of augmentation were extremely rapid, with augmented release evident 1 min after addition of the stimuli. Serum requirements for preparation of Zx were consistent with involvement of C3b in the enhancement. Failure to demonstrate participation of a soluble component(s) indicated that Zx enhanced release by a direct effect, presumably mediated by a C3b receptor, on the basophil membrane. It is suggested that occurrence of a similar enhancement in vivo may contribute to an amplification of allergic reactions that are superimposed upon an inflammatory state.
The Nature of the Stimulatory Cell in Human Allogeneic and Autologous MLC Reactions; Role of Isolated IgM-Bearing B Cells
Authors: Gottlieb Alice Bendix , Fu Shu Man , Yu David T. Y. , Wang Chang Yi , Halper James P. , Kunkel Henry G.
Published: December 2022

Highly purified surface IgM-bearing B cells were isolated from T cell- and monocyte-depleted mononuclear cells by their ability to rosette with purified anti-IgM antibodies covalently linked to bovine red blood cells. Surface IgM-bearing B cells from both peripheral blood and tonsil were found to be excellent stimulator cells in unidirectional MLC reactions. In the majority of cases, the surface IgM-bearing B cells had a higher stimulatory capacity in the allogeneic MLC than non-T cells lacking surface IgM. Monocytes were also demonstrated to be good stimulators in the allogeneic MLC when a low number of cells was used. At high numbers, inhibition of tritiated thymidine uptake was observed, which appeared to be partly due to the release of prostaglandins and cold thymidine.

The population of non-T cells lacking surface IgM also showed stimulation that was especially strong in certain individuals. Because of the heterogeneity of this fraction, the primary cell involved could not be determined. It is of interest that this population was a more potent stimulator in the autologous MLC than was the surface IgM-bearing B cell fraction, although the latter was clearly active. Results from these studies also indicated that monocytes were relatively poor stimulators in the autologous MLC. Isolated Fab′ fragments of a rabbit anti-human Ia antiserum caused significant inhibition of both the allogeneic and autologous MLC.
A New Mouse Myeloma Cell Line that Has Lost Immunoglobulin Expression but Permits the Construction of Antibody-Secreting Hybrid Cell Lines
Authors: Kearney John F. , Radbruch Andreas , Liesegang Bernhard , Rajewsky Klaus
Published: December 2022

We have isolated a subclone of the mouse myeloma cell line P3-X63-Ag8 that does not express immunoglobulin heavy or light chains. This clone X63-Ag8.653 can be used for efficient fusion with antibody-forming cells to obtain hybrid cell lines producing pure monoclonal antibodies. Screening of hybrid cell lines for specificity and immunoglobulin classes was done with a modified enzyme-linked immunosorbent assay.
Differential Activation of Cytotoxic and Suppressor T Cells against Syngeneic Tumors in the Mouse
Authors: Yamauchi Katsumi , Fujimoto Shigeyoshi , Tada Tomio
Published: December 2022

The cytotoxic T cell against a methylcholanthrene-induced sarcoma, S1509a, was induced in syngeneic mice by deliberate immunization with mitomycin C (MMC)-treated live tumor cells. The soluble tumor antigen (STA) extracted from the same tumor by 3 M KCl was, however, unable to induce the cytotoxic T cell upon immunization, although it was able to activate predominantly the suppressor T cell that then specifically suppressed the effect of the cytotoxic T cell against the homologous tumor. The suppressor T cell generated by STA had the same characteristics as those found in tumor-bearing animals: 1) The suppressor T cell has a very strict specificity against individual tumors; 2) The cell expresses cell surface determinants controlled by genes in the I-J subregion of the mouse H-2 complex.

The activity of the cytotoxic T cell was completely inhibited by live tumor cells but not by STA, whereas that of the suppressor T cell was neutralized by STA. The results that cytotoxic and suppressor T cells are activated under different conditions, and that the antigenic determinants recognizable by these two cell types are not the same. The soluble extract contains only the determinants recognizable by the suppressor T cell, and the cytotoxic T cell can be activated only by the determinants associated with self antigen present on the surface of live tumor cells.