bluuebunny/crossref_metadata_2025 huggingface dataset:
Cellular and growth factor requirements for activation of human T lymphocytes by neuraminidase and galactose oxidase-treated lymphoid cells.
Authors: Lipkowitz S , Rubin A L , Stenzel K H , Novogrodsky A
Published: December 2022

Proliferation of peripheral blood mononuclear leukocytes (PBL) can be stimulated by irradiated autologous PBL that have been treated with neuraminidase and galactose oxidase (NAGO). We determined the types of cells that stimulate and respond by isolating populations of cells enriched for macrophages, B lymphocytes, and T lymphocytes. With PBL as responding cells, proliferation was stimulated by irradiated, NAGO-treated cells with the following hierarchy: macrophage greater than PBL greater than B cells greater than T cells. Irradiated NAGO-treated macrophages and PBL induced proliferation of isolated T cells greater than or equal to the proliferation they stimulated in PBL, indicating that T cells are the predominant responding cell type. Irradiated, NAGO-treated B cells or T cells were weak stimulators of isolated T cell proliferation when compared to their ability to stimulate PBL. The ability of irradiated NAGO-treated B cells or T cells to stimulate isolated T cell proliferation was greatly enhanced by the addition of untreated macrophages or by the addition of conditioned media from mitogen-activated PBL. Biologic, biochemical, and biophysical characterization of the conditioned media revealed it contained both lymphocyte polypeptide growth factors, interleukin 1 and interleukin 2. Semipurified preparations of either of these growth factors were capable of enhancing T cell proliferation stimulated by irradiated NAGO-treated B or T cells. These data indicate T cell proliferation induced by irradiated, NAGO-treated cells requires the aldehyde-bearing cells for the induction of soluble growth factor production and for the induction of putative membrane receptors for these growth factors.
The role of interleukin 1 in human B cell activation: inhibition of B cell proliferation and the generation of immunoglobulin-secreting cells by an antibody against human leukocytic pyrogen.
Authors: Lipsky P E , Thompson P A , Rosenwasser L J , Dinarello C A
Published: December 2022

The role of factors released by monocytes (M phi) in the activation of human B lymphocytes was examined by studying the effect of an antiserum against human leukocytic pyrogen (LP) on mitogen-stimulated B cell proliferation and the generation of immunoglobulin-secreting cells (ISC) by peripheral blood mononuclear cells (PBM). Antiserum against LP was obtained from rabbits immunized with LP-containing human M phi supernatants. The globulin fraction of this antiserum inhibited pokeweed mitogen- (PWM) stimulated B cell proliferation and the generation of ISC in a concentration-dependent manner, with 50% inhibition of responsiveness observed with 10 micrograms/ml. By contrast, PWM-induced T cell [3H]thymidine incorporation was not inhibited by concentrations of anti-LP as great as 2000 micrograms/ml. The F(ab')2 fraction of anti-LP also inhibited the generation of ISC in response to both PWM and formalinized Staphylococcus aureus, but required 50 micrograms/ml to achieve 50% inhibition. Anti-LP inhibited the generation of ISC only if present during the first 24 hr of a 6 to 7-day incubation; later addition was not inhibitory. Inhibition was more marked in cultures partially depleted of M phi than in whole PBM cultures. Whereas absorption of the anti-LP with PBM failed to remove the capacity to inhibit the generation of ISC, anti-LP-mediated inhibition of responsiveness could be reversed by the addition of crude M phi culture supernatants or a variety of highly purified interleukin 1 (IL 1) preparations, but not by T cell supernatants. These results indicate anti-LP inhibits human B cell activation by removing the requisite M phi-derived factor IL 1 and also confirm that IL 1 plays an essential role in B cell proliferation and the generation of ISC in man.
T cell lines and variant subclones with differing susceptibility to tumor promoters.
Authors: Di Pauli R , Rassat J
Published: December 2022

The effect of tumor-promoting phorbol esters on cloned cytotoxic T cell (CTL) lines, H-2 restricted and specific for minor histocompatibility antigens, was studied. We found that two of the three CTL lines tested can be induced by 12-O-tetradecanoylphorbol 13 acetate (TPA or PMA) to synthesize DNA. One of the lines did not respond. TPA concentrations in the range of 1 nmol were active, and induction of DNA synthesis was independent from cell densities. Testing of other phorbol esters showed correlations with their tumor-promoting activity, because TPA and phorbol 12,13-didecanoate were active, whereas phorbol and 4 alpha phorbol 12,13-didecanoate had no effect. Interestingly, two subclones isolated from one of the lines responding to TPA behaved differently: one of the subclones synthesized DNA when incubated with TPA, whereas the other remained completely negative. This difference was reflected also in the dissimilarity of morphologic alterations (observed by scanning electron microscopy) occurring in the responding cells. The nonresponsiveness, in terms of DNA synthesis, by some of the CTL lines is not due to a lack of interaction between TPA and the cells. When incubated simultaneously with TPA and T cell growth factor (IL 2), all lines responded with greater DNA synthesis than when incubated with IL 2 or TPA alone, thus demonstrating a synergistic action between TPA and IL 2. We have also done experiments to test whether TPA induces the synthesis and secretion of a factor leading to proliferation of the cells. From bystander experiments it appears that the action of TPA is not mediated by a secreted product but is a direct one.
The ability of lymphokine and lipopolysaccharide to induce procoagulant activity in mouse macrophage cell lines.
Authors: Farram E , Geczy C L , Moon D K , Hopper K
Published: December 2022

A comparison was made of the abilities of mouse macrophage cell lines and cultured bone marrow-derived macrophages to respond to lymphokine and lipopolysaccharide (LPS) in the macrophage procoagulant assay (MPCA). Mouse macrophage cell lines PU5 and WEHI 265 responded to low concentrations of lymphokine and LPS by increased procoagulant activity; the activity of PU5 cells was comparable to that of TG-PEC. Approximately one-half of the MPCA induced by lymphokine after 20 hr culture was detectable after 2 hr with either cell line. J774 was unresponsive to LPS but responded weakly to lymphokine, and P388D1 and WEHI 274 were unresponsive to both stimuli as determined by the MPCA test. Addition of purified T cells (five T cells to one macrophage) to the unresponsive cell lines induced procoagulant activity in the presence of either stimulus, which suggests that cell contact is necessary in some cases. Only J774 cells responded to lymphocyte-derived chemotactic factor (LDCF) or endotoxin-activated mouse serum (EAMS) in chemotaxis assays. Immature bone marrow cells responded to chemotactic stimuli, and procoagulant activity was induced with lymphokine after 4 days in culture. Cells cultured for 7 days had about twice as much MPCA, but further culturing did not result in cells with enhanced activity. The procoagulant responsiveness of different macrophage cell lines to lymphokine or LPS may reflect differences in the maturation and differentiation states of these cells.
Two surface antigens expressed on proliferating mouse T lymphocytes defined by rat monoclonal antibodies.
Authors: Takei F
Published: December 2022

A hybrid cell line resulting from the fusion of a Con A-activated normal mouse spleen cell and a transformed mouse T cell (EL-4BU) has been used to prepare and select rat monoclonal antibodies reactive with molecules expressed on the surface of proliferating, as opposed to resting, mouse T cells. In this report, the characterization of two such antigens identified in this way is described. One antigen is a membrane component common to mitogen-activated T and B cells, some bone marrow cells, and various transformed cell lines but is not detectable on either normal thymocytes or the majority of spleen cells by radioimmunoassay or FACS analysis. It has a m.w. of approximately 200,000 daltons under nonreducing conditions and 100,000 daltons under reducing conditions. Antibodies to this antigen precipitate cell-bound transferrin but do not react directly with transferrin itself. It would thus appear that the antigen is the transferrin receptor molecule. The second antigen is not detectable on normal thymocytes, spleen cells, bone marrow cells, or mitogen-stimulated spleen cells but is expressed at high levels on some transformed T cell lines. It, too, appears to be a dimer, with a m.w. of 95,000 daltons under nonreducing conditions, decreasing to 50,000 daltons under reducing conditions. Although the function of the 95,000-dalton antigen is not yet known, its lack of expression on adult T cell populations both before and after activation suggests either a short-lived role at a very early stage of T cell development and/or an association with T cell transformation.
Decreased C5b67-inhibitor activity in two families with hereditary functional deficiency of the eighth component of complement.
Authors: Zeitz H J , Zeff R A , Gewurz H , Lint T F
Published: December 2022

In addition to its role in hemolysis and host defense against Neisseria infection, the eighth component of human complement (C8) is one of several plasma proteins that are C5b67-inhibitors (C5b67-INH). The recent identification in our laboratory of two new families with hereditary deficiency of C8 provided an opportunity to study further the role of C8 as a C5b67-INH. Based on mixing and reconstitution experiments, the deficiency of C8 seemed to be due to a selective lack of the C8 beta-chain in one family and the C8 alpha-gamma subunit in the other family. Sera from individuals homozygous for the C8 abnormality were substantially deficient in C5b67-INH activity as well as totally deficient in hemolytic activity. Sera from control individuals possessed approximately 2500 C5b67-INH U/ml, whereas sera from the C8-deficient individuals had markedly depressed C5b67-INH activity, with a mean of only 428 U/ml. C5b67-INH activity was completely reconstituted in C8-deficient serum by the addition of purified human C8. We conclude that C8 constitutes the substantial majority of the C5b67-INH activity of normal human serum.
Cell-bound C4b resists reduction by reducing agents: analysis by chain structure and by hemolytic activity.
Authors: Ejzemberg R , Brown E J , Ohanian S H , Hammer C H , Borsos T
Published: December 2022

EA3H-C4hu treated with 2-mercaptoethanol (MSH) or dithiothreitol (DTT) were analyzed for residual chains and hemolytic activity. MSH treatment resulted in no loss of the chains of C4b and no loss of reactivity with a polyclonal anti-human C4 rabbit antibody, and the cells did not lose their ability to generate SAC4b2a. DTT-treated cells lost about 70% of the gamma-chain and over 95% of C4b activity; there was no loss of alpha' and beta chains or the ability to react with anti-C4 antibody. Kinetic studies indicated that the remaining 30% of the gamma-chains could not be removed by prolonged incubation with DTT, implying heterogeneity of cell bound C4b. The data also imply that the gamma-chain is important in the generation of SAC4b2a.
Characterization of mast cell precursors by physical means: dissociation from T cells and T cell precursors.
Authors: Yung Y P , Wang S Y , Moore M A
Published: December 2022

Murine bone marrow precursors (MCP) differentiate into mast cells and proliferate in response to mast cell growth factor (MCGF). An assay system based on the incorporation of [3']thymidine by proliferating mast cells in response to substantially purified MCGF was used to titrate MCP. Murine bone marrow was separated into fractions by Percoll density gradient centrifugation or velocity sedimentation at unit gravity. Individual fractions were analyzed to determine the relative concentrations of MCP. To determine if mast cells were derived from precursors different from those for T lymphocytes and granulocytes and macrophages, separated cells were also analyzed for responsiveness to interleukin 2 (IL 2) in standard [3H] thymidine incorporation assays and to granulocyte-macrophage colony stimulating factors (CSF) in agar cloning (CFU-GM) assays. Cultured marrow cells from Dexter type long-term marrow cultures (LMC) were included as a source of marrow cells devoid of mature T cells, so as to dissociate MCP from pre-T cells. MCP were readily dissociable from T cells and/or pre-T cells present in fresh, as well as cultured, marrow cells by either density or velocity of sedimentation separation techniques. MCP were, however, not readily separable from CFU-GM. Nevertheless, MCP appeared not to share a common precursor with pure macrophage type colony-forming cells (CFU-M phi). The implications of the findings on the lineage origin of mast cells were discussed.
The malaria merozoite surface: a 140,000 m.w. protein antigenically unrelated to other surface components on Plasmodium knowlesi merozoites.
Authors: Hudson D E , Miller L H , Richards R L , David P H , Alving C R , Gitler C
Published: December 2022

We previously identified three proteins on the surface of merozoites (140,000, 105,000 and 75,000 m.w.). To determine if 140,000 m.w. protein was related to other surface proteins, we immunized mice with liposomes containing merozoite proteins from the 140,000 m.w. region of the polyacrylamide gel. The immune sera reacted with the surface of viable merozoites and acetone-fixed schizonts by immunofluorescence. The sera immunoprecipitated only the 140,000 m.w. protein from surface-labeled merozoites. We demonstrated that monoclonal antibody 13C11 immunoprecipitated a 250,000 m.w. protein from metabolically labeled schizonts and bound to the merozoite surface. This monoclonal antibody immunoprecipitated the 75,000 and lower m.w. proteins from surface-labeled merozoites but did not bring down the 140,000 m.w. protein. Because the mouse immune sera did not immunoprecipitate the 250,000 m.w. protein from metabolically labeled schizonts or proteins other than the 140,000 m.w. protein from surface-labeled merozoites, we conclude that the 140,000 m.w. protein is unrelated to other merozoite surface antigens identified to date. The mouse immune sera against the 140,000 m.w. protein on the merozoite surface did not immunoprecipitate a 140,000 m.w. protein from metabolically labeled schizonts. Instead, the major protein immunoprecipitated had a m.w. of 144,000. By analogy to the 250,000 m.w. protein and its cleavage products, we propose that the 140,000 m.w. protein on the merozoite surface is a cleavage product of the higher m.w. protein.
A unique antigen expressed on myeloid cells and acute leukemia blast cells defined by a monoclonal antibody.
Authors: Ball E D , Graziano R F , Fanger M W
Published: December 2022

A murine hybridoma-derived monoclonal antibody, PM-81, was obtained from a fusion of cells of the NS-1 myeloma cell line with cells from a mouse immunized with the HL-60 promyelocytic leukemia cell line. This cytotoxic IgM monoclonal antibody was specific for myeloid cells. Employing indirect immunofluorescence and flow cytometry, we determined that this antibody reacts strongly with normal human granulocytes, eosinophils, and monocytes but not lymphocytes (including phytohemagglutinin-activated lymphocytes), null cells, red blood cells, or platelets. Moreover, the PM-81 antibody reacts with leukemia cells from 19 of 22 patients with acute myelocytic leukemia of all FAB subclasses, three of three patients with common acute lymphocytic leukemia, four of four patients with chronic myelocytic leukemia (CML) in myeloid blast crisis (terminal transferase (TdT)-negative) but did not react with cells from two patients with CML in lymphoid blast crisis (TdT-positive) or five patients with chronic lymphocytic leukemia. The myeloid cell lines HL-60, K562, KG-1, and U937 were all reactive with PM-81. The lymphoid lines CCRF-CEM and Daudi did not express PM-81 but HSB-2 was positive. The PM-81 antigen was absent on myeloid and erythroid progenitor cells as determined by their insusceptibility to complement-dependent lysis. In addition, only PM-81-unreactive cells were capable of colony formation. Furthermore, the PM-81 antibody does not appear to induce modulation of the antigen to which it binds. Thus, this monoclonal antibody appears to fulfill several criteria for clinical utility in the diagnosis and treatment of both acute myelocytic and acute lymphocytic leukemia.
Characterization of cloned cytotoxic lymphocytes with NK-like activity.
Authors: Acha-Orbea H , Groscurth P , Lang R , Stitz L , Hengartner H
Published: December 2022

Cloned H-2b-restricted male antigen-specific cytotoxic T cells were kept in culture in the presence of IL 2 for over 2 yr. During this time, they lost their antigenic specificity and now exhibit high NK-like activity. Three separate clones HY 1, HY 2, and HY 3 and their subclones, when tested on a panel of NK-sensitive and NK-insensitive target cells, showed typical lysis patterns of NK cells. Three cytotoxicity-loss mutants were obtained by subcloning. At least one of these cytotoxicity-loss mutants still exhibited binding to NK-sensitive target cells. The various HY clones were compared with cloned alloreactive cytotoxic T cells of B10 anti-B10.D2 specificity with respect to surface markers, target specificity, kinetics of lysis, morphology, and influence of monoclonal antibodies and chemicals on cytotoxicity. Only one major difference between cloned NK-like and alloreactive T cells was detected: monoclonal anti-Lyt-2 antibodies inhibited cytotoxic activity of alloreactive T cells but not of NK-like cells, although both expressed Lyt-2 on their surface. Morphologically, HY cells were characterized by numerous lysosomal granules in their cytoplasm. Clones with NK-like activity differed from cytotoxicity-loss mutants by both ultrastructural appearance and enzyme histochemistry of lysosomes. Interaction of NK-like cells with YAC-1 cells induced membrane lesions of variable size in target cells. Effects of chemicals influencing lysosomal pathways such as monensin, NH4Cl, and chloroquine on cytotoxicity were investigated. Monensin as well as NH4Cl inhibited cytotoxicity of NK as well as alloreactive clones, whereas chloroquine did not. Spleen cells from acutely virus-infected mice with high NK reactivity showed the same behavior except for chloroquine, which was slightly inhibitory. The addition of monensin to the cells resulted in morphologic changes in the lysosomal granules. These data strongly suggest that NK-like killing is triggered by unknown substance(s) released from lysosomes of effector cells. Cloning of antigen-specific cytotoxic T cells under conditions of limiting dilution and continuous growing in medium containing Con A supernatant but not the relevant antigen often leads to loss of specificity and appearance of NK-like activity. The results presented are representative of a great amount of work in this and probably many other laboratories attempting to clone cytotoxic effector T cells.
Studies on the mechanism of natural killer cytotoxicity. III. Activation of NK cells by interferon augments the lytic activity of released natural killer cytotoxic factors (NKCF).
Authors: Wright S C , Bonavida B
Published: December 2022

The mechanism by which interferon (IFN) pretreatment of effector cells augments natural killer (NK) cell-mediated cytotoxicity (CMC) was examined by determining whether IFN has any effect on the production of natural killer cytotoxic factors (NKCF). NKCF are released into the supernatant of co-cultures of murine spleen cells and YAC-1 stimulator cells, and their lytic activity is measured against YAC-1 target cells. It was demonstrated that pretreatment of effector cells with murine fibroblast IFN or polyinosinic-polycytidylic acid (pIC) resulted in the release of NKCF with augmented lytic activity. Evidence indicated that the IFN-induced augmentation of NKCF activity required protein synthesis during the IFN pretreatment period, because concurrent pretreatment with both IFN and cycloheximide abrogated the IFN effect. Protein synthesis, however, is not required for the production of base levels of NKCF because emetine pretreatment of normal spleen cells did not result in a decrease in NKCF production. Furthermore, substantial levels of NKCF activity could be detected in freeze-thaw lysates of freshly isolated spleen cells. Cell populations enriched for NK effector cells, such as nylon wool-nonadherent nude mouse spleen cells, produced lysates with high levels of NKCF activity, whereas lysates of CBA thymocytes were devoid of NKCF activity. Pretreatment of spleen cells with either IFN or pIC resulted in an augmentation of the NKCF activity present in their cell lysates. Taken altogether, these findings suggest that freshly isolated NK cells contain preformed pools of NKCF. Pretreatment of these cells with IFN causes de novo synthesis of additional NKCF and/or activation of preexisting NKCF. According to our model for the mechanism of NK CMC, target cell lysis is ultimately the result of transfer of NKCF from the effector cell to the target cell. The evidence presented here suggests that the IFN-induced augmentation of NK activity could be accounted for by an increase in the synthesis, activation, and/or release of NKCF.
Alterations in lymphocyte homing patterns within mice exposed to ultraviolet radiation.
Authors: Spangrude G J , Bernhard E J , Ajioka R S , Daynes R A
Published: December 2022

This report presents the results of an investigation designed to establish whether exposure of mice to ultraviolet radiation (UVR) is capable of influencing the factors that control the distribution of lymphoid cells in vivo. We found that such exposure resulted in a dramatic and long-lasting increase in the tropism of peripheral lymph nodes for circulating lymphoid cells. Termination of UVR exposure did not result in a reversal of this phenomenon. Since an increase in lymphocyte migration into the lymph nodes of UVR-exposed mice was apparent within 2 hr of infusion of the radiolabeled cells, we conclude that the homing assay data reflect a relatively increased binding of circulating lymphocytes to high endothelial venules (HEV) within the lymph nodes of irradiated animals. A histologic analysis of skin from UVR-exposed mice established that the dermal microvasculature had expanded in terms of size and number of vessels, a condition that also does not completely reverse after the termination of treatments. In spite of the increase in dermal microvasculature, very few inflammatory cells were detected in the irradiated skin site. These observations support our conclusion that the enhanced traffic of lymphocytes into peripheral lymph nodes of UVR-exposed mice occurs primarily via lymphocyte-HEV interactions rather than afferent drainage of the irradiated skin.
Augmentation of syngeneic tumor-specific immunity by semiallogeneic cell hybrids.
Authors: Toffaletti D L , Darrow T L , Scott D W
Published: December 2022

Hybrid cell lines were established from fusions between lipopolysaccharide- (LPS) stimulated C57BL/6J spleen cells and MPC-11 tumor cells (45.6TG1.7, abbreviated M45), and were tested for their ability to immunize semiallogeneic mice against a parental tumor challenge. These hybrids were tumorigenic in syngeneic (BALB/c X C57BL/6J) F1 (CB6F1) mice but did not grow in semiallogeneic (BALB/c X A/J) F1 (CAF1) mice. All hybrids express both parental major histocompatibility antigens (H-2b and H-2d) as detected by indirect immunofluorescence and by their ability to function as either stimulators or targets for allogeneic cytotoxic lymphocytes (CTL). M45 tumor-associated antigens (TAA) were expressed on the hybrid surface as shown by their ability to act as either stimulators or targets for syngeneic CTL specific for M45 TAA. Immunization of semiallogeneic CAF1 mice with the hybrids i.p. followed by a challenge with M45 tumor cells resulted in extended survival when compared to untreated mice or animals immunized i.p. with M45 tumor cells. This immunity was specific and was not due to an allogeneic effect; immunization with an unrelated H-2bd tumor, 70Z/3, or H-2bd B6D2F1 spleen cells or with semiallogeneic spleen cells plus M45 did not protect mice from M45 challenge. Interestingly, prophylactic priming with semiallogeneic hybrid tumor cells or parental myeloma cells led to M45-specific CTL and "help" for an in vitro CTL response; however, the degree of CTL priming by hybrid tumors was not augmented when compared to the level of CTL achieved with parental tumor alone. Hence, stimulation of CTL activity per se by hybrid tumor cells cannot explain the protective effect of hybrid tumor immunization. These studies nevertheless confirm that semiallogeneic hybrids, which we show express TAA and alloantigens, can be used to immunize mice against a lethal syngeneic myeloma tumor challenge.
Parenteral immunization causes antigen-specific cell-mediated suppression of an intestinal IgA response.
Authors: Koster F T , Pierce N F
Published: December 2022

Rats immunized s.c. with cholera toxin (CT) or toxoid (CTd) show antigen-specific suppression of the jejunal IgA anti-CT response to subsequent enteric doses of CT. We determined whether this effect was partly cell-mediated and studied the suppressor cell response involved. Spleen cells from s.c.-immunized rats suppressed the jejunal anti-CT response in adoptive recipients when cell transfer was at, or 4 days before, intraduodenal priming with CT. Transferred suppression was antigen-specific, and the suppressor cells were nylon wool-nonadherent. Increasing the s.c. dose of CT from 0.1 to 40 micrograms, and the interval between immunization and cell harvest from 2 to 16 wk, each increased the suppressive effect of a constant spleen cell inoculum. After s.c. immunization, suppressor cells were present in the spleen within 1 to 2 wk, among TDL within 4 to 8 wk, and in Peyer's patches and thymus within 8 to 16 wk; this sequence suggested they arose in the spleen and later migrated to mucosae and the thymus. Prior splenectomy did not alter the suppressive effect of s.c. CT, however, nor did it prevent suppressor cell appearance among TDL, indicating that suppressor cells also arose from nonsplenic sites. Transferred suppressor cells acted by interfering with the development of specific immunologic memory within Peyer's patches during enteric priming; transfer of suppressor cells at the time of enteric boosting had no effect upon the secondary mucosal anti-CT response. We conclude that the suppressive effect of s.c. immunization on a specific mucosal IgA response is due largely to the action of systemically derived suppressor cells upon the primary mucosal immune response within Peyer's patches. This sequence resembles the mirror image of oral tolerance, which involves the suppression of systemic IgG and IgM responses by suppressor cells that arise in Peyer's patches and migrate to the spleen after antigen feeding.
Polyvalent epitope inhibits but added with anti-idiotype antibody enhances idiotype suppression.
Authors: Kim B S , Massey P B
Published: December 2022

When BALB/c spleen cell cultures were preincubated with polyvalent PC-antigens, regardless of their immunogenicity, cultures became resistant to id suppression induced by subsequent treatment with anti-id antibody and immunogenic PC-antigen. In addition, incubation with both anti-id and any of these antigens assisted the induction of rapid, irreversible B cell tolerance. These results suggest that B cells bearing receptors for an epitope undergo an initial triggering in response to polyvalent epitope regardless of the T dependence and/or immunogenicity of the antigen. An additional signal must then facilitate the differentiation of the partially activated B cells to produce antibody. Therefore, a nonimmunogenic polyvalent epitope-bearing molecule can deliver either an initial triggering signal or a tolerogenic signal, depending on the presence of anti-id antibody, and it is as efficient as a complete immunogen.
Possible involvement of the T4 molecule in T cell recognition of class II HLA antigens: evidence from studies of proliferative responses to SB antigens.
Authors: Biddison W E , Rao P E , Talle M A , Goldstein G , Shaw S
Published: December 2022

The T4 molecule has been identified as a marker of human T cell differentiation, but the function of this molecule remains to be defined. We have investigated its possible functional involvement in T cell proliferative responses to class II HLA antigens encoded by the recently described SB locus. The responses of SB-primed cells (specific for each of four different SB antigens) were studied with the use of two proliferation-inducing stimuli, SB antigen or TCGF. The proliferative responses to both stimuli were found to be mediated by T4+, T8- cells. Monoclonal antibodies against some epitopes on the T4 molecule (OKT4A and OKT4B) substantially blocked antigen-stimulated proliferative responses; antibodies against other epitopes of the T4 molecule (OKT4, T4C, T4D) blocked less well. Inhibition of SB-specific proliferation by antibodies to the T4 molecule was maximal only when the antibodies were incubated with the responder cells before the addition of stimulator cells. Proliferative responses of SB-primed cells stimulated with TCGF alone were not inhibited by any of the OKT4-related antibodies, but were completely inhibited by the anti-Tac monoclonal antibody, which reacts with the TCGF receptor. These results lend further support for the hypothesis that the T4 molecule is involved in T cell recognition of and/or activation by class II HLA antigens. We suggest that 1) the T4 molecule binds a nonpolymorphic epitope on class II HLA molecules, and 2) this interaction may facilitate, but not be an obligate requirement for, T cell activation by class II antigens.
The interaction of lymphocyte surface-bound immune complexes and neutrophils.
Authors: Archibald A C , Cheung K , Robinson M F
Published: December 2022

Neutrophils can be stimulated directly by a variety of stimulants resulting in the production of highly reactive oxygen derivatives. Included in these stimulants are peripheral blood lymphocytes with bovine serum albumin-anti-bovine serum albumin immune complexes (BSA-IC) or aggregated gamma-globulin (AHG) bound to their surface receptors. Through the use of chemiluminescence (CL) studies, we found that B lymphocytes preincubated with AHG stimulated neutrophils to a much greater extent than similarly preincubated T lymphocytes. Preincubated Raji cells (a B lymphoblastoid cell line) were also capable of stimulating neutrophils. We further demonstrated that after periods of mixed incubation with neutrophils, lymphocytes with surface-bound AHG did show an abnormal proliferative response to pokeweed mitogen (PWM), but not to phytohemagglutinin (PHA). This was not due to loss of cell viability, as judged by chromium release cytotoxicity assays and trypan blue exclusion, or to neutrophil enzyme release. The data suggest that neutrophils, stimulated with lymphocyte surface-bound immune complexes, are capable of producing an environment of in vitro oxidant stress. This stress, although not great enough to cause a significant decrease in lymphocyte viability, can cause impaired lymphocyte function. Physiologically, this may relate in the long term to immunologic malfunction observed in patients with high levels of circulating immune complexes.
The T lymphocyte response to cytochrome c. IV. Distinguishable sites on a peptide antigen which affect antigenic strength and memory.
Authors: Hansburg D , Fairwell T , Schwartz R H , Appella E
Published: December 2022

The murine T cell proliferative response to the carboxyl terminal cyanogen bromide cleavage fragment 81-104 of pigeon cytochrome c (cyt) has been studied. Two interesting properties of this response have been previously described. First, T cells from B10.A mice primed with pigeon cyt 81-104 show more vigorous proliferation when restimulated with moth cyt 81-103 than when stimulated with pigeon cyt 81-104; that is, the B10.A T cell response to pigeon shows heteroclitic restimulation by moth. Second, T cells primed with the acetimidyl derivative (Am) of pigeon cyt 81-104 did not cross-react with the unmodified cyt fragments, but Am-moth cyt 81-103 still stimulated Am-pigeon cyt 81-104 primed T cells better than the Am-pigeon cyt 81-104 fragment. These results raised the issue of whether the antigenic sites on the fragments responsible for the specificity of T cell priming in vivo differed from the residues that contributed to the heteroclitic response of pigeon (or Am pigeon)-primed T cells to moth cyt c fragments. In this paper, synthetic peptide antigens were tested in order to identify which residues caused the heterocliticity of the moth fragment and which residues were involved in the antigenic differentiation of native and derivatized fragments. The heterocliticity of the T cell response to moth fragment 81-103 was found to be due to the deletion of the penultimate residue (Ala103) from the pigeon fragment. However, the ability to cause heterocliticity was not uniquely a property of this deletion. T cells from animals primed with peptides containing substitutions at positions 100 or 102 were also heteroclitically stimulated by the moth-like antigen. The observation that T cells could not be primed for recognition of the changes in peptide sequence that caused heteroclitic stimulation suggests that T cells do not directly recognize determinants in this region. The antigenically significant site of derivatization for T cell priming was found to be Lys99. Furthermore, substitution of a Gln at position 99 also resulted in elicitation of yet a third set of T cell clones specific for the presence of that residue. That is, the specificity of the primed T cell population was found to be altered by changes at residue-99, but no such alterations in specificity were demonstrable when T cells primed with peptides altered at residue-103, residue-102, or residue-100 were compared. Overall, the results demonstrate that the antigen can be divided into two functionally distinct sites that are in close physical proximity.
Rapid turnover of HLA proteins in quiescent lymphocytes: proposed connection with immunologic surveillance.
Authors: Monos D S , Cooper H L
Published: December 2022

Current immunologic theory defines a role for HLA-A, -B, -C proteins in cells presenting foreign antigens to cytotoxic T lymphocytes. No clear function, however, is assigned to the prominently represented HLA molecules on the T cells themselves. To investigate this question, the synthesis and turnover of HLA-A, -B, -C molecules in human peripheral lymphocytes was studied. Newly synthesized HLA-A, -B, -C and beta 2-microglobulin molecules were identified among total lymphocyte proteins by two-dimensional gel electrophoresis and specific immunoselection. In resting T lymphocytes, the polymorphic set of HLA-A, -B, -C proteins was among the most prominently synthesized and rapidly turned over of total cell proteins. Its half-life was 6 to 7 hr, which is considerably shorter than that of total cell protein. HLA and beta 2M were the major proteins of the plasma membrane undergoing active synthesis and turnover in resting T lymphocytes. After growth stimulation, the rate of HLA synthesis increased to a lesser degree than total protein synthesis. Survival of newly synthesized HLA was increased, however, indicating reduction in HLA turnover. The result was the accumulation of HLA in the plasma membrane. It is suggested that continuous degradation and replacement reflects the biochemical nature of the participation of HLA molecules in immunologic surveillance by quiescent lymphocytes. Cessation of HLA turnover, with accumulation of surface HLA molecules, is a biochemical adjustment related to T cell activation.
Characterization of the oligosaccharides from the invariant chain associated with murine Ia antigens.
Authors: Swiedler S J , Hart G W , Freed J H
Published: December 2022

The oligosaccharide side chains of the murine invariant chain (Ii) glycoprotein associated with the Ia antigens have been characterized. Affinity chromatography using a monoclonal antibody column was employed to purify the I-Ak antigen from lysates of the AKTB-1b B cell lymphoma. The invariant chain isolated by this procedure was subsequently digested with proteases, and the resulting glycopeptides were fractionated by reverse-phase high-pressure liquid chromatography (HPLC). The invariant chain appears to contain two glycosylation sites, both of which carry high-mannose oligosaccharides, each with a restricted size distribution and an average composition of Man6GlcNAc2 as judged by gel filtration and alpha-mannosidase digestion. These data, together with the observation that the invariant chain cannot be labeled metabolically with 3H-glucosamine, 3H-fucose, or 3H-galactose, allow the conclusion that the murine invariant chain does not contain complex oligosaccharides and is not O-glycosylated during its association with the Ia antigens.
Idiotypic and fluorometric analysis of the antibodies that distinguish the lesion of the I-A mutant B6.C-H-2bm12.
Authors: Melino M R , Epstein S L , Sachs D H , Hansen T H
Published: December 2022

The serologic lesion of the I-A mutant mouse strain, bm12, was investigated with the use of monoclonal anti-Iab antibodies and anti-idiotypic (Id) reagents produced against these antibodies. In a fluorometric analysis, three different monoclonal anti-Iab antibodies (25-9-17, 34-5-3, 28-16-8) failed to bind bm12 cells, whereas two anti-Iab antibodies (25-5-16 and 17/227), which bound bm12 cells, showed about one-half the fluorescence intensity that they showed in binding to Iab antigens. Of the three monoclonal antibodies that failed to react with bm12 cells, two antibodies (25-9-17 and 34-5-3) were found to bind the same steric site on Iab molecules (cluster I). In contrast, the antibodies (25-5-16 and 17/227) that reacted with both Iab and Iabm12 antigens were found to bind a second distinct site (cluster II). The binding of antibody 28-16-8 to Iab antigens inhibited reciprocally the binding of cluster I and II anti-Iab antibodies, suggesting a possible third site, sterically located intermediate between the other two sites. To assess the relatedness of the antibodies defining the serologic lesion of bm12 mice, xenogeneic and syngeneic anti-Id reagents were produced against antibodies 25-9-17 and 28-16-8. By using these anti-Ids in a binding site-related inhibition assay, a cross-reactive idiotype was detected that is shared by 25-9-17 and 34-5-3 antibodies; thus these two monoclonal antibodies share several features, including 1) idiotypic determinants, 2) failure to bind bm12 cells, 3) binding the same spatial Iab site, and 4) having indistinguishable serologic fine specificity that corresponds with a previously defined predominant alloantigenic determinant recognized in the bm12 anti-Iab humoral response. Therefore, several parameters of antibody recognition of Ia can now be correlated with structural changes in Ia molecules. These findings will potentiate future studies of the T cell recognition of these same Ia epitopes.
The fate of IgE bound to rat basophilic leukemia cells. II. Endocytosis of IgE oligomers and effect on receptor turnover.
Authors: Isersky C , Rivera J , Segal D M , Triche T
Published: December 2022

We have assessed the internalization of variously sized oligomers of IgE bound to rat basophilic leukemia (RBL) cells by measuring their accessibility to the extracellular environment, and by direct visualization of the radiolabeled ligands. We also followed the fate of the internalized ligands and their receptors, as well as the fate of the free receptor on cells internalizing oligomers. In contrast to monomeric IgE, surface-bound oligomeric IgE was internalized. Notably, dimers provided an effective signal for internalization, although larger oligomers seem to be internalized more efficiently. In our experiments, 48% of the cell-bound dimers and 67% of the trimers were eliminated from the cell surface in 180 min. One-half of the maximal internalization observed with dimers and trimers occurred in 25 and 11 min, respectively. Release of radioactivity into the supernatant followed internalization; the released radioactivity did not bind to fresh cells and was only partially TCA-precipitable. Radioactive ligands remaining associated with the cells were unchanged as judged by m.w; they also were shown to remain receptor-bound. During either internalization or release of substantial amounts of the originally cell-bound oligomers, there was no increase in IgE-binding activity. In contrast, there was a transient drop (25%) in the number of free surface receptors suggesting internalization of the free receptors together with the oligomer-occupied receptor. Cells that failed to release histamine (RBL-I) processed dimeric and trimeric IgE similarly to histamine-releasing (RBL-2H3) cells. We conclude that dimeric and trimeric IgE are internalized by RBL cells and later are released to the medium in a partially degraded form. The ligand-bound receptor seems to be internalized with the ligand, along with some free receptor, and does not appear to be reusable or to recycle rapidly to the cell surface.
Models of T cell deficiency in listeriosis: the effects of cortisone and cyclosporin A on normal and nude BALB/c mice.
Authors: Schaffner A , Douglas H , Davis C E
Published: December 2022

It is often difficult to test the role of T lymphocytes in resistance to infection because most models of T cell deficiency are associated with altered nonspecific resistance. In an attempt to address this problem, we compared the effects of cyclosporin A (CyA), cortisone (CA), and the athymic state on the course of murine listeriosis. We chose listeriosis because resistance to Listeria monocytogenes occurs in two phases. Bacterial multiplication is controlled by nonspecific defense mechanisms in the early phase and by acquired T cell-dependent immunity in the second phase. Mice treated with CA died during the early phase, probably because of inhibition of the antimicrobial activity of nonimmune macrophages. Accordingly, the immunosuppressive effect of CA was similar in athymic and normal mice. Untreated nude mice developed chronic low grade infection, probably because of heightened activity of nonimmune macrophages. In contrast, immunosuppression with CyA did not affect early resistance but induced overwhelming, fatal disease in the later phase when control mice began to acquire resistance. CyA did not change the course of listeriosis in nude mice, confirming its specificity for T cell-dependent immunity. Thus, this study shows that CyA is a potent and specific inhibitor of T cell-mediated immunity and that T cell-dependent resistance is essential for survival from listeriosis, a conclusion that could not have been established by studies of the nude mouse or immunosuppression by CA.
Establishment of a continuous T cell line capable of suppressing anti-tumor immune responses in vivo.
Authors: Roberts L K , Spellman C W , Warner N L
Published: December 2022

This report describes the induction, phenotypic characteristics, and functional properties of a continuous suppressor T cell line. This cell line, UV1, is capable of suppressing anti-tumor immune responses both in vivo and in vitro. The UV1 cell line was derived from a T cell-enriched (nylon wool nonadherent, Ia-negative panned fraction) spleen cell population from a ultraviolet radiation-(UV) exposed BALB/c Wehi mouse. By using an in vivo functional assay designed to demonstrate tumor-specific UV-induced suppressor T lymphocyte (Ts cell) activity, it was found that UV1 cells were capable of rendering normal syngeneic mice susceptible to the growth of UV-induced regressor tumors. In addition to their suppressive activity in vivo, UV1 cells displayed in vitro suppressive activity by blocking the differentiation of cytotoxic T cells from the draining lymph nodes of UV-tumor immunized animals. By flow cytometric analysis it was determined that UV1 cells expressed a number of T lymphocyte differentiation antigens and did not express any detectable amounts of surface immunoglobulin, I-A or E/C antigens, Fc receptors, or macrophage antigens. These data suggest that the UV1 cell line may be representative of the UV-induced Ts cell population and provide a potential means for studying UV-induced immunoregulatory mechanisms in greater detail.
Immune regulation by monoclonal antibodies: failure to enhance mouse skin allografts.
Authors: Lems S P , Tax W J , Capel P J , Koene R A
Published: December 2022

Monoclonal anti-H-2k alloantibodies were analyzed for their capacity to enhance the survival of B10.A skin grafted onto B10.D2 recipients. Included were five anti-class I and four anti-class II antibodies. In contrast to conventional B10.D2 anti-B10.A serum, none of the individual anti-class I or anti-class II monoclonal antibodies induced enhancement. The same negative results were obtained with various mixtures of anti-class I, anti-class II, or anti-class I + anti-class II antibodies. The failure to induce enhancement was not due to inefficient antigen binding in vivo, because monoclonal antibodies were as effective as conventional B10.D2 anti-B10.A serum in the induction of acute antibody-mediated graft rejection, and in the opsonization of 51Cr-labeled B10.A leukocytes injected into B10.D2 recipients pretreated with antibody. These results demonstrate that monoclonal antibodies cannot always substitute for conventional sera, at least not in immune regulation. They also show that although opsonization may be a prerequisite for the induction of enhancement, it does not guarantee that enhancement will invariably occur.
Functional studies on B cell hybridomas with B cell surface antigens. II. Ia molecules on the cell membrane and the differentiative response to anti-mu antibody.
Authors: Hamano T , Leiserson W M , Asofsky R
Published: December 2022

TH 2.52, a subline of the B cell hybridoma with Iab, Iad, and IgM molecules on the cell membrane, was treated with F(ab')2 fragments of purified goat anti-mouse mu antibody (anti-mu), and the change in the expression of surface Ia molecules was determined by microcytotoxicity assays, quantitative absorption tests, and analyses of flow microfluorometery (FMF). We have previously reported that TH 2.52 cells can markedly generate IgM after stimulation with anti-mu without T cell factors. In the present studies, it was shown that the expression of surface Iab molecules on TH 2.52, originated from normal B cells of C57BL/6 (B6) mice, significantly decreased after treatment with anti-mu. In contrast, Iad molecules derived from M12.4.1 lymphomas did not change under the same conditions. These results indicate that cross-linking of anti-mu with surface IgM molecules on TH 2.52 provides signals for differentiation into IgM-secreting cells; this is followed by a decrease in the expression of Iab molecules on the cell membrane. Furthermore, monoclonal anti-Ia antibody (anti-Ia) did not inhibit the generation of IgM-secreting cells by TH 2.52 cells treated with anti-mu. In addition, la- sublines of TH 2.52 obtained after mutagenesis with ethyl methanesulfonate (EMS), as well as the original TH2.52, could differentiate into IgM-secreting cells in the presence of anti-mu. These findings suggest very strongly that la molecules on the cell membrane are not required for the induction of IgM secretion by B cells treated with anti-mu.