bluuebunny/crossref_metadata_2025 huggingface dataset:
The dual interaction of prostaglandin E2 (PGE2) and interferon (IFN) on NK lytic activation: enhanced capacity of effector-target lytic interactions (recycling) and blockage of pre-NK cell recruitment.
Authors: Targan S R
Published: December 2022

In this study we addressed the question of whether the known NK activators prostaglandin E2 (PGE2) and interferon (IFN) can synergistically activate NK lysis and whether they potentiate the same or different subsets of NK cells. We demonstrated that the activation of NK lysis can be enhanced as well as blocked by the interaction of these 2 activators. The enhancement of activation was not due to recruitment of separate subsets of pre-NK cells, but rather the synergistic enhancement of the recycling capabilities of the same set of NK cells. There was a temporal relationship to this enhancement. That is, if PGE2 was added during cellular processing required for full expression of NK activation induced by IFN, pre-NK lytic potential was inhibited. If, on the other hand, PGE2 is added after completion of full expression of the IFN-induced activated lytic process, there is enhancement of recycling capabilities of the same number of NK cells.
Epidermal cell (keratinocyte)-derived thymocyte-activating factor (ETAF).
Authors: Luger T A , Stadler B M , Katz S I , Oppenheim J J
Published: December 2022

In order to determine whether keratinocytes play a role in the modulation of the immune response, we investigated the murine keratinocyte cell line Pam 212. In culture these cells generate a substance with a biologic activity that greatly enhances phytohemagglutinin-induced thymocyte proliferation. We have, therefore, called this substance epidermal cell thymocyte-activating factor (ETAF). This keratinocyte-derived supernatant activity is mainly produced at the onset of the logarithmic growth phase and is directly mitogenic for murine thymocytes. Although ETAF by itself exhibits no T cell growth factor activity, ETAF enhances Interleukin 2 production by mitogen-stimulated murine spleen cells. Murine ETAF is not genetically restricted and lacks species specificity since it decreases lectin-induced proliferation of human peripheral blood lymphocytes (as well as murine spleen cells) and also enhances the production of human Interleukin 2. The factor has a m.w. between 15,000 and 25,000 as determined by gel filtration and elutes as a single peak from anion exchange chromatography columns. The activity is maintained mainly at alkaline pH and is rapidly destroyed at temperatures above 60 degrees C. These observations suggest that epidermal cells may interact with the immune system by elaborating nonspecific factors that modulate lymphocyte proliferation and augment lymphokine production.
Requirement of adherent/phagocytic cells for the induction of F1 anti-parent cell-mediated lympholysis. I. Parental adherent splenocytes serve as stimulators.
Authors: Freimuth W W , Cudkowicz G
Published: December 2022

Primary F1 hybrid anti-parent T cell-mediated lympholysis (F1 alpha P CML), induced in mixed spleen cell cultures, is an immune response specific for self H-2 antigens. The cellular requirements for induction were investigated by depleting or enriching stimulator splenocytes of adherent/phagocytic cells. Relatively large numbers of parental cells that are adherent to Sephadex G-10 particles or to plastic surfaces, and phagocytic for carbonyl iron, were required for stimulation (approximately 10(5) purified stimulators for 3.5 X 10(6) responders). Such parental cells are resistant to gamma radiation (in vivo and in vitro) and hydrocortisone (in vivo), negative for Thy-1, and positive for I-A and I-E/C surface antigens. This subpopulation of macrophages includes the predominant or sole immunogenic parental cell type of the spleen for the F1 alpha P response. Whereas removal of adherent/phagocytic cells from parental splenocytes entailed loss of stimulatory activity for F1 responder T lymphocytes, this was not the case for allogeneic responders. The ability to induce allogeneic CML was, in fact, fully retained by depleted parental cells. Adherent/phagocytic cells of F1 responder origin were available for cellular interactions in all mixed spleen cultures, yet they were competent for presentation of alloantigens, but not of self antigens borne by parental cells other than macrophages. In conclusion, the cellular requirements for stimulation differ quantitatively and qualitatively in F1 alpha P CML as compared to allogeneic and other CML systems. The distinct requirements presumably relate to the generation of specific effectors that are autoreactive as opposed to reactive with foreign antigens.
Binding reactivity of C-reactive protein for polycations. II. Modulatory effects of calcium and phosphocholine.
Authors: Potempa L A , Siegel J N , Gewurz H
Published: December 2022

C-reactive protein previously was shown to selectively and reversibly precipitate with certain small polymers of arginine and lysine. In the present report, calcium was shown to inhibit this reactivity in a dose-dependent manner, in direct contrast to the requirement for calcium for precipitation of CRP with C-polysaccharide. However, in the presence of phosphocholine, CRP rapidly precipitated and formed stable complexes with the polycationic polymers in the otherwise inhibitory calcium concentrations. alpha-Glycerol-phosphocholine, unlike phosphocholine, did not reverse this inhibitory effect. These results extend the characterization of the binding reactivity of CRP for polycations and suggest a relationship between this binding site and the sites for calcium and phosphocholine. We propose that CRP-polycation interactions in the presence of phosphocholine may have physiologic significance during the acute inflammatory process.
Monoclonal anti-IgM can separate T cell from B cell proliferative responses in the frog, Xenopus laevis.
Authors: Bleicher P A , Cohen N
Published: December 2022

The role of SIgM+ lymphocytes in frog mitogen responses and mixed lymphocyte reactions was examined. A murine monoclonal antibody with specificity for Xenopus laevis IgM was produced. The anti-IgM activity was not inhibited by glycoproteins other than Xenopus IgM, but could be inhibited with periodate-treated frog IgM. Fluorescent microsphere-coupled anti-IgM was used to show that adult and larval thymocytes had few sIgM+ lymphocytes, whereas spleens contained 19 to 34% sIgM+ lymphocytes. The spleens of larvally thymectomized adults were greatly enriched for sIgM+ cells. Lymphocyte suspensions were depleted of sIgM+ cells by incubation of spleen cells on plastic Petri dishes coated with anti-IgM monoclonal antibody. Compared with unseparated controls, the nonadherent cells cultured in serum-free medium were enriched for Con A and PHA mitogen responses and mixed lymphocyte culture (MLC) reactivity. Nonadherent cells were partially depleted of LPS mitogen responsiveness. Depletion or enrichment of the mitogen response was not a result of changes in kinetics or dose-response characteristics of the cells. In 1% FCS-supplemented cultures, the LPS response was not depleted, whereas the PHA response was still enriched. Thus, thymus-dependent and thymus-independent mitogen and MLC responses can be separated by the criterion of sIgM positivity in this anuran species.
Limitations of recognition of modified-self by alloantigen-activated cytotoxic T lymphocytes.
Authors: Ballas Z K
Published: December 2022

Alloantigen-activated CTL from CBA (H-2k), but not from C57BL/6 (H-2b), mice were able to lyse autologous cells modified by incubation with TNP-HSA. Moreover, allogeneic CTL of the H-2k haplotype lysed syngeneic cells modified with high or low concentrations of TNBS. Allogeneic CTL of the H-2b haplotype, however, did not lyse autologous cells modified with low concentrations of TNBS, even though they lysed syngeneic cells modified with high concentrations of TNBS. These limitations of recognition by the different H-2 haplotypes paralleled the restrictions evident for MHC-restricted, TNBS-specific CTL. It was thus apparent that alloantigen-activated CTL are governed by the same limitations of recognition operant for the MHC-restricted, TNBS-specific CTL.
Genetics of susceptibility to pristane-induced plasmacytomas in BALB/cAn: reduced susceptibility in BALB/cJ with a brief description of pristane-induced arthritis.
Authors: Potter M , Wax J S
Published: December 2022

BALB/cAn and BALB/cJ inbred strains of mice were separated 43 yr ago, but are still very closely related because they carry the same allelomorphs at many loci. When injected 3 times i.p. with 0.5 ml of pristane (2, 6, 10, 14 tetramethylpentadecane) 11% of strain BALB/cJ develop plasmacytomas in contrast to 61% in BALB/cAn sublines. BALB/cJ first injected with pristane as neonates also developed a reduced number of plasmacytomas as compared with BALB/cAn. About 20% of pristane-treated BALB/cAn and 70% of pristane-treated BALB/cJ develop arthritis, which first appeared in the ankle joints after a latent period of 4 mo or more.
Opsonization of pneumococci. II. Metabolic effects of a "third" human serum activity that mediates intracellular killing.
Authors: Guckian J C , Cavallo T , Fine D P
Published: December 2022

We investigated the mechanisms by which a serum activity, neither complement nor immunoglobulin, mediates killing of pneumococci by polymorphonuclear leukocytes (PMN). Electron microscopy revealed type 25 pneumococci to be within PMN when incubated in normal serum, in serum absorbed twice at 0 degrees C with type 25 pneumococci, or in absorbed plus heat-inactivated serum. Uptake of radiolabeled bacteria, and activation of oxygen consumption and of the hexose monophosphate shunt by PMN with pneumococci, were similar in normal serum, absorbed serum, or the combination of absorbed and heat-activated serum. Reduction of nitroblue tetrazolium (NBT) and of cytochrome c by PMN in the presence of type 25 pneumococci and absorbed serum, with or without heat-inactivated serum were one-third and one-half, respectively, of those in normal serum. Likewise, protein iodination was one-half that in normal serum. Reduction of cytochrome c by cytochalasin B-treated PMN was the same in normal, absorbed, or absorbed plus heat-inactivated serum. Furthermore, release of beta-glucuronidase from PMN after ingestion of pneumococci in 10% normal, absorbed, or absorbed plus heat-inactivated serum was identical. These data indicate that the "third" serum activity is not necessary for attachment of pneumococci to or ingestion by PMN, nor is it necessary for stimulation of the plasma membrane oxidase. Rather, it functions somehow in intracellular killing.
Spontaneous release of a factor with properties of T cell growth factor from a continuous line of primate tumor T cells.
Authors: Rabin H , Hopkins R F , Ruscetti F W , Neubauer R H , Brown R L , Kawakami T G
Published: December 2022

A continuous lymphoid cell line had been previously established from a gibbon with spontaneous lymphosarcoma. This cell line, designated as MLA144, when tested after several years in culture was shown to release spontaneously a factor biologically and biochemically similar to human T cell growth factor (TCGF). Conditioned media (CM) from MLA144 cells support growth and DNA synthesis of T cells from humans, several other species of primates, and also from mice and rabbits. The activity in the MLA144 CM is resistant to 60 degrees C and to low and high pH, has a m.w., as determined by gel filtration, of 21,500, elutes from DEAE-cellulose at 0.04 to 0.06 M sodium phosphate buffer, pH 7.6, and has an isoelectric point of about 6.45. Surface-marker analysis of MLA144 cells by rosetting techniques indicates that they are T cells lacking in the receptor for the Fc portion of IgG. The release of TCGF by MLA144 cells should have practical value in terms of ease of TCGF production and should be of great help in the facilitation of studies on the cell biology and molecular biology of TCGF production.
Studies on natural killer (NK) cells. II. Serologic evidence for heterogeneity of murine NK cells.
Authors: Burton R C , Bartlett S P , Kumar V , Winn H J
Published: December 2022

NK cells are divided into 2 subclasses on the basis of cell surface antigens: NKA cells, which preferentially lyse lymphoma cells and which are readily destroyed by treatment with anti-H-2, rabbit anti-mouse, or anti-NK specific antibodies in the presence of C; and NKB cells, which preferentially lyse nonlymphoma tumor cells and which are resistant to anti-NK specific antibodies, partially sensitive to anti-H-2 specific antibodies, and only fully sensitive to rabbit anti-mouse serum and C. Thus, suspensions of spleen cells that were treated with anti-NK-1.2 antibodies and C lost virtually all lytic activity for 6 lymphoma cell lines, whereas lytic activity for 2 nonlymphoma cell lines remained largely intact. It must be noted, however, that unlike NK-1.2 expression, target cell preference is not an absolute indicator of NK cell subclass. Cytotoxic T cells were readily distinguished from NKA and NKB cells on the basis of their sensitivity to monoclonal anti-Thy-1.2 antibodies and resistance to anti-NK-1.2 antiserum and C.
Biochemical characterization of Ia alloantigens in guinea pigs. II. Comparative peptide mapping of Ia antigens from B cells, T cells, and macrophages.
Authors: Lyons C R , Cook R G , Tucker T F , Uhr J W
Published: December 2022

Radioactive Ia.4 molecules were prepared from 3H- or 14C-labeled splenocytes, selected PEL, or bronchoalveolar macrophages (M phi). Studies in the accompanying paper indicated that incorporation into Ia.4 in these 3 populations is due to B cells, T cells, and macrophages, respectively. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was used to isolate the 58,000 m.w. form of Ia.4. The eluted molecule was then reduced, and the resultant alpha and beta subunits were isolated by a second separation on SDS-PAGE. Alpha (or beta) chains from 1 population labeled with 3H-amino acids was mixed with alpha (or beta) chains obtained from a population representing a different cell type that was labeled with 14C-amino acids and the mixture was digested with trypsin. Double-label (3H/14C) comparative peptide mapping was performed using high-pressure liquid chromatography to separate the peptides. Eighteen to 20 peaks of radioactivity were resolved from alpha chains, and 12 to 15 from beta chains. No reproducible differences were observed when comparing alpha or beta chains of T cells and macrophages, or those of B cells and macrophages. These results indicate that the primary structure of Ia.4 molecules is identical on the 3 cell types in question. The implications of having a T cell bearing the same Ia that it recognized on a macrophage in conjunction with antigen is discussed.
Resistance to immune cytolysis induced by 4NQO and MNNG: a possible mechanism involved in chemical carcinogenicity.
Authors: Okada H , Tanaka H , Cyong J C
Published: December 2022

L1210 cells were cultured for 20 hr in the presence of 4NQO or MNNG. Both agents rendered L1210 cells resistant to immune cytolysis by rabbit antibody and guinea pig complement. This resistance to immune cytolysis was not due to a loss or a distribution change of antigenic determinants to rabbit antibody, since the treated cells retained the capacity to consume guinea pig C4 in the presence of rabbit antibody to the same extent as untreated L1210 cells. Because acyltransferase activity to incorporate free fatty acids into phospholipids and neutral lipids of L1210 cells increased after the 4NQO and MNNG treatments, it was suggested that there might be a close relationship between the increased resistance of L1210 cells to immune cytolysis and the increase in their acyltransferase activity.
Monoclonal antibodies recognizing normal human T lymphocytes and malignant human B lymphocytes: a comparative study.
Authors: Martin P J , Hansen J A , Siadak A W , Nowinski R C
Published: December 2022

We have undertaken a direct comparison of 8 different murine monoclonal antibodies that recognize antigens shared by human T cells and certain malignant B cells. Immune precipitation, lysostripping, and competitive binding experiments indicate that the antigenic determinants detected by antibodies Leu 1, T101, 17F12, SC1, A50, OKT1, and 10.2 are closely associated on the same molecular species and may in fact be identical. The antigenic determinant recognized by antibody 12.1, which has properties similar to the 1 detected by other antibodies, is present on a distinct cell-surface molecule. Our results emphasize the importance of a quantitative direct comparison of different monoclonal antibodies in distinguishing newly reported antibodies from those already described. Such a comparison can resolve apparent discrepancies that arise either from methodologic variations or from differences in antibody avidity.
Stimulation of mouse lymphocytes by a mitogen derived from Mycoplasma arthritidis. I. Transformation is associated with an H-2-linked gene that maps to the I-E/I-C subregion.
Authors: Cole B C , Daynes R A , Ward J R
Published: December 2022

Cell-free supernatant derived from broth cultures of Mycoplasma arthritidis functions as an effective T cell mitogen for lymphocytes from H2k and H2d strain mice. Lymphocytes from animals of the H2b, H2q, and H2s haplotypes failed to respond to this mitogen. By employing lymphocytes derived from congenic and recombinant strains we established the H2 linkage and mapped the controlling gene to the I-E/I-C subregion. In addition, these studies suggested that a correlation existed between responsiveness to this microbial product and the presence of the Ia7 specificity, a product of the I-E subregion.
Hydrogen peroxide and cytolytic factor can interact synergistically in effecting cytolysis of neoplastic targets.
Authors: Adams D O , Johnson W J , Fiorito E , Nathan C F
Published: December 2022

Two secretory products from activated macrophages, which are thought to be involved in the lysis of tumor cells, are H2O2 and cytolytic factor (CF). To examine the potential of interaction between these 2 mediators of cytolysis, we first exposed MCA-I sarcoma targets to the 2 lytic agents. MCA-I targets, which have been previously shown to be lysed by CF, could also be lysed by H2O2 (the concentration of H2O2 causing 50% lysis of targets was approximately 1.4 x 10(-3) M). Exposure of the MCA-I targets to a nonlytic concentration of H2O2 for 1 hr potentiated the subsequent lytic effects of CF and did so synergistically. Exposure of targets to CF, however, did not potentiate the subsequent lytic effects of H2O2, even when the targets were treated with a lytic concentration of CF for 4 hr. Similar results were seen with P-388 lymphoma targets. The P388 cells were quite sensitive to the lytic effects of H2O2 (the concentration of H2O2 causing 50% lysis of these targets was approximately 2.0 x 10(-5) M) but proved relatively resistant to CF in comparison to the MCA-I targets. The data suggest that H2O2 and CF can interact synergistically to produce cytolysis of neoplastic targets if the targets are exposed to the 2 substances in a defined sequence.
Defective suppressor cell generation in juvenile onset diabetes.
Authors: Lederman M M , Ellner J J , Rodman H M
Published: December 2022

Lymphocytes of juvenile onset, insulin-dependent diabetics failed to generate suppressor cell activity after preincubation with concanavalin A (Con A). The mean suppression of the autologous proliferative response to phytohemagglutinin (PHA) was 6.1 +/- 7.8% (mean +/- SEM) in the diabetics and 34.0 +/- 4.9% in 9 age-matched healthy controls (p less than 0.01). Cell mixing experiments identified the defect to be in the generation of suppressor cells rather than in the responses to their action. Plasma of insulin-dependent diabetics had no effect on the generation of suppressor activity. In contrast to our findings in insulin-dependent diabetics, Con A preincubation of lymphocytes from 4 maturity onset diabetics induced normal suppression of PHA responses. Defective immunoregulation is present in insulin-dependent diabetes and may underlie autoimmunity.
T cell growth factor-enhanced PHA response of human thymus cells: requirement for T3+ cells.
Authors: Hayward A R , Kurnick J T , Clarke D R
Published: December 2022

Human thymocytes normally proliferate poorly in PHA-stimulated cultures, as measured by thymidine uptake. Their response increases when supernatant factors from mitogen-stimulated human lymphocyte cultures (SUP) are added. The cells that proliferate in PHA + SUP-stimulated cultures continue to divide if they are resuspended with fresh SUP every 3 to 4 days. PHA-stimulated thymocytes are deficient in the production of the T cell growth factor interleukin 2 (IL 2). The phenotype of thymocytes prior to stimulation was 90% T6+, 80% T4+, 83% T8+, and 12% T3+, whereas that of the cells after 3 days in culture with PHA + SUP was 1% T6+, 18% T4+, 66% T8+, and 80% T3+. Lysis of T3+ cells from the thymocyte preparation before culture greatly reduced the proliferative response to PHA + SUP. This suggests that the PHA-induced proliferation of more mature thymus cells is restricted by a lack of essential growth factors (either IL 1 or IL 2) and that the co-stimulating activity of SUP on human cells is predominantly on thymus cells that already are T3+.
Cyclophosphamide pretreatment induces eosinophilia to nonparasite antigens.
Authors: Vadas M A
Published: December 2022

Eosinophilia is usually regarded as a T cell-dependent hemopoietic response, although in most instances of immune responses by T cells there is no accompanying eosinophilia. BALB/c mice immunized with keyhole limpet hemocyanin (KLH) in complete Freund's adjuvant (CFA) were found not to develop eosinophilia, whereas pretreatment of such mice with 150 mg/kg cyclophosphamide (CY) induced marked eosinophilia in the blood (1000 to 3000/mm3) and bone marrow. CY as well as CY and KLH in CFA cause a leukocytosis and neutrophilia, but significant eosinophilia was seen only in mice receiving both CY and KLH-CFA. The eosinophilic response depended on the dose of CY and peaked 13 days after immunization, and was preceded by a bone marrow eosinophilia that peaked 8 days after immunization. No eosinophilia occurred in BALB/c nu/nu mice after CY and KLH-CFA but did occur in the nu/+ littermates. This model in mice of eosinophilia in response to CY followed by a nonparasite antigen should be useful in the analysis of T cell signals that might regulate eosinophilia.
Alloreactive T cells from individual soft agar colonies specific for guinea pig Ia antigens. II. Cellular and molecular requirements for optimal proliferative responses.
Authors: Malek T R , Shevach E M
Published: December 2022

We have investigated the cellular and molecular requirement for optimal proliferative responses of several alloreactive T cell lines that were derived from individual soft agar colonies and were specific for guinea pig Ia antigens. Optimal proliferation of several colonies was observed in cultures containing purified allogeneic macrophages and growth factor(s) present in supernatant fluids of Con A-activated T cells (Con A-S). Significant proliferative responses of these alloreactive T cell colonies were also routinely detected in cultures only supplemented with unfractionated irradiated allogeneic peritoneal exudate cell (PEC). The T cell component of the stimulator cell population was crucial for these responses by producing necessary growth factor(s) endogenously in the culture. Thus, 2 signals, allogeneic Ia antigens and growth factor(s), were required for optimal proliferative responses of these alloreactive T cell colonies. Furthermore, macrophage-associated Ia antigen was more efficient than B cell-associated Ia for these responses. The requirement for allogeneic Ia antigen was not absolute, since the colonies could easily be expanded when the cultures were supplemented with irradiated syngeneic PEC and the T cell mitogens, Con A or PHA. The effect of the mitogen was mediated via the T cells in the irradiated PEC, since removal of the T cells from these PEC markedly reduced the responses. Thus, it is likely that a nonspecific signal(s) presumably from T cells can promote proliferation of alloreactive T cell colonies in the absence of allogeneic Ia antigen. These results suggest 2 mechanisms of activation of these alloreactive T cells.
Induction of indigenous immunoglobulin-containing cells by gamma-irradiation and lymphoid cell transfer in mice.
Authors: Tseng J
Published: December 2022

Mice irradiated with gamma-ray develop large numbers of Ig-containing cells of various isotypes in all the lymphoid tissues examined except the thymus. The maximal dose of irradiation is approximately 500 R/mouse. These indigenous Ig-containing cells start to appear at day 3 post-irradiation, peak at day 6, and subside thereafter. The expression of the indigenous Ig-containing cells can be enhanced in the spleen of recipient mice by passive transfer of various lymphoid cells, except thymocytes. Lymphoid cells from histocompatible donors are more effective enhancers than those from nonhistocompatible donors. Cells involved in the enhancement are primarily T cells. In contrast, when T cells are eliminated from the lymphoid cell inocula, the indigenous Ig-containing cells are reduced to a level lower than that in the irradiated controls. This suppression appears to be mediated by macrophages, since recovery was found when macrophages were removed from these T cell-depleted inocula.
Interferon acts directly on human B lymphocytes to modulate immunoglobulin synthesis.
Authors: Härfast B , Huddlestone J R , Casali P , Merigan T C , Oldstone M B
Published: December 2022

At different times of exposure, interferon (IFN) enhanced and suppressed pokeweed mitogen- (PWM) induced IgG synthesis by human peripheral blood lymphocytes (PBL). Pretreatment of PBL and IFN frequently increased antibody production by more than 100% when compared with that by untreated PBL. Results of experiments in which PBL were separated into T and B subpopulations indicated that IFN preparations acted directly on B cells. Thus, mixtures of IFN-treated B cells and untreated T cells from 5 of 7 persons tested produced 81% to 500% more IgG than untreated, matched control cells. However, IFN-treated monocytes mixed with untreated B and T cells or IFN-treated T cells mixed with untreated B cells failed to enhance IgG production significantly in similar assays. In contrast to the pretreatment protocol, when IFN was present in the incubation mixture throughout the PWM assay, IgG production decreased. Sephadex chromatography of the IFN and tests of the resulting fractions indicated that the IgG production-enhancing activity was located in the fraction carrying the antiviral activity.
Cellular and antigenic requirements for production of mixed leukocyte reaction suppressor factor.
Authors: Dennison D K , Rich S S , Rich R R
Published: December 2022

When murine spleen cells, alloantigen-sensitized previously in vivo, are incubated with spleen cells bearing the sensitizing alloantigens, a supernatant factor is produced that inhibits 3H-thymidine incorporation by responding lymphocytes in the mixed leukocyte reaction. This study evaluates the cellular and antigenic requirements during restimulation for elaboration of this suppressor factor, MLR-TsF. BALB/c spleen cells, sensitized to C57BL/6 (B6) alloantigens in vivo, produced MLR-TsF when cultured with B6 spleen cells in vitro, despite depletion of Sephadex G-10-adherent cells from factor-producing cells, stimulator cells, or from both populations. T cells were not required within the stimulating population, but a requirement for viable stimulator cells was demonstrated when heat-killed or glutaraldehyde-fixed stimulator cells failed to induce MLR-TsF production. The alloantigenic requirements for MLR-TsF production were addressed by 2 approaches. Treatment of stimulator cells with appropriate anti-I region antisera and complement did not affect MLR-TsF production, demonstrating that an absolute requirement for cells expressing I region determinants did not exist. However, spleen cells primed against entire H-2 haplotype differences produced significant quantities of MLR-TsF when they were restimulated with spleen cells homologous to the priming cells in only the I region, in the K and D regions, or in the D region alone. The additive nature of subregion-specific restimulation suggests that distinct subpopulations of K, I, and D region-specific MLR-Ts comprise the MLR-Ts population primed to entire H-2 haplotype differences.
Influenza virus-specific cytotoxic T cells are restricted by multiple HLA-A3-related self antigens: evidence for recognition of distinct self structures in conjunction with different foreign antigens.
Authors: Biddison W E , Shearer G M , Shaw S
Published: December 2022

In the experiments described in this report we examined the similarities and differences in target cell recognition by influenza-immune CTL that share HLA-A3 but no other defined HLA-A, -B, or -C specificity with each of a panel of virus-infected target cells. Virus-immune CTL populations obtained from selected HLA-A3-positive donors can distinguish between the virus-infected target cells of unrelated donors, as assessed by direct cytotoxicity and cold target inhibition. The CTL restriction antigens that are associated with HLA-A3 in the population are encoded by gene(s) that segregate with HLA-A3 in an HLA-A/C recombinant family. These results suggest that HLA-A3-positive cells from unrelated individuals may differ with respect to 1 or more individual CTL restriction antigens associated with their HLA-A3 molecules, and that each of these distinct restriction antigens is recognized by a different subpopulation of virus-immune CTL. Analysis of the self specificity of T cells from individual donors sensitized against 2 noncross-reacting viruses, type A and type B influenza, indicated that CTL can recognize different foreign antigens in conjunction with different HLA-A3-related self antigens. These results are consistent with a model that proposes that each HLA-A and -B molecule possesses multiple CTL restriction antigens, each of which may function as self recognition structures for CTL that respond to different foreign antigens.
Regulation of influenza virus-specific cytotoxic T cell responses by monoclonal antibody to a human T cell differentiation antigen.
Authors: Biddison W E , Shearer G M , Chang T W
Published: December 2022

The results in this report indicate that the OKT3 monoclonal antibody, which is specific for a human T cell differentiation antigen present on 90 to 95% of peripheral T cells, can exert several effects that regulate the generation and expression of human influenza virus-immune cytotoxic T lymphocytes (CTL). The OKT3 antibody, but not OKT1 or OKT11 (which bind to all peripheral T cells), is able to inhibit anti-influenza CTL effector cell activity. An F(ab')2 preparation of OKT3 IgG were as effective as whole IgG for the inhibition of CTL effectors, indicating that the inhibitory activity of the antibody was not a function of the Fc portion of the molecule. OKT3 IgG and OKT3 F(ab')2 fragments (but not OKT4, OKT8, or OKI were able to inhibit the generation of anti-influenza CTL. The culture of human lymphoid cells with OKT3 in the presence or absence of influenza virus induced radioresistant cells that could suppress the CTL response of fresh autologous lymphocytes to influenza. These results suggest that T cell functions can be regulated by signals that are initiated by the binding of antibody to cell surface molecules that may not be related to the T cell antigen-specific receptor(s).
Nonspecific immunosuppressive factors in malignant ascites: further characterization and possible relationship to erythrocyte receptors of human peripheral T cells.
Authors: Oh S K , Moolten F L
Published: December 2022

Nonspecific suppressor factor has been purified from malignant human ascites through partial delipidation, ammonium sulfate precipitation, ACA-22 gel filtration followed by goat anti-human IgG immunoadsorbent chromatography. Molecular weight estimated by SDS-PAGE under mild reducing conditions is 50,000 and 25,000 under vigorous reducing conditions. It inhibits PHA, Con A, and PWM responses of peripheral lymphocytes by 50% at microgram concentrations per milliliter in vitro and inhibits plaque-forming cell response to SRBC at 100 microgram/mouse in vivo. It also inhibits 2-way mixed lymphocyte reactions and natural killer cell activities. It inhibits SRBC rosette formation of peripheral T cells and appears to be an erythrocyte receptor of peripheral T cells.
Human C5a des Arg increases vascular permeability.
Authors: José P J , Forrest M J , Williams T J
Published: December 2022

C5a were generated in human plasma by incubation with zymosan in the presence of a carboxypeptidase B inhibitor. The carboxypeptidase inhibitor was added to prevent cleavage of the carboxyl terminal arginine from C5a and enabled it to be purified on the basis of spasmogenic activity on the guinea-pig isolated ileum. When injected into rabbit skin, purified C5a induced marked plasma leakage over a 30-min period, but only if the substance was first mixed with a vasodilator substance such as prostaglandin (PG)E2. The responses to C5a + PGE2 did not appear to be related to anaphylatoxic, histamine-releasing activity because an antihistamine, mepyramine, had only a small effect on plasma leakage. Further, removal of the carboxyl terminal arginine by carboxypeptidase B abolished activity on the ileum but not in the skin. The observation that both human C5a and C5a des Arg were able to increase vascular permeability in vivo suggested a parallel with leukotactic activity in vitro. In support of this, no responses to C5a + PGE2 were obtained in rabbits depleted or circulating polymorphonuclear leukocytes. Thus, inflammatory edema resulting from extravascular complement activation may be dependent on 2 components: a leukocyte/endothelial cell interaction triggered by C5a, and the concomitant generation of a vasodilator, prostaglandin.
The transition from specific to nonspecific desensitization in human basophils.
Authors: MacGlashan D W , Lichtenstein L M
Published: December 2022

Human basophils can be desensitized to IgE-mediated stimuli either specifically (to the desensitizing antigen only) or nonspecifically (to all antigens). It has been suggested that the specificity of desensitization depends on the number of membrane-bound, antigen-specific IgE antibody molecules per basophil. We have varied the number of IgE antibody molecules/basophil by passive sensitization of mixed leukocyte preparations with increasing concentrations of purified IgE anti-penicillin (BPO) antibody. The cells were then desensitized with penicillin-human serum albumin (BPO-HSA). Desensitization was specific (lack of response to BPO-HSA only) with 1000 specific antibody molecules/basophil, and increasingly nonspecific (greater than 70% desensitization to rechallenge with anti-IgE and ragweed antigen E as well as lack of response to BPO-HSA) as the number of antibody molecules was increased to 14,000. This formally established that the number of specific IgE antibody molecules/basophil determines the mode of desensitization.
Translation of lymphocyte mRNA into biologically-active Interleukin 2 in oocytes.
Authors: Bleackley R C , Caplan B , Havele C , Ritzel R G , Mosmann T R , Farrar J J , Paetkau V
Published: December 2022

A variant line of murine T lymphoma EL4 produces high levels of the lymphokine Interleukin 2 (IL 2) when it is stimulated with phorbol myristate acetate. We have extracted poly A+ RNA from the stimulated cells and injected it into Xenopus laevis oocytes. The injected oocytes synthesize a material with biologic and biochemical properties of murine IL 2. Namely, it stimulates the continued growth of a cloned, cytotoxic T cell line (the T cell growth factor assay) and it chromatographs on a gel filtration column (G-100) with IL 2 produced by the stimulated EL4 line. The RNA responsible for the biologic activity sediments with markers of 11 to 12S in a sucrose gradient. The IL 2 produced by injected oocytes from a given preparation of mRNA is about 1% of the amount produced by the EL4 cells stimulated originally with phorbol myristate acetate. When RNA is extracted and purified from unstimulated EL4 cells it does not induce IL 2 production in oocytes. We conclude that IL 2 is essentially protein in nature, that the protein is coded for by poly A+ mRNA, and that the amount of this mRNA increases significantly after stimulation of the variant EL4 cells with the inducer phorbol myristate acetate.
Diseases caused by reactions of T lymphocytes to incompatible structures of the major histocompatibility complex. V. High titers of IgG autoantibodies to double-stranded DNA.
Authors: van Elven E H , van der Veen F M , Rolink A G , Issa P , Duin T M , Gleichmann E
Published: December 2022

IgG autoantibodies to double-stranded DNA (dsDNA), reaching a maximal serum titer of 1 in 2560, were spontaneously produced by (C57BL/10 x DBA/2)F1 mice injected with T lymphocytes from strain DBA/2. Anti-dsDNA antibodies were detected by the indirect immunofluorescence technique applying the kinetoplast dsDNA of Crithidea luciliae as antigenic substrate. The anti-dsDNA antibodies belonged to the IgG1, IgG2, IgM, and IgA (sub)classes. That of least some of them were produced by B cells of the F1 recipients was shown by the presence of F1-derived Ig-1b allotypic markers on such antibodies. In the F1 recipients, an incompatibility at H-2 was required for the formation of anti-dsDNA. These findings are consistent with the concept that the mechanism underlying the formation of SLE-like autoantibodies in this model is an abnormal cooperation between alloreactive donor T cells, presumably helper T (TH) cells, and H-2-incompatible F1 B cells. It is known from the literature that abnormal T-B cell cooperation is a unique tool for inducing vigorous primary antibody responses to antigens that carry repeating antigenic determinants on a rigid backbone and are poor immunogens or tolerogens in the absence of abnormal T-B cell cooperation. We suggest that the self-antigen dsDNA falls into the same category of structurally and immunologically peculiar antigens.
The syngeneic mixed leukocyte reaction represents polyclonal activation of antigen-specific T lymphocytes with receptors for self-Ia antigens.
Authors: Dos Reis G A , Shevach E M
Published: December 2022

We used a combination of positive and negative selection techniques to evaluate the relationship between the T cell that responds to self-Ia antigens in the SMLR and the T cells that respond to foreign antigen in association with self-Ia. BudR and light treatment of T cells proliferating in the SMLR abolished their subsequent primary and secondary proliferative response to antigen but had little effect on their response to allogeneic Ia. F1 cells that had been positively selected in the SMLR with stimulator cells from 1 parental strain demonstrated a markedly enhanced proliferative response to antigen in association with the parental strain used in the SMLR selection culture. The observed genetic restriction on the antigen responses of SMLR primed F1 T cells was not the result of haplotype-specific help or suppression and most likely represents direct activation of antigen-specific T cell clones by unmodified Ia antigens.
Cellular dissemination of priming for a mucosal immune response to cholera toxin in rats.
Authors: Pierce N F , Cray W C
Published: December 2022

Using CT as the test antigen, we sought 1) to learn whether primary immunization at 1 mucosal site caused priming of distant nonstimulated mucosae, 2) to study the role of migrating memory cells in the dissemination of mucosal priming, and 3) to compare disseminated priming with priming that occurs at the site of initial immunization. CT given i.c. or i.d. caused priming in tracheal and nonexposed enteric mucosae; i.t. immunization, however, did not cause detectable enteric priming. Adoptive transfer of immune TDLs showed that priming was conveyed by migrating memory cells. These appeared to be of 2 types: those that recirculated briefly before settling in MALT, and those that continued to recirculate until recruited by antigen to the site of mucosal challenge. Both types were required for secondary responses at mucosae distant from the site of priming. The time-course of disseminated mucosal priming resembled that of priming at the site of initial CT exposure, both lasting at least 16 wk. Disseminated priming persisted better in jejunal than tracheal mucosa, suggesting that the subgroup of memory cells that did not continue to recirculate settled preferentially in jejunal MALT. Disseminated priming supported smaller challenge responses than priming at the site of initial CT exposure did, suggesting that sessile memory cells also contributed to the latter process. These observations extend the concept of a "common mucosal immune system" to include cellular dissemination of mucosal priming, but also show quantitative differences between local and disseminated priming that probably reflect the patterns of distribution of migrating and sessile memory cells.
Mitogenic and co-mitogenic properties of hemin.
Authors: Stenzel K H , Rubin A L , Novogrodsky A
Published: December 2022

We report here that hemin, an agent known to induce cellular differentiation in Friend erythroleukemia cells, induces mitogenesis in human T cells and enhances mitogenic responses to supraoptimal concentrations of Con A. Mitogenesis induced by suboptimal or optimal concentrations of Con A is not affected by concentrations of hemin that markedly potentiate responses to supraoptimal amounts of Con A. Maximum mitogenic responses are reached 4 to 5 days after initiation of the cultures. The mitogenic response is macrophage dependent, whereas enhancement of responses to supraoptimal concentrations of Con A is not. Indeed, a greater degree of enhancement is observed after removal of adherent cells. Responses to other mitogens (phytohemagglutinin, wheat germ agglutinin, and sodium periodate) are also enhanced by hemin. Compounds that are metabolically or structurally related to hemin are neither mitogenic nor co-mitogenic. Protoporphyrin IX, which is nonmitogenic, is rendered mitogenic by the chemical insertion of iron to form hemin. Hemin does not affect Con A binding to lymphocytes. These findings indicate that hemin is a macrophage-dependent T cell mitogen with the unusual and remarkable property of overcoming the inhibition of mitogenesis induced by high concentrations of Con A. Since hemin has a limited number of metabolic effects, it provides a useful probe for determination of molecular events associated with lymphocyte activation.
Early effects of low-doses exposure to asbestos on local cellular immune responses in the lung.
Authors: Rola-Pleszczynski M , Massé S , Sirois P , Lemaire I , Bégin R
Published: December 2022

We recently developed an animal model to study the pulmonary reactions after exposure to environmental pollutants. In this model, sheep received monthly intratracheal instillations of a suspension of varying amounts of chrysotile asbestos in saline. All animals were studied by pulmonary function tests, transbronchial biopsies, and bronchoalveolar lavage. During the 6 mo of the study, no changes were seen in the 2 former tests, but bronchoalveolar lavage yielded leukocytes that responded with enhanced proliferative activity to concanavalin A, pokeweed mitogen, and staphylococcal lysate. The response to phytohemagglutinin was poor but also enhanced in the asbestos-exposed animals. Enhanced responsiveness was dose-related, except for the highest dose used, 128 mg, which was associated with a less enhanced proliferative response or none. Our data suggest that the initial response to low-dose exposure to asbestos consists in a stimulation of lymphocyte function; the possibility that this phenomenon may be linked to the eventual fibrogenic response of asbestosis warrants further studies.
Phenotypes of human natural killer cell populations detected with monoclonal antibodies.
Authors: Zarling J M , Clouse K A , Biddison W E , Kung P C
Published: December 2022

In our recent studies, human natural killer (NK) cell activity was found to be decreased 2- to 4-fold after treatment of monocyte-depleted peripheral mononuclear cells with monoclonal antibody OKM1 and complement (C). The present study was undertaken to determine whether there is an additional population of NK cells that is OKM1-, since treatment with OKM1 and C decreased, but did not eradicate, NK cell activity. Treatment of lymphocytes with monoclonal antibody OKT11A, which reacts with all sheep red blood cell rosetting lymphocytes, and C also decreased NK cell activity. Although approximately 90% of OKT11A+ cells are OKT3+, NK cell activity resides within the OKT11A+ cell population, which is OKT3- since OKT3-cell depletion fails to decrease NK cell activity. Double fluorescence analysis of OKT3-depleted lymphocytes revealed that 54% of the OKM1+ cells are OKT11A- and 45% of the OKT11A+ cells are OKM1-, thus demonstrating that within the OKT3-depleted population, approximately one-half the OKM1+ cells are OKT11A- and vice versa. Treatment of lymphocytes with OKM1 together with OKT11A and C decreased NK cell activity against 3 NK-sensitive leukemia lines--K562, MOLT-4, and HSB-2--more than did treatment with either antibody alone; virtually no lytic activity was retained after elimination of OKM1+ and OKT11A+ cells. The results thus provide strong evidence that there is at least 2 populations of human NK cells; one is OKM1+ and the other is OKT11A+
Specific anti-influenza virus antibody production in vitro by human peripheral blood mononuclear cells.
Authors: Yarchoan R , Murphy B R , Strober W , Schneider H S , Nelson D L
Published: December 2022

Cultures of human peripheral blood mononuclear cells stimulated in vitro with influenza virus were found to produce anti-influenza virus antibody. An enzyme-linked immunosorbent assay was used to measure cumulative antibody secretion into culture supernatants. Optimal antibody production was obtained with 2 x 10(6) peripheral blood mononuclear cells cultured in flat-bottomed wells for 12 days in 2 ml of RPMI 1640 medium supplemented with 10% fetal calf serum, antibiotics, and 4 mM L-glutamine. Antibody could be detected in cultures stimulated with live A/Hong Kong/68 (H3N2), purified formalin-inactivated A/Aichi/68 (H3N2), or live B/Hong Kong/68 influenza virus. Antibody of both IgG and IgM isotypes was produced. Under optimum conditions, the secreted IgG antibody was found to be directed preferentially at antigens on the stimulating virus. Antibody production was shown to require the cooperative interaction of B and T cells. Poisson analysis of B cell precursor frequency showed that approximately 1 in 40,000 B cells could be induced to produce IgG anti-influenza virus antibody in virus-stimulated cultures. Further investigation of this in vitro response should allow dissection of the cellular mechanisms involved in the regulation of antibody production in man.