bluuebunny/crossref_metadata_2025 huggingface dataset:
Immunologic tolerance after oral administration of reovirus: requirement for two viral gene products for tolerance induction.
Authors: Rubin D , Weiner H L , Fields B N , Greene M I
Published: December 2022

We have demonstrated that reovirus type 1, but not type 3, generates serotype-specific immunologic tolerance for DTH responses following oral administration of UV-inactivated virus. As shown by adoptive transfer experiments, the tolerance is secondary to the generation of viral-specific suppressor T cells that are present in spleen and mesenteric lymph nodes. No tolerance was generated when live virus was administered orally. Using recombinant viral clones, it was found that 2 viral gene products were required for tolerance induction; serotype specificity is a property of the viral hemagglutinin (the product of the S1 dsRNA segment). The inability of type 3 reovirus to induce suppression following oral administration is a property of the mu 1C polypeptide (the product of the M2 dsRNA genome segment).
Characterization of the recognition of target cells sensitive or resistant to cytolysis by activated rat peritoneal macrophages.
Authors: Hamilton T A , Fishman M
Published: December 2022

The interaction between tumoricidal rat peritoneal exudate macrophages and normal or neoplastic target cells has been examined both with respect to the sensitivity of the target cells to macrophage-mediated cytolysis and the ability of such targets to compete with one another for the lytic activity of macrophages. The results demonstrate that, in confirmation of earlier work from other laboratories, target cells are either sensitive or largely resistant to lysis, and such sensitivity correlates well with the neoplastic phenotype of the target cell. However, in target cell competition experiments all cultured cells as well as Con A-stimulated splenocytes and freshly isolated thymocytes exhibit the capacity to block cytolysis of sensitive indicator targets. In contrast, freshly prepared non-mitogen-stimulated splenic or lymph node lymphocytes are completely devoid of competitive activity. These results show that macrophages can recognize and interact with both normal and tumorigenic cells and that such target recognition can be distinguished from target cytolysis. Furthermore, the evidence suggests that at least 1 target cell feature that correlates with recognition is cell cycle traverse.
Antibody-independent interactions between Escherichia coli J5 and human complement components.
Authors: Betz S J , Isliker H
Published: December 2022

With the characterization of an increasing number of molecules that are capable of activating the 1st component of the classical pathway of complement (C), the possibility that some Gram-negative bacteria may activate C1 independent of naturally occurring antibody has been reexamined. We have confirmed a previous report that purified C1 (the activated form of C1) can bind to certain strains of bacteria and it retains its enzymatic activity when thus bound. The availability of purified C1 in its precursor form has allowed us to extend these observations to the native C1 molecule. Using a semirough mutant of Escherichia coli, the galactose epimerase-deficient strain E. coli J5, we have examined the binding and activation of radiolabeled C1. J5 bound radiolabeled C1 in a dose-dependent manner and essentially all of the bound C1 was activated as judged by SDS-PAGE. The bacteria-C1 complex consumed purified C4 and C2 and the consumption of C2 was proportional to the C4 concentration. Subsequent addition of terminal C components C3-9 supplied as serum-EDTA caused a highly significant decrease in bacterial viability. These results demonstrate that C1 may bind to the bacterial membrane in such a manner as to initiate a bactericidal reaction. Therefore, antibody-independent binding and activation of C1 must be considered in the assessment of serum sensitivity of Gram-negative bacteria.
Definitive evidence that natural killer (NK) cells inhibit experimental tumor metastases in vivo.
Authors: Hanna N , Burton R C
Published: December 2022

Effector lymphoid cells that inhibit hematogenous tumor metastasis in normal unimmunized mice were studied. Mice pretreated with cyclophosphamide (Cy) exhibited a diminished ability to destroy circulating tumor cells, which, in turn, caused an increased incidence of experimental pulmonary metastasis. Adoptive transfer of spleen cells from unimmunized syngeneic donors abrogated the Cy-induced enhancement of metastasis formation. The effector cell active in these adoptive transfer experiments was sensitive to treatment with specific anti-NK-1.2 antibodies and complement. This treatment was shown by in vitro tests to deplete or inactivate NK cells selectively. These findings provide direct evidence that NK cells play a significant role in the inhibition of hematogenous tumor metastases in vivo.
The role of H-2 in T cell recognition of Mls.
Authors: Gress R E , Wesley M N , Hodes R J
Published: December 2022

The role of H-2 was evaluated in T cell recognition of Mls-encoded antigens during primary mixed lymphocyte responses (MLR). Mlsc was used as a stimulating determinant in MLR and its recognition by T cells was assessed by linear regression analysis under culture conditions in which (A x B)F1 responder cell number was the factor limiting total response. Results of such experiments indicated the presence of distinct (A x B)F1 responder T cell subpopulations capable of differentially recognizing the foreign Mls antigen in association with one or the other parental H-2 haplotype. These findings demonstrate that T cells do not recognize Mlsc products in isolation, but rather are restricted to recognition of Mlsc in the context of "self" H-2 determinants.
The functional helper T cell repertoire specific for L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT).
Authors: Speck N A , Maurer P H , Pierce S K
Published: December 2022

Experiments have been carried out to examine the potential helper T cell repertoire specific for the random terpolymer GAT on responder, nonresponder, and (responder x nonresponder)F1 murine strains. The ability of GAT-MBSA immunized T cells to collaborate with DNP-specific primary and secondary B lymphocytes of each strain in response to the antigen DNP-GAT was tested with the splenic fragment culture system. The results of these experiments show that there are GAT-specific T lymphocytes in the responder, nonresponder, and F1 strains but that these 3 GAT-specific T cell populations differ in their collaborative potential. In sum, these findings present new evidence that the nonresponder status to the terpolymer GAT is due, in part, to a functional deletion of helper T cells capable of recognizing the antigen in the context of the nonresponder haplotype. Further, a new responsive phenotype is evidenced when F1 secondary B cells are stimulated in nonresponder GAT-MBSA-primed recipients. In this case, rather than the IgG1 responses observed in such strain combinations to other antigens such as DNP-Hy or DNP-Gl phi 9, only IgM responses were obtained. This new phenotype may be the result of GAT-specific suppression of isotype switching by B cells bearing the nonresponder cell surface alloantigens.
Natural cytotoxicity and interferon production in human cancer: deficient natural killer activity and normal interferon production in patients with advanced disease.
Authors: Kadish A S , Doyle A T , Steinhauer E H , Ghossein N A
Published: December 2022

Natural cytotoxicity was measured in 51 adult patients with solid epithelial malignant tumors and in 27 normal subjects. Peripheral blood leukocytes (PBL) from 31% of the patients and 7% of the controls failed to kill target cells (K562) in a short-term chromium-release assay. When patients were classified according to clinical stage, PBL from 12% of patients with localized cancers, but 50% of patients with advanced disease, failed to exhibit cytotoxicity within the normal range. Pretreatment of PBL with interferon alpha (IFN alpha) or with Newcastle Disease Virus (NDV), a potent inducer of IFN alpha, enhanced cytotoxicity from all normal subjects. Of patients whose PBL lacked spontaneous cytotoxicity, half were able to kill normally after pretreatment of PBL with IFN alpha or NDV. Virtually all the patients whose PBL were unable to kill despite pretreatment with IFN alpha or virus had disseminated malignancies. IFN alpha production by PBL exposed to NDV and to K562 cells was normal in all the patients regardless of stage of disease or ability to kill K562 cells. The observed defect in natural cytotoxicity is thus unlikely to be due to a failure of PBL to produce IFN alpha.
Impaired regulation of Epstein-Barr virus-induced lymphocyte proliferation in rheumatoid arthritis is due to a T cell defect.
Authors: Depper J M , Bluestein H G , Zvaifler N J
Published: December 2022

The rate of outgrowth of EBV-infected B lymphocytes is regulated by normal T lymphocytes. Removal of T cells from normal whole lymphoid populations (PBM) markedly shortens the outgrowth time of the remaining B lymphocytes. There is little difference in the much more rapid outgrowth of rheumatoid PBM after the removal of T cells, which suggests that RA lymphoid cells are unable to regulate this process. To determine whether RA T cells are defective, or EBV-infected RA B cells are unresponsive to regulatory signals, EBV-induced outgrowth in autologous and allogeneic mixtures of RA and normal B and T cells was evaluated, employing morphologic criteria and 3H-thymidine incorporation. The difference in outgrowth between RA and normal PBM was reproduced by reconstitution of EBV-infected B cells with mitomycin-treated autologous T cells. In cell-mixing experiments, normal T cells appropriately regulated both normal and RA B cells similarly, whereas RA T cells were defective in regulating either B cell population. Thus, the rapid outgrowth of EBV-infected rheumatoid lymphoid cells is due to defective T cell regulation. Moreover, normal regulation does not require cell proliferation.
Moculating effect of the late-acting components of the complement system on the bactericidal activity of human polymorphonuclear leukocytes on E. coli 0111:B4.
Authors: Tedesco F , Rottini G , Patriarca P
Published: December 2022

The effect of complement (C) components on the intracellular killing of E. coli 0111:B4 by human PMN was studied. Various intermediate bacteria were prepared by opsonizing IgM-coated 0111:B4 with yeast cell-treated human serum (BAC1-3), a C6-deficient human serum (BAC1-5), a C8-deficient human serum (BAC1-7), a C8-deficient human serum, and with partially purified C8 (BAC1-8). All these bacterial preparations were phagocytosed by human PMN, but only BAC1-8 and, to a lesser extent, BAC1-5 were killed. Similar results were obtained when the 400 x G postnuclear supernatant (PNS) of PMN homogenate was used instead of intact leukocytes. The C9 nature of the killing factor in the PMN homogenate was ruled out by its inability to lyse EAC1-8 and by the finding that the killing of BAC1-8 by the PMN factor was not inhibited by the antiserum against human C9. The anti-C5 and anti-C8 antisera were unable to inhibit the killing by the PNS of BAC1-5 and BAC1-8, respectively, suggesting that bound C5 and C8 do not provide a binding site for the killing factor.
Ly-18, a new alloantigen present on cytotoxic T cells and controlled by a gene(s) to the Igh-V locus.
Authors: Finnegan A , Owen F L
Published: December 2022

Antiserum raised in allotype-congenic mice recognizes a T cell differentiation antigen designated Ly-18. This antiserum was raised by immunization of (CB.20 x CBA-Igh-1b)F1 mice with secondary BALB/c anti-CBA allospecific cytotoxic cells. This antigenic specificity is expressed on cytotoxic T cells but not on cells proliferating in an MLR, plaque-forming cells specific for SRBC, or resting lymphoid cells. The expression of Ly-18 is controlled by gene(s) linked to the VH gene complex of Igh-1 allotype strains. There is no evidence at present for MHC gene control. Unlike other putative antibodies against antigen receptors, anti-Ly 18 does not block cytotoxicity. In addition, Ly-18 expression is not altered by the antigenic specificity of the Igh-1 effector. We conclude Ly-18 is a differentiation antigen present on a subpopulation of activated T cells. The gene(s) that codes for expression of Ly-18 is linked to the expression of Igh-1 genes.
Preparation and characterization of 4-azido-2-nitrophenyl human serum albumin as an antigen for covalent cross-linking of immune complexes.
Authors: Mannik M , David K A , Gauthier V J
Published: December 2022

Human serum albumin (HSA) was conjugated with 4-fluoro-3-nitrophenyl azide to yield varying density of the 4-azido-2-nitrophenyl haptenic group, useful for covalent cross-linking in the antibody-combining site. The epitope density of the antigen influenced several examined biologic properties. Precipitation in gel diffusion occurred when the average epitope density was 13 or above. Complement (C) activation was not found by incubation with guinea pig C, by binding to human Clq, or by conversion of the electrophoretic mobility of human C3 with epitope densities up to 13. Upon i.v. injection, rapid removal of the conjugated HSA occurred when more than seven 4-azido-2-nitrophenyl groups were present. This rapid removal was in part due to hepatic uptake. These studies point out the epitope density-dependent alterations of biologic properties of an antigen useful for preparation of immune complexes covalently cross-linked in the antibody-combining site.
Covalently cross-linked immune complexes prepared with multivalent cross-linking antigens.
Authors: Mannik M , David K A
Published: December 2022

Covalently cross-linked immune complexes were prepared with multivalent antigens, obtained by coupling varying numbers of 4-azido-4-nitrophenyl groups (NAP) on human serum albumin as the carrier molecule (NAPn . HSA). In this system the haptenic group served to bind the antigen to the antibody (antibodies to NAP) and to form covalent bonds upon photoactivation. The covalently cross-linked immune complexes contained around 30% of antibodies that were dissociable from complexes by SDS polyacrylamide gel electrophoresis. A comparable portion of antibody-combining sites were accessible to the free hapten (NAP . lysine) in molar excess by equilibrium dialysis. The stable, covalently cross-linked complexes with NAP7.0 . HSA and NAP12.9 . HSA were prepared and separated into complexes with varying degrees of lattice by sequential steps of gel filtration. Ag1Ab1 complexes were obtained with reasonable homogeneity. Other preparations contained successively higher lattices but were not homogeneous. When these complexes were injected into mice, the increasing lattice of complexes resulted in increasingly rapid removal of the complexes from the circulation. The antigen, independent of lattice, also contributed to removal of complexes from circulation. NAP12.9 . HSA alone was removed from circulation faster than NAP7.0 . HSA, and Ag1Ab1 complexes with NAP12.9 . HSA were removed faster than Ag1Ab1 complexes with NAP7.0 . HSA. The studied system adds covalently cross-linked immune complexes with multivalent antigens to the armamentarium of covalently cross-linked complexes that previously were obtained only with bivalent affinity labels.
Differentiation of T cells from immature precursors in murine T cell colonies.
Authors: Ching L M , Muraoka S , Miller R G
Published: December 2022

We describe a system for growing murine colonies containing subsets of functional T cells derived from immature T cell precursors. In contrast with previously described systems in which colony formation appears to depend upon activation and clonal expansion of mature T cells, we obtain comparable or increased numbers of colonies after depletion of T cells from spleen or bone marrow, or on culturing spleen or bone marrow from athymic nude mice. The proportion of cells expressing the T cell surface markers Thy-1, Lyt-1, and Lyt-2 is highly variable from colony to colony, consistent with the colonies arising from immature T cells, which differentiate in the colonies to give rise to various subsets of mature T cells.
Natural killer activity in the rat. II. Analysis of surface antigens on LGL by flow cytometry.
Authors: Reynolds C W , Sharrow S O , Ortaldo J R , Herberman R B
Published: December 2022

With the use of monoclonal antibodies and continuous flow microfluorometry, we have examined the cell surface antigenic characteristic of rat large granular lymphocytes (LGL) and compared them with monocytes, T cells, and polymorphonuclear leukocytes (PMN). The antigenic profile of rat LGL was of particular interest because these cells have been shown to be highly associated with natural killer (NK) activity. This analysis of enriched populations demonstrated that rat LGL are in antigenically distinct population of cells that share some characteristics with monocytes, T cells, and PMN. Essentially all of the cells in the enriched LGL population expressed with W3/13, OX-8, leukocyte-common (L-C), and asialo GM1 antigens. A portion of these cells were also positive with a monoclonal Ab against the ART-1a antigen. In contrast, few LGL expressed the Ia, W3/25, surface immunoglobulin (sIg), or Thy 1.1 antigens. Monocytes demonstrated a pattern of reactivity similar to LGL but were found to by OX-8 negative. As previously reported, a portion of the T cells were found to express ART-1a, W3/25, and/or OX-8. These results demonstrate that rat LGL are not typical T cells, B cells, monocytes, or PMN. However, the sharing of antigenic markers by LGL with some T cells and monocytes raises the possibility that LGL may be in the development lineage of 1 of these populations.
Inhibition of lectin-induced lymphocyte activation by 2-cyclohexene-1-one: decreased intracellular glutathione inhibits an early event in the activation sequence.
Authors: Fischman C M , Udey M C , Kurtz M , Wedner H J
Published: December 2022

The importance of normal intracellular glutathione (GSH) levels in the activation of human peripheral blood lymphocytes (PBL) by mitogenic lectins was explored using 2-cyclohexene-1-one (2-CHX-1), an agent that selectively decreases the levels of intracellular GSH. PBL incubated with mitogenic lectins and graded doses of 2-CHX-1 showed a dose-dependent inhibition of activation assayed by [3H]-thymidine uptake and percentage of blast transformation. 2.5 x 10(-5) M 2-CHX-1, a concentration that decreases GSH levels to less than 20% of control, caused an 80 to 90% suppression of both [3H]-thymidine uptake and blast transformation; lower concentrations were less effective. Time course studies showed that 2-CHX-1 was maximally effective only if added during the first 4 hr of culture. At the concentrations used, 2-CHX-1 was not cytotoxic to lectin-stimulated or unstimulated PBL and did not interfere with mitogen-lymphocyte interaction. These results suggest that adequate levels of glutathione are necessary for lymphocyte activation and that the glutathione requirement is exerted in the early activation sequence.
Antigens displayed on murine B lymphocyte precursors.
Authors: Kincade P W , Lee G , Watanabe T , Sun L , Scheid M P
Published: December 2022

The surface antigen phenotype of the immediate precursors of clonable B lymphocytes was investigated with conventional alloantisera and monoclonal antibodies directed by B lineage antigens. Ia was demonstrable on B cells, but not their immediate precursors in adult marrow. Adult, but not fetal, B cell precursors were susceptible to lysis with anti-Lyb-2 or anti-Qa. A panel of monoclonal rat antibodies was prepared and placed into categories on the basis of recognition patterns obtained with established cell lines. Of 2 groups that are described here, 1 (typified by antibodies from clone 14.8) detect an antigen that is preferentially expressed on B cells and their precursors, a proportion of antibody-secreting cells, and a subpopulation of peripheral T lymphocytes. Cells that did not display demonstrable amounts of antigen include brain, granulocytes, macrophages, mastocytoma cells, and erythroleukemia cells. A 2nd category of antibodies revealed an antigen that was more widely distributed on hemopoietic cells. Cells capable of quickly maturing into functional, colony-forming B lymphocytes in culture or after transfer to irradiated recipients specifically adhered to 14.8 antibody-coated, polystyrene petri dishes in the cold. Reductions in numbers of stem cells (CFU-s) and myeloid progenitors (CFU-c) by this treatment were minimal. Of particular importance was the fact that these antibodies recognized cells in embryonic liver as well as in adults that were destined to become B lymphocytes. These observations provide new perspective on B lineage precursor heterogeneity and suggest ways of localizing and dissecting some of the earliest events that are critical to development of the humoral immune system.
A monoclonal antibody specific for immature human hemopoietic cells and T lineage cells.
Authors: Bodger M P , Francis G E , Delia D , Granger S M , Janossy G
Published: December 2022

An anti-human monoclonal antibody (RFB-1) has been produced that reacts with a group of hemopoietic precursor cells in human bone marrow. These include terminal deoxynucleotidyl transferase-positive (TdT+) cells and myeloid colony-forming unit cells, functionally identifiable progenitor cells of the granulocytic-monocytic series. The expression of RFB-1 antigen on myeloid cells decreases as the cells become more mature; myeloblasts are weakly RFB-1+ but most promyelocytes are RFB-1-. RFB-1 also reacts with TdT+ thymic blast cells and cortical thymocytes but is unreactive with TdT- medullary thymocytes, although the majority of peripheral T cells are weakly RFB-1+. RFB-1 is the first anti-precursor cell reagent that labels human TdT+ cells in both the thymus and bone marrow as well as hemopoietic precursor cells but is unreactive with pre-B blasts and B lymphocytes.
Qualitative characteristics of histamine release from human basophils by covalently cross-linked IgE.
Authors: Kagey-Sobotka A , Dembo M , Goldstein B , Metzger H , Lichtenstein L M
Published: December 2022

We have studied the effects of permanent oligomers of human IgE produced using the cross-linking reagent, dimethyl suberimidate, on histamine release from human basophils. IgE dimers were found to be sufficient stimuli for both release and desensitization of these cells; monomeric IgE had no effect. Histamine release was augmented by deuterium oxide (D2O) in the medium, but D2O was not an absolute requirement to observe release. Desensitization by the dimeric IgE was specific in that the response to anti-IgE was not affected by preincubation of the leukocytes with the IgE dimer under suboptimal releasing conditions. IgE trimers and higher oligomers of IgE also caused both release and desensitization. IgE trimers were 3- to 4-fold more effective than IgE dimers with regard to the amount required for 50% histamine release. Dilution studies with monomeric IgE suggested that the difference was due to the presence of more "active" dimers in the trimeric IgE fractions. We conclude that dimeric IgE, by juxtaposing 2 receptors on the basophil membrane, is the "unit signal" for both release and desensitization of these cells.
T cell recognition in the mixed lymphocyte response. II. Ia-positive splenic adherent cells are required for non-I region-induced stimulation.
Authors: Ahmann G B , Nadler P I , Birnkrant A , Hodes R J
Published: December 2022

Ia-positive splenic adherent cells (SAC) have been shown to be the predominant stimulators of a mixed lymphocyte response (MLR) to whole H-2 differences, in which most of the proliferative response is directed against I region-encoded determinants. The present studies were undertaken to examine the ability of several purified lymphoid subpopulations to activate T cells in response to the non-H-2-linked MIs products or to products of the K or D regions of H-2. The results demonstrated that adherent cell-depleted populations of T and B cells were nonstimulatory, whereas SAC were potent stimulators for responses involving each of these genetic differences. Treatment of these SAC with anti-Ia and C abrogated their MLR-stimulating ability. In contrast, whereas treatment of SAC with anti-Ia and C abrogated their ability to stimulate an MLR directed against K or D region-encoded determinants, this treatment had no effect on their ability to generate a cytotoxic T lymphocyte response against these same determinants. These findings suggest that in addition to presenting allogeneic I region-encoded determinants, Ia-positive SAC also play a unique role in the presentation of non-I region-encoded alloantigens to proliferating T cells.
T lymphocyte immunity to reovirus: cellular requirements for generation and role in clearance of primary infections.
Authors: Letvin N L , Kauffman R S , Finberg R
Published: December 2022

The cellular requirements for the development of an immune response to reovirus type 1 and the role of such a response in the clearance of a primary infection with that virus were explored. An Ia-bearing antigen-presenting cell requirement is demonstrated for the in vitro generation of secondary anti-reovirus cytolytic T lymphocytes (CTL). It is then shown that mice whose spleens are depleted of Ia-bearing adherent cells by exposure in vivo to ultraviolet (UV) radiation exhibit depressed priming for reovirus-specific T lymphocyte function-CTL generation, delayed-type hypersensitivity reactivity, and T cell proliferative responsiveness. These UV-irradiated mice clear primary systemic reovirus infections as readily as normal mice. Further, athymic 'nude' mice show no defect in their ability to clear a reovirus infection. The implications of these findings for our understanding of the role of virus-specific T cell function in the clearance of systemic viral infections are discussed.
Experimental cutaneous leishmaniasis. I. Nonspecific immunodepression in BALB/c mice infected with Leishmania tropica.
Authors: Scott P A , Farrell J P
Published: December 2022

Leishmania tropica in BALB/c mice causes a fatal infection accompanied by the development of multiple metastatic lesions. Spleen cells from these mice were shown to have depressed proliferative responses to concanavalin A (Con A), phytohemagglutinin (PHA), and lipopolysaccharide (LPS). Coinciding with this immunodepression was the development of a cell population capable of suppressing normal spleen cell responses to Con A. This suppressor cell activity was first observed at 6 wk and was present throughout the remainder of the infection. At 12 wk the suppressor cells could be removed by Sephadex G-10 passage or carbonyl iron treatment; however, Sephadex G-10 passage could not reverse the suppression at 18 wk. Indomethacin, a prostaglandin synthetase inhibitor, was found to abrogate the activity of the adherent suppressor cell, suggesting that prostaglandin production may be involved in the immunosuppression seen in these mice. In addition, Sephadex G-10 passage and indomethacin were found to markedly augment spleen cell responses to leishmanial antigen, indicating that the adherent suppressor cell is capable of regulating specific immunologic responses.
Determination of surface antigens on highly purified human NK cells by flow cytometry with monoclonal antibodies.
Authors: Ortaldo J R , Sharrow S O , Timonen T , Herberman R B
Published: December 2022

We examined highly purified populations of NK cells, T cells, monocytes, and PMN with a variety of monoclonal reagents presumed specific for T cells and monocytes. The enriched NK cells showed a pattern of reactivity that was distinct from that of the other 3 populations, sharing antigens with T cells but also sharing antigens with monocytes and/or PMN. A portion of the enriched NK cells expressed OKT8, OKT10, and Lyt-3 antigens that were reported previously as T cell-associated antigens, but no reactivity was seen with other T cell-associated antigens (OKT3, OKT4, OKT6, and OKT11). In addition a portion of the enriched NK cells were positive for OKM1, asialo-GM1, and Ia antigens, which are undetectable on typical T cells but are present on PMN and/or monocytes. By depletion of antigen-positive cells on antibody monolayers, it was found that the activity of the enriched NK population was associated mainly with cells reactive with OKT10, OKM1, and Lyt-3. Although considerable evidence exists to support the T lineage of NK cells, the sharing of some antigens by NK cells and monocytes has suggested a possible relationship of NK cells to cells of the monocyte lineage. A possibility certainly exists that NK cells are not in either T cell or monocyte lineages and may represent a separate lineage of lymphoid cells.
Allogeneic mixed lymphocyte reactions in humans: pretreatment of either the stimulator or the responder cell population with monoclonal anti-Ia antibodies leads to an inhibition of cell proliferation.
Authors: Accolla R S , Moretta A , Cerottini J C
Published: December 2022

We analyzed the effect of 2 hybridoma monoclonal antibodies (Mab), D1-12 and D4-22, with specificity for common determinants of human Ia molecules, on the mixed lymphocyte culture (MLC) response. The results show that addition of either of the 2 Mab as late as day 3 after the onset of the culture completely inhibits the proliferative response generated in MLC. Because the antigenic determinants recognized by the 2 Mab that were used in this study have been shown to belong to distinct Ia molecules, it appears the inhibitory effect observed in MLC containing such Mabs cannot be explained simply by the masking of Ia molecules on the stimulator cell population. In agreement with previous studies by other investigators, treatment of a leukocyte population with the cytolytic D1-12 Mab plus complement strongly reduced its ability to stimulate in MLC. More importantly such a treatment also decreased the ability of a leukocyte population to respond in MLC. In the latter case, the inhibitory effect appears to be directed against T cells since highly purified E-rosetting cells treated with D1-12 plus complement were unable to respond in MLC. The possible implications of these results are discussed.
A monoclonal antibody discriminating between subsets of T and B cells.
Authors: Bruce J , Symington F W , McKearn T J , Sprent J
Published: December 2022

A description is given of a rat anti-mouse hybridoma antibody, JIId, which reacts with erythrocytes, neutrophils, greater than 90% of thymus cells, and most B cells. JIId does not have detectable activity for mature T cells, pluripotential stem cells, platelets, or cells of the monocyte-macrophage lineage. Although the JIId antigen is present on 90 to 95% of typical small B lymphocytes, pretreatment of spleen cells with JIId plus complement has no effect on secondary IgG antibody responses; by contrast, primary IgM responses and proliferative responses to lipopolysaccharide are substantially reduced. Unlike the precursors of IgG antibody-producing cells (AFC), IgG AFC per se are strongly JIId-positive. Rapid acquisition of the JIId antigen also applied to the early progeny of pluripotential stem cells.
Circadian fluctuations in the activity of phagocytic cells in blood, spleen, and peritoneal cavity of mice as measured by zymosan-induced chemiluminescence.
Authors: Knyszynski A , Fischer H
Published: December 2022

Circadian variations in the phagocytic activity of mouse whole blood, spleen, and peritoneal cells were studied using the zymosan-induced chemiluminescence assay as a measure of phagocytosis. On a regimen providing for light from 7:00 to 19:00 alternating with darkness, the phagocytic activity of mouse blood, spleen, and peritoneal cells was high around 10:00 and low around 22:00, the integrated counts of chemiluminescence being 82.33 x 10(5) and 52.76 x 10(5) for peritoneal cells, 83.3 x 10(5) and 32.2 x 10(5) for spleen cells, and 12.33 x 10(5) and 3.99 x 10(5) for blood cells. Variations of a similar tendency were also found in blood leukocyte and granulocyte counts, the counts being again higher at 10:00 compared with the blood samples withdrawn at 22:00. In contrast to the differences in the intensity of the zymosan-induced chemiluminescence, the shapes of the curves (Fig. 6) of each cell preparation were similar, irrespective of the time period of the day the cells were prepared. Comparison of the zymosan-induced chemiluminescence curves of the 3 cell suspensions studied, prepared at the same period of the day, revealed some similarity between the kinetics of blood and spleen samples; the intensity, however, of zymosan-induced chemiluminescence emitted by spleen cells was much higher. The kinetics of zymosan-induced chemiluminescence curves of peritoneal cells differed from the other 2, being slower at the onset, the chemiluminescence lasting for a longer time and declining more slowly. We have shown here circadian variations in the activity of mouse phagocytic cells. The simple and rapid method of chemiluminescence measurements used in this study appears to be a powerful tool for the further investigation of such circadian variations.
Binding of C-reactive protein to human lymphocytes. I. Requirement for a binding specificity.
Authors: James K , Hansen B , Gewurz H
Published: December 2022

Our laboratory previously reported that C-reactive protein (CRP) binds selectively to T lymphocytes and inhibits certain of their reactivities in vitro. However, these findings could not be repeated using more highly purified CRP preparations even under a variety of experimental conditions. Purified CRP alone did not bind to peripheral blood lymphocytes (PBL); however, in the presence of a ligand such as pneumococcal C-polysaccharide (CPS), CRP binding was readily detectable both by immunofluorescence and by a radioassay established for this purpose. The optimal concentration of CRP, ratio of CRP:CPS, and time and temperature for reactivity were determined using both assays. A markedly enhanced rate of binding was observed after pre-equilibration of CRP with calcium. A small percentage (mean 3.0%; range 0.5 to 8.0%) of PBL bound complexed CRP, and saturation was reached with 200 microgram CRP/ml. Reactivity of CRP with a multimeric form of phosphocholine (PC) (KLH-PC44) led to binding comparable to that observed with CPS, whereas monomeric PC inhibited the binding. Thus, in the presence of a multimeric binding specificity, CRP binds to a small fraction of peripheral blood lymphocytes, which are characterized in the accompanying paper.
Immunoregulatory function of T cells activated in the autologous mixed lymphocyte reaction.
Authors: James S P , Yenokida G G , Graeff A S , Elson C O , Strober W
Published: December 2022

This study was undertaken to define the functional properties of T cells stimulated in the autologous mixed lymphocyte reaction (MLR) by purified B cells or macrophages. In preliminary experiments, it was found that T cells that had been cultured with autologous non-T cells inhibited pokeweed mitogen- (PWM) stimulated immunoglobulin synthesis by autologous B cells. In addition, the T cell-mediated suppression was eliminated by x-irradiation and hydrocortisone treatment, was mediated by a mechanism that occurred early in the PWM-stimulated cultures, and did not involve killing of mature immunoglobulin-secreting cells. T cells were then cultured with either autologous B cells or macrophages in order to determine whether such autoreactive T cells had a similar capacity to regulate PWM-induced immunoglobulin synthesis. Although T cell populations stimulated either by B cells or by macrophages suppressed proliferative responses and immunoglobulin synthesis, both these populations of autoreactive T cells provided help for immunoglobulin synthesis that was not significantly different from that provided by fresh T cells. These results suggest that the predominant functional consequence of activation of T cells in the autologous MLR is the generation of suppressor T cells capable of inhibiting immunoglobulin synthesis. Thus, the autologous MLR may represent a negative feedback mechanism for the regulation of the immune response.
Immunogenic and antigenic epitopes of immunoglobulins. II. Antigenic differences between secreted and membrane IgG demonstrated using monoclonal antibodies.
Authors: Partridge L J , Lowe J , Hardie D L , Ling N R , Jefferis R
Published: December 2022

Twenty-three monoclonal antibodies with specificity for epitopes in the Fc fragment of IgG have been used to investigate antigenic differences between secreted and membrane forms of IgG produced by 2 human B lymphoblastoid cell lines (LCL). All of the monoclonals reacted with IgG secreted by the cell lines, as demonstrated by their ability to agglutinate SRBC coated with immunoglobulin isolated from culture supernatants. Membrane IgG expression was studied using direct and indirect rosette assays with antibody-coated ORBC. A surprisingly high number of antibodies, 13 on EB2 and 9 on EB4, did not bind to the cell surface immunoglobulin. These included antibodies with specificities for both C gamma 3 and C gamma 2 domain determinants. Similar results were obtained with an indirect radiobinding assay, indicating that negative results with the rosette test were not due to steric hindrance by the red cell carrier. Their performance in indirect hemagglutination indicated that most of the antibodies that did not bind to membrane IgG were of high avidity. It is concluded that the epitopes for which these antibodies are specific are not available on the cell surface. Possible explanations for the apparent antigenic differences between secreted and membrane forms of IgG are discussed against the background of previous work on the structure and mode of insertion of cell surface immunoglobulin.
The high background immune reactivity of mice to polymorphic determinants on xenogeneic erythrocytes: theoretical and practical implications.
Authors: Mosmann T R , Longenecker B M
Published: December 2022

Background responses have been assessed by fusing lipopolysaccharide- (LPS) stimulated spleen cells from unimmunized mice with MOPC 315.43 myeloma cells and screening the hybrids for the production of antibody against chicken red blood cells (CRBC). Clones specific for CRBC represented about 1% of total hybrid clones (1000 to 5000 clones were obtained per mouse). The majority of the anti-CRBC clones (greater than 95%) secreted antibody against polymorphic CRBC determinants (present on CRBC from some but not all chickens) rather than species-specific determinants present on all CRBC. Some of the polymorphic determinants were linked to the B locus (the MHC of the chicken) and some were non-B antigens. The relative amount of these 2 categories varied slightly according to the mouse strain. These results agree well with the specificities of natural mouse antibody and rosette-forming spleen cells. The response of immunized mice against CRBC and human RBC was also selective for polymorphic determinants. These results have considerable importance for the use of xenogeneic RBC as "standard" antigens, and are interpreted in terms of a model for the advantages of genetic polymorphism as a protection against antigen mimicry by parasites.
Human T cell subpopulations defined by a monoclonal antibody. I. A small subset is responsible for proliferation to allogeneic cells or to soluble antigens and for helper activity for B cell differentiation.
Authors: Corte G , Mingari M C , Moretta A , Damiani G , Moretta L , Bargellesi A
Published: December 2022

A monoclonal antibody (5/9) was obtained that reacts with the majority of alloactivated human T cells but only with 15 to 20% peripheral T cells. The populations reactive with the 5/9 antibody were separated from the remaining T cells by rosetting techniques using 5/9-coated ORBC. 5/9+ and 5/9- populations were analyzed for different in vitro activities. The helper activity of PWM- driven B cell differentiation appeared to be restricted and highly enriched in the 5/9+ population. In addition, 5/9+ cells contained all the cells capable of proliferation to tetanus toxoid and allogeneic cells. Thus, in vitro activities commonly used for evaluation of T cell function seem to be confined to a small fraction of peripheral T cells.
Regulation of cell-mediated immunity in cryptococcosis. I. Induction of specific afferent T suppressor cells by cryptococcal antigen.
Authors: Murphy J W , Moorhead J W
Published: December 2022

By injecting cryptococcal antigen i.v. into CBA/J mice, we have simulated a level of antigenemia that is known to occur naturally in murine and human cryptococcosis. The cryptococcal antigen induced a population of lymph node (LN) cells that, upon adoptive transfer, could specifically suppress the delayed-type hypersensitivity (DTH) response to cryptococcal antigen and the splenic T lymphocytes responsible for in vitro inhibition of growth of Cryptococcus neoformans. The cryptococcal-antigen-induced suppressor cell could be detected in the LN as early as 5 days and was still in significant numbers 7 days after antigen injection. Suppressor cells were not found in the spleens of tolerized mice, nor could serum from these animals adoptively transfer suppression. The suppressor cells in this model were shown to be T lymphocytes, which exerted their effect on the afferent limb of the DTH response; therefore, they were similar to the Ts1 cell in the ABA-DTH, NP-DTH, and NP cutaneous sensitivity models and to the afferent suppressor cells in the DNFB contact sensitivity system. The percent suppression of the cryptococcal DTH response was dependent on the amount of antigen injected. Approximately, 35 micrograms of cryptococcal antigen per ml of serum was sufficient to induce significant suppression of the cell-mediated immune response.
Effect of cyclosporin A on human lymphocyte responses in vitro. III. CsA inhibits the production of T lymphocyte growth factors in secondary mixed lymphocyte responses but does not inhibit the response of primed lymphocytes to TCGF.
Authors: Hess A D , Tutschka P J , Santos G W
Published: December 2022

The mechanism whereby Cyclosporin A (CsA) inhibits secondary mixed lymphocyte responses was assessed. CsA added to secondary MLR cultures inhibited proliferation and induction of cytolytic lymphocyte activity. This inhibition was found to be associated with the inhibition of T lymphocyte stimulating growth factor(s) (TCGF) production in the supernatants of secondary MLR cultures. As little as 1.0 micrograms/ml of CsA added to secondary MLR cultures resulted in no measurable TCGF activity. In contrast, moderate doses of CsA (1.0, 2.5 micrograms/ml), which completely inhibited the secondary MLR response to alloantigen, did not inhibit the proliferative and CML response of alloantigen-primed lymphocytes to these stimulating growth factors. Even at high doses of CsA (20 micrograms/ml), substantial levels of proliferation (50% of control response) and CML induction (60% of control response) were observed when the primed cells were exposed to secondary MLR supernatants containing TCGF activity. It was concluded that inhibition of secondary mixed lymphocyte responses by CsA may be due in part to the inhibition of TCGF production rather than the inhibition of the effect of TCGF on mature cytotoxic T lymphocytes.
Further biochemical characterization of the human thymocyte differentiation antigen T6.
Authors: van Agthoven A , Terhorst C
Published: December 2022

Charge heterogeneity of the human thymocyte antigen T6 was studied by isoelectric focusing and two-dimensional gel electrophoresis. The larger subunit of T6 (a 49,000 m.w. glycoprotein) contained several oligosaccharide side chains bearing up to 12 terminal sialic acids. When T6 antigens from 19 individual thymuses were analyzed, no differences in the isoelectric focusing patterns of the larger subunit could be detected. The larger subunit of the T6 antigen from MOLT-4 cells (52,000 m.w.) contained an extra oligosaccharide if compared with the T6 antigen from thymus or three other T leukemic cell lines. Two types of small subunits of T6 were found. In addition to a protein of m.w. 12,000, pI 5.5 identified as beta 2-microglobulin, a (m.w. 12,000, pI 7.0) nonglycosylated protein, was detected on two dimensional gels. This protein does not cross-react with beta 2-microglobulin, and its amount varied in different T6 preparations. The tissue distribution, the m.w. the association with beta 2-microglobulin, and the limited structural heterogeneity of T6 support the idea that T6 is the human homologue of the murine thymus leukemia antigen (TL).
The H-2 haplotype of a thymus graft influences the Ir gene regulated IgG3, IgG1, IgG2b, and IgG2a anti-(T,G)-A--L antibody responses of high-responder F1.nude mice.
Authors: Press J L , Haber J , Wortis H H
Published: December 2022

In order to examine whether the H-2 haplotype of a thymus graft influences the levels of IgG3, IgG1, IgG2b, and IgG2a antibodies produced in an in vivo response to antigens under immune response (Ir) gene control, genotypic high-responder (H-2b X H-2k)F1.nude mice were grafted with thymuses from irradiated, neonatal low-responder (H-2k) or high-responder (H-2b or (H-2k X H-2b)F1) mice and immunized with (T,G)-A--L. All IgG antibody responses to (T,G)-A--L in high-responder mice were shown to be thymus dependent. The majority of F1.nude mice grafted with thymuses from high-responder haplotype donors produced high-responder levels of IgG anti-(T,G)-A--L antibodies. Conversely, the majority of F1.nude mice grafted with thymuses from low-responder haplotype donors gave low-responder phenotypic patterns. The modulation of the Ir gene phenotype was not restricted to a particular IgG isotype, but affected IgG3, IgG1, IgG2b, and IgG2a. The F1 . nude mice grafted with low-responder haplotype thymuses were able to produce IgG1, IgG3, and IgG2b antibodies to sheep erythrocytes, a thymus-dependent (TD) antigen not under overt Ir gene control. Circulating peripheral T cells were shown to be of host origin. By these criteria, the thymus grafts did enable F1.nude mice to respond to a TD antigen. These results support the concept that thymic H-2 determinants are involved in at least the selection of H-2 restricted T cell subsets, if not also the derivation of the T cell receptor repertoire for self and/or antigen recognition.
Biologic activity of an idiotype-bearing suppressor T cell factor produced by a long-term T cell hybridoma.
Authors: Takaoki M , Sy M S , Whitaker B , Nepom J , Finberg R , Germain R N , Nisonoff A , Benacerraf B , Greene M I
Published: December 2022

Biologic activities and immunochemical characteristics of an azobenzenearsonate- (ABA)specific suppressor T cell factor produced by a longterm T cell hybridoma, F12, were studied. In vivo administration of F12 culture supernatant resulted in the suppression of ABA-specific delayed-type hypersensitivity (DTH) responses and the inhibition of priming for ABA-specific cytotoxic T lymphocyte responses. Moreover, F12 induced a second set of suppressor cells that act in the efferent phase of DTH. The active material in the F12 culture supernatant expressed major cross-reactive idiotypic (CRI) determinants of anti-ABA antibodies of A/J mice and I-J subregion-coded specificities but not express determinants of immunoglobulin constant regions. These results demonstrated that F12 is a functioning hybrid cell line of the first-order suppressor T cell subset.
Expression of T cell differentiation antigens and Ia on rat cytotoxic T lymphocytes.
Authors: Duarte A J , Carpenter C B , Strom T B
Published: December 2022

Study of lymphocyte subsets has been facilitated by the detection of various surface markers that are related to distinct functions. We have explored the use of affinity methods for separation of large quantities (10(8)) of rat cells and used this technique to identify surface markers of cytotoxic T lymphocytes. Splenocytes from animals bearing skin allografts for 7 days were depleted of B cells and were separated into T cell subsets by using W3/25 and OX8 monoclonal antibodies, which mark two nonoverlapping populations of peripheral T cells. Our results demonstrate that the cytotoxic T cells are W3/25- and OX8+. They also bear minimal or no Ag-F alloantigen, a peripheral T cell marker, and they express Ia-like products analogous to those of the mouse I-A subregion but not of the I-E/C subregion.