bluuebunny/crossref_metadata_2025 huggingface dataset:
Antigen-specific suppressor T cell interactions. I. Induction of an MHC-restricted suppressor factor specific for L-glutamic acid50-L-tyrosine50.
Authors: Kapp J A , Araneo B A
Published: December 2022

The synthetic polymers L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT) and L-glutamic acid50-L-tyrosine50 (GT) stimulate specific suppressor T cells in certain strains of mice. Extracts from these T cells contain factors (TsF) that inhibit GAT- or GT-specific antibody responses by normal spleen cells or proliferative responses by primed T cells. We constructed T cell hybridomas that constitutively produce GAT-TsF or GT-TsF, which functionally and serologically are identical to factors extracted from suppressor T cells. In this report we demonstrate that monoclonal GT-TsF can induce specific unresponsiveness in vivo or in vitro and that this unresponsiveness is due to development of second-order antigen-specific suppressor T cells. T cell hybridomas were constructed by fusion of BW5147 with GT-TsF1 induced second-order suppressor T cells and clones that produced suppressor factor (GT-TsF2) were isolated and characterized. GT-TsF2 differs from the GT-TsF1 used to induce it in that GT-TsF1 acts across allogeneic barriers whereas GT-TsF2 does not. This restriction is controlled by genes in the H-2 gene complex and maps to the I-J subregion. GT-TsF2 is antigen-specific in suppressive activity and also in its antigen-binding site(s). Thus, GT-TsF2 closely resembles the carrier-specific, I-J+, genetically restricted factor described by Tada and his colleagues. Because GT-TsF2 was induced by GT-TsF1, we suggest cells producing GT-TsF1 are an early cell in the pathway of suppression, and that this cell is required for the activation of antigen-specific, MHC-restricted TsF.
T cell clones restricted to "hybrid" HLA-D antigens?
Authors: Hansen G S , Svejgaard A , Claësson M H
Published: December 2022

The genetics of T lymphocyte/accessory cell interactions were studied by using antigen-specific monoclonal T cell lines from human peripheral blood and X-irradiated non-T cells (E rosette-depleted) as accessory cells. The T cell lines were obtained by expanding single colonies from soft agar-cloned E rosette-positive lymphocytes, previously incubated for 3 days in liquid culture with 30% autologous X-irradiated non-T cells and antigen (PPD). When investigated with both autologous and allogeneic accessory cells of various HLA-D genotype, different sets of PPD-specific T cell lines were obtained from HLA-D heterozygous individuals; two sets restricted to each one of the two HLA-D antigens, and a third set responding to PPD only when presented by accessory cells carrying both HLA-D antigens possessed by the T cell donor. A few cell lines responded to PPD only in the presence of autologous accessory cells.
Identification of the T cell subset that produces human gamma interferon.
Authors: O'Malley J A , Nussbaum-Blumenson A , Sheedy D , Grossmayer B J , Ozer H
Published: December 2022

Positive and negative selection procedures combined with cytofluorographic analysis and lysis with monoclonal antibodies were utilized to identify the T lymphocyte subset that produces human gamma interferon (gamma-IFN) (formerly referred to as "immune" or "type II" interferon) in response to mitogen stimulation. Lymphocytes were separated on the basis of their Fc receptors for IgG or IgM, their nonreactivity with IgM or IgG antibodies, and their reactivity with the monoclonal antibodies OKT4, OKT8, OKT11a, and OKM1. Isolated T cell subsets were incubated with the gamma-IFN inducer, phytohemagglutinin. Three days after induction, the cell supernatants were harvested and assayed for interferon. The T cell subset that produces gamma-IFN was identified as E rosette positive with the phenotype: T gamma, T non-micro, OKM1+, OKT4-, OKT8- and OKT11a+. gamma-IFN production by cells was resistant to doses of x-irradiation that abrogate mitogen-induced T suppressor function but was highly sensitive to low doses of 4-hydroperoxycyclophosphamide. These data demonstrate that gamma-IFN is produced by the T gamma, OKM1+ lymphocyte subset, but these cells may also require the presence of accessory monocytes for elaboration of gamma-IFN. The anti-proliferative activity of gamma-IFN may be responsible for the previously described suppressor function of this subset, and gamma-IFN production by T gamma cells may distinguish this subset from the suppressor/cytotoxic functions of the OKT8+ subset or the mitogen-induced OKT4+ suppressor.
Antigen receptors on murine T lymphocytes in contact sensitivity. II. Presentation and characterization of syngeneic anti-idiotype serum against DNFB-sensitized T cells.
Authors: Moorhead J W , Sy M S
Published: December 2022

This manuscript describes the preparation and characterization of a syngeneic antiserum raised in BALB/c mice against nylon wool-purified lymph node (LN) T cells from 2,4-dinitrofluorobenzene- (DNFB) sensitized BALB/c mice. This antiserum (anti-T DH-DNP) plus complement blocks the ability of LN or purified T cells from DNFB-sensitized donors to transfer immunity to naive recipients. The inhibitory activity of the serum is due to antibodies; these antibodies have specificity for idiotype (Id) determinants as shown by their ability to bind to purified mouse anti-DNP antibodies but not to rabbit anti-DNP or normal mouse Ig. Inhibition of transfer of immunity by the anti-Id serum is not restricted by either H-2 gene products or Igh-1 allotypes. The antiserum is antigen specific when tested on LN cells from oxazolone-sensitized mice, but has a low level of cross-reactivity against T cells from 2,4,6-trinitrochlorobenzene- (TNCB) sensitized mice. In addition, it was shown that naturally occurring anti-Id antibodies, epsilon-DNP-L-lysine, or DNP-protein (but not NIP-protein) and the anti-Id in syngeneic anti-T DH-DNP serum compete for similar binding sites on DNFB-immune T cells. We interpret these findings to indicate that immunization of mice with T cells from DNFB-sensitized syngeneic mice induces the production of anti-Id antibodies. These anti-Id antibodies are specific for determinants expressed on the T cell receptor or receptor subsite that is specific for the hapten DNP.
Acceptor site(s) for T cell-replacing factor (TRF) on B lymphocytes. II. Activation of B cells by cross-linkage or aggregation of the TRF acceptor molecule.
Authors: Tominaga A , Takatsu K , Hamaoka T
Published: December 2022

An in vitro experimental system was established to demonstrate the TRF-substituting activity of an alloantiserum raised in TRF low-responder (DBA2/2Ha x BALB/c)(DC)F, male mice against TRF high-responder parental BALB/c B cells. The TRF substituting activity of the antiserum was apparent in that anti-Thy-1 plus C-treated, DNP-primed B cells from TRF high-responder mice were effectively stimulated, whereas B cells from TRF low-responder DBA/2Ha mice were not, as evidenced by the induction of secondary anti-DNP IgG PFC responses. The specificity of the reaction of the antibody with a component present on the TRF high-responder B cells was also substantiated by the fact that antibody activity was virtually eliminated by absorption with B cells from various TRF high-responder mice but not by TRF low-responder DBA/2Ha B cells. The IgG fraction of antiserum and the F(ab')2 and Fab' fragments of the antibody, which possess a comparable reactivity in regard to the TRF acceptor site(s), were prepared, and analysis of the B cell-triggering mechanism by the antibody was carried out. The results revealed that both the IgG fraction and F(ab')2 fragment, but not the monovalent Fab' fragment, demonstrated effective TRF-substituting activity, indicating that cross-linkage or aggregation of the TRF acceptor site(s) may give rise to differentiation signals to the B cells.
Detection of a tumor-associated antigen on the surface of human myelogenous leukemia cells.
Authors: Malcolm A J , Shipman R C , Levy J G
Published: December 2022

Membrane extracts of human acute myelogenous leukemia cells were passed over immunoadsorbent columns containing antibodies directed to equivalent preparations of pooled normal peripheral blood leukocytes. This material, when compared to equivalent normal cell preparations on nonreducing polyacrylamide gels, demonstrated the presence of four unique bands, not found in normal cell preparations. With the use of a previously described leukemia-specific heteroantiserum, these bands, after elution, were tested for reactivity with the antiserum. The eluted material containing reactive antigen was used to immunize rabbits. The resulting antiserum showed strong reactivity with myelogenous leukemic cells extracts and none with normal materials. A series of bone marrow samples from patients with a variety of lymphoproliferative and myeloproliferative disorders were tested with this antiserum in the fluorescence-activated cell sorter. In every instance, bone marrow cells from patients with acute myelogenous leukemia and chronic myelogenous leukemia showed strong positive fluorescence with this antiserum. However, no bone marrow cells from patients with lymphoproliferative disorders or patients with other leukemias showed any fluorescence above background with this antiserum.
Eosinophil peroxidase-mediated inactivation of leukotrienes B4, C4, and D4.
Authors: Henderson W R , Jörg A , Klebanoff S J
Published: December 2022

The slow-reacting substance (SRS) bioactivity of leukotrienes C4 (LTC4) and D4 (LTD4) was rapidly decreased by incubation with eosinophil peroxidase (EPO), H2O2, and iodide, bromide, or to a lesser degree, chloride, LTB4 chemotactic activity was also decreased by the EPO-H2-H2-halide system, although at a slower rate. Myeloperoxidase could substitute for EPO in these reactions. Leukotriene inactivation was greatly decreased or abolished by deletion of any of the components of the system or by the addition of the hemeprotein inhibitors, azide, cyanide, or aminotriazole, indicating a requirement for peroxidase. The H2O2 concentration employed in the above studies was 10(-4) M. H2O2 at higher concentrations (5 x 10(-4) to 10(-2) M) inactivated LTC4 and LTD4 in the absence of EPO and a halide but had no effect on the chemotactic activity of LTB4. We have previously shown that horse eosinophils stimulated with the calcium ionophore A23187 generate SRS. In the present study, eosinophils stimulated in this way were found to release extracellularly both H2O2 and EPO. Incubation of eosinophils with azide that inhibits EPO, and catalase that degrades H2O2, significantly increased the amount of SRS activity detected in the extracellular medium after A23187 stimulation. These findings suggests eosinophils may play an important modulating role in hypersensitivity reactions both by the production of leukotrienes and by their inactivation through the release of H2O2 and EPO.
H-2 effects on cell-cell interactions in the response to single non-H-2 alloantigens. V. Effects of H-2Kb mutations on presentation of H-4 and H-3 alloantigens.
Authors: Wettstein P J
Published: December 2022

The effects of a panel of H-2Kb mutants on the presentation of the non-H-2 histocompatibility (H) antigens H-4.2 and H-3.1 to antigen-specific T cells were examined. Kb-restricted cytotoxic effector T cells were presented with H-4.2 and H-3.1 in the context of mutant Kb alleles. H-4.2 was differentially presented by Kb mutants to Kb-restricted cytotoxic effectors. Cold target inhibition analysis demonstrated that Kb-restricted, H-4.2-specific cytotoxic effectors recognized H-4.2 in the context of at least three restriction sites on the Kb molecule. One site is specific for Kb alone, whereas two additional sites are differentially expressed by Kb mutant molecules. The effect of Kb mutations on the presentation of H-4.2 and H-3.1 was H antigen-specific. Identical, respective patterns of presentation of H-4.2 and H-3.1 by Kb mutants were observed with Kb-restricted T cells proliferating in secondary mixed lymphocyte culture. Further, individual mice varied in their preference for H-3.1 presented in the context of different Kb mutant molecules. These observations demonstrate that the presentation of H antigens by mutant Kb molecules is antigen-specific; analysis of presentation of H antigen by mutants to wild-type Kb-restricted effectors allows the identification of multiple restriction sites on the Kb molecule.
A new murine cell surface differentiation antigen (Leugp90) defined by a rat monoclonal antibody: cellular distribution and biochemical characterization.
Authors: Moldenhauer G , Haustein D , Hüppe T , Wagner A , Hartmann K U
Published: December 2022

Monoclonal antibodies to murine lymphocyte differentiation antigens were generated by fusing the mouse myeloma cell line X63-Ag8.653 with spleen cells derived from Lewis rats hyperimmunized with lymphoid cells from nude (C57BL/6) mice. One of these antibodies--designated 3MB1--recognizes a previously undescribed cell surface antigen. This antigen is expressed on 62% of spleen cells, 9% of thymus cells, and 98% of peritoneal macrophages. Virtually all LPS- and Con A-induced lymphoblasts carry this newly found antigen. The 3MB1 determinant is limited in its expression to leukocytes. It is not found in other tissues such as brain, liver, kidney, or on erythrocytes. Biochemical analysis reveals that the 3MB1 target antigen consists of a single chain glycoprotein with an approximate m.w. of 90,000.
Ultrastructure of human natural killer cells: nature of the cytolytic contacts in relation to cellular secretion.
Authors: Carpen O , Virtanen I , Saksela E
Published: December 2022

The ultrastructure of human NK cells, NK/target cell conjugates, and the effect of monensin, a secretion inhibitor, were studied. Human peripheral blood lymphocytes, highly enriched (75 to 85%) in large granular lymphocytes (LGL), which are known to be the mediators of human NK activity, were used as effectors, and K562 cells as targets. LGL-type of cells were characterized in electron microscopy by a low nucleocytoplasmic ratio, indented nucleus, several large mitochondria, prominent Golgi apparatus, and cytoplasmic osmiophilic granules bound by a unit membrane. Their surface was of intermediate villous type. Two types of effector/target contacts were seen: either effector cell protrusions were pushed deep into pouches and lacunae of the target cell surface, or a wide area of intimate cell-to-cell contact was formed. The contact formation was followed by polarization of the effector cell Golgi apparatus towards the contact area. Monensin, a carboxylic ionophore, which interrupts the vesicular traffic of Golgi-derived vacuoles to the cell surface, caused an accumulation of cytoplasmic vesicles in the LGL but had no effect on the effector/target contact formation. Monensin inhibited the NK lysis apparently via cessation of secretion by the LGL. It did not affect the viability of the effector cells. The lytic steps of human NK cells thus involve binding to the target cell, polarization of cytoplasmic organelles towards the target, and apparently a precisely directed exocytosis of secretory material.
Isolation and characterization of circulating feline leukemia virus-immune complexes from plasma of persistently infected pet cats removed by ex vivo immunosorption.
Authors: Snyder H W , Jones F R , Day N K , Hardy W D
Published: December 2022

IgG and circulating IgG immune complexes (CIC) were purified from plasma of three pet cats persistently infected with feline leukemia virus (FeLV) by adsorption to, and elution from, Staphylococcus aureus Cowan I. CIC were then separated from free IgG by sucrose gradient ultracentrifugation and were analyzed for the presence of FeLV structural proteins and corresponding specific antibodies. Radioimmunoprecipitation analysis indicated that FeLV envelope (gp70) and major core (p30) proteins, along with cat IgG heavy and light chains, were present in the CIC from all three cats. Further analysis of the CIC from one of the cats also revealed the presence of FeLV core proteins p15 and p12. IgG purified from isolated CIC was also shown to bind specifically to purified FeLV gp70, p30, and p15. These data provide direct evidence for FeLV-specific CIC in the plasma of persistently viremic pet cats, and suggest these animals are immunologically response to the virus even though free antibodies against the major structural proteins cannot be demonstrated in standard assays.
Heterogeneity of the natural humoral anti-tumor immune response in mice as shown by monoclonal antibodies.
Authors: Colnaghi M I , Menard S , Tagliabue E , Torre G D
Published: December 2022

Four anti-tumor cytotoxic antibodies, namely, A6 of the IgG2 class and B3, C2, and D1 of the IgM class, were obtained in monoclonal form, from hybridization of mouse myeloma cells with spleen cells of normal untreated mice selected for high natural anti-tumor immune responses. The specificity of the four monoclonals was tested by complement-dependent cytotoxicity assay on the reference EL4 lymphoma at different days of in vivo transplant, on normal adult and fetal tissues, on the SC-1 fibroblastic cell line uninfected or infected with a murine ecotropic type-C virus, and on a panel of murine lymphomas of different origin. The four antibodies had different specificities: the A6 and B3 recognized virus-related structures, the C2 a structure expressed on fetal cells, and the D1 a normal component of fibroblasts. The different classes and specificities of the four monoclonals, as well as their in vitro-demonstrated synergistic cooperation, give support to the hypothesis that the natural humoral anti-tumor cytotoxic immune response could be the result of the cooperative activity of an array of heterogeneous antibody molecules.
Two isoallergens of short ragweed component Ra5.
Authors: Roebber M , Klapper D G , Marsh D G
Published: December 2022

Two isoallergenic forms of short ragweed (Ambrosia elatior) allergen Ra5 were isolated in ultrapure form by a combination of molecular sieving (through Amicon hollow-fiber cartridges), cation-exchange chromatography, and gel filtration. The two forms, Ra5A and Ra5B, exhibited charge differences by CM-Sephadex chromatography and agarose electrophoresis at pH 8.5. The A form as identical to conventional Ra5 by its physical and chemical properties and was more cationic than the previously undescribed B form. Ra5A and Ra5B had identical m.w. of 5000 by SDS-PAGE. Their amino-terminal sequences of 30 residues showed no differences from each other or from the previously published sequence of the major form of Ra5. The two forms were antigenically indistinguishable when using hyperimmunized animal antisera to Ra5 and crude ragweed extract. Although the IgG and IgE ab of most Ra5-responder patients could not discriminate between the two Ra5 forms, significant small differences in binding to Ra5A and Ra5B were consistently observed in 10 to 15% of the patients studied. Further analysis of the Ra5A preparation suggested a purity of at least 99.5% and probably greater than or equal to 99.9%.
Serum IgM-bound secretory component (sIgM) in liver diseases: comparative molecular state of the secretory component in serum and bile.
Authors: Delacroix D L , Jonard P , Dive C , Vaerman J P
Published: December 2022

The sera of 15 patients with liver disease and high serum concentrations of secretory component (SC) were analyzed by density gradient ultracentrifugation and radioimmunoassays for SC, IgA, and IgM to determine the molecular state of SC and IgA. Results in serum were compared to those obtained by simultaneous analysis of bile in five of the patients, and to those in serum and bile of a case of total IgA deficiency. Free SC was virtually not found in sera, although it was well represented in all bile, up to 88 and 97% of total bile SC in a case of complete biliary obstruction and in IgA deficiency, respectively. IgM-bound SC was found in all sera, amounting to 91% and 100% of total serum SC, respectively, in a case of acute hepatitis with a very high serum IgM concentration and in IgA deficiency. The proportions of SC bound to IgM and to polymeric IgA (p-IgA) in the sera correlated with their IgM/p-IgA molar ratio. This suggests that during liver disease, the hepatobiliary tissues could release free SC into the circulation, where it binds to p-IgA and IgM according to their respective concentrations and affinities for SC. The proportion of p-IgA in serum was not increased in four cases of biliary obstruction, in contrast to our cases of cirrhosis and acute hepatitis, indirectly supporting a minor transfer of p-IgA from blood to bile in humans, in contrast to rats and rabbits.
Differential interference by azathioprine and 6-mercaptopurine with antibody-mediated immunoregulation: synergism of azathioprine and antibody in the control of an immune response.
Authors: Medzihradsky J L , Klein C , Elion G B
Published: December 2022

Comparative studies on the effects of immunosuppressive agents azathioprine (AZ) and 6-mercaptopurine (6-MP) in antibody-mediated immunoregulation are reported. To determine the degree of interference of these agents with the generation of antibody molecules controlling an anti-SRBC response, the donors of mouse anti-SRBC sera were given daily doses of the drugs. Primary antisera from AZ-treated donors were not significantly inhibited by daily doses of 180, 270, and 360 mumol/kg in their ability to suppress the direct PFC response; on the other hand 6-MP was inhibitory at the two higher equimolar dose levels. When the antisera were used in the antibody-dependent cellular cytotoxicity (ADCC) and phagocytosis (AD phi) assays, similar differential effects of the two agents were found. Multiple doses of AZ and 6-MP given to the donors of ADCC effector cells enhanced the capacity of these cells to lyse antibody-sensitized SRBC target cells. The enhancement of ADCC with 6-MP was less manifest than that with AZ. When injected before the antigen, AZ acted synergistically with the antibody in the control of the direct PFC response. Possible mechanisms by which AZ can interact with antibody-mediated immune regulation are discussed.
VH families in the antibody response to p-azophenylarsonate: correlation between serology and amino acid sequence.
Authors: Milner E C , Capra J D
Published: December 2022

Anti-arsonate hybridoma antibodies of A/J mice have been grouped into 3 VH families by amino acid sequence analysis. One of these, the 91A3 family, which includes all CRI-positive molecules and some CRI-negative molecules, has been serologically defined by the presence of a VH marker (91A3 marker). Although the 91A3 family of antibodies is present at low levels in preimmune A/J serum and represents a substantial component of the A/J response to p-azophenylarsonate, the 91A3 family has not been detected serologically in the anti-Ar response of any other inbred strain tested. Analysis of sera from AXC Igh recombinant mice maps the 91A3 marker to the left of Igh-Dex, consistent with the location of the VH genes controlling the arsonate CRI. These studies establish a serologic assay that detects all members of a family of anti-arsonate antibodies defined previously by primary sequence analysis. This assay makes it possible to monitor in its entirety the expression of the products of a single or closely related group of genes.
Use of H-2:H-7 congenic mice to study H-2-mediated resistance to Friend leukemia virus.
Authors: Wettstein P J , Blank K J
Published: December 2022

Congenic and double congenic mice expressing different H-7 alleles with varying H-2 haplotypes on the C57BL/10 (B10) background were tested for their susceptibility to Friend virus (FV) infection. Mice expressing the H-7b allele derived from BALB/c were susceptible to FV infection whereas mice expressing the H-7a allele of B10 were absolutely resistant. Coexpression of H-7b and the H-2a and H-2d haplotypes determined high susceptibility; mice expressing both H-7b and H-2b were relatively resistant compared to H-7b mice expressing H-2a or H-2d. Parallel experiments with BALB/c and BALB.B recipients did not demonstrate differences in susceptibility associated with H-2d and H-2b. Mapping experiments with the H-2h4 and H-2i5 recombinants indicated that the gene determining relative resistance to FV-induced spleen focus formation mapped to the K-end (KbAb) of H-2b. Female recipients expressing H-7b, regardless of their H-2 haplotype, were more susceptible to FV infection than their syngeneic male counterparts. These experiments demonstrate the utility of congenic and double congenic strains that define H-7 and Fv-2 on different H-2 haplotype backgrounds for the analysis of Fv-2:H-7:H-2 interactions in determining susceptibility to FV infection.
T cell-mediated immunity in malaria. I. The Ly phenotype of T cells mediating resistance to Plasmodium yoelii.
Authors: Jayawardena A N , Murphy D B , Janeway C A , Gershon R K
Published: December 2022

CBA mice that recover from Plasmodium yoelii 17X infection are resistant to reinfection. T cells from these mice transfer immunity to nonimmune recipients. To analyze the nature and mode of action of these T cells, we transferred selected subsets into T cell-deprived recipients. Treatment of the T cells with anti-Ly-1 but not anti-Ly-2 serum and C abrogated their ability to transfer immunity. A mixture of anti-Ly-1 and anti-Ly-2 serum-treated cells (i.e., a population devoid of Ly-123 cells) transferred a level of immunity comparable to that of unselected T cells. Hence, the T cells mediating resistance to P. yoelii 17X were primarily of the Ly-1+23- phenotype. T cell-deprived mice reconstituted with these immune Ly-1 cells developed a) high levels of IgM and IgG antibodies, b) DTH responses to parasitized RBC, and c) enhanced blood monocyte responses. The addition of immune B cells to the Ly-1 population dramatically increased its ability to transfer immunity and induce antibody production. B cells from immune CBA/N mice had no such effect. Thus, the transfer of optimal protective immunity against malaria stems from an interaction between Ly-1 cells and a select B cell subset that CBA/N mice lack. This "selective synergy" is a protective mechanism against pathogens that has not been previously appreciated.
Phorbol ester tumor promoter modulation of alloantigen-specific T lymphocyte responses.
Authors: Andreotti P E
Published: December 2022

The phorbol diester tumor promoter PMA was studied for its effects on T lymphocyte activation in allogeneic human mixed lymphocyte cultures. Lymphocyte proliferation was significantly enhanced in cultures with PMA at concentrations greater than or equal to 12.5 ng/ml, and interferon-gamma production was synergistically increased in cultures with PMA at concentrations greater than or equal to 5 ng/ml. In contrast, the generation of HLA-specific cytotoxic T lymphocyte activity was completely inhibited in cultures with PMA at concentrations greater than or equal to 5 ng/ml. Fluorescence-activated cell sorter analysis with OKT mouse monoclonal antibodies indicated these PMA effects on lymphocyte activation are associated with an increase in the number of activated OKT4-positive T lymphocytes.
Production of T cell-activating monokine of guinea pig macrophages induced by MDP and partial characterization of the monokine.
Authors: Iribe H , Koga T , Onoue K
Published: December 2022

The production of T cell-activating factor(s) by macrophages stimulated with muramyl dipeptide (MDP) was studied. By MDP stimulation, a rapid increase in intracellular activity of the T cell-activating factor was induced, which preceded an increase of the activity in the extracellular medium. The rapid appearance of the activity in the cell and that in the medium were both inhibited by cycloheximide or puromycin. These results demonstrated that MDP stimulated rapid production of T cell-activating factors by inducing de novo synthesis of the factors, and that these newly formed factors are rapidly secreted. The activities of intracellular and extracellular secreted factors were both found in high m.w. (50,000 to 90,000) and low m.w. (10,000 to 30,000) fractions by gel filtration. The secreted high m.w. factor migrated as a single peak and did not dissociate into smaller components in SDS-PAGE analysis, indicating that the high m.w. factor is neither a complex of low m.w. factor with other proteins nor an aggregate of low m.w. factors. The properties are similar to those of our previously reported factor that helped antigenic activation of T cells for lymphokine production.
Cell-cell cooperation in lymphocyte colony formation: studies in human allogeneic marrow transplantation.
Authors: Donnenberg A D , Elfenbein G J , Santos G W
Published: December 2022

The pace of restoration of phytohemagglutinin- (PHA) stimulated lymphocyte colony-forming response was investigated in 50 bone marrow transplant (BMT) recipients (49 allogeneic, 1 syngeneic) studied on 86 occasions. On the majority of occasions (28 of 48), patients studied early (3 to 8 wk) after BMT failed to make detectable lymphocyte colonies, whereas patients studied late (greater than 6 mo) after BMT displayed responses comparable to those of healthy adult volunteers. The sequential study of 10 of the 15 patients with positive lymphocyte colony-forming responses early after BMT revealed that this response is short-lived. The results of recipient donor mixing experiments and experiments involving the addition of the lymphokine interleukin 2 (IL 2) to recipient cells indicated that lymphocytes obtained from unresponsive recipients evaluated early after BMT can be driven to colony formation, and suggest that the major limiting component in such subjects is functional T help.
Functionally different T lymphocyte subpopulations determined by their sensitivity to complement-dependent cell lysis with the monoclonal antibody 4A.
Authors: Morishima Y , Kobayashi M , Yang S Y , Collins N H , Hoffmann M K , Dupont B
Published: December 2022

A human T lymphocyte antigen 4A (40,000 daltons) is defined by a monoclonal, complement- (C) fixing, hybridoma-produced antibody (moAb 4A). This antigen, which is identical to the previously reported antigen 3A 1, is expressed at quantitatively different levels on functional subsets of peripheral T lymphocytes as determined by the cell sensitivity to antibody and C. Peripheral T lymphocytes can be divided into two populations: 4A high-density T cells (4A increases), which are killed in vitro by moAb 4A + C, and 4A low-density T cells (4A decreases), which are not affected in vitro by moAb 4A + C treatment. The helper/inducer T cell lineage, defined by moAb Leu 3a, and the cytotoxic/suppressor T cell lineage, defined by moAb Leu 2A, contain both 4A increases and 4A decreases T cell populations. Functional studies by moAb 4A + C treatment show that 1) the 4A increases T cell population contains T helper cells, which are necessary for in vitro antibody production against red cell-bound determinant; 2) the 4A decreases T cell population contains the precursor T cells, which proliferate in vitro in MLC, and the precursor T cells, which are necessary for the in vitro generation of the alloreactive cytotoxic T cells; and 3) the cytotoxic activity of alloreactive T cells generated in CML assay is abrogated by moAb 4A + C treatment; 4) the activation of T cells by PHA and Con A increases the quantitative expression of 4A antigen.
Distribution of T lymphocyte subsets in human colostrum.
Authors: Richie E R , Bass R , Meistrich M L , Dennison D K
Published: December 2022

Lymphocytes were isolated from human colostrum and peripheral blood collected within 7 days postpartum. A 20-fold enrichment of lymphocytes from colostrum was achieved by centrifugal elutriation followed by equilibrium density gradient centrifugation. The distribution of T lymphocyte subsets was determined by immunofluorescence analysis of reactivity with monoclonal antibodies of the OKT series. In confirmation of previous reports employing sheep erythrocyte rosette assays, the majority of colostral lymphocytes reacted with OKT3 monoclonal antibody, a reagent that detects a surface antigen on mature T cells. Neither colostral nor blood lymphocytes from postpartum donors expressed the OKT6 antigen that is found in immature thymocytes. A novel finding is that the T lymphocyte population in colostrum contains both OKT4 (helper/inducer phenotype) and OKT8- (cytotoxic/suppressor phenotype) positive subsets. The relative ratio of OKT4 to OKT8-positive T cell subsets in colostrum, however, is generally lower than that observed for peripheral blood T cells.
Characterization of a soluble suppressor of human B cell function produced by a continuous human suppressor T cell line. II. Evidence for suppression through a direct action of CTC-SISS-B on human B cells.
Authors: Greene W C , Fleisher T A , Depper J M , Leonard W J , Stanton G J , Waldmann T A
Published: December 2022

CTC-SISS-B is an antigen-nonspecific suppressive lymphokine elaborated by an interleukin 2-dependent suppressor T cell line that produces noncytotoxic inhibition of human B cell but not T cell function. Like SISS-B, a soluble suppressive lymphokine present in the supernatants of Con A-activated peripheral blood T cell cultures, CTC-SISS-B is of 60,000 to 90,000 m.w., and its action is blocked by the simple sugar L-rhamnose. CTC-SISS-B inhibits human B cell Ig production and proliferation through a direct interaction with human B cells rather than through indirect effects on immunoregulatory T cells or monocytes. CTC-SISS-B suppression occurs through inhibition of an early event(s) in B cell activation since proliferation and Ig production by established human B cell lines are not inhibited by this lymphokine. Despite sharing many biochemical and biologic properties, CTC-SISS-B and gamma-interferon appear to be distinct mediators.
A self determinant recognizing T cell hybridoma.
Authors: Juy D , Barbier E , Le Guern C , Cazenave P A
Published: December 2022

A T cell hybridoma (53(113)) obtained by fusion of BALB/c spleen cells and the BW 5147 lymphoma T cell line is described. This hybridoma recognizes mouse RBC (MRBC) and rat RBC, but not human, rabbit, guinea pig, or SRBC. The culture supernatant possesses hemagglutinating activity for the same indicator RBC. In addition to this, 53(113) cells are able to form protein A plaques in the presence of guinea pig complement and normal mouse serum (NMS) or purified mouse immunoglobulins (Ig). Because mouse Ig as well as sonicates from MRBC are able to inhibit the rosettes between the hybridoma cells and the MRBC, and because the sonicates inhibit protein A plaque formation, it seems likely that the same product can recognize a similar determinant expressed on MRBC and mouse Ig. The hypothesis that a 53(113) structure recognizes identical or cross-reactive carbohydrate determinants shared by murine Ig and C is considered.
Class I major histocompatibility gene products of the brown Norway rat display two major antigenic regions.
Authors: Klinman D M , Smilek D E , McKearn T J
Published: December 2022

Sixteen Lewis anti-Brown Norway monoclonal antibodies and sera from alloimmunized Lewis rats were used to study the topographic relationships of antigenic determinants on class I major histocompatibility gene products. Only two independent antigenic regions were identified in competition binding assays. The first region is composed of a set of overlapping epitopes that are conserved in class I major histocompatibility products of mice and humans, as well as rats. In contrast, the second antigenic region appears in a restricted number of inbred rat strains and is not detected in other species. The data provide serologic confirmation, at the monoclonal and serum alloantibody level, of conserved polymorphisms in the major histocompatibility gene products of different species, a finding that is consistent with the amino acid sequences of these molecules.
Encephalitogenic activity of guinea pig myelin basic protein in the SJL mouse.
Authors: Pettinelli C B , Fritz R B , Chou C H , McFarlin D E
Published: December 2022

GPBP was shown to be encephalitogenic in SJL mice by direct challenge and in experiments in which an adoptive transfer system was employed. The three fragments obtained by treating GPBP with pepsin were assessed in the same manner. The encephalitogenic activity resided in the C terminal half of the molecule (residues 89-169). LNC also proliferated to the same fragment in vitro. Fragments 1-37, and, to a lesser extent, 44-48 stimulated sensitized LNC to proliferate but did not induce disease.
Biochemical analysis of glucocorticoid-induced inhibition of IgE-mediated histamine release from mouse mast cells.
Authors: Daëron M , Sterk A R , Hirata F , Ishizaka T
Published: December 2022

Pretreatment of mouse mast cells with 10(-7) to 10(-6) M dexamethasone (DM) during overnight sensitization with mouse IgE antibody resulted in inhibition of antigen-induced histamine release and degranulation. The inhibition of both degranulation and histamine release increased linearly with the duration of the treatment; maximal inhibition was obtained after approximately 16 hr with DM. The addition of DM to sensitized mast cells immediately before antigen challenge did not affect the antigen-induced histamine release. DM interacted directly with mast cells by binding to DM-specific cytoplasmic receptors. The treatment of mast cells with DM did not affect the binding of IgE to mast cells or intracellular cAMP levels. Bridging of cell-bound IgE anti-DNP antibody on mouse mast cells either by multivalent DNP-HSA or by anti-IgE induced phospholipid methylation at the plasma membrane and Ca++ influx into the cells. Pretreatment of mast cells with DM inhibited the antigen-induced phospholipid methylation and Ca++ uptake but failed to affect histamine release by Ca++ ionophore A23187. The results suggest that DM treatment inhibits histamine release by the inhibition of the early stage of biochemical processes leading to opening Ca++ channels but does not affect the process distal to Ca++ influx or the binding of IgE molecules to IgE receptors.
Microinjection of macromolecules into normal murine lymphocytes by means of cell fusion. II. Enhancement and suppression of mitogenic responses by microinjection of monoclonal anti-cyclic AMP into B lymphocytes.
Authors: Ohara J , Sugi M , Fujimoto M , Watanabe T
Published: December 2022

Reproducible methods are now available for introducing protein molecules such as antibodies into normal murine lymphocytes by fusion with protein molecule-containing erythrocyte ghosts. Monoclonal antibodies against cyclic AMP were raised by hybridoma technique and packed into erythrocyte ghosts. Then, monoclonal anti-cyclic AMP containing ghosts were fused with splenic B lymphocytes by polyethylene glycol-mediated fusion at various intervals after LPS stimulation. This method made it possible for us to quantitatively microinject antibodies into B lymphocytes. Microinjection of anti-cyclic AMP antibody molecules into lymphocytes at a very early stage of LPS stimulation resulted in a marked enhancement of DNA synthetic responses as well as increased numbers of plaque-forming cells. Intracellular cyclic AMP levels were found to be markedly decreased after microinjection of monoclonal anti-cyclic AMP, suggesting that lowering the intracellular cyclic-AMP level in the B lymphocytes at an early stage of stimulation might have induced the enhanced proliferative as well as differentiative responses to LPS. Similar enhancing effects on cell proliferation were obtained when antibodies were injected 18 hr after stimulation. Microinjection of anti-cyclic AMP at 12 hr after culture, however, inhibited the DNA synthetic responses, and induction of plaque-forming cells was suppressed when anti-cyclic AMP was injected 6 hr after LPS stimulation. The present data suggest the biphasic regulatory roles of cyclic AMP at the early stage of B lymphocyte activation. This approach may be useful in identifying regulatory molecules in B lymphocyte induced by mitogenic or antigenic stimulation.
Humoral immune response in human syphilis to polypeptides of Treponema pallidum.
Authors: Hanff P A , Fehniger T E , Miller J N , Lovett M A
Published: December 2022

A molecular characterization of the polypeptide antigens of Treponema pallidum reactive with sera from patients with different stages of syphilis is described. Polypeptides of motile, virulent T. pallidum, purified from rabbit testes, were separated on SDS polyacrylamide gels and electrophoretically transferred to nitrocellulose for antigenic analysis ("Western blotting"). Serum IgG from uninfected individuals reacts weakly with three polypeptides of 45,000, 33,000, and 30,000 m.w. In this study patients with primary syphilis have IgM antibody, and all patients with syphilis have IgG antibody to at least four polypeptides of 45,000, 33,000, 30,000, and 15,500 m.w. Antibody to polypeptides of 42,000 and 16,500 m.w. appear to be markers for nonprimary syphilis. These six polypeptides have been termed the major antigenic proteins (MAP) of T. pallidum. Those patients studied with secondary and early latent syphilis acquire antibody to a set of 16 additional polypeptide antigens. Those patients studied with late latent or late syphilis have antibody to a much smaller set of five or four antigens, respectively, in addition to MAP. The results suggest that a correlation exists between acquisition of antibody and the development of "chancre immunity." Additionally, the loss of antibody that characterizes late latent and late syphilis may be associated with the potential development of destructive late syphilis.
Characterization of a progressive tumor from C3H fibroblasts transformed in vitro with SV40 virus. Immunoresistance in vivo correlates with phenotypic loss of H-2Kk.
Authors: Gooding L R
Published: December 2022

Cultured SV40-transformed fibroblasts from C3H mice (SV-C3H) were "adapted" to in vivo growth by serial passage through sublethally irradiated, syngeneic recipients. After four in vivo passages, a population of cells was obtained (V4) that was weakly oncogenic in nonirradiated mice. Cells isolated from large V4 tumors (V5) were found to be highly oncogenic, producing lethal tumors at doses of less than 10(3) cells. V5 is insensitive to SV40-specific transplantation immunity in syngeneic animals but can be rejected completely by H-2 allogeneic mice. In vitro studies revealed that although V4 and the parent SV-C3H cells can induce SV40-specific cytotoxic T cells (CTL) in vitro and are lysed by these CTL, V5 does neither. The failure of V5 to interact with CTL was traced to the loss of H-2Kk antigen expression on these cells. The correlation between H-2Kk loss and immunoresistance in vivo suggests a central role for the cytotoxic T cell in in vivo tumor elimination in this system.
Suppressor T cells are activated in vivo in patients with multiple sclerosis coinciding with remission from acute attack.
Authors: Huddlestone J R , Oldstone M B
Published: December 2022

By a variety of assays, investigators (1-4) from several laboratories previously observed changes in numbers of thymus-derived suppressor lymphocytes (Ts) in patients with acute active multiple sclerosis (MS). Changes in numbers of Ts in the peripheral blood compartment of such patients closely follow disease activity. Extending these observations we now report Ts are activated in the peripheral blood of certain patients with MS. This activation occurred in vivo in the majority of patients with active disease, but only during remission after an acute episode of MS. In contrast, activated Ts were found infrequently in patients with chronic progressive or stable MS, with neurologic diseases other than MS, or healthy individuals. The suppressor activity found in peripheral blood T cells during the acute remitting stage of disease was associated with elevated numbers of lymphocytes bearing Fc receptors for IgG and/or OKT8 markers.
RIIIS/J mice lack a gene influencing the capacity to make an antibody response to helper T cell-independent antigens.
Authors: Hiernaux J , Rollwagen F M , Rudbach J A , Baker P J
Published: December 2022

RIIIS/J mice lack an autosomal dominant gene(s) that has a decisive influence in determining the capacity to make an IgM antibody response to several representative helper T cell-independent antigens; these mice appear to possess functionally active helper T cells because of their ability to make a reasonably good antibody response to SRBC, a helper T cell-dependent antigen. The genetic defect involved cannot be attributed to the absence of B cells capable of giving a mitogenic response to bacterial LPS.
Qa-1 and Qa-2 expression on CFU-s.
Authors: Lynes M A , Tonkonogy S , Flaherty L
Published: December 2022

Multipotential stem cells can be detected in bone marrow cells preparations by injecting these cells into syngeneic irradiated hosts. Colonies (CFU-s) are visible macroscopically on the spleen after 8 days. Pretreatment of A-Tlab (Qa-1.2) bone marrow with A-anti-A-Tlab serum (anti-Qa-1.2), in the presence of complement, reduced CFU-s by 80% whereas A-Tlab anti-A serum (anti-Qa-1.1) treatment had no effect. Pretreatment of A(Qa-1.1) bone marrow with A-Tlab anti-A serum reduced macroscopic splenic colonies by 78%. Similar results were obtained by using C57BL/6 (Qa-1.2) and B6-Tlaa (Qa-1.1) bone marrow. Thus, both Qa-1.1 and Qa-1.2 are expressed on CFU-s. Qa-2 expression on CFU-2 was examined in the same manner. B6.KI anti-B6 (anti-Qa-2,3) serum reduced CFU-s numbers by 87% on B6 (Qa-2,3+) while having little or no effect on B6.KI (Qa-2,3-). Treatment with D3.262, a monoclonal anti-Qa-2 antibody, reduced B6 CFU-s by 80%, and had no effect on B6.KI CFU-s. Detection of Qa-1 and Qa-2 on CFU-s underscores the wide distribution of the Qa antigens in the hematopoietic system.
Lectin-induced regulation of human thymus-independent anti-TNP responses in vitro.
Authors: Golding B , Burger D R , Rittenberg M B
Published: December 2022

Recently we showed that human hapten-specific T cell-independent (TI) antibody responses could be elicited in vitro with the antigen trinitrophenylated Brucella abortus (TNP-Ba). Although by definition T cells are not required for TI responses, they have also shown capable of regulating such responses in the mouse. In this study we examine the ability of the T cell lectins Con A and PHA to modulate TI responses of human tonsil cells. Addition of the lectins to cultures on day 0 or day 3 resulted in inhibition (greater than 90%) or enhancement (greater than 150%) of the anti-TNP PFC response, respectively. The inhibition was only apparent if the lectins were added at the onset of culture but not 24 hr later, and was T cell-dependent, because T cell-depleted cultures (less than 0.5% E-rosetting cells) could not be inhibited. The enhancement observed was even greater in T cell-depleted cultures and was antigen-specific because TNP-SRBC but not PC-SRBC targets were lysed and the PFC were inhibited by soluble TNP (greater than 70%). This finding suggested that anti-TNP PFC precursors were proliferating more vigorously or that additional, previously antigen-insensitive cells were being recruited in the presence of lectin.