bluuebunny/crossref_metadata_2025 huggingface dataset:
Enhanced TNP-reactive helper T cell activity and its utilization in the induction of amplified tumor immunity that results in tumor regression.
Authors: Fujiwara H , Moriyama Y , Suda T , Tsuchida T , Shearer G M , Hamaoka T
Published: December 2022

The present study was designed to investigate the generation of trinitrophenyl (TNP)-reactive helper T cell activity potent enough to induce the regression of a syngeneic tumor; this occurs by augmenting antitumor-specific immunity through T-T cell interaction. Mice whose skin was painted with trinitrochlorobenzene (TNCB) exhibited a variety of anti-TNP T cell responses, including delayed-type hypersensitivity (DTH) and cytotoxic T cell responses, as well as helper T cell activity. Pretreatment of C3H/He mice with TNP-conjugated copolymer of D-glutamic acid and lysine (TNP-D-GL) or cyclophosphamide, which have been shown, respectively, to inactivate TNP-specific suppressor T cells or suppressor T cells in general, exhibited a slight or marginal augmentation of DTH and cytotoxic potentials when tested 5 wk after TNCB painting. In contrast, the same pretreatment regimens induced an appreciably amplified generation of anti-TNP helper T cell activity. This amplified TNP-helper T cell activity was demonstrated to enhance cytotoxic responses to antigens other than TNP in an antigen-nonspecific way. In fact, such helper T cells enhanced antitumor CTL responses when co-cultured with spleen cells from syngeneic X5563 plasmacytoma-bearing mice in the presence of TNBS-modified X5563 tumor cells. This amplified TNP-helper cell system was utilized for its immunotherapeutic potential. When TNCB was injected into X5563 tumor mass of syngeneic C3H/He mice in which the amplified TNP-helper T cell activity had been generated, an appreciable number of growing tumors was observed to regress. This contrasted with the low incidence of tumor regression observed in mice in which TNP-helper activity had been induced by TNCB painting without inactivation of suppressors. Thus, the present model provides an effective immunotherapeutic manipulation for eliciting enhanced in vivo tumor regression, and emphasizes a role of helper T cells in augmentation of syngeneic tumor immunity.
Antigen-reactive cloned helper T cells. I. Unresponsiveness to antigenic restimulation develops after stimulation of cloned helper T cells.
Authors: Wilde D B , Fitch F W
Published: December 2022

Murine helper T lymphocyte (HTL) clones reactive to ovalbumin (OVA) were maintained in continuous culture in vitro. Clones were propagated by weekly stimulation in the presence of irradiated splenic filler cells, antigen, and supernatant fluid (SF) containing IL 2. By varying the quantity of these reagents in cultures of HTL cells, the reactivity to antigen of the cloned cells was altered markedly. After stimulation by antigen or SF, HTL clones became profoundly unresponsive to antigenic restimulation. Cells remained unresponsive for 2 to 9 days after stimulation, depending on the culture conditions that were chosen for their maintenance. The addition of SF containing a high concentration of IL 2 prolonged the duration of unresponsiveness by 3 days, and the presence of a non-T splenic filler cell increased the period of unresponsiveness by an additional 4 days. The use of a high concentration of OVA in cell cultures also prolonged the time of unresponsiveness. The results described here demonstrate that the response to antigen of HTL cells is down-regulated after stimulation, and appears to be correlated with exposure to SF that contains IL 2.
A reappraisal of "T-independent" antigens. II. Studies on single, hapten-specific B cells from neonatal CBA/H or CBA/N mice fail to support classification into TI-1 and TI-2 categories.
Authors: Nossal G J , Pike B L
Published: December 2022

Spleen cells from adult CBA/H or CBA/N mice, or from neonatal CBA/H mice, were fractionated on thin layers of fluorescein (FLU)-gelatin to yield FLU-specific B lymphocytes. A single cell, or small numbers ranging from 1 to 10, were cultured in 10-microliter microcultures together with various antigens and mitogens. The results were compared with those of bulk culture or limiting dilution cultures supported by thymus filler cells. B cell growth and differentiation-promoting conditioned media (BGDA) were added to some cultures. The CBA/N results gave no support to the commonly used classification of T cell-independent (TI) antigens into TI-1 and TI-2 categories. A typical supposed TI-1 antigen, FLU-LPS, strongly stimulated normal adult single FLU-specific B cells to proliferate and form antibody, but virtually failed to trigger CBA/N B cells of comparable antigen-binding avidity. The same was true of LPS or LPS plus dextran sulfate acting as mitogens. The allegedly TI-2 antigen FLU-Ficoll, although still triggering comparatively poor responses, was actually marginally more active than FLU-LPS. FLU-Brucella abortus (FLU-BA) + BGDA gave the best results with single CBA/N B cells, but still induced only 1.27% of cells to develop into antibody-forming clones vs 12.2% with CBA/H cells. The results obtained with single neonatal B cells also lent no support to the distinction between TI-1 and TI-2. Both "TI-1" and "TI-2" stimuli caused adequate proliferation, one "TI-2" antigen stimulating 23.2% of the cells. None of the antigens caused good antibody formation, however, probably because multivalent antigens can deliver signals impeding the differentiation of immature B cells. It is therefore suggested that the classification of TI-1 antigens into two subcategories be abandoned, at least for the time being.
Avian thymic accessory cells.
Authors: Oliver P D , LeDouarin N M
Published: December 2022

On the basis of morphologic criteria and ingestion of latex particles, two basic types of accessory cells can be identified from quail and chick thymuses, dendritic cells, and macrophages. By using embryonic grafting techniques, we show that cells of this lineage enter the thymus during the initial colonization of the epithelial thymic rudiment by hemopoietic cells, and within a few days differentiate into cells exhibiting properties of glass adherence, Ia expression, and formation of rosettes with thymocytes. It appears that the precursors of this lineage undergo extensive, but finite, proliferation and are eventually replaced by further influx of the accessory cell lineage. In chimeric grafts, quail thymocytes were seen forming rosettes with chick accessory cells, and vice versa, indicating, as in the interaction between the epithelial cells and thymocytes, that the molecules involved in thymocyte-accessory cell association can interact across species barriers in our system.
Deoxyadenosine modulates human suppressor T cell function and B cell differentiation stimulated by Staphylococcus aureus protein A.
Authors: Cohen A H , Bluestein H G , Redelman D
Published: December 2022

Adenosine deaminase (ADA) deficiency and the resultant accumulation of deoxyadenosine (AdR) are associated with profound T cell dysfunction and variable B cell dysfunction in vivo. We examined the effects of AdR on the in vitro function of normal human peripheral blood B and T lymphocytes whose ADA activity was inhibited by 2'-deoxycoformycin. We found that OKT8+ T cell-mediated suppression of SPA-induced Ig production was markedly reduced by concentrations of AdR (3 to 10 microM) that did not affect helper T cell function. Because the lectin-induced proliferative responses of OKT8+ T cells and OKT8- T cells were equally susceptible to AdR, modulation of in vitro immune responses by low-dose AdR probably reflected different proliferative requirements for the expression of T cell helper or suppressor functions. Although low doses of AdR did not inhibit Ig production in SPA-stimulated cultures, we found that T cell-dependent, SPA-stimulated B cell proliferation was blocked by 3 to 10 microM AdR. Therefore, it appeared that B cell proliferation was not required for the induction of Ig synthesis in this system. Higher doses (30 to 100 microM) of AdR did block the induction of Ig synthesis, presumably by interfering with T-helper functions via a mechanism other than inhibition of proliferation and/or by inhibiting B cell differentiation events.
Lymphocyte ecto-5'-nucleotidase activity in infancy: increasing activity in peripheral blood B cells precedes their ability to synthesize IgG in vitro.
Authors: Bastian J F , Ruedi J M , MacPherson G A , Golembesky H E , O'Connor R D , Thompson L F
Published: December 2022

Ecto-5'-nucleotidase activity was measured in peripheral blood lymphocytes isolated from serial specimens from nine healthy full-term infants and two premature infants at 0, 2, 4, and 6 mo of age. The postnatal nadir in activity was 7.1 +/- 2.0 nmol/hr/10(6) cells, which is the same as the activity in cord blood lymphocytes (7.0 +/- 2 nmol/hr/10(6) cells). The activity rose twofold to 13.2 +/- 3.8 nmol/hr/10(6) cells at 6 mo of age (p less than 0.001, paired t-test), which is similar to the activity in adult peripheral blood lymphocytes (14.1 +/- 6.3 nmol/hr/10(6) cells). This increased activity in total lymphocytes reflects increased activity in the B cell population. B cell ecto-5'-nucleotidase activity in two infants at 12 to 13 mo of age was 19.3 and 25.2 nmol/hr/10(6) cells, values that are four-to fivefold higher than for cord blood B cells (5.6 +/- 2.8 nmol/hr/10(6) cells) and within the normal range for adult B cells (27.9 +/- 12 nmol/hr/10(6) cells). In spite of a greatly expanded peripheral blood B cell population, studies of immunoglobulin biosynthesis in vitro demonstrated that infant peripheral blood B cells are functionally immature with no synthesis of IgG in response to Epstein Barr virus. Thus, the increase in peripheral blood B lymphocyte ecto-5'-nucleotidase activity in infants precedes their acquisition of a capacity for IgG synthesis in vitro. Data from a hypogammaglobulinemic infant revealed a persistently low ecto-5'-nucleotidase activity over a 10-mo period until at 14 mo of age the activity was 8.8 nmol/hr/10(6) cells in total lymphocytes and 13.0 nmol/hr/10(6) cells in B cells, which correlated with in vivo and in vitro evidence of delayed B cell maturation. Thus, ecto-5'-nucleotidase activity may be a useful cell surface marker in studies of human postnatal B cell maturation.
Differential inhibition of human antigen-specific T cell clone proliferative responses by distinct monoclonal anti-HLA-DR antibodies.
Authors: Triebel F , Missenard-Leblond V , Couty M C , Charron D J , Debre P
Published: December 2022

The inhibition of diphtheria toxoid and varidase-specific T cell clones from a single DR 6/7 donor by eight distinct monoclonal anti-HLA-DR antibodies was tested in proliferative assays. These moAb were selected because they had been previously defined for their ability: 1) to react with Ia molecules; 2) to recognize similar or different epitopes; 3) to share or not share idiotypic specificities. Our results show a distinct inhibition pattern for each clone tested. Furthermore, the various moAb could be classified into three groups according to their inhibitory effect on T cell proliferation. These data suggest: a) an epitopic restriction by class II antigens of antigen-specific human T cell clone proliferation; and b) the recognition of functional epitopes on the human Ia-like antigens by some but not all moAb studied.
Lysosomal enzyme release from polymorphonuclear leukocytes induced by immune complexes of IgM and of IgG.
Authors: Lucisano Y M , Mantovani B
Published: December 2022

Lysosomal enzyme release by rabbit polymorphonuclear leukocytes induced by immune complexes of IgM or IgG antibodies and ovalbumin was studied by in vitro experiments. It was shown that IgM immune complexes in the form of precipitates were able to stimulate the release of the lysosomal enzymes beta-glucuronidase and alkaline and acid phosphatases, in contrast to heat-aggregated IgM, which had only a small effect. This stimulation was dependent on the presence of Ca2+ in the extracellular medium; Mg2+ showed a slight effect as a substitute for Ca2+. IgG precipitated immune complexes prepared from different regions of the precipitation curve were tested. It was shown that immune precipitates from the equivalence region were the most effective; those at antibody excess have a smaller but comparable capacity, whereas the precipitated immune complexes at antigen excess were the least effective. Soluble immune complexes of IgG with an excess of antigen concentration in the range of 2.5 to 5 times that of equivalence were also able to stimulate the enzyme release, but that capacity decreased rapidly with the increase in antigen concentration. The enzyme release induced by IgM immune complexes was not inhibited by competition with free immunoglobulin in the medium, either IgM or IgG in approximate physiologic concentrations; contrarily, with IgG immune complexes, free monomeric IgG (but not IgM) can completely block the process. The possible implications of these findings in the immune complex diseases are discussed.
Suppression of NK-mediated natural resistance by interferon treatment of murine lymphomas.
Authors: Greenberg A H , Miller V , Jablonski T , Pohajdak B
Published: December 2022

The observation that interferon (IFN) can suppress the NK lytic sensitivity of murine lymphomas in vitro led us to examine the consequences of this treatment on tumor behavior in vivo. Preincubation in IFN suppressed natural resistance to two lymphomas in syngeneic DBA/2 and semisyngeneic BDF1 mice in a dose-dependent manner, measured by the retention of (131I)dUrd-labeled tumor. Poly I:C enhancement of NK-mediated natural resistance in the lung, liver, and peritoneal cavity was also abolished by IFN pretreatment. IFN was, however, ineffective in altering the elimination of the IFN-resistant L1210R lymphoma when compared to its IFN-sensitive variant, L1210S. In DBA/2 mice that were made NK-deficient by treatment with cyclophosphamide or rabbit anti-asialo GM1 antiserum, or in congenitally NK-deficient bg/bg strain mice, IFN-treated tumor and control tumor were rejected equally well. This indicated that the effects of IFN were dependent on the presence of NK cells in these mice, and suggests that the IFN suppressed the sensitivity of the lymphomas to NK cell-mediated host resistance.
Phorbol ester increases the level of interleukin 2 mRNA in mitogen-stimulated human lymphocytes.
Authors: Hirano T , Fujimoto K , Teranishi T , Nishino N , Onoue K , Maeda S , Shimada K
Published: December 2022

TPA alone did not induce the production of IL 2 in human tonsillar lymphocytes but enhanced the PHA-induced IL 2 production by seven-fold. That the effect of TPA was due to an increase in IL 2 mRNA was demonstrated by examining the amount of IL 2 mRNA translatable in Xenopus laevis oocytes, and by Northern blotting analysis using IL 2 cDNA as a probe. In these ways, it was shown that TPA alone did not induce any significant IL 2 mRNA synthesis, but when added together with PHA it increased the level of IL 2 mRNA by at least 10-fold, as compared with that induced by PHA alone.
Phosphorylcholine on Streptococcus pneumoniae R36a is responsible for in vitro polyclonal antibody secretion by human peripheral blood lymphocytes.
Authors: Beckmann E , Levitt D
Published: December 2022

We have shown that Pc on the C-polysaccharide of Streptococcus pneumoniae R36a is responsible for polyclonal PFC responses induced in vitro by this bacterium in humans. R36a grown in media containing EA instead of CL, and therefore having phosphorylethanolamine instead of Pc in their C-polysaccharide, were unable to induce substantial PFC responses. When EA-substituted bacteria were chemically conjugated with Pc, their ability to induce polyclonal PFC was restored. Specific removal of Pc from the surface of the bacteria by the use PLC also resulted in abrogation of the polyclonal antibody response. These data are consistent with our hypothesis that polyclonal activation resulting from R36a stimulation may be mediated by a recently described Pc-binding receptor that is distributed on the surface of a subpopulation of B lymphocytes in humans and mice.
The effect of deoxyguanosine on human lymphocyte function. I. Analysis of the interference with lymphocyte proliferation in vitro.
Authors: Spaapen L J , Rijkers G T , Staal G E , Rijksen G , Wadman S K , Stoop J W , Zegers B J
Published: December 2022

The effect of deoxyguanosine on mitogen- and antigen-induced proliferation of peripheral blood lymphocytes from healthy donors was studied. Deoxyguanosine was found to inhibit the proliferative response to mitogens and antigens. Concentrations of deoxyguanosine causing 50% inhibition of the proliferation proved to be dependent on the activity of catabolic enzymes, such as purine nucleoside phosphorylase (PNP), in sera used in the culture media. The inhibitory effect of deoxyguanosine on phytohemagglutinin (PHA)-induced cell proliferation was prevented by deoxycytidine as well as by hypoxanthine. These findings were analyzed further by determination of intracellular (deoxy)-nucleotide levels. Stimulation of lymphocytes by PHA in the presence of deoxyguanosine leads to intracellular accumulation of dGTP. The presence of hypoxanthine in addition to deoxyguanosine abolished the inhibitory effect but did not prevent dGTP accumulation. On the other hand, the addition of deoxycytidine in combination with deoxyguanosine did not lead to intracellular accumulation of detectable amounts of dGTP, but only gave partial protection against the toxic effect. Furthermore, guanosine inhibited mitogen-induced cell proliferation to the same extent as did deoxyguanosine provided that the culture media were supplemented with pretreated fetal calf serum. Peripheral blood lymphocytes of a PNP-deficient or a HGPRT-deficient patient in cultures stimulated with PHA or pokeweed mitogen were resistant to the inhibitory effects of guanosine and were less sensitive to deoxyguanosine than cells of normal donors. The present results clearly show the involvement of two pathways contributing to deoxyguanosine-mediated inhibition of the proliferation of normal lymphocytes, i.e., on the one hand degradation of deoxyguanosine by PNP, salvage of guanine by HGPRT, and (possibly) phosphorylation of GMP eventually leading to GTP, and on the other hand formation of dGTP by direct phosphorylation of deoxyguanosine by deoxycytidine kinase.
Partial immunochemical characterization of the Ro and La proteins using antibodies from patients with the sicca syndrome and lupus erythematosus.
Authors: Elkon K B , Culhane L
Published: December 2022

Ro and La are intracellular ribonucleoproteins that are frequent targets for autoantibodies in the sicca syndrome and in lupus erythematosus. We analyzed the m.w. of the protein (antigen) moieties of Ro and La in saline cell extracts of human spleen by SDS-PAGE and immunoblotting, gel filtration, and sucrose density ultracentrifugation with radioimmunoassay. pI values for Ro and La proteins were established by isoelectric focusing on thin layer agarose gels and immunoblotting on nitrocellulose transfers. The La protein had an m.w. of approximately 43 kd and was heterogeneous in charge, with pI values from 4.2 to 4.8. Composite two-dimensional maps developed by immunoblotting revealed a characteristic set of seven dots of m.w. 43 kd. Ro determinants were identified on polypeptides of 50 and/or 57 kd. Antigenic activity was also detected in the void volume of spleen extract fractionated by Sephadex G-200 and in 8 to 9S and greater than 19S regions of sucrose gradients, suggesting either aggregation of the Ro protein or participation in protein-protein complexes. pI values of 4.3 to 5.5 were obtained for the Ro antigen, and two-dimensional maps revealed that the 57 kd polypeptide had a similar charge heterogeneity to the La protein, whereas the 50 kd polypeptide had a different fingerprint. Immunoblotting of extracts from bovine, rabbit, and dog extracts showed that antibodies to Ro and La reacted with a limited number of polypeptides (m.w. 50 and/or 57 kd for anti-Ro and 43 or 50 kd for anti-La). These studies support the physical independence of the isolated Ro and La polypeptides, although a precursor product or functional relationship in vivo is possible. These studies also suggest that, in addition to Western blotting, techniques involving immunodeletion, isoelectric focusing with capillary immunoblotting, and 2D immunoblotting provide useful approaches to characterize saline-soluble cellular antigens.
Suppression of human peripheral blood lymphocytes by Fusobacterium nucleatum.
Authors: Shenker B J , DiRienzo J M
Published: December 2022

Fusobacterium nucleatum has been implicated in the pathogenesis of several diseases, including urinary tract infections, bacteremia, pericarditis, otitis media, and disorders of the oral cavity such as pulpal infections, alveolar bone abscesses, and periodontal disease. In this study, we examined sonic extracts of F. nucleatum strain FDC 364 for its ability to alter human lymphocyte function. We found that the soluble cytoplasmic fraction (CF) of the sonic extract was able to cause a dose-dependent inhibition of human lymphocyte responsiveness to Con A, PHA, PWM, and the recall antigen SKSD. Suppression involved altered DNA, RNA, and protein synthesis; there was no effect on cell viability. The suppressive activity is nondialyzable and heat labile. To achieve maximal suppression in 96-hr cell cultures, the CF had to be added to cells during the first 24 hr of incubation. Inhibition was reduced when the CF was added at 48 hr, and no suppression was observed when addition was at 72 or 96 hr (along with [3H]TdR). Furthermore, cells could be protected from the suppressive effects of the CF by washing within 24 hr of exposure. Suppression did not involve nonspecific effects on thymidine utilization. Although the mechanism of action of the F. nucleatum immunosuppressive activity has not yet been determined, we can rule out a requirement for monocytes/macrophages and activation of T suppressor cells. It has been proposed that impaired host defense may play a pivotal role in the pathogenesis of many diseases. The data presented in this paper suggest that local and/or systemic immunosuppression could be initiated by F. nucleatum. This immunosuppression may alter the nature and consequences of host-parasite interactions, thereby enhancing the pathogenicity of F. nucleatum itself or that of some other opportunistic organism.
The immunopathology of experimental allergic encephalomyelitis. I. Quantitative analysis of inflammatory cells in situ.
Authors: Sobel R A , Blanchette B W , Bhan A K , Colvin R B
Published: December 2022

Acute experimental allergic encephalomyelitis (EAE) is a T cell-mediated, neurologic disease that is under immunogenetic control. We systematically analyzed the quantity and distribution of T cells, B cells, and macrophages in the central nervous system (CNS) of susceptible and resistant guinea (GP) with a panel of seven monoclonal antibodies by using the avidin-biotin complex (ABC) immunoperoxidase technique and alpha-naphthyl-butyrate esterase (ANBE) staining. Adult EAE-susceptible strain 13 GP immunized with isogeneic spinal cord homogenate (SC) or with myelin basic protein (MBP) developed clinical signs (paralysis, weight loss, etc.) in 2 to 3 wk. T cells were present in all CNS inflammatory foci and comprised 44% of the perivascular mononuclear cells. T cells diffusely infiltrated the neuropil away from inflammatory cell aggregates. These T cells were judged to be extravascular by the lack of an associated identifiable vessel in counter-stained sections, and by their persistence following exhaustive perfusion of the brains. In routine sections, mononuclear cells could be detected only in perivascular aggregates. IgM+ B cells comprised 9% of the perivascular infiltrates and did not diffusely infiltrate the parenchyma. ANBE+ macrophages comprised the remaining 47% of the identified perivascular cells. SC- and MBP-immunized GP showed equivalent numbers of inflammatory foci, T cells, and macrophages, but SC-immunized GP had more IgM+ cells in the meninges and choroid plexus (p less than 0.001, p less than 0.02, respectively). Virtually all cells in perivascular locations were Ia+. Ia+ mononuclear cells were also present in the neuropil. EAE-resistant strain 2 GP immunized with SC developed no clinical signs. These GP had fewer perivascular foci than strain 13 GP but, when present, the cellular composition, including the density of diffuse parenchymal T cell infiltrates, was indistinguishable. Significantly fewer parenchymal mononuclear cells in the strain 2 GP, however, displayed Ia, both in perivascular and diffuse infiltrates (p less than 0.001). We conclude that T cell migration into the CNS parenchyma is a characteristic feature of acute EAE in the GP, but that T cells can occur in this pattern without clinical signs of disease. The two features that distinguish susceptible and resistant strains were the frequency of perivascular infiltrates and the expression of Ia on parenchymal mononuclear cells, which probably reflects their enhanced immunologic activation in situ.
Modulation of class I HLA antigens on HL-60 promyelocytic leukemia cells by serum-free medium: re-induction by gamma-IFN and 1,25-dihydroxyvitamin D3 (calcitriol).
Authors: Ball E D , Guyre P M , Glynn J M , Rigby W F , Fanger M W
Published: December 2022

During studies on the effect of different nutrient media on the growth and differentiation of the HL-60 promyelocytic leukemia cell line, we found that the density of the class I HLA antigens is profoundly decreased on cells cultured in a serum-free medium. The ability of recombinant DNA-derived interferons (IFN) and a number of myeloid differentiating agents to induce re-expression of class I HLA antigens and beta-2-m was therefore studied. All three classes (alpha, beta, and gamma) of IFN were capable of re-inducing HLA and beta-2-m, although gamma-IFN was more potent. Of a variety of chemical differentiating agents, only 1,25-dihydroxyvitamin D3 (calcitriol) was found to induce HLA class I antigens. These results suggest that HLA and beta-2-m are not necessarily constitutive cell surface proteins, but instead that their expression is highly inducible.
Two-dimensional gel analysis of a second family of class II molecules by polymorphic HLA-DR4,5, and w9 monoclonal antibodies.
Authors: Maeda H , Hirata R , Okuyama M , Thompson A , Tohyama H
Published: December 2022

Human Ia-like, class II molecules were isolated by immunoprecipitation with monoclonal antibodies from various HLA-D/DR homozygous cell lines and were analyzed by two-dimensional gel electrophoresis. The monoclonal antibody PLM12 reacted with B cells carrying DR4, DR5, DRw6.2, and DRw9 phenotypes, and its reactivity perfectly correlated with the previously defined TB21 (MB3-like) specificity. Class II molecules detected by PLM12 were structurally distinct from those precipitated by the anti-DR monoclonal antibody NC1 on all HLA-DR4, DR5, DRw6.2, and DRw9 homozygous cell lines and showed polymorphism in heavy and light chains among these cell lines. The monoclonal antibodies PLM2 and PLM9 only reacted with B cells carrying DR5 and DRw6.2 and also detected a distinct set of class II molecules from those precipitated by NC1 but identical to those of PLM12. Thus, PLM2 and PLM9 serologically detected a new subtypic antigen of the PLM12-reactive class II molecules. Furthermore, the antibody NC1 precipitated two light chains and one heavy chain from HLA-DRw6.2 homozygous cell line EBV-Sh. The result indicated the presence of three sets of class II molecules: two in a DR family and another carrying the polymorphic determinants detected by PLM2, PLM9, and PLM12 in a second family.
Monoclonal antibodies against SSEA-1 antigen: binding properties and inhibition of human natural killer cell activity against target cells bearing SSEA-1 antigen.
Authors: Harris J F , Chin J , Jewett M A , Kennedy M , Gorczynski R M
Published: December 2022

We describe the properties of three monoclonal antibodies (Mab) against stage-specific embryonic antigen-1 (SSEA-1) in terms of their binding activity to HL60, K562, OTF9, and SOTF9 tumor target cells and their functional activity in modulating human natural killer (NK) cytotoxicity assays in vitro against these target cells. Indirect binding, competition, and Western blot analyses indicate that the Mab AEC3A1-9 (3A1), ASSEA-1, and AECAB1-32 (AB1) recognize cell-defined SSEA-1 antigen with activity characteristic of the cell source (HL60 greater than OTF9 greater than K562 much greater than SOTF9). The addition of anti-SSEA-1 Mab to the NK cytotoxicity assay resulted in an inhibition of LU per 1 X 10(6) PBL that correlated closely with the expression of SSEA-1 antigen on the target cell. No significant inhibition was seen for seven other Mab. Inhibition of NK activity (greater than 30%) was observed in the presence of anti-SSEA-1 Mab for 18 of 21 and 6 of 7 human donors examined for HL60 and OTF9 target cells, respectively. The pretreatment of fixed competing cells with anti-SSEA-1 Mab reduced the efficacy of those cells to act as cold competitors in a standard NK cytotoxic assay. Taken together these data suggest that SSEA-1 determinants are important at some stage in the cytolysis produced by NK cells.
Activation of rat serosal mast cells by chymase, an endogenous secretory granule protease.
Authors: Schick B , Austen K F , Schwartz L B
Published: December 2022

Chymase, the major neutral protease of the rat serosal mast cell (RMC) secretory granule, causes RMC to release their secretory granules and to oxidatively metabolize endogenous arachidonic acid to prostaglandin D2 (PGD2). The granule markers, endogenous beta-hexosaminidase and exogenously added [3H]serotonin, were released from 2.5 X 10(5) RMC in 50 microliters in parallel and in dose-response fashion, reaching a maximum net percent release of approximately 50% with 0.5 to 1.0 units chymase (15 U/mg)/ml. With incremental concentrations of chymase, the release of granule markers occurred with a shorter lag period and in a greater maximal net percent, whereas the release of PGD2 was dose-related without a reduction in latency to detectable generation. Inhibition of the esterase activity of chymase with lima bean trypsin inhibitor decreased the subsequent mast cell response, indicating that the active site of chymase was required to initiate granule secretion and PGD2 generation. The monophasic indomethacin-resistant rise in cellular cAMP at 15 to 45 sec coincident with the onset of chymase-induced mediator release and PGD2 secretion is similar to that observed with IgE receptor-initiated coupled activation-secretion. The ability of heparin to block the activation function of chymase without inhibition of esterase activity reveals a possible physiologic regulatory mechanism for limiting the potential action of secreted chymase.
An immunoprotective monoclonal antibody to lipopolysaccharide.
Authors: Kirkland T N , Ziegler E J
Published: December 2022

The ability of antisera against lipopolysaccharide (LPS) raised by immunization with gram-negative bacteria to prevent LPS toxicity and death from gram-negative bacteremia is well established. To demonstrate conclusively that the protective antibody is specific for LPS, we tested an anti-LPS monoclonal antibody (mAb) in three animal models. 7G is an IgG3 mAb directed against an oligosaccharide side chain determinant of LPS from E. coli 0111:B4. This anti-LPS mAb increased the LD50 of 0111:B4 LPS in mice and protected rabbits against the dermal Shwartzman reaction elicited by 0111:B4 LPS. 7G mAb also protected mice against lethal infection with mucin-enhanced E. coli 0111:B4. Pretreatment with 250 micrograms of 7G increased the LD50 by more than 1.5 logs. These studies prove that oligosaccharide side chain-specific antibody to LPS confers protection against LPS toxicity in vivo and against experimental gram-negative infection. In addition, these studies suggest the potential of anti-LPS monoclonal antibody as therapy for gram-negative infection.
Genetic control of the induction of cytolytic T lymphocyte responses to AKR/Gross viral leukemias. II. Negative control by the Fv-1 locus in AKR mice of responder H-2b haplotype.
Authors: Green W R
Published: December 2022

To assess whether the presence of a responder H-2b haplotype would be sufficient to allow mice of nonresponder "high leukemic" phenotype to generate syngeneic anti-AKR/Gross virus cytolytic T lymphocytes (CTL), the AKR.H-2b strain was examined. Although capable of mounting vigorous apparent anti-minor histocompatibility-specific CTL responses, AKR.H-2b mice failed to produce anti-viral CTL after a variety of stimulation protocols. In contrast, the "doubly congenic" AKR.H-2b:Fv-1b strain was able to respond with substantial levels of H-2-restricted anti-AKR/Gross virus CTL activity. These results indicated that Fv-1n alleles could exert negative epistatic control over responder H-2b-encoded gene(s). Because the B6.Fv-1n congenic was also able to generate anti-viral CTL indistinguishable from the prototype B6 strain, however, it was apparent that other genes of AKR background were required for the Fv-1n-mediated inhibition in AKR.H-2b mice. The mechanism by which Fv-1 intereacted with other genes to override positive H-2b control appeared to be related to the expression of the CTL-defined, virus-associated antigens by normal AKR.H-2b cells. Thus, AKR.H-2b spleen cells but not thymus cells were able to stimulate the production of B6 anti-AKR/Gross virus CTL and were recognized as target cells by such anti-viral CTL. In contrast, both spleen cells and thymocytes from AKR.H-2b:Fv-1b mice were negative when tested as stimulator or target cells in these assays. In addition, AKR.H-2b but not AKR.H-2b:Fv-1b spleen cells were shown to display serologically defined gp70 determinants and the Gross cell surface antigen. Taking these data together, it appeared that the inhibition of anti-viral CTL responsiveness might be due to tolerance induced by the cell surface expression of virus-associated antigens by normal AKR.H-2b cells. Widespread display of viral antigens, in turn, may have been due to the permissive effects of Fv-1n on the spread of the early arising N-ecotropic, endogenous AKR leukemia virus controlled by other background genes. In this context, the implications of the multi-gene control of anti-AKR/Gross virus CTL production are discussed with respect to the induction of spontaneous leukemia in the high incidence AKR strain.
Expression and regulation of two idiotype families and subsets within an idiotype family among BALB/c antibodies against p-azophenylarsonate.
Authors: Brown A R
Published: December 2022

The expression and regulation of the two different idiotype (id) families associated with the anti-p-azophenylarsonate (Ar) antibodies of BALB/c mice were examined. Both families ( 5AF6 and 3C6 ) represented cross-reactive idiotypes (CRI) expressed in the anti-Ar of most individual BALB/c mice. In response to keyhole limpet hemocyanin-Ar, an average of about 28% of BALB/c anti-Ar had 5AF6 family idiotopes, while 3C6 family was expressed on about 16% of BALB/c anti-Ar antibodies. Suppression induced by anti-idiotype treatment against one family did not suppress the expression of the other family, suggesting that the two families were regulated independently. However, the relative expression of one family could influence the expression of the other, because depression of the 5AF6 family tended to increase the expression of the 3C6 family of anti-Ar. Analysis of the 5AF6 family showed that a majority of BALB/c mice produced antibodies bearing most or all of the idiotopes associated with the family, but that a subset of about 35% of the antibodies synthesized lacked idiotopes associated with a monoclonal anti-Ar member of this family, 2.4. Treatment of mice with anti-idiotypes prepared against two different monoclonal anti-Ar of the 5AF6 family produced different effects: one enhanced while the other suppressed idiotype expression, suggesting that there are differences in the idiotopes associated with these two regulatory pathways. Additionally, results indicated that subsets of antibodies within the 5AF6 idiotype family could be regulated independently of each other.
B lymphocyte precursors in embryonic and adult W anemic mice.
Authors: Landreth K S , Kincade P W , Lee G , Harrison D E
Published: December 2022

Mice homozygous for mutations at the dominant spotting or W locus on chromosome 5 have been extensively used as models of severe macrocytic anemia caused by defective hemopoietic stem cells. We examined cells of the developing B lineage in adult and embryonic W anemic mice both by phenotypic analyses and by three distinctly different functional assays for B lymphocyte precursors. Adult W/Wv mice had normal numbers of B cells in the spleen and bone marrow, and normal numbers of pre-B cells and cells identified by a monoclonal antibody directed to a B lineage cell surface antigen (14.8) in the bone marrow. Embryonic W/Wv and Wx/Wx mice had hypoplastic liver development at 16 days gestation with a corresponding reduction in absolute numbers of pre-B cells, 14.8+ cells, and clonable granulocyte-macrophage progenitor cells, although their frequencies were normal. As expected, spleen colony-forming units were greatly reduced both in absolute number and frequency. Adult bone marrow cells and fetal liver cells from W anemic mutants generated B cells in vitro as well as did cells from normal littermates, but W anemic cells failed to generate B lymphocytes as well in vivo. These observations likely reflect differences in precursor cells that contribute to B cell formation in these assays, and suggest that early B lineage precursors are reduced or defective in W anemic mice.
Capacity of different cell types to stimulate cytotoxic T lymphocyte precursor cells in the presence of interleukin 2.
Authors: Dröge W , Moyers C , Wehrmaker A , Schmidt H , Panknin S , Männel D , Falk W
Published: December 2022

Plastic-adherent cells enriched for dendritic cells (AC) were found to be among the most potent stimulator cells for the activation of cytotoxic T lymphocytes (CTL) in vitro in the presence of interleukin 2 (IL 2) and a constant second set of allogeneic stimulator cells. Concanavalin A-activated nylon wool-nonadherent spleen cells ( CNWT ), concanavalin A-activated unfractionated spleen cells ( Cspl ), and some variants of the ESb T lymphoma line were equally effective as stimulator cells, however, and provoked a substantial cytotoxic response at concentrations of 10(4) cells per culture or less. In contrast, nonactivated nylon wool-nonadherent spleen cells ( NWT ) or unfractionated spleen cells (Spl) and cells of the P815 mastocytoma, the Meth A fibrosarcoma, and the T cell lymphomas Ly 5178 Eb and ESb did not stimulate cytotoxic responses at these cell concentrations. The strong stimulatory potential of the Cspl preparation was reduced by treatment with anti-Thy-1 antibody plus complement, whereas the stimulatory activity of the AC preparation was resistant to this treatment. All cell types tested expressed class I major histocompatibility antigens. Nonactivated NWT cells, in contrast to the CNWT preparation, showed no detectable staining with anti-I-E or anti-I-A antibodies and also a slightly weaker staining with class I antisera. Experiments with the tumor cell lines revealed, however, that there was no strict correlation between stimulatory potential and density of class I alloantigens or the expression of I-E determinants. Experiments on primary cytotoxic responses in vivo gave similar results. Experiments in cultures with a single set of stimulator cells and I region-compatible responder cells indicated that AC and Cspl or CNWT also have a markedly stronger capacity than NWT to induce IL 2-dependent DNA synthesis.
Tolerance to Mls-disparate cells induces suppressor T cells that act at the helper level to prevent in vivo generation of cytolytic lymphocytes to hapten-altered self.
Authors: Vargo P M , Battisto J R
Published: December 2022

We have been examining the mechanisms that control in vivo development and down regulation of cytolytic T lymphocytes (CTL) to trinitrophenyl (TNP)-altered self antigens. In vivo generation of hapten-specific CTL requires an auxiliary antigenic stimulus, which can be provided by H-2 compatible but Mls-disparate cells. These experiments were designed to study the effect of tolerization with such Mls-disparate cells on CTL development. C3H/HeN (H-2k, Mlsc ) mice sensitized in the footpads with C3H-TNP spleen cells plus CBA/J (H-2k, Mlsd ) spleen cells develop CTL in the draining lymph nodes that will lyse 51Cr-labeled TNP-modified C3H targets. However, we have found that if C3H/HeN mice are given tolerizing doses of CBA/J spleen cells 5 to 7 days before sensitization, a splenic suppressor T cell (Ts) appears. This Ts will suppress CTL development in its tolerant host, and can be transferred adoptively to function in naive mice. Ts and its precursor are cyclophosphamide insensitive and therefore different from the naturally existing suppressor cell present in mice. When triggered by cells with Mlsd , the Ts produces a factor (TsF) that hinders helper factors from functioning in an in vitro CTL assay. Furthermore, TsF acts to prevent utilization of IL 2 by an IL 2-dependent cell line. Thus, evidence has been provided that the in vivo generation of CTL toward hapten-altered self can be down regulated at the level of helper signals by a Ts. The latter is inducible by the Mls-disparate cells that are needed at a different site to trigger the helper factors in this CTL system.
Interleukin 2 enhancement of lymphokine secretion by T lymphocytes: analysis of established clones and primary limiting dilution microcultures.
Authors: Kelso A , MacDonald H R , Smith K A , Cerottini J C , Brunner K T
Published: December 2022

The effect of exogenous interleukin 2 (IL 2) on lymphokine production by T lymphocytes was examined in two systems: the secretion of macrophage-activating factor (MAF) and interferon (IFN) by cloned long-term T cell lines, and a limiting dilution system for estimating the frequency of precursors of MAF-secreting cells in normal spleen. An IL 2-containing, MAF- and IFN-free supernatant from the EL-4 thymoma (EL-4 SN) significantly enhanced release of MAF and IFN by mitogen- or antigen-stimulated, cytolytic or noncytolytic T lymphocyte clones directed against alloantigens or Moloney leukemia virus-associated antigens. Highly purified IL 2 produced equivalent enhancement as EL-4 SN in cultures of alloreactive clones stimulated with concanavalin A. Kinetics experiments showed that EL-4 SN increased both the rate and duration of MAF release by T cell clones. EL-4 SN also increased MAF production when added during restimulation of limiting dilution cultures of positively selected Lyt-2+ and Lyt-2- C57BL/6 splenic T lymphocytes activated against DBA/2 alloantigens. This enhancement resulted in a threefold increase in the apparent precursor frequency of MAF-secreting cells among Lyt-2+ lymphocytes, but did not affect the frequency among Lyt-2- cells. Additional analysis indicated that average MAF production in cultures of Lyt-2-+ cells was sixfold lower than in cultures of Lyt-2- cells, and hence that EL-4 SN allowed detection of a significant proportion of Lyt-2+ cell cultures secreting low levels of MAF. Under these improved conditions, the MAF assay detected the majority of responding Lyt-2+ and Lyt-2- lymphocytes.
Immunohistochemical distribution of the antigenic determinants detected by monoclonal antibodies to carcinoembryonic antigen.
Authors: Yachi A , Imai K , Fujita H , Moriya Y , Tanda M , Endo T , Tsujisaki M , Kawaharada M
Published: December 2022

By using four distinct monoclonal antibodies to CEA, the molecular profile of which was clarified in our accompanying companion paper, immunohistochemical distribution of the antigenic determinants on both cancerous and noncancerous tissues as well as fetal tissues was studied with the use of the immunoperoxidase method. All of the monoclonal antibodies recognize different antigenic determinants on the tissue section. None of the antibodies stained granulocytes in the peripheral blood or in the normal liver tissues tested. Three of our monoclonal antibodies stained columnar epithelial cells in morphologically normal colonic mucosa; however, monoclonal antibody YK024 did not stain them. This antibody was also found to be unreactive with intestinal metaplasia lesions of the stomach, but reacted with a 16-wk-old fetal stomach as well as with cancerous parts of the colon and of the stomach. Moreover, it was found that this monoclonal antibody mainly reacted with moderately or poorly differentiated adenocarcinoma lesions of the colon and the stomach. Periodic acid treatment in this study, together with trypsin treatment on the antigen as described in our accompanying companion paper, may suggest that this antibody recognizes the carbohydrate antigenic determinant in nature.
Streptococcal Fc receptors. II. Comparison of the reactivity of a receptor from a group C streptococcus with staphylococcal protein A.
Authors: Reis K J , Ayoub E M , Boyle M D
Published: December 2022

The reactivity of a soluble Fc receptor from a group C streptococcus ( FcRc ) was compared antigenically and functionally with the staphylococcal Fc receptor, protein A. Protein A and FcRc were found to inhibit each others' binding to the Fc region of human IgG, indicating that they bind to sites that are in close proximity on the Fc region of human IgG. The two bacterial Fc receptors were antigenically unrelated. Differences were observed in the species and subclass reactivity of the two receptors. The patterns of binding of protein A and FcRc under various conditions suggested that these receptors reacted with distinct regions on the Fc region of immunoglobulins. FcRc bound more efficiently to goat, sheep, and cow IgG, protein A bound more efficiently to dog IgG, and neither receptor bound to rat IgG. Differences were also observed in the reactivity towards human IgG subclasses. The FcRc bound to all samples of the four human IgG subclass standards. Protein A bound to IgG1, IgG2, and IgG4, and to one of two IgG3 myeloma proteins tested. The reactivity of our soluble FcRc corresponds to a type III streptococcal Fc receptor classified by the reactivity of intact bacteria.
Treatment of experimental visceral leishmaniasis with lymphokine encapsulated in liposomes.
Authors: Reed S G , Barral-Netto M , Inverso J A
Published: December 2022

Highly susceptible mice were infected with Leishmania donovani chagasi and were treated with supernatants, free or encapsulated in liposomes, from concanavalin A-stimulated or unstimulated mouse spleen cell cultures. Treatment consisted of multiple i.v. injections beginning 2 days before to 2 days after infection. Mice treated with lymphokine-rich supernatants encapsulated in liposomes had significantly fewer liver parasites than the control groups, demonstrating in vivo activity of lymphokine against an infectious organism.
Intracellular destruction of Leishmania donovani and Leishmania tropica amastigotes by activated macrophages: dissociation of these microbicidal effector activities in vitro.
Authors: Hockmeyer W T , Walters D , Gore R W , Williams J S , Fortier A H , Nacy C A
Published: December 2022

Resident peritoneal macrophages were exposed to amastigotes of Leishmania donovani and L. tropica and treated with soluble products of antigen or mitogen-stimulated lymphocytes (lymphokines, LK). Macrophages infected with L. donovani were responsive to lower concentrations of LK for elimination of this parasite than cells infected with L. tropica, and the disappearance of L. donovani amastigotes from activated macrophages preceded that of L. tropica by at least 24 hr. That the differential elimination of amastigotes from LK-treated macrophages may reflect different intracellular killing mechanisms for the two parasites is supported by studies with macrophages activated in vivo; macrophages from BCG-treated C3H/HeJ mice that were incapable of eliminating intracellular L. tropica killed amastigotes of L. donovani. Whether the effector mechanism(s) for intracellular destruction of the two Leishmania are qualitatively or only quantitatively different is as yet unknown.