Showing papers on "Antigen published in 1978"

T Cell Growth Factor: Parameters of Production and a Quantitative Microassay for Activity

Abstract: Several soluble factors have recently been associated with the proliferation and differentiation of thymus-derived lymphocytes. One of these factors present in medium conditioned by T cell mitogen-stimulated lymphocytes has the ability to promote the long-term culture of normal and antigen-specific cytotoxic T cells. We report a method to test for this proliferative stimulus in the form of a sensitive microassay based upon the tritiated thymidine incorporation of continuous murine tumorspecific cytotoxic T cell lines (CTLL). The microassay requires microliter quantities of sample fluid and is amenable to quantitative analysis. This highly reproducible, quantitative assay for T cell growth factor (TCGF) has allowed investigation as to the kinetics of TCGF generation and has revealed that T lymphocytes are required for its production. Further investigation has supported the notion that this nonspecies-specific factor is actively removed from tissue culture medium by the proliferation of either T cell mitogen-activated lymphocytes or CTLL.

The lectins : carbohydrate-binding proteins of plants and animals

Abstract: Publisher Summary Lectins play an important role in the development of immunology. Lectins also find application in serological laboratories for typing blood and determining secretor status, separating leucocytes from erythrocytes, and agglutinating cells from blood in the preparation of plasma. They serve as reagents for the detection, isolation, and characterization of carbohydrate-containing macromolecules, including blood-group antigens. In their interaction with saccharides, lectins serve as models for carbohydrate-specific antibodies, with the important advantage to purify lectins in gram quantities. Lectins are classified according to their carbohydrate-binding specificity that includes D-mannose(D-glucose)-binding lectins and 2-acetamido-2-deoxy-D-glucose-binding lectins. The chapter considers only those lectins that have been purified to homogeneity, and studied with regard to their biophysical, biochemical, and carbohydrate-binding specificity. The chapter also describes the cell-binding and biological properties of lectins. The chapter concludes with the description of several glycopeptide structures showing the carbohydrate-binding loci with which various lectins interact.

Monoclonal antibody defining a stage-specific mouse embryonic antigen (SSEA-1).

Abstract: A monoclonal antibody derived by fusion of mouse myeloma cells with spleen cells from a mouse immunized with F9 teratocarcinoma cells is described. This antibody, which reacts with embryonal carcinoma cells of mouse and human origin and with some preimplantation stage mouse embryos, defines an embryonic stage-specific antigen. This stage-specific antigen (SSEA-1) is first detected on blastomeres of 8-cell stage embryos. Trophectodermal cells are transitorily positive; however, each cell in the inner cell mass eventually expresses this antigen.

Natural Cell-Mediated Immunity

Abstract: Publisher Summary The recognition of the existence of natural cell-mediated immunity, particularly natural cell-mediated cytotoxicity, has significantly altered the concepts concerning the potential mechanisms for in vivo resistance against tumor growth and for in vitro cell-mediated immune reactions. When cytotoxic reactions are measured, the evaluation of the role of natural killer (NK) cells and other more well-known mechanisms of cytotoxicity must be done carefully. This chapter focuses on natural cell-mediated cytotoxicity. It summarizes the known information on the expression of natural cytotoxicity in rodents and in man, its specificity, the nature of the effector cells and their relationship to other immune mechanisms, and the possible in vivo relevance of natural cytotoxicity, particularly in regard to resistance against tumor growth. The basic observation that initiated the studies of natural cell mediated cytotoxicity includes that lymphoid cells from some normal mice, rats, and human donors, which are not inoculated with tumor cells or other sources of antigen, exhibit significant levels of cytotoxic reactivity against certain syngeneic or allogeneic tumor cells.

Microdroplet testing for HLA-A, -B, -C, and -D antigens. The Phillip Levine Award Lecture.

Abstract: The microdroplet lymphocyte cytotoxicity test is universally accepted as the standard test for HLA antigen determination. An update of the technical details of the test is given, based on the authors’ testing 160,000 persons. Methods for quality control of the test as well as reproducibility data are provided. International standardization of the specificities has been accomplished by seven international workshops and a continuous cell exchange involving testing of 108 cells since 1974 by as many as 180 laboratories. The test has recently become applicable to the detection of HLA-D determinants, previously thought to be detectable only by lymphocyte-determined (LD) tests. Purified peripheral blood lymphocytes are reacted with antisera from which HLA-A, -B, and -C antibodies have been removed. B lymphocytes were found to be smaller than T lymphocytes by Coulter counter sizing. The purity of cell suspensions enriched for B lymphocytes can be individually monitored, as shown by the reactions produced by 126 test samples. HLA-D antigens have a linkage disequilibrium with certain HLA-A and -B specificities as demonstrated by population and family studies. Haplotypes found in 34 parents of 18 families demonstrate the new haplotypes, which now consist of four antigens per haplotype. Studies of HLA-D frequencies in Caucasians, Negroes, and Orientals show a distinctive distribution in the races. B lymphocytes also appear to have an autoantigen against which autoantibodies are readily produced. The autoantibodies are more active against B than T lymphocytes and act most effectively at 5 C. Although they appear in many diseases, most notably in systemic lupus erythematosus, they also occur in 10% of normal males and females. In patients awaiting kidney transplants, antibodies against B lymphocytes are often found. Patients with cold B-cell antibodies (autoantibodies) were shown to have higher transplant survival rates than those with warm B-cell antibodies (allogeneic). Thus, in performing crossmatch tests it is now necessary to distinguish antibodies against T and B lymphocytes and those that react in cold and in warm conditions.

Autoantibody to a Nuclear Antigen in Proliferating Cells

Abstract: This study reports the identification of an autoantibody in the sera of some patients with systemic lupus erythematosus that reacted with nuclear antigen(s) of proliferating cells. The autoantibody was initially detected by the observation that it did not react in immunofluorescence with nuclei of renal tubular or glomerular cells, nor with hepatic parenchymal cells, but only reacted with scattered cells in the interstitial tissue of these two organs. In contrast, many lymphocytes in lymph node follicles, spleen, and thymus reacted with this antinuclear antibody. Normal peripheral blood lymphocytes did not show nuclear staining but after mitogenic stimulation, 20% of cells became positive. Nuclear staining was not restricted to lymphocytes but was also observed in several tissue culture cells lines such as Hep-2 cells (human epithelial carcinoma), Ehrlich ascites tumor cells, and baby hamster kidney cells. The reactive nuclear antigen(s) was soluble in physiologic saline and reacted with serum autoantibody to give a precipitin line in immunodiffusion that was immunologically distinct from DNA and other known nuclear antigen-antibody precipitating systems. Autoantibodies to proliferating cell nuclear antigen(s) might serve as useful biologic markers to study stimulated lymphocytes and other proliferating cells.

On the thymus in the differentiation of "H-2 self-recognition" by T cells: evidence for dual recognition?

Abstract: In the thymus, precursor T cells differentiate recognition structures for self that are specific for the H-2K, D, and I markers expressed by the thymic epithelium. Thus recognition of self-H-2 differentiates independently of the T cells H-2 type and independently of recognition of nonself antigen X. This is readily compatible with dual recognition by T cells but does not formally exclude a single recognition model. These conclusions derive from experiments with bone marrow and thymic chimeras. Irradiated mice reconstituted with bone marrow to form chimeras of (A X B)F1 leads to A type generate virus-specific cytotoxic T cells for infected targets A only. Therefore, the H-2 type of the host determines the H-2-restricted activity of killer T cells alone. In contrast, chimeras made by reconstituting irradiated A mice with adult spleen cells of (A X B)F1 origin generate virus-specific cytotoxic activity for infected A and B targets, suggesting that mature T cells do not change their self-specificity readily. (A X B)F1 leads to (A X C)F1 and (KAIA/DC) leads to (KAIA/DB) irradiation bone marrow chimeras responded against infected A but not B or C targets. This suggests that cytotoxicity is not generated against DC because it is abscent from the host's thymus epithelium and not against DB because it is not expressed by the reconstituting lymphoreticular system. (KBIB/DA) leads to (KCIC/DA) K, I incompatible, or completely H-2 incompatible A leads to B chimeras fail to generate any measurable virus specific cytotoxicity, indicating the necessity for I-specific helper T cells for the generation of killer T cells. Finally adult thymectomized, irradiated and bone marrow reconstituted (A X B)F1 mice, transplanted with an irradiated thymus of A origin, generate virus-specific cytotoxic T cells specific for infected A targets but not for B targets; this result formally demonstrates the crucial role of thymic epithelial cells in the differentiation of anti-self-H-2 specificities of T cells.

Use of Radiolabeled Antibodies to Carcinoembryonic Antigen for the Detection and Localization of Diverse Cancers by External Photoscanning

Abstract: To determine whether tumors containing carcinoembryonic antigen could be detected by administration of a Radio-labeled, affinity-purified, goat IgG having 70 per cent immunoreactivity against carcinoembryonic antigen, 18 patients with a history of cancer of diverse histopathology received an average total dose of 1.0 mCi of 131l-labeled IgG. Total-body photoscans were performed with a gamma scintillation camera at various intervals after administration of the radioactive antibody. Ordinary photoscans proved difficult to interpret because of blood-pool background radioactivity, thus necessitating the computer subtraction of radioactive blood-pool agents from the antibody's 131I activity. Tumor location could be demonstrated at 48 hours after injection in almost all cases studied. The scans were negative in patients without demonstrable tumors or with tumors apparently devoid of carcinoembryonic antigen. Circulating antigen levels of up to 350 ng per milliliter did not prevent successful tumor imag...

Immunologic functions of Ia-bearing epidermal Langerhans cells

Abstract: Langerhans cells (LC) constitute a morphologically well-characterized minor subpopulation of the mammalian epidermis whose functional role is still a matter of conjecture. The hypothesis that LC represent an epidermal equivalent to cells of the monocyte-macrophage-histiocyte series is supported by the recent observations that in humans and guinea pigs LC are the only epidermal cells that express Fc-IgG receptors, C3 receptors, and Ia antigens. Using inbred strain 2 and strain 13 guinea pigs, we investigated in this study whether LC can mediate the same immunologic functions as Ia-bearing macrophages. LC-enriched and LC-depleted epidermal cells were prepared by separation of Fc-IgG rosetting epidermal cells on density gradients. When both populations were tested for the biosynthesis of alloantigens by immunoprecipitation techniques, Ia antigen synthesis was restricted to the LC-enriched fraction. Functional studies demonstrated that antigen-pulsed LC-enriched epidermal cells induce a proliferative response in immune T cells that is comparable in magnitude to that seen with macrophages. Moreover, effective presentation of immunologically relevant antigen requires syngeneity between LC-enriched epidermal cells and responder lymphocytes. In the mixed leukocyte reaction (MLR), LC-enriched epidermal cells were as effective stimulators as macrophages. LC-depleted epidermal cells, by contrast, induced little or no stimulation in both assay systems. Both the antigen-presenting and the MLR-stimulatory capacities of LC-enriched epidermal cells could be abrogated by pretreatment with anti-Ia sera and complement. The presence in the epidermis of Ia-bearing LC, capable of mediating the immunologic functions of Ia-bearing macrophages, has important clinical implications with regard to the role of LC as sensitizing cells in both contact hypersensitivity and skin graft rejection.

Persisting oncogenic herpesvirus induced by the tumour promoter TPA

Abstract: PERMISSIVE cell systems for the replication of the Epstein–Barr virus (EBV) and some of the related EBV-like agents (for example, Herpesvirus papio and herpesvirus pan) have not yet been established1. EBV is regularly demonstrated in a small number of cells from lymphoblastoid lines of B-cell origin which have been derived from certain lymphoma patients, most frequently from Burkitt's lymphoma or patients with infectious mononucleosis, but also from healthy EBV-reactive individuals. EBV DNA usually persists in cells of these lines in multiple copies2–4, but spontaneous induction of viral antigens and particle synthesis occurs in a majority of the lines at a low rate. To some extent, the percentage of induced cells seems to be cell line-specific1, and two lines which are rather high producers of EBV are the P3HR-I line of BL origin5 and the marmoset B95-8 line isolated from lymphocytes transformed with EBV of infectious mononucleosis origin6. At any time during cultivation, 2–10% of the cells reveal viral capsid antigen synthesis as determined by indirect immunofluorescence7. All attempts to increase the virus yield in these lines by temperature shifts or chemical or physical inducers (IUdR, mitomycin C, X rays) have usually resulted in only barely significant enhancement of viral replication8–10. We now report on a very efficient induction of EBV and other oncogenic herpesviruses by the well established cocarcinogen and tumour promoter 12-O-tetra-decanoylphorbol-13-acetate (TPA)11. In addition some TPA-treated cells show cytopathogenic changes, including polyploidisation. These observations may enable a more complete investigation of the virus-cell-gene balance hypothesis12.

Monoclonal xenogeneic antibodies to murine cell surface antigens: identification of novel leukocyte differentiation antigens

Abstract: Hybrid myeloma cell lines secreting monoclonal antibodies to mouse cell surface antigens have been prepared. Spleen cells from a DA rat immunized with B10 mouse spleen cells that had been enriched for T cells were fused to cells from a nonsecreting mouse myeloma line (NSI). The presence in the culture supernatants of antibodies binding to mouse spleen cells was tested by a binding assay with 125I-labeled anti-rat IgG. From a large number of positive cultures, ten independent hybrid clones were purified, each secreting a different antibody. Each antigenic target was analyzed by (a) gel electrophoresis of immunoprecipitated 125I-labeled cell surface molecules, (b) heat stability, (c) strain and species distribution and (d) cross-inhibition of binding of different monoclonal antibodies. It was concluded that the ten monoclonal antibodies regognized four types of antigen. One was the heterophile, heat-stable, Forssman antigen. The second (mol.wt. 210 000) appears to be a major 125I-labeled lymphoid cell surface protein. The third, a minor component of spleen cells, was precipitated as two polypeptides of mol.wt. 190 000 and 105000. Five IgG-secreting clones identify the fourth antigen, a heat-stable, possibly glycolipid component expressed on mouse red blood cells and also on thymocytes. Cross-inhibition studies suggest that these last monoclonal antibodies bind to overlapping, but not identical, determinants. The class and chain composition of the monoclonal antibodies were studied by gel electrophoresis, isoelectric focusing and ability to lyse red blood cells and thymocytes.

Lethal graft-versus-host disease after bone marrow transplantation across minor histocompatibility barriers in mice. Prevention by removing mature T cells from marrow.

Abstract: In two situations, transfer of normal unsensitized bone marrow cells into heavily irradiated H-2-identical allogeneic mice caused a high incidence of lethal chronic graft-versus-host disease (GVHD), i.e. mortality occuring between days of 20 and 80 postirradiation. Minor histocompatibility determinants appeared to be the main target for eliciting GVHD. Removing mature T cells from the marrow with anti-Thy 1.2 serum and complement before injection prevented GVHD. On the basis of adding purified T cells to T-cell-depleted marrow cells, it was concluded that contamination of the marrow with as few as 0.3% T cells was sufficient to cause a high incidence of lethal GVHD in certain situations. No GVHD was found with the injection of non-T cells (Thy 1.2-negative cells) or with tolerant T cells. Irradiated recipients of T-cell-depleted marrow cells remained in good health for prolonged periods. These mice showed extensive chimerism with respect to the donor marrow, normal numbers of T and B cells and were immunocompetent. The data provide no support for the view that chronic GVHD developing after bone marrow transplantation in man is the result of an attack by the progeny of the donor stem cells. The results imply that mature T cells contaminating marrow inocula are probably the main cause of GVHD seen in the clinical situation.

An efficient method of antibody elution for the successive or simultaneous localization of two antigens by immunocytochemistry.

Abstract: The method described in this paper is available for removal of antibodies retained by tissue antigen after immunohistochemical staining. Its application to the antehypophysis has allowed the successive or the simultaneous localization of two different hormones.

A hypothesis to relate the specificity of T lymphocytes and the activity of I region-specific Ir genes in macrophages and B lymphocytes.

Abstract: Ever since the discovery of specific immune response (Ir) genes in the I regions of the major histocompatibility complex (MHC) of mammals (1), certain characteristic features of the phenomena they control presented a considerable challenge: 1)H-linked Ir gene control has been observed only for thymus-dependent antigens that are proteins or polypeptides, and is concerned with T cell responses such as the stimulation of delayed-type hypersensitivity (DTH) and specific helper T cells (1, 2). 2)The specificity of Ir gene function is remarkable and distinguishes readily between antigens with different amino acid primary sequences (3–6). This has been demonstrated for synthetic antigens and also for native antigens. It is nevertheless remarkably distinct from the specificity of the T cell responses themselves (7), a matter of considerable concern to me when we initially proposed that the Ir gene product could be the elusive T cell receptor (1).

Method of producing antibodies

Abstract: Continuous cell lines of genetically stable fused cell hybrids capable of producing large amounts of monoclonal antibodies against specific viruses and their antigenic determinants have been developed. The cell lines are fused cell hybrids between viral antibody producing cells and myeloma cells. Fused cell hybrids between influenza virus-primed mouse spleen cells and mouse myeloma cells can be maintained indefinitely in culture and continue to produce large amounts of anti-influenza antibody.

Subclass restriction of murine anti-carbohydrate antibodies.

Abstract: Examination of the subclass distribution of murine antibodies directed against groups A and C streptococcal carbohydrate, alpha-(1 leads to 3) dextran and phosphocholine yields the surprising observation that these carbohydrate antigens stimulate IgG responses largely restricted to the rare IgG3 subclass This subclass restriction is particularly impressive in light of the low circulating levels of IgG3 in nonimmune mouse serum and the failure of a variety of other antigens including proteins and aromatic haptens to stimulate IgG3 antibody production Attempts to alter the subclass restriction of antibodies with carbohydrate specificity by immunization with carbohydrate-coupled protein have been unsuccessful and indicate that immunoregulation of subclass expression probably occurs at the level of the antibody forming (B) cell It is therefore conceivable that VH regions of murine immunoglobulins may be restricted to particular IgG subclasses A similar type of subclass restriction has been reported in human and rat anti-carbohydrate antibodies This recruitment of a minor immunoglobulin isotype by carbohydrate antigens in several species further supports the concept of immunoregulation at the level of subclass, and suggests that these and other mammals may share a structurally similar isotype with perhaps a common evolutionary origin

Type B Hepatitis after Transfusion with Blood Containing Antibody to Hepatitis B Core Antigen

Abstract: We tested the hypothesis that donor blood containing antibody to hepatitis B core antigen (anti-HBc) but lacking detectable hepatitis B surface antigen (HBsAg) and antibody (anti-HBs)might transmit Type B hepatitis by examining donor and recipient serums from a Veterans Administration study of post-transfusion hepatitis. Donor blood was available from three patients with Type B hepatitis and from one patient with hepatitis B virus infection (development of anti-HBs and anti-HBc) without symptomatic disease. All four had received 1 unit of blood with high titer of anti-HBc but lacking HBsAg and anti-HBs. In contrast, no such units had been transfused into nine patients with "immunization-like" response (development of anti-HBs without anti-HBc) or into 26 control patients. These data stress the importance of anti-HBc as an indicator of hepatitis B virus infection and support the hypothesis that high-titer anti-HBc-positive blood might be infectious.

Pneumocystis carinii Infection: Evidence for High Prevalence in Normal and Immunosuppressed Children

Abstract: Using Pneumocystis carinii organisms propagated through three passages in embryonic chick epithelial lung cultures, specific antigens and antisera were prepared for use in counterimmunoelectrophoresis and indirect immunofluorescent antibody techniques. These methods proved to be specific and sensitive for the detection of P. carinii antigen and antibody, respectively, in sera, and were applied to the study of cancer patients with P. carinii pneumonitis (PCP), cancer patients without pneumonitis, and normal children. Antigenemia was detected in 95% of patients with PCP, in 15% of cancer patients without pneumonitis, and in none of the normal children tested. In cross-sectional and longitudinal studies of normal infants and children, acquisition of serum antibody to P. carinii was demonstrated to occur progressively with increase in age. By 4 years of age two thirds of the normal children were found to have antibody to P. carinii in titers of 1:16 or greater. These studies indicate that subclinical P. carinii infection is highly prevalent in normal children, analogous to other opportunistic infections where active disease is manifest predominantly in the compromised host.

The mouse gut T lymphocyte, a novel type of T cell. Nature, origin, and traffic in mice in normal and graft-versus-host conditions.

Abstract: Lymphocytes of the mouse intestinal mucosa, identified in tissue sections or purified suspensions of intraepithelial lymphocytes as T cells (gut T lymphocytes [GTL]), were studied in normal mice or in beige mice (the equivalent of the Chediak-Higashi syndrome in man, characterized by giant granules in various cell types, including mast cells). Mice were studied in normal or in germ-free conditions, or during a graft versus host (GVH) reaction resulting from the injection of parental thymocytes into lethally irradiated F1 mice, a condition leading to massive accumulation of T lymphocytes of donor origin in the host gut mucosa. In normal as well as in GVH conditions, a high percentage of the gut IE lymphocytes contain granules (up to 80% in the beige mouse). These granules have ultrastructural, hostochemical and other features resembling those of mast cell granules; in beige mice, up to 50% of them can be shown to contain histamine. Granulated T cells are also found in the lamina propria. It appears that the GTL may progressively lose their surface T antigens when the granules become more developed. Kinetics of [3H]TdR labeling of the GTL, transfer experiments with T cells of various origins, selective [3H]TdR labeling and selective irradiation of the Peyer's patches (PP), and effect of thoraic duct (TD) drainage led to the conclusion that GTL are the progeny of T cells stimulated to divide in the PP microenvironment, which endows them with a gut-homing tendency. From the PP, these cells follow a cycle, migrating to the TD and to the blood to colonize the whole intestinal mucosa, the majority of them as dividing cells undergoing a single round of traffic, with some probably able to recirculate and becoming a more long-lived variety. Antigenic stimulation within the PP is necessary for the emergence of GTL progenitors, but their gut-homing property is unrelated to the antigen as shown with fetal gut grafts, notably in GVH where grafts syngeneic to the host or donor become similarly infiltrated by GTL. On the basis of their properties and of further evidence to be reported elsewhere, it is proposed that GTL belong to a special class of T lymphocytes, related to the immune defenses of the mucosal systems in general, and capable of acting as progenitors of mucosal mast cells.

Immunoregulatory circuits among T-cell sets. I. T-helper cells induce other T-cell sets to exert feedback inhibition.

Abstract: These experiments test the hypothesis that cells carrying the Ly1+23- surface phenotype are programmed exclusively for helper and not suppressive activity regardless of external conditions such as the mode or type of antigen stimulation. To this end, we have stimulated purified populations of Ly1 cells with antigen in vitro using conditions devised to induce unselected T cells to express optimal levels of antigen specific T-suppressor activity. We find that after such stimulation, Ly1 cells generate SRBC-specific T-helper activity but not T-suppressive activity. These findings establish that the Ly1.2+,2.2/3.2- surface phenotype is a stable, and probably invariant, marker of T cells that are programmed to express only helper activity and have lost the capacity to directly suppress the antibody response. These findings support the concept that the genetic program for a single differentiated set of cells combines information for cell surface phenotype and function. We also demonstrate that antigen-stimulated Ly1 cells, in addition to inducing B cells to secrete antibody, can induce or activate other sets of resting T cells to develop profound suppressive effects. The surface phenotype of this feedback suppressive T-cell set is shown to be: Ly1+2+3+Qa1+. These findings, taken together, indicate that activation of resting Ly123 cells by immune Ly1 TH cells may represent an important homeostatic immunoregulatory mechanism.

Determinant selection and macrophage function in genetic control of the immune response

Abstract: The immune response to insulin, in both mouse and guinea pig, is under control of H-linked immune response genes. When immunized with either pork or beef insulin in CFA, both strain 2 and 13 guinea pigs respond by antigen-specific lymphocyte proliferation and synthesis of specific antibody. The specificities of the elicited antibodies and indistinguishable between these inbred strains. By constrast, strain 2 T cells recognized a distinct region of the A chain alpha loop consisting of amino acid residues 8, 9 and 10, while strain 13 T cells see an as yet undefined region of the B chain. H2b (A chain alpha loop responder) and H2d (B chain responder) mice similarly discriminate which areas of the molecule are recognized by their T lymphocytes. The function of the Ir gene in both the guinea pig and mouse appears to be an intramolecular selection of discrete regions within the antigen for recognition by the T cell. The data presented suggest that this function operates at the level of the macrophage.

Peripheral human T cells sensitized in mixed leukocyte culture synthesize and express Ia-like antigens.

Abstract: We have studied the modulation of Ia-like antigens on the surface membrane of human T cells responding in a one-way mixed leukocyte culture. A heterologous antiserum, (anti-p23,30), which is specific to HLA-D-related antigens and which is unreactive with normal peripheral T cells or thymocytes, was found to bind significantly to all T cells transformed in mixed leukocyte culture (MLC) as determined by indirect immunofluorescence on a fluorescence-activated cell sorter 1. Furthermore, cytotoxic T cells responsible for cell-mediated lympholysis were shown to react with anti-p23,30, whereas their unactivated progenitors did not. Immunoprecipitation and sodium dodecyl sulfate polyacrylamide gel electrophoresis of a radioactive 29,000 and 34,000 dalton complex from MLC-primed T cells labeled with [35S]methionine indicated that allosensitized T cells synthesized these HLA-D-related antigens.

Murine B Cell Leukemia Line with Inducible Surface Immunoglobulin Expression

Abstract: A chemically induced leukemia of BDF 1 mice, designated 70Z/3, was adapted to tissue culture in our laboratory A minority of these cells displayed surface immunoglobulin (sIg) as detected by immunofluorescence with rhodamine-labeled class (IgM)-specific antibodies Addition of the B cell mitogen, LPS, to the cultures induced sIg expression on all of these cells The kinetics of this transition were dependent on the dose of LPS As little as 01 µg/ml induced sIg on >97% of the cells within 36 hr DxS also induced sIg whereas other mitogens (Con A, PPD) or inducing agents (DMSO, dimethyl formamide, butyric acid), tested over a wide range of concentrations, failed to induce sIg expression The cells bear H-2 antigens but lack IgD, IgG, IgA, Ia, and Thy 12 Exposure to LPS had no effect on the presence or absence of these structures A small percentage of cells possessed receptors for complement and for the Fc portion of immunoglobulin Cytoplasmic IgM was detectable within all of the cells, and constitutive production of small quantities of IgM was confirmed by SDS-polyacrylamide gel electrophoresis of serologic precipitates of cell lysates after pulsing with radioactive amino acids Using similar methods, we failed to detect active secretion of immunoglobulin Thus, this cell line has properties similar to cytoplasmic Ig + , surface Ig - cells found in immature tissues and in bone marrow of mice and humans that are thought to be immediate precursors of sIg + B lymphocytes It may provide a model for studying the mechanism of LPS activation and the molecular events associated with externalization of cell surface receptors

Major histocompatibility complex-linked immune-responsiveness is acquired by lymphocytes of low-responder mice differentiating in thymus of high-responder mice.

Abstract: Female murine T cells can respond to the Y antigen of male cells by generating cytotoxic T-killer lymphocytes. Responsiveness is linked to several H-2 genes. Two types of low responders can be distinguished: the B10.A(5R) (H-2i5) strain, a low responder because it lacks Y-specific precursor T cells able to differentiate into cytotoxic T-killer cells; and the CBA/J (H-2k) strain, a low responder because it lacks Y-specific T-helper cells able to support differentiation of T-killer cell precursors. B10.A(5R) stem cells differentiating in an x-irradiated (CBA/J X C57BL/6) (H-2k X H-2b)F1 host respond to Y antigen by generating T-killer cells whereas CBA/J stem cells do not. The results are consistent with the hypothesis that diversity of T-cell receptors is generated by somatic mutation of germ-line genes encoding specificity for self-H-2. A detailed account of this hypothesis is presented.

T-T-cell interactions during the vitro cytotoxic allograft responses. I. Soluble products from activated Lyl+ T cells trigger autonomously antigen-primed Ly23+ T cells to cell proliferation and cytolytic activity.

Abstract: Secondary murine cytotoxic T lymphocyte responses from alloantigen-primed T cells can be induced in vitro by apparently unrelated regimens, such as addition of either concanavalin A (Con A), conditioned medium from Con A stimulated lymphocyte cultures, conditioned medium from secondary mixed lymphocyte cultures (MLC), or stimulator cells sharing only the I-region with the stimulating cells used for primary sensitization. We now report that upon polyclonal (Con A), or antigen-specific (MLC) stimulation, Lyl+ T cells release a factor, which in turn triggers alloantigen primed Ly23+ T cells to proliferation and cytolytic activity. The secondary cytotoxic T lymphocyte inducing factor (SCIF) is produced within 24 h. For its production, an intact protein metabolism, not DNA metabolism, is required. Once induced, the functional activity of SCIF is nonspecific and not H-2 restricted. SCIF allows exponential growth and long-term propagation of cytolytic Ly23+ T cells with specificity to alloantigens used for primary sensitization. SCIF induced activation of alloantigen primed Ly23+ T cells does not require the presence of alloantigens. The results therefore reveal a process by which Lyl+ T-cell-derived nonspecific factor(s) induce autonomously Ly23+ T-cell-mediated, antigen-specific, cytotoxic T lymphocyte responses.

H-2 antigens of the thymus determine lymphocyte specificity.

Abstract: After immunization, normal H-2 heterozygous mice (for example H-2(b) × H-2(d)) generate two populations of cytotoxic effector T cells, one specific for target cells expressing H-2(b)-plus-antigen and the other specific for H- 2(d)-plus-antigen. With a multideterminant antigen, these two populations have about the same activity. We show here that the H-2 type of resident cells in the thymus determines the H-2 preference of cytotoxic T lymphocytes. F(1)(B 10 × B 10.D2) (H-2(b) × H-2 (d)) mice were thymectomized, lethally irradiated, and reconstituted with T-cell-depleted syngeneic hematopoietic cells. Groups of such ATXBM mice were grafted subcutaneously with neonatal thymus lobes from parental mice, either B10 (H-2 (b)) or B10.D2 (H-2(d)). 2-3 mo later, the mice were immunized against the minor histocompatibility antigens on F(1)(BALB/c × BALB.B) cells and assayed for cytotoxic T-cell activity. H-2(b) × H-2(d) ATXBM mice with H-2(b) thymus grafts responded to antigen-plus-H-2(b) much better than to antigen-plus-H-2(d), and vice versa for the mice with H-2(d) thymus grafts. As judged by antiserum treatment, the effector cells were of F(1) origin. To explore the possibility that the “thymus preference” may have been due to suppression of T-cell activity, nonimmune spleen and lymph node cells from normal H-2(b) × H-2(d) mice and cells from H-2(b) × H-2(d) mice bearing a homozygous thymus were mixed 1:1 and immunized in adoptive transfer. The mixture responded to antigen-plus-H-2(b) and antigen-plus-H-2(d) equally well, demonstrating that the cells that showed a “thymus preference” could not suppress a response to antigen in association with the nonthymic H-2 type. We conclude from these and other experiments that H-2 antigens present on resident cells of the thymus determine the spectrum of specificity of T cells which mature in that thymus and eventually make up the peripheral T- cell pool.

Two distinct types of helper T cells involved in the secondary antibody response: independent and synergistic effects of Ia- and Ia+ helper T cells.

Abstract: We have described here two distinct types of carrier-specific helper T cells which act independently and synergistically to augment the B-cell response to a hapten. They are separable by passage through a nylon wool column. The first type of helper T cell, which we designate as Th1, is nylon nonadherent, and can help the response of hapten-primed B cells only if the haptenic and carrier determinants are present on a single molecule (cognate interaction). The second type of helper T cell, Th2, adheres to the nylon wool column, and can help the B-cell response to a hapten coupled to a heterologous carrier upon stimulation with unconjugated relevant carrier (polyclonal interaction). The addition of a small number of Th2 to the mixture of Th1 and B cells significantly augmented the net response to the hapten carrier conjugate. Both Th1 and Th2 cells belong to the Lyt-1+,2-,3- subclass. Th1 has no detectable Ia antigen, whereas Th2 is killed by certain anti-Ia antisera and complement. The Ia antigen detected on Th2 was found to be controlled by a locus in the I-J subregion. The results clearly established the fact that there are two distinct pathways in the T- and B-cell collaboration, which involves two different subsets of carrier-specific helper T cells.

Expression and function of idiotypes of lymphocytes.

Abstract: Publisher Summary This chapter summarizes a series of new procedures for the production of anti-idiotypic reagents. It discusses the serological analysis of antigen receptors on lymphocytes and discusses the functional aspects of lymphocyte receptor idiotypes within an immune network. Cross-idiotypic specificity was observed between human cold agglutinins of different individuals between a mouse myeloma protein with specificity for phosphorylcholine and antibodies induced in mice with this antigen among antibodies to streptococcal carbohydrates from different rabbits and between human anti-γ-globulins. Idiotypic cross-reactions between immunoglobulin molecules from more than one source promoted a series of studies on the structural requirements for the expression of idiotypic determinants. Idiotypic determinants of the antigen-specific receptors of lymphocytes are discussed in this chapter with respect to their structural and serological definition as well as to their possible functional role in immune regulation. The serological basis of idiotypes on lymphocytes is established with anti-idiotypic antisera to total “idiotypes” as well as to idiotypic subspecificities such as V H - and V L -associated idiotypic determinants and binding site- or framework-associated idiotypic determinants. Lymphocyte receptor idiotypes play a fundamental role in immune regulation. Within one individual's immune system, spontaneous auto-anti-idiotypic antibodies can be observed.

Characterization of two types of human papillomaviruses in lesions of epidermodysplasia verruciformis

Abstract: Human papillomaviruses (HPVs) found in lesions of 11 patients suffering from epidermodysplasia verruciformis were compared to HPV type 1 (HPV-1) and HPV type 2 (HPV-2) previously characterized in plantar and common warts, respectively. Complementary RNAs (cRNAs) to HPV-1, HPV-2, and viruses obtained from two patients with epidermodysplasia verruciformis (J.D. HPV and J.K. HPV) were used in cRNA·DNA filter hybridization experiments. No sequence homology was detected between HPV-1 or HPV-2 DNAs and DNAs obtained from the 11 epidermodysplasia verruciformis HPV isolates. Furthermore, with J.D. and J.K. HPV cRNAs, epidermodysplasia verruciformis HPV DNAs fell into two groups showing little, if any, sequence homology. A lower extent of annealing was observed for the DNAs of some isolates showing a genetic heterogeneity within each of the two groups. Almost no antigenic crossreaction was detected by immunodiffusion and indirect immunofluorescence tests, either between epidermodysplasia verruciformis HPVs and HPV-1 or HPV-2 or between J.D. and J.K. HPVs. Viruses belonging to the same group have common antigenic properties, but antigenic differences were observed when two of the viruses sharing only partial DNA sequence homology were compared. Viruses related to J.D. HPV were preferentially associated with flat wart-like lesions of epidermodysplasia verruciformis and were further found in the lesions of five patients bearing multiple flat warts. Viruses related to J.K. HPV were found in morphologically distinct lesions (red spots) present in some patients with epidermodysplasia verruciformis. Thus, we propose to distinguish two other types of HPVs designated provisionally as HPV type 3 (HPV-3) and HPV type 4 (HPV-4), with J.D. and J.K. HPVs as prototypes, respectively. Malignant conversion of some epidermodysplasia verruciformis lesions is more frequently associated with HPV-4 than with HPV-3 infection.