bluuebunny/crossref_metadata_2025 huggingface dataset:
Contact hypersensitivity reactions to dinitrofluorobenzene mediated by monoclonal IgE anti-DNP antibodies.
Authors: Ray M C , Tharp M D , Sullivan T J , Tigelaar R E
Published: December 2022

Several studies have suggested a possible role for IgE antibodies in the pathogenesis of cutaneous hypersensitivity reactions that reach maximum intensity 24 to 48 hr after antigen challenge. The recent availability of murine monoclonal IgE anti-hapten antibodies has made possible the direct examination of the range of cutaneous inflammatory reactions that can be mediated by such antibodies. We have examined the effects of passively sensitizing BALB/c mice with monoclonal IgE anti-dinitrophenyl (DNP) antibody 48 hr before antigen challenge. Inflammatory responses were assessed by measuring ear swelling in mice challenged on the ears with the reactive hapten 2,4-dinitrofluorobenzene (DNFB). Compared with unsensitized controls, the ears of mice passively sensitized with IgE anti-DNP displayed a biphasic pattern of ear swelling after DNFB challenge. An early, transient response (present within 15 to 30 min of challenge and returning to control levels within 4 to 9 hr) was followed by a second, more persistent increase in ear swelling that peaked 24 to 48 hr after challenge. This biphasic pattern of ear swelling seen in IgE-sensitized mice was temporally indistinguishable from that observed in mice conventionally sensitized for allergic contact dermatitis reactions by epicutaneous application of DNFB 5 days before DNFB ear challenge. Antigen specificity of the IgE-mediated contact hypersensitivity reactions was demonstrated by the failure of mice passively sensitized with IgE anti-DNP to display early or delayed ear swelling greater than unsensitized controls when challenged with either of two noncross-reacting haptens, fluorescein isothiocyanate or oxazolone. Mice passively sensitized with a monoclonal IgA anti-DNP antibody (MOPC 315) 48 hr before DNFB challenge failed to display early or delayed ear swelling greater than unsensitized controls. Heat inactivation of the IgE anti-DNP ascitic fluid at 56 degrees C for 30 min completely abolished its capacity to passively sensitize mice for contact hypersensitivity reactions after DNFB challenge. These results document the existence of an antigen-specific, IgE-mediated, delayed-in-time cutaneous hypersensitivity response that can be elicited by epicutaneous challenge (contract) with a reactive hapten.
A partial characterization of suppressor cells in the spleens of mice conditioned with fractionated total lymphoid irradiation (TLI).
Authors: May R D , Slavin S , Vitetta E S
Published: December 2022

Total lymphoid irradiation (TLI) is a highly effective modality for inducing immunosuppression and transplantation tolerance. The cellular basis for this immunosuppression is not clear, although T cells have been implicated. To study further the effect of TLI on the immune system, we have examined the B cells and suppressor cells in the spleens from TLI-conditioned mice. Our results indicate that after TLI, the spleen is rapidly repopulated with many large, immature cells. The probable source of these cells is the shielded bone marrow (BM). The B cells from TLI-conditioned mice are transiently immature and hyporesponsive in vitro to a T-independent antigen. Spleen cells from TLI-conditioned mice nonspecifically suppress the in vitro T-independent anti-TNP response of normal B cells. The suppressor cells lack both B and T cell markers and adhere to Sephadex G-10. The suppressor cells in spleens from TLI-treated mice bear a number of similarities to those present in normal BM. When normal BM cells were analyzed by indirect immunofluorescence for the presence of the Mac-1 antigen, two populations of suppressor cells could be identified: one was Mac-1+ and the other was Mac-1-. These data are consistent with the possibility that a subpopulation of the suppressor cells found in normal BM and in the spleens from TLI-conditioned mice are immature cells of the monocytic/granulocytic lineage.
Monoclonal antibodies to streptococcal group A carbohydrate. II. The VK1GAC light chain is preferentially associated with serum IgG3.
Authors: Fulton R J , Nahm M H , Davie J M
Published: December 2022

A kappa-light chain variable region (V kappa) dominantly employed in the serum antibody response of A/J mice to streptococcal group A carbohydrate (GAC) has been termed VK1GAC. Examination of in vitro recombinants between the isolated heavy and light chains of VK1GAC+ and VK1GAC-anti-GAC hybridomas and non-GAC-binding myeloma proteins indicated that two antisera (anti-Id5 and anti-Id20) recognized the VK1GAC light chain when it was free in solution or paired with several heterologous heavy chains. Screening of a panel of A/J anti-GAC monoclonal antibodies with these antisera showed almost complete concordance between Id5 and Id20 expression and the presence of VK1GAC light chain as detected by its unique isoelectric focusing spectrotype. These antisera were used to examine serum expression of the VK1GAC light chain in normal and hyperimmune serum of A/J mice. Normal A/J serum contained from 20 to 100 micrograms Id5/ml serum, whereas only 1 to 10 micrograms Id20/ml serum was detected. The levels of both VK1GAC idiotypes increased dramatically 10- to 20-fold after hyperimmunization of mice with group A vaccine. When serum IgG from normal and immune mice was fractionated into the IgG subclasses (IgG1, IgG2a, and IgG3), it was found that the VK1GAC light chain does not pair randomly with heavy chains of the IgG subclasses, but rather is associated preferentially with heavy chains of the IgG3 subclass whether or not it is associated with antibodies to GAC. These results suggest that the heavy chain pairing exhibited by this VK product may not be random.
Serologic relatedness between Thy-1.2 and actin revealed by monoclonal antibody.
Authors: Dales S , Fujinami R S , Oldstone M B
Published: December 2022

Monoclonal antibodies with affinity for Thy-1.2 on thymocytes also can bind to actin within marsupial, murine, and human cells. A similar cross-reactivity between Thy-1.1 and vimentin was revealed by Dulbecco and co-workers employing monoclonals. A computer-assisted analysis of the amino acid composition provided suggestive evidence for the occurrence of sequence homology between Thy-1.1 or Thy-1.2, actin, and vimentin that likely accounts for the serologic relatedness detected by hybridoma antibodies.
Description of a new Qa antigenic specificity, "Qa-m9," whose expression is under complex genetic control.
Authors: Sutton V R , Hogarth P M , McKenzie I F
Published: December 2022

The "Qa-m9" specificity has been defined by a monoclonal antibody, and evidence for three-gene control of its expression has been demonstrated. The Qa allocation was made on the basis of H-2 linkage (H-2 congenic strains, Rl lines), differential reaction with B6.K1 and B6.K2, and biochemical analysis (two chains: 39,000 Mr and 12,000 Mr). "Qa-m9" is expressed on T and B cells in spleen and lymph node; it is present on a small population of bone marrow cells but is absent from thymocytes. The three genes involved in the expression of the antigen are: a) an H-2 linked Qa gene (possibly encoded by a gene different from Qa-2-6); b) an H-3 linked gene--probably beta 2m; and c) an H-2D linked gene that controls the amount of antigen present. It is likely that the determinant is of the "combinatorial" or conformational type, whereby the C57BL/6-type Qa heavy chain and the appropriate beta 2m light chain must both be in association. Thus, the study provides evidence for a multi-gene control of antigen expression and demonstrates the potential for generating new antigens with diversity at a post-translational level.
Elimination of complement channels from the plasma membranes of U937, a nucleated mammalian cell line: temperature dependence of the elimination rate.
Authors: Ramm L E , Whitlow M B , Koski C L , Shin M L , Mayer M M
Published: December 2022

We have studied the release of radiolabeled small markers from nucleated cells carrying complement channels in order to determine the life-span of these channels at various temperatures. U937 cells, a human histiocytic cell line, were labeled with 14C-aminoisobutyric acid or 86RbCl, and treated with sublytic doses of C to form transmembrane channels. The cells were then incubated at various temperatures, and the persistence of channels was evaluated by measuring the release of the intracellular markers through the remaining channels. The results indicate that the life-span of the C channels in the plasma membranes of these cells varies markedly with temperature. Thus, at 2 degrees C, the half-life of the channels was about 2 hr, whereas at 37 degrees C, the half-life was estimated to be approximately 1 min. The rapid elimination of the transmembrane channels from the plasma membranes of these nucleated cells contrasts sharply with the long persistence of C channels in the membranes of erythrocytes or erythrocyte ghosts. It is likely that the multi-hit requirement recently reported for lysis of nucleated mammalian cells by C is due, at least in part, to the rapid disappearance of channels.
Immunologic response of athymic rats to Schistosoma mansoni infection. II. Antibody-dependent mechanisms of resistance.
Authors: Capron M , Capron A , Abdel-Hafez S K , Bazin H , Joseph M , Phillips S M
Published: December 2022

The responses of congenitally athymic rats to Schistosoma mansoni were compared to thymic reconstituted, heterozygous littermate controls, and inbred Fischer rats. The mechanisms of the impaired resistance of athymic rats to initial exposure and re-exposure to S. mansoni were investigated by the study of various parameters of antibody response. The uninfected athymic animals demonstrated normal levels of total IgM but reduced levels of total IgG2a and IgE. After infection with S. mansoni, the immunoglobulin increases in athymic rats were less than those observed in heterozygote control rats. In addition, the level of anti-S. mansoni IgG antibody, utilizing ELISA assay, was reduced. Furthermore, the functional avidity of the IgG2a antibody, which was produced by the athymic animals, was significantly lower than that of control heterozygote and Fischer animals. Similarly, the levels of IgE and IgG2a anaphylactic antibodies were reduced in the congenitally athymic animals. After thymic reconstitution and exposure to S. mansoni of the congenitally athymic animals, all of these parameters became similar to the analogous value obtained from exposed heterozygous and homozygous animals. In vitro studies of antibody-dependent cell-mediated cytotoxicity (ADCC) activity indicated that the antibody response of the congenitally athymic animals was characterized by significant reductions in IgE-macrophage-mediated, IgG-eosinophil-mediated, and IgE-eosinophil-mediated cytotoxicity directed against schistosomula. These results, coupled with previously reported in vivo observations, that athymic animals produced antibody that was less capable of transferring resistance in adoptive-challenge experiments, suggest that the mechanisms of impaired resistance in the congenitally athymic rat may involve the failure to develop adequate, functional ADCC mechanisms. As such, these studies suggest a relationship between in vivo resistance and possible in vitro mechanisms of that resistance.
Infection with vaccinia favors the selection of hybridomas synthesizing autoantibodies against intermediate filaments, one of them cross-reacting with the virus hemagglutinin.
Authors: Dales S , Fujinami R S , Oldstone M B
Published: December 2022

Immunization of mice with lysates from infected cells and pure vaccinia virus generated hybridomas, a large fraction of which make autoantibodies binding to intermediate filaments (greater than IF) and vimentin. Antibodies that can be detected in preimmune mice as weakly binding to the cytoskeleton (greater than CS) are elevated in potency after immunization with either active or UV-inactivated pure vaccinia, suggesting that virus replication may not be obligatory for activation of greater than CS lymphocyte clones. Demonstration of a cross-reactivity of one IF monoclonal antibody with virally coded hemagglutinin, expressed at the surface, raises questions about the development of pathologic conditions associated with virus-mediated autoimmunity.
Monoclonal antibodies to murine leukemia virus gp 70 recognize the same T lymphoma cell surface molecules as anti-immunoglobulin.
Authors: Baird S , Lesley J , Majda J , Raschke W
Published: December 2022

Chicken antisera to mouse immunoglobulin (anti-Ig) detect molecules on T lymphocytes of mice consisting of "heavy chains" similar in size to IgM mu chains and "light chains" similar in size to lambda or kappa chains. Because these T cell-derived proteins react with anti-Ig and are composed of Ig-like heavy and light chains, these molecules have been viewed as candidates for "IgT," the putative T lymphocyte receptor for antigen. We have demonstrated, by immunoprecipitation and two-dimensional gel analysis, that the lymphoma surface molecule precipitated by chicken anti-Ig is identical to the viral 70,000 dalton glycoprotein (gp 70) expressed by that lymphoma and is unassociated with any "light chain" or equivalent. For each of the three lymphomas analyzed, the gp 70 two-dimensional gel pattern was individually distinctive, and, in each case, the molecules precipitated by anti-Ig exhibited the same pattern as the gp 70. The viral gp 70 does have cross-reacting determinants with mouse IgM as seen by the chicken antiserum. We have substantiated these results by demonstrating that monoclonal antibodies to viral gp 70 precipitate the same molecules as chicken anti-Ig. These findings demonstrate that these T lymphoma cell surface molecules do not represent the T cell immunoglobulin receptor for antigen unless gp 70 itself is an antigen receptor.
Gangliotriaosylceramide (asialo GM2), a glycosphingolipid marker for cell lines derived from patients with Hodgkin's disease.
Authors: Kniep B , Monner D A , Burrichter H , Diehl V , Mühlradt P F
Published: December 2022

Three cell lines, isolated from different patients with Hodgkin's disease, were analyzed for their neutral glycosphingolipids by high performance liquid chromatography. On two of these lines gangliotriaosylceramide (asialo GM2) was identified by the comparison of retention time to authentic standards and by degradation with specific glycosidases. This glycosphingolipid was also serologically detected with a specific monoclonal antibody of the IgM class. The third line, which according to other criteria such as the Epstein Barr virus nuclear antigen differs phenotypically from the first two, did not exhibit gangliotriaosylceramide. This glycosphingolipid is an unusual marker for human cells and could not be detected on the peripheral blood leukocytes of healthy donors.
Specific stimulation of human T lymphocytes by substance P.
Authors: Payan D G , Brewster D R , Goetzl E J
Published: December 2022

Substance P (SP) and SP(4-11) stimulated significantly the proliferation of human T lymphocytes in vitro at respective concentrations of 10(-11) M to 10(-7) M and 10(-9) M to 10(-7) M, as assessed by the uptake of [3H]thymidine and [3H]leucine. The proliferative response of human T lymphocytes to PHA was augmented by 10(-9) M to 10(-8) M SP and 10(-10) M to 10(-7) M SP(4-11). The inactive analogue [D-Pro2, D-Phe7, D-Trp9]-SP inhibited the effects of SP on T lymphocytes. The specific stimulation of T lymphocytes by SP suggests a unique mechanism for the regulation of immunologic responses by peripheral nerve-derived factors.
An IL 2-secreting T cell hybridoma that responds to a self class I histocompatibility antigen in the H-2D region.
Authors: Endres R O , Marrack P , Kappler J W
Published: December 2022

A self-reactive T cell hybridoma that secretes IL-2 in response to H-2d haplotype cells resulted from a fusion of BALB/cBy lymph node cells with the AKR thymoma BW5147. The lymph node cells used had been enriched for cells reactive to (TG)-A--L, but neither this antigen nor fetal calf serum were required for stimulation of the hybridoma designated 3DT52.5. The gene product responsible for stimulation mapped to the H-2D region. Allogeneic cells of the b, f, k, q, and s haplotypes failed to stimulate. Not all H-2d haplotype cells were effective stimulators of 3DT52.5. Peritoneal cells and splenic B cells were much more stimulatory than splenic T cells. Most tumor cell lines of H-2d derivation and of B cell or macrophage/monocyte lineage were stimulatory, whereas H-2d T cell lines were not. The capacity to stimulate 3DT52.5 did not correlate with the ability to stimulate I region-restricted hybridomas, or with the ability to be induced to stimulate such hybridomas. Stimulatory cell lines did not apparently produce a soluble factor required for stimulation, and negative cell lines were not inhibitory. The monoclonal antibody 27-11-13, which reacts with H-2D of the b, d, and q haplotypes, inhibited stimulation of 3DT52.5 but did not inhibit stimulation of the sibling hybridoma 3DT18.11, which responds to (TG)-A--L plus I-Ad. Conversely, the monoclonal anti-I-Ad antibody MK-D6 inhibited stimulation of 3DT18.11 but not 3DT52.5. Although it is clear that 3DT52.5 recognizes a class I antigen coded for in the H-2D region, the precise molecular nature of the antigen is unknown. The structure of the antigen receptor on this hybridoma may prove to be of interest when it can be compared with receptors found on T cell hybridomas restricted by class II histocompatibility antigens.
Unresponsiveness in hapten-specific cytotoxic T lymphocytes. IV. Functional deletion of T cell subsets required for trinitrophenyl (TNP)-specific cytotoxic T cell generation after acute or chronic treatment with trinitrobenzene sulfonic acid (TNBS).
Authors: Butler L D , Chiller J M
Published: December 2022

The generation of cytotoxic T cell responses requires the interaction of a network of immunocompetent cells. Within this network, a dynamic balance of positive and negative immunoregulatory cells and their products modulate the cytotoxic T cell response. We have developed protocols which shift the balance to favor negative immunoregulation and thus result in antigen-specific unresponsiveness. In this report, a variety of approaches have been utilized to examine the effects of acute and chronic tolerance protocols on specific T cell subsets, namely amplifier helper T cells and precursor cytotoxic T cells. The results to be detailed show differential effects on these T cell subsets which are related to the tolerance protocol employed and the strains of mice tested.
T cell proliferative responses to a mitogen derived from Mycoplasma arthritidis are controlled by the accessory cell.
Authors: Lynch D H , Gurish M F , Cole B C , Daynes R A
Published: December 2022

Cell-free supernatants from broth cultures of Mycoplasma arthritidis (MAS) induce vigorous proliferative responses in thymus-derived T lymphocytes from H2k or H2d strains of mice. Populations of lymphoid cells from mice of H2b, H2q, or H2s haplotypes do not respond to this stimulus. Previous studies with lymphoid cells from congenic and recombinant strains of mice indicate that the T cell proliferative response induced by MAS is controlled by a gene(s) that maps to the I-E/C subregion of the murine major histocompatibility complex (MHC). The T cell proliferative response induced by MAS is dependent upon the presence of a population of la+, radioresistant accessory cells (AC). Data presented here demonstrates that responder strain AC that have been pulsed with MAS (followed by extensive washing) induced vigorous proliferative responses in subsequently added T cell populations. Pulsing of T cells with MAS, followed by the addition of AC, however, did not result in T cell proliferation. MAS was found to stimulate (responder X nonresponder) F1 T cells to proliferate if the MAS was presented in the context of either responder or (responder X nonresponder) F1 AC; nonresponder strain AC were ineffective in this regard. Nonresponder strain T cells were found to be capable of responding to MAS if it was presented in the context of responder strain AC, even if the T cells and AC were completely allogeneic. Thus, nonresponder strain T cells mounted vigorous proliferative responses if the MAS was presented in the context of responder strain AC. Conversely, responder strain T cells did not respond to MAS presented in the context of nonresponder strain AC. In addition, lymphoid cells from a B10 leads to B6AF1 radiation bone marrow chimera were also found to be capable of responding to MAS, but only in the presence of AC that expressed cell surface determinants controlled by the I-E/C subregion. The data presented here indicate that MAS-induced T cell proliferative responses are controlled at the level of the AC by a gene(s) that maps to the I-E/C subregion of the MHC.
Evidence for a nonoxidative mechanism of human natural killer (NK) cell cytotoxicity by using mononuclear effector cells from healthy donors and from patients with chronic granulomatous disease.
Authors: Kay H D , Smith D L , Sullivan G , Mandell G L , Donowitz G R
Published: December 2022

In vitro natural killer (NK) activity expressed by blood mononuclear cells from patients with chronic granulomatous disease of childhood (CGD) was equivalent to that expressed by cells from normal, healthy volunteers. Because neutrophils and monocytes from these same donors exhibited extremely depressed oxidative functions, our data could be interpreted to show that a) NK cells derived from a unique and separate cellular lineage unaffected by the disease-related oxidative defect, or b) the in vitro cytolytic mechanism(s) of NK cells were not dependent on oxygen metabolites. These hypotheses were examined by using as NK effector cells large granular lymphocytes (LGL) from healthy donors whose monocytes and neutrophils had normal oxidative functions. Such functions were measured in the nitroblue tetrazolium dye reduction assay, which is a qualitative measurement of superoxide anion production; by reduction of ferric cytochrome c, a more specific and quantitative measurement of superoxide anion production; and in the luminol-enhanced chemiluminescence assay, an extremely sensitive measure of several reactive oxygen radicals, including superoxide anion, hydroxyl radical, and singlet oxygen. Whereas monocytes and neutrophils from healthy donors were readily stimulated with zymosan or phorbol myristate acetate (PMA) in each of these assays. LGL produced no detectable amounts of oxygen metabolites when co-incubated either with K562 erythroleukemia cells, PMA, E. coli endotoxin, or the calcium ionophore A23187. Thus, because NK cell activity is normal in CGD patients with major oxidative defects, and because no reactive oxygen metabolites could be detected in LGL that simultaneously exhibited potent NK activity, we conclude that in vitro NK activity by human mononuclear cells involves a lytic mechanism(s) independent of oxygen metabolites.
The modeled structure of the IgG Gar VL region and its implications for anti-flavin and anti-DNP fine specificities.
Authors: Kiefer C R , McGuire B S , Osserman E F , Garver F A
Published: December 2022

The variable domain (V) sequence of the lambda-light chain of the Gar IgG2 human myeloma protein was determined from enzymatically generated peptides that were isolated, characterized, and ordered by overlap and/or by subgroup homology. The sequence consisted of the amino terminal 107 residues of the light chain, and shared subgroup V lambda III-specific residues as well as a pattern of CDR deletions unique to most V lambda III sequences. The structural role of the light V region in the high affinity for flavins and the low affinity for 2,4-dinitrophenyl haptens characteristic of the intact IgG molecule was coarsely assessed by way of model building and by comparison with the refined combining site model of MOPC 315, an Ig with relative affinity values for DNP and riboflavin the converse of those observed in Gar.
Activation of C1 by soluble IgG aggregates as detected by a novel one-step hemolytic assay that specifically measures the proenzyme form of C1s.
Authors: Doekes G , van Es L A , Daha M R
Published: December 2022

A new hemolytic assay is described that specifically measures the precursor form of the C1s subcomponent of the complement system. The assay employs a C1s-depleted reagent obtained by immunoadsorption of fresh human plasma on immobilized goat anti-human C1s antibodies. Linear Z plots are obtained with nanogram levels of precursor C1s, whereas C1s completely fails to induce hemolysis in the assay. Because low concentrations of C1s do not interfere with the activity of precursor C1s, the assay can be used for the stoichiometric measurement of C1 activation. The precursor C1s assay was applied to the study of C1 binding and activation by soluble aggregates of human IgG (AIgG). Incubation of purified human C1 with AIgG caused a temperature-independent consumption of whole C1 hemolytic activity, indicating binding of C1, but almost no consumption of the total (precursor + activated) C1s activity. On the other hand, activation of C1, measured as the time- and temperature-dependent consumption of precursor C1s, could greatly exceed the binding of C1. These findings can be explained by using recent findings concerning the association-dissociation equilibrium between C1q and the tetrameric complex of C1r and C1s.
Analysis of the effect of mast cell granules on lymphocyte blastogenesis in the absence and presence of mitogens: identification of heparin as a granule-associated suppressor factor.
Authors: Frieri M , Metcalfe D D
Published: December 2022

To explore the effect of mast cell products on lymphocytes, the blastogenic response of lymphocytes to mast cell granules in the absence and presence of mitogens was determined. Membrane-free granules (MFG) and dialyzed, washed histamine-free granules (DWG) in the absence of mitogens did not induce significant blastogenesis in peripheral blood mononuclear cells (PBMC) before or after partial depletion of macrophages (5 to 10%). In contrast, concanavalin A- (Con A) (5 micrograms/ml), pokeweed mitogen- (PWM) (1/200 dilution), tetanus toxoid-, and Candida-, but not phytohemagglutinin-, induced blastogenesis in macrophage-depleted PBMC were inhibited by both MFG and DWG. Fractionation of solubilized DWG by anion exchange Dowex chromatography into protein- and heparin-rich fractions revealed the inhibitory activity resided in the heparin-containing fractions. The ability of commercial porcine heparin glycosaminoglycan as well as native rat heparin proteoglycan to inhibit lectin-induced blastogenesis in a dose-response fashion confirmed the previous data with the use of fractionated DWG. Histamine and commercial porcine heparin glycosaminoglycan were not additive in inhibiting lectin-induced lymphocyte blastogenesis. The inhibitory effect of heparin on Con A-induced blastogenesis could be overcome by adding excess Con A to macrophage-depleted mononuclear cell preparations and could be reproduced by using over-sulfated chondroitin 6-sulfate. This is the first examination of the ability of mast cell granules to influence lymphocyte function and the first demonstration of the ability of native heparin proteoglycan to modulate lymphocyte blastogenesis.
Modulation of the biologic activities of IgE-binding factor. IV. Identification of glycosylation-enhancing factor as a kallikrein-like enzyme.
Authors: Iwata M , Munoz J J , Ishizaka K
Published: December 2022

Stimulation of normal rat splenic T cells with pertussigen (lymphocytosis-promoting factor, LPF, from Bordetella pertussis) resulted in the release of a soluble factor that enhanced the glycosylation of IgE-binding factors during their biosynthesis. The soluble factor was detected by the ability of a culture filtrate of LPF-stimulated spleen cells to switch a T cell hybridoma, 23A4, from the formation of unglycosylated IgE-binding factor to the formation of glycosylated IgE-binding factor. The glycosylation-enhancing factor (GEF) had affinity for D-galactose, and the binding of the factor to hybridoma cells via a cell surface galactose was essential for modulation of IgE-binding factors. The GEF was inactivated by irreversible inhibitors of serine proteases such as phenylmethylsulfonyl fluoride, diisopropylfluorophosphate, and p-nitrophenyl ethylpentylphosphonate but was not affected by nonphosphorylating analogues of the organophosphorus compounds. Benzamidine, a competitive and reversible inhibitor of trypsin, also inhibited the glycosylation of IgE-binding factors by GEF. The factor could be purified by absorption to p-aminobenzamidine agarose followed by elution with benzamidine. The capacity of GEF to enhance the glycosylation of IgE-binding factors was inhibited by synthetic substrates of trypsin but not by substrates of chymotrypsin, indicating that GEF is a trypsin-like enzyme. Indeed, trypsin, plasmin, and kallikrein enhanced the glycosylation of IgE-binding factors during their biosynthesis. An inhibitor of trypsin-like enzyme(s), N-alpha-p-tosyl-L-lysine chloromethylketone (TLCK), inhibited trypsin and plasmin but not kallikrein, and TLCK failed to inhibit the GEF-mediated enhancement of glycosylation. It was also found that bradykinin, the biologically active product of cleavage of kininogen by kallikrein, enhanced the glycosylation of IgE-binding factors. The results indicate that GEF is a kallikrein-like enzyme.
Monoclonal anti-parasite and anti-RBC antibodies produced by stable EBV-transformed B cell lines from malaria patients.
Authors: Lundgren K , Wahlgren M , Troye-Blomberg M , Berzins K , Perlmann H , Perlmann P
Published: December 2022

To produce human monoclonal antibodies associated with infectious disease, peripheral blood lymphocytes (PBL) from patients with Plasmodium falciparum malaria were transformed with EB-virus in vitro. To enrich for malaria-specific B cells, PBL were incubated for 3 days with unsoluble P. falciparum antigen before EBV-transformation. Furthermore, cyclosporin A was added during and after transformation to eliminate T cell suppression of B cell growth. Microcultures were screened for antibodies against blood stage antigens of P. falciparum or of noninfected erythrocytes by ELISA and indirect immunofluorescence. Cultures producing anti-P. falciparum and/or anti-erythrocyte antibodies were developed from the lymphocytes of eight patients, including some individuals with their first infection. Positive cultures were cloned and propagated for several weeks. Seven of 15 clones producing antibody at a stable rate have now been kept in cultures for more than 1 yr. Of six cultures analyzed in detail, all produced IgM antibodies of either K or lambda isotype. Although three clones were monoclonal after one cloning, three were oligoclonal. Of the former, two produced P. falciparum-specific antibodies directed to an antigen associated with the surface of merozoites. One of the oligoclonal cultures produced anti-erythrocyte antibodies, and it was probably reacting with spectrin.
Secondary in vitro generation of CTL specific for MM antigen by stimulation with somatic tumor hybrid, but not with parental tumor cells.
Authors: Ishikawa S , Tachibana T
Published: December 2022

Somatic tumor hybrid cells (L-FM3A#2), obtained by hybridization of MM tumor cells (FM3A/R) with HGPRT-less L cells, could induce CTL directed against MM antigen, a tumor-associated transplantation antigen that is expressed on some ascitic mammary tumor cell lines of C3H/He mice; parental tumor cells (FM3A/R) could not produce such CTL in syngeneic mice. In this study, the mechanisms of the generation of CTL by stimulation with L-FM3A#2 hybrid cells were investigated. In the secondary in vitro stimulation system, L cell component(s) that were introduced into hybrid cells by cell fusion play a role in the induction of CTL, as shown by the fact that stimulation with a mixture of L and FM3A/R cells could induce MM antigen-specific CTL, and that the killer helper effect of L cells could be replaced by cell free culture supernatants obtained from co-cultures of unprimed C3H spleen and L cells. IFN activity, but no IL 2 activity, was detected in the culture supernatants. Both IFN and killer helper activity were lost with pH 2 treatment; furthermore, CTL were generated by stimulation of primed spleen cells with FM3A/R cells in the presence of mouse beta-IFN, but not in its absence. These results suggest that IFN liberated by the helper cells that recognize L cell component(s) on the surfaces of tumor hybrid cells plays an essential role in the generation of CTL specific for MM antigen.
The use of specific helper T cell clones to study the regulation of isotype expression by antigen-stimulated B cell clones.
Authors: Teale J M
Published: December 2022

In order to determine the mechanism by which helper T cells regulate the production of the various immunoglobulin (Ig) classes, a number of helper T cell clones specific for keyhole limpet hemocyanin (KLH) were generated. These helper T cell clones then were used in a modified splenic fragment system whereby cloned helper T cells and a source of B cells were limit-diluted into naive, lethally irradiated recipients. The B cell clones that were subsequently stimulated in such an assay system by the addition of the antigen 2,4-dinitrophenol (DNP)-KLH then were tested for the various isotypes produced. The results of these studies indicate that the use of a single helper T cell clone could result in the production of all known Ig isotypes including IgE. Moreover, the use of a single helper T cell clone could result in multiple isotype production by a single B cell clone. However, a comparison of the isotypes secreted by a number of different B cell clones that were stimulated with the same helper T cell clone indicated that a variety of isotypic patterns could be obtained. In addition, it was found that the majority of B cell clones produced in the presence of T cell clones secrete fewer numbers of different isotypes compared with B cell clones generated with a heterogeneous population of T cells. Finally, no evidence could be found for isotype-specific helper T cell clones, although a few of the T cell clones appeared to induce a somewhat restricted isotype pattern in which only two or three different isotypes were observed.
Regulation of the hapten-specific T cell response. I. Preferential induction of hyporesponsiveness to the D-end of the major histocompatibility complex in the hapten-specific cytotoxic T cell response.
Authors: Scott C F , Tsurufuji M , Benacerraf B , Sy M S
Published: December 2022

In this paper we have examined the phenomenon of hapten-specific tolerance in the cytolytic T lymphocyte (CTL), using the trinitrophenyl (TNP) and azobenzenearsonate haptens. We found that the H-2 K and H-2 D-end restricted CTL in H-2a mice are differentiable in the ease with which they are tolerized to the TNP hapten. With TNP modified syngeneic spleen cells (TNP-SC), or low amounts of trinitrobenzylsulfonic acid as tolerogen, preferential hyporesponsiveness of D-end restricted CTL can be observed. Larger doses of hapten, e.g. a higher amount of trinitrobenzylsulfonic acid, will tolerize both K- and D-end restricted TNP-specific CTL in H-2a mice. The phenomenon of preferential D-end restricted CTL hyporesponsiveness is not observed in H-2d, H-2k, or H-2b mice, nor is it observed in H-2a mice with respect to the azobenzenearsonate hapten. We have also shown that the clones of CTL responsible for lysis of TNP-modified allogeneic targets (cross-reactive lysis) in H-2a mice probably overlap with the D-end restricted TNP-specific CTL since D-end restricted hyporesponsiveness induced by intravenous injection of TNP spleen cells also results in the elimination of cross-reactive lysis of TNP-modified allogeneic targets. The possible mechanisms of preferential D-end hyporesponsiveness to the TNP hapten in the H-2a mice as well as its significance and relationship to previous work in this area are discussed in this paper.
Lymphokine-mediated activation of a T cell-dependent IgA antipolysaccharide response.
Authors: Kagnoff M F , Arner L S , Swain S L
Published: December 2022

Immune responses to bacterial polysaccharides are important to host immunity at mucosal surfaces. We previously showed that BALB/c mice produce substantial T cell-dependent IgA responses to alpha (1,3) glucan determinants on the bacterial capsular polysaccharide dextran B1355. The data in this study demonstrate that the requirement for T cells for the activation of the IgA anti-alpha (1,3) dextran B1355 response can be replaced by T cell-derived nonantigen specific helper factors that appear to act during the late stages of B cell differentiation. Supernatants from the activated T cell lines cr-15 and (DL)C.C3.11.75, which contain interferon and late-acting T cell replacing factor activity, supported terminal differentiation of dextran-stimulated B cells to IgA anti-alpha (1,3) glucan antibody-forming cells and substantially increased IgM anti-alpha (1,3) glucan responses in culture. Although supernatants with interleukin 2 activity did not support optimal antigen-driven plaque-forming cell responses, they synergized with supernatants having interferon and T cell replacing factor activity in the production of IgA and IgM anti-alpha (1,3) glucan responses and IgM anti-SRBC responses. Supernatants from the T cell lines B6.11 and (DL)A.4 contained B cell growth factor activity but did not support activation of IgA anti-alpha (1,3) glucan PFC. These studies suggest that interferon and/or T cell replacing factor play an important role in the antigen-driven differentiation of B cells of the IgA and IgM isotypes to antibody-forming cells.
Genetic analysis of the inheritance of B cell hyperactivity in relation to the development of autoantibodies and glomerulonephritis in NZB x SWR crosses.
Authors: Eastcott J W , Schwartz R S , Datta S K
Published: December 2022

Autoimmune NZB mice have a primary B cell abnormality manifested by spontaneous hypersecretion of IgM in short-term cultures. The purpose of this study was to characterize the genes that specify this intrinsic B cell defect and determine their relationship to "autoimmunity" genes. The increase in IgM secretion was shown to result from two independently segregating genetic traits: a) an increased number of IgM-containing cells, and b) an increased rate of secretion of IgM per cell. In this study, approximately 600 NZB, SWR, F1, F2, and reciprocal back-cross mice were hemisplenectomized at 5 to 6 months of age to determine the number of IgM-containing cells and the secretion of IgM in 4-hr cultures. These animals were followed prospectively for the development of anti-DNA (native and denatured) and anti-red blood cell autoantibodies, and proteinuria, and studied at autopsy for the development of glomerulonephritis. The severity and incidence of renal lesions was influenced to some extent by the presence of B cell hyperactivity. However, a proportion of F2 and backcross progeny mice that did not show B cell hyperactivity eventually developed autoantibodies and autoimmune disease.
Relationship of germinal centers in lymphoid tissue to immunologic memory. VI. Transfer of B cell memory with lymph node cells fractionated according to their receptors for peanut agglutinin.
Authors: Coico R F , Bhogal B S , Thorbecke G J
Published: December 2022

We isolated germinal center B cells by exploiting their high affinity for peanut agglutinin (PNA). The PNA+ and PNA- B cells, fractionated by panning on PNA-coated petri dishes, were examined for their ability to transfer memory responses to irradiated recipients at various times after priming. With such fractionated B cells from lymph nodes taken at the peak of germinal center formation, the largest response was obtained in recipients of the PNA+ B cell population. At 4 to 5 wk after priming, and 10 days after challenge with an unrelated antigen, memory responses were approximately equal in recipients of PNA+ or PNA- B cells. At 14 wk after priming, memory responses were found only in recipients of the PNA- B cell population. Memory B cells from the spleen, taken from mice primed in the footpad 8 wk earlier, were also PNA-. Finally, we show that boosting with a TNP-conjugate in the footpad, 6 mo after priming in the same footpad, induced the reappearance of marked memory responsiveness in the PNA+ B cell fraction of the draining node.
A new mediator (suppressor cell induction factor) activating T cell-mediated suppression: characterization of suppressor cells, kinetics of their generation, and mechanism of their action.
Authors: Kasakura S , Taguchi M , Murachi T , Uchino H , Hanaoka M
Published: December 2022

Human T cells precultured in MLC supernatant (SIF) could suppress responder cells in MLC. SIF-activated cells expressed suppressor effects equally on both autologous and allogeneic responder cells in MLC. The suppression observed was not due to cell crowding, cell death, depletion of nutrients, absorption of IL 2, alternation in the kinetics of MLC response, production of soluble inhibitory factor, or generation of cytotoxic cells. In this system, suppressor cells were generated after 5 days of culture, and Ia-positive cells were required in the induction of suppressor cells. Suppressor cells themselves, however, were characterized as Ia-negative subsets of T cells; they could be derived from both Fc-IgG receptor-positive and -negative subsets of T cells. Suppressor activity of SIF-treated cells on the MLC response was sensitive to mitomycin C treatment. They apparently exerted their suppressor effect on some early phase of the MLC response. This early event was almost complete within 48 hr after the initiation of MLC. The possibility that SIF may be involved in T-T interactions required in the generation of effective suppression is discussed.
NK cell function in severe combined immunodeficiency (SCID): evidence of a common T and NK cell defect in some but not all SCID patients.
Authors: Peter H H , Friedrich W , Dopfer R , Müller W , Kortmann C , Pichler W J , Heinz F , Rieger C H
Published: December 2022

The immunologic work-up of eight infants with the clinical diagnosis of severe combined immunodeficiency (SCID) was performed with special emphasis on natural killer (NK) cell function and ontogeny. Contrary to previous reports, our study shows that not all SCID patients lack NK activity; some may even express very high NK- and antibody-dependent cellular cytotoxicity (ADCC). The present group of eight SCID infants was homogeneous with respect to normal levels of the purine metabolism enzymes adenosine deaminase (ADA) and purine nucleoside phosphorylase (PNP). They all had low serum Ig levels and were defective for specific antibody formation against BSA and diphtheria toxin (DiT). None of the infants' peripheral blood mononuclear cells (PBMC) proliferated significantly upon in vitro stimulation with PHA, concanavalin A (Con A), pokeweed mitogen (PWM), and irradiated allogeneic lymphocytes. Seven of eight patients, however, responded significantly to mitogenic factors present in a lectin-free interleukin 2 (IL 2) preparation, and two exhibited a positive costimulation as well with simultaneous exposure to IL 2 + Con A. The lymphocyte marker analysis revealed high percentages of OKT10+ cells in seven of eight infants, whereas peripheral T cells (OKT3+) with suppressor/killer (OKT8+) or helper/inducer (OKT4+) phenotypes were abnormally low in all infants with one exception. The PBMC of two patients formed low to normal percentages of E rosettes but expressed no B cell markers (B-/SCID). The six other infants had high percentages of B cells (B+/SCID) but lacked E rosette-forming cells. High NK and ADCC activity was found in the two B-/SCID patients. The B+/SCID infants either totally lacked NK and ADCC function (four of six) or expressed low to normal NK activity together with some T cell markers as revealed by monoclonal antibody staining but not by E rosette formation (two of six). From the data presented, an ontogenic model is proposed that assumes the status of an independent cell lineage in between T cells and monocytes for human NK cells, or that places these cells in close proximity to early differentiation steps of the T cell lineage. In any case, NK cell function clearly constitutes an additional parameter of heterogeneity in the immunologic analysis of SCID.
OKT8 antibody inhibits OKT3-induced IL 2 production and proliferation in OKT8+ cells.
Authors: Welte K , Platzer E , Wang C Y , Rinnooy Kan E A , Moore M A , Mertelsmann R
Published: December 2022

We studied IL 2 production and proliferation induced by OKT3 mitogenic monoclonal antibody in the OKT8+ T cell subset. OKT3 antibody induced IL 2 production and proliferation in OKT8+ cells in a typical time-dependent manner: maximal IL 2 levels were found in 24 hr culture supernatants; maximal proliferation was found on day 3. OKT3 antibody was mitogenic over a wide range of concentrations (0.125 to 500 ng/ml). The presence of OKT8 antibody (greater than or equal to 100 ng/ml) in these cultures resulted in almost complete inhibition of IL 2 production and proliferation. Kinetic studies demonstrate that OKT8 antibody suppresses both IL 2 production and response to exogenous IL 2 in OKT8+ cells when added within the first 2 hr of culture. After 14 to 20 hr of culture, addition of OKT8 only blocks IL 2 production but not the IL 2 response of activated OKT8+ cells. The specificity of inhibition by OKT8 antibody of OKT3 mitogenicity on OKT8+ cells was confirmed by the failure of Leu-I and OKT4 antibody to produce the same effect and by the lack of inhibition by OKT8 antibody of OKT3-induced IL 2 production and proliferation in OKT4+ cells.
Capacity of human large granular lymphocytes (LGL) to produce multiple lymphokines: interleukin 2, interferon, and colony stimulating factor.
Authors: Kasahara T , Djeu J Y , Dougherty S F , Oppenheim J J
Published: December 2022

Human large granular lymphocytes (LGL), which are known to be responsible for natural killer (NK) cell activity, also produced a variety of lymphokines including interleukin 2 (IL 2), colony stimulating factor (CSF), and interferon (IFN) in response to phytohemagglutinin (PHA) or concanavalin A (Con A). Human peripheral blood LGL, which were purified by removal of monocytes adhering to plastic flasks and nylon columns, followed by separation on a discontinuous Percoll gradient, and additional treatment with anti-OKT3 and Leu-M1 plus complement, were more potent producers of these lymphokines than unseparated mononuclear cells (MNC), nylon column-eluted cells, or purified T lymphocytes. Moreover, IL 2 production by LGL could be further distinguished in that it was not enhanced by the addition of macrophages or macrophage-derived factor, i.e., IL 1, whereas addition of macrophages did potentiate IL 2 production by T lymphocytes. Further analysis of cells in the LGL population using various monoclonal antibodies revealed that removal of cells with OKT11 or AF-10, a monoclonal antibody against human HLA-DR antigen, decreased IL 2 production, whereas removal of OKT8+, OKM1+, Leu-M1+, or Leu-7+ cells led to enhanced IL 2 production. The LGL population is therefore heterogeneous and includes at least three functionally and phenotypically distinct subsets. An atypical T cell subset (OKT3-, Leu-1-, OKT11+) rather than the myeloid subset of LGL (Leu-M1+ or OKMI+) was the source of LGL-derived IL 2, whereas the latter subset and/or another subset of OKT8+ cells appear to regulate this IL 2 production. In addition to performing NK activity, LGL on a per cell basis seem to be more effective than T lymphocytes in producing lymphokines, namely, IL2, CSF, and IFN.