bluuebunny/crossref_metadata_2025 huggingface dataset:
mu Polypeptide phenotypes of lymphoid and myeloid cell lines containing RNA transcripts of the immunoglobulin C mu gene.
Authors: Walker I D , Harris A W
Published: December 2022

We have examined whether polyadenylated-RNA molecules encoded by the immunoglobulin C mu gene (C mu RNA) that have recently been shown to occur in mouse T lymphoma, myeloid tumor, and pre-B lymphoma cell lines are translated. Three T lymphoma, 1 myeloid tumor, and 4 Abelson lymphoma cell lines, all known to contain C mu RNA molecules, were examined for evidence of synthesis of mu polypeptides using radioimmunoassay, biosynthetic peptides were detected in any of the T lymphoma or myeloid cell lines, although mu-chains were readily detected in 2 B lymphoma lines known to contain small amounts of mu mRNA. Detection failure is unlikely to be the result of assay insensitivity and was not caused by lack of an appropriate antiserum reactivity or a requirement for mu-chains to be associated with immunoglobulin light chains. Proteolytic activity that might degrade newly synthesized mu polypeptides in these cell lines was not detected. In 3 of the 4 Abelson lymphoma cell lines containing C mu RNA, corresponding polypeptides were absent, although in the 4th they were easily detected, indicating that transcription of the C mu gene is not the sole determinant of whether a cell will synthesize the corresponding polypeptide. The parallel between the C mu RNA+, mu polypeptide- Abelson cell lines and the T lymphoma cell lines suggests that the onset of C mu transcription may occur in both T and B lineages before translational competence is acquired.
In vitro growth of murine T cells. VI. Accelerated skin graft rejection caused by adoptively transferred cells expanded in T cell growth factor.
Authors: Rosenstein M , Eberlein T , Kemeny M M , Sugarbaker P H , Rosenberg S A
Published: December 2022

The ability of adoptively transferred cytotoxic T cells expanded in T cell growth factor (TCGF) to induce accelerated graft rejection of B10.BR skin grafts from normal B6AF1 recipient mice was tested. In vitro sensitized cells expanded in regular TCGF were capable of accelerating graft rejection when adoptively transferred i.v. 1 day after grafting. Cells expanded in LF-TCGF (lectin-free) were more effective in accelerating graft rejection than cells expanded in lectin containing TCGF. Nonsensitized cells or cells sensitized to irrelevant B10.D2 antigenic determinants in vitro and subsequently expanded in LF-TCGF were unable to induce accelerated rejection of the B10.BR grafts. Expanded cytotoxic effector cells known to be immunologically active in vivo were also unable to accelerate graft rejection if irradiated with 2000 R before adoptive transfer. The ability of cells to lyse the appropriate targets in 4-hr 51Cr release assays correlated poorly with the effectiveness of these cells after in vivo injection. These studies are the first to demonstrate that cytotoxic cells expanded in TCGF are capable of mediating a cytotoxic function in vivo.
Discordant expression of 2 Epstein-Barr virus-associated antigens, EBNA and RANA, in man-rodent somatic cell hybrids.
Authors: Slovin S F , Glassy M C , Dambaugh T , Catalano M A , Curry R A , Ferrone S , Kieff E , Vaughan J H , Carson D A
Published: December 2022

The Epstein Barr nuclear antigen (EBNA) and the rheumatoid arthritis nuclear antigen (RANA) develop in human B lymphocytes that have been infected and transformed by Epstein Barr virus (EBV). Antibodies to RANA and EBNA are found only in individuals with prior exposure to EBV. The purpose of the present studies was to determine the relation of the 2 antigens to each other and to EBV genetic material, in human-rodent somatic cell hybrids. Cultured human B lymphoblastoid cells, Raji, Daudi, and RPMI 4098 were fused with thymidine kinase-deficient mouse or hamster fibroblasts. After selection and cloning in ouabain-HAT medium, the hybrid nature of the surviving cells was confirmed by isozyme analysis. The hybrid clones were analyzed for EBNA by anti-complement im,munofluorescence, and for RANA by anti-immunoglobulin immunofluorescence and immunodiffusion. The results showed that RANA and EBNA segregated entirely independently of each other in the hybrid clones. Two methods were used to detect the presence of EBV DNA sequences in the intracellular DNA of hybrid clones. The 1st method relied on the hybridization of labeled cRNA prepared from virion DNA with DNA from 8 hybrid clones affixed to nitrocellulose filters. The 2nd approach was to hybridize labeled intracellular DNA from 3 hybrid clones to Southern blots of cloned fragments of EBV DNA. These results suggested that the presence of EBV DNA was not sufficient for the expression of either antigen. One stable RANA-positive hybrid cell line contained at least 80% of the EBV genome in the absence of detectable EBNA.
A murine stage-specific embryonic antigen (SSEA-2) is expressed on some murine SV40-transformed cells.
Authors: Shevinsky L H , Knowles B B , Howe C , Aden D P , Solter D
Published: December 2022

A stage-specific embryonic antigen-2 (SSEA-2) found on murine preimplantation embryos is maximally expressed on 4- to 8-cell stage embryos and is present in decreasing amounts of morulae and blastocysts. This antigenic determinant is also expressed on murine teratocarcinoma cells, sperm, and some, but not all, SV40-transformed mouse cell lines. Analysis of solubilized immunoprecipitates by SDS-gel electrophoresis indicates that this cell surface molecule is not the 54,000 m.w. protein shared by teratocarcinoma and SV40-transformed cell lines.
In vitro antibody response to influenza virus. II. Specificity of helper T cell recognizing hemagglutinin.
Authors: Anders E M , Katz J M , Jackson D C , White D O
Published: December 2022

Intraperitoneal immunization of mice with liver influenza virus was shown to induce helper T (TH) cells with specificity for the hemagglutinin (HA). The interaction of virus-primed TH cells with purified HA was studied independently of B cell reactivity to the same antigen by using the generation of nonspecific help as an index of activation of HA-specific TH cells. TH cells from mice primed with any of the H3 viruses A/Aichi/68 X A/Bel/42 (H3N1), A/Memphis/102/72 X A/Bel/42 (H3N1) or A/Port Chalmers/73 (H3N2) were strongly cross-reactive towards HA of other strains within the H3 subtype. In addition, several examples of cross-reactivity towards HA of a different subtype were observed, usually of a lower magnitude. TH cells from mice primed to any of the H3 viruses above or to A/Bel/42 (H1N1) virus cross-reacted with the HA of A/Japan/305/57 (H2); furthermore, priming with A/Bel/42 or with A/Jap/305/57 X A/Bel/42 (h2N1) virus yielded TH cells that cross-reacted with certain of the H3 HA preparations. The cross-reactivity observed between subtypes was not due to the common chicken host carbohydrate component of HA, since no response to the purified type A HA preparations was obtained with T cells from mice primed with egg-grown influenza B/Hong-Kong/8/73 virus. The results indicate that HA of different subtypes may share cross-reactive antigenic determinants recognized by TH cells. Within a subtype, HA are highly cross-reactive with respect to tH cell recognition.
Analysis of the cytotoxic T cell response to H-Y in CBA/H mice.
Authors: Brenan M , Müllbacher A
Published: December 2022

The dose response, kinetics, and target cell specificity of the H-Y immune Tc-cell response of fp-primed CBA/H mice was analyzed. Non-MHC genes were shown to influence responsiveness in fp-primed H-2k strains of mice. The possibility that dominant T-suppressor cells mask responsiveness in i.p. primed CBA/H mice was not confirmed in mixing experiments. Female spleen-adherent cells injected with male spleen cells i.p. elicited H-Y immune Tc-cell responses.
Helper cells in the autologous mixed lymphocyte reaction. I. Generation of cytotoxic T cells recognizing modified self H-2.
Authors: Wolos J A , Smith J B
Published: December 2022

The autologous mixed lymphocyte reaction (AMLR) in mice measures the proliferative response of T cells to determinants on syngeneic non-T spleen cells. Normally, cytotoxic T lymphocytes (CTL) are not generated in this reaction. However, the addition of trinitrophenyl-modified mitomycin C-treated syngeneic T cells (TNP-Tm) to the AMLR results in the generation of TNP-specific CTL but does not alter the proliferative response. Significant cytotoxic activity is not detectable against TNP in association with Ia unless TNP is present on cells bearing those determinants. Thus, if unselected spleen cells are TNP-modified and used as stimulators in the AMLR, the proliferative response is enhanced and CTL are generated that recognize TNP in association with K, D, and I region-encoded determinants. The CTL generated in the AMLR are H-2 restricted and dependent on the presence of adherent cells in the sensitization cultures. The experiments presented here suggest that the AMLR can provide the help necessary for generating cytotoxic T cells in vitro.
Hapten-specific T cell response to 4-hydroxy-3-nitrophenyl acetyl. X. Characterization of distinct T cell subsets mediating cutaneous sensitivity responses.
Authors: Sunday M E , Dorf M E
Published: December 2022

NP-O-Succinimide-induced cutaneous sensitivity (CS) responses can be adoptively transferred by NP-primed lymphoid cells into naive K-, I-, or D-compatible recipients. The distinct fine specificities of I- versus D-restricted T cell clones from various strains of mice suggested that presence of different idiotypic receptors on these T cell subsets. We now demonstrate that NP-O-Su immune lymphoid cells are composed of 2 T cell subsets with distinct antigen recognition patterns, as well as different Lyt 2 phenotypes. Thus, I-restricted cells respond to NP-coupled to a protein carrier but not to NP-coupled cells, whereas D-restricted clones react to NP-cells, but not to NP-protein conjugates.
T lymphocyte-mediated cytolysis. I. A common mechanism for target recognition in specific and lectin-dependent cytolysis.
Authors: Berke G , Hu V , McVey E , Clark W R
Published: December 2022

In this and the accompanying paper we examine the nature of the interactions between effector cells and target cells leading to lysis in T cell-mediated cytolysis reactions. In the first paper, we re-examine the role of lectin (Con A) in the process of lectin-dependent cell-mediated cytotoxicity (LDCC). Lectin has generally been thought to act simply as a bridge between the cytotoxic effector cell (EC) and the target cell (TC), thus bypassing the need for receptor-antigen interaction and accounting for the nonspecific character of this lytic reaction. A role for lectin in the activation of cytotoxic function in the EC has also been suggested. In this paper we confirm that LDCC occurs when TC alone are pretreated with lectin, but not when EC alone are pretreated with lectin, unless free lectin is also present in the assay mixture. In addition, we demonstrate that the failure of lectin-pretreated EC to bind to and lyse TC is not caused by agglutination or self-destruction of the EC. Moreover, lectin-pretreated EC are not inherently deficient in the ability to bind or lyse TC, if the latter are pretreated with lectin, or if free lectin is present in the assay. Finally, when EC are used as both effectors and targets in LDCC, lysis is observed only in the direction of the lectin-pretreated partner. From these results we conclude that in LDCC, as in specific CTL cytolysis, CTL-TC recognition proceeds through interaction of an EC surface receptor (distinct from the lectin receptor), and a TC antigen perhaps modified by, but distinct from, the lectin itself. In the accompanying paper we present evidence that this receptor-determinant interaction involves TC MHC antigen.
T lymphocyte-mediated cytolysis. II. Role of target cell histocompatibility antigens in recognition and lysis.
Authors: Berke G , McVey E , Hu V , Clark W R
Published: December 2022

Although compelling genetic and serologic evidence implicate target cell (TC) MHC antigens in specific cytotoxic T lymphocyte (CTL)-TC interaction leading to lysis, it is not entirely clear whether TC recognition through an MHC determinant(s) is a prerequisite for lysis to occur. In fact the finding that both specific and nonspecific TC are lysed equally well in lectin-dependent cell-mediated cytotoxicity (LDCC) challenges the necessity for TC MHC involvement in the cytolytic process beyond providing the basis for specificity in direct (nonlectin-dependent) CTL-mediated lysis. In the present paper we present evidence suggesting that even in nonspecific LDCC, as well as in nonspecific lymphocyte-mediated lysis of TC oxidized by periodate treatment or by galactose oxidase (ODCC), TC MHC components are required for lytic interactions with cytotoxic effector cells. This conclusion is based on 3 types of experimental evidence: 1) cells displaying reduced amounts of MHC proteins are poor targets in LDCC; 2) removal of H-2 by papain renders murine target cells refractory to lysis in LDCC, even though Con A binding is only slightly reduced; 3) antisera to target cell H-2-coded products block lysis in both LDCC and ODCC, whereas antisera to other cell surface antigens do not. A theory explaining nonspecific effector target interaction leading to lysis based on involvement of CTL receptor(s) and TC MHC components is presented.
Long-term in vitro culture of murine mast cells. II. Purification of a mast cell growth factor and its dissociation from TCGF.
Authors: Yung Y P , Eger R , Tertian G , Moore M A
Published: December 2022

Procedures are described for the isolation of a mast cell growth factor (MCGF) from medium conditioned by mitogen-activated splenic leukocytes (CM). Although optimal conditions for the production of MCGF in CM are identical to those for the production of T cell growth factor (TCGF), MCGF can be dissociated from TCGF after the first stage of purification on a DEAE-cellulose column. MCGF elutes from the column in the breakthrough fraction, whereas TCGF binds avidly to DEAE and is eluted only at high salt concentration. MCGF also differs from TCGF with respect to m.w. (as estimated by Sephadex G-150 chromatography) and sensitivity to trypsin. In addition, MCGF is produced by the murine myelomonocytic leukemia WEHI-3 and the radiation induced thymic lymphoma LBRM-33 cells, whereas TCGF is produced only by the latter in the presence of a mitogen. Another hemopoietically active factor, granulocyte colony-stimulating factor (G-CSF) present in media conditioned by WEHI-3 and LBRM-33 cells, however, shares a number of properties with MCGF. Although studies with purified or partially purified MCGF have thus far failed to reveal a correlation between MCGF and G-CSF, further biochemical analyses are necessary to dissociate MCGF from G-CSF.
Immunotherapy of murine leukemias by monoclonal antibody. I. Effect of passively administered antibody on growth of transplanted tumor cells.
Authors: Kirch M E , Hammerling U
Published: December 2022

The objective of the present study was to establish a model system for the evaluation of passive immunotherapy of murine leukemias. Monoclonal antibodies directed at T lymphocyte differentiation antigens (Thy 1 and Lyt 2) were tested for their effect on tumors that were grown in hosts congenic for the target antigen. Tumor challenges were selected that were at least 500 times the dose that was lethal in 50% of untreated controls. The A strain leukemia, ASL.1, was transplanted subcutaneously into a/Thy 1.1 congenic hosts. Intraperitoneal administration of anti-Thy 1.2 monoclonal antibodies of the IgG3 and IgM classes reduced tumor growth. Up to 90% of the mice receiving antibody of the IgG3 subclass failed to develop tumors, whereas IgM antibodies prolonged survival time, but the mice eventually died of tumors. Antibody was most effective if administered within 24 hr of tumor inoculation; delay of antibody injection for 48 hr prolonged host survival but did not eradicate cells at the injection site or prevent metastases. The C57BL/6-derived tumors, ERLD and EL4, were evaluated for susceptibility to treatment with antibody directed at the Lyt 2.2 alloantigen using the protocol that was effective in treating aSL.1. Monoclonal antibody of the IgG2a subclass was effective in the case of C57BL/6/Lyt 2.1 congenic mice bearing ERLD, but caused a decrease in survival time of mice bearing the transplanted EL4 tumor. Thus, antibody of the appropriate immunoglobulin subclass can be effective in controlling tumor growth if administered in the optimal treatment regimen, but inherent features of the tumor cell ultimately determine whether abrogation or enhancement of growth will occur.
Human T lymphocyte/monocyte interaction in response to lectin: kinetics of entry into the S-phase.
Authors: Maizel A L , Mehta S R , Hauft S , Franzini D , Lachman L B , Ford R J
Published: December 2022

PHA-mediated mitogenesis of human peripheral blood T lymphocytes was studied by using highly purified cell populations. The kinetics of human, mature T cell [3H]-Tdr incorporation were examined with respect to those elements necessary and sufficient for the progression of the activated T cell into the S-phase of the cell cycle. These experiments indicated that although a lectin may independently initiate morphologic T cell blastogenesis, this event is not associated with significant progression through the G1 phase of the cell cycle. This blastogenic response is associated with the subsequent T cell receptivity to monocyte-initiated cell cycle progression, and the effect of monocytes can be substituted by partially purified Interleukin 1 (IL-1). Progression of a lectin exposed T cell into the S-phase of the cell cycle could also be achieved by exposing the activated T cell to partially purified Interleukin 2 (IL-2). Given the prior demonstrations that IL-1 functions to induce the T cell-dependent production of the IL-2, it appears that IL-2 is the requisite signal necessary for the activated human lymphocyte to actually progress through the prereplicative phase of the cycle into the S-phase.
Demonstration of a C1q receptor on the surface of human endothelial cells.
Authors: Andrews B S , Shadforth M , Cunningham P , Davis J S
Published: December 2022

A receptor for C1q on the surface of human endothelial cells has been demonstrated. This receptor is present on the surface of viable cultured endothelial cells derived from human umbilical veins, and C1q binding can also be demonstrated to the endothelial lining cells of human umbilical artery and vein on frozen tissue sections. Receptors for the complement components C3b and C3d were not detected on tissue sections or endothelial cells in suspension. Endothelial cell C1q receptors are discussed in relationship to possible immune complex localization in vivo.
Immunoregulatory changes induced by total lymphoid irradiation (TLI). I. Appearance of a population of cells bearing the thymus leukemia (TL) surface antigen in the lymph nodes and spleen.
Authors: King D P , Strober S , Kaplan H S
Published: December 2022

BALB/c mice were treated with total lymphoid irradiation (TLI; 3400 rads) or single-dose, whole-body irradiation (WBI; 550 rads), and the lymph nodes and spleen were assayed for the level of H-2, Ia, Thy-1.2, Lyt-1.2, Lyt-2.2,3.2, and TL-bearing cells by an in vitro cytotoxicity assay. The level of cells bearing all these markers, except for TL, was similar to that observed in normal unirradiated mice. The level of TL+ cells was increased 10- to 20-fold in the TLI-treated mice as compared to WBI-treated mice or normals. The increased number of TL+ cells persisted for at lest 200 days after radiotherapy, and was not dependent upon irradiation of the thymus. However, thymectomy before TLI prevented the subsequent development of peripheral TL+ cells. The findings suggest that this abnormal subpopulation of T cells develops as a result of changes in the peripheral lymphoid tissue microenvironment induced by TLI.
Stimulation of anti-influenza cytolytic T lymphocytes by CNBr cleavage fragments of the viral hemagglutinin.
Authors: Wabuke-Bunoti M A , Fan D P , Braciale T J
Published: December 2022

Purified hemagglutinin of influenza virus, subtype A/Jap/305/57 (H2/N2) was cleaved with CNBr, and the resulting fragments were capable of stimulating H-2 restricted cytolytic T lymphocytes in vitro from spleen cells of mice immunized previously with A/Jap/305/57. When the peptides were separated by high-performance liquid chromatography, stimulatory activity was confined primarily to a series of peptides contained within a limited chromatographic range. The majority of the lytic activity was specific for cells infected with A/Jap virus, although a small amount of cross-reactive killing of A/PR/8/34-(H0/N1) infected cells could not be excluded. These results demonstrated that H-2-restricted influenza viral-specific CTL could be generated by fragments of a complex polypeptide antigen, the influenza hemagglutinin, and offer the possibility of analyzing specific antigenic determinants recognized by cytolytic T cells.
A serum C3-activating factor: its characterization and its presence in glomerular deposits.
Authors: Roberts J L , Levy M , Chioros P G , Bartlow B G , Forristal J , West C D , Habib R , Lewis E J
Published: December 2022

A non-IgG C3 activating factor (C3AF) isolated from the serum and plasma of a patient with membranoproliferative glomerulonephritis with subendothelial and subepithelial deposits (type III) was studied to define its molecular structure, functional characteristics, and presence in renal biopsies. Alternative complement pathway (ACP) activation of C3 was comparable to that of zymosan, but significantly less than that of IgG C3 nephritic factor (C3NeF). Functionally active isolate contained beta 1H globulin (beta 1H) and the C3AF protein, which had a gamma-electrophoretic migration. beta 1H present in the isolate exhibited an altered gamma-beta electrophoretic mobility. The function of this complex is heat labile. Time/temperature studies demonstrated that inactivated preparations of this isolate show a return to the normal beta migration of beta 1H followed by the development of an alpha 2 electrophoretic migration of the previously gamma-migrating material of functionally active isolate. Normal human serums contain an alpha 2 protein that shows complete antigenic identity with the gamma protein of C3AF, but this alpha 2 protein lacks C3 activating function. On polyacrylamide gel electrophoresis, the C3AF isolate contains a high m.w. protein of 500,000 to 590,000 that is composed of 2 major subunits. Carbohydrate is not demonstrable, and the protein is antigenically distinct from plasma fibronectin. Immunofluorescence studies of renal biopsies of patients with types I and III MPGN revealed the presence of C3AF antigen in granular deposits in 10 of 16 cases. The presence of this antigen in glomerular deposits was associated with C3 and beta 1H, but was independent of Ig. These studies suggest that the mechanism of ACP activation by C3AF results from interference with beta 1H function. The presence of this material in renal biopsies from some MPGN patients further suggests a pathogenetic role in mediating glomerular pathology.
Perturbation of early development of the immune system by normal adult lymphoid cells.
Authors: Raybourne R , Arnold L W , Haughton G
Published: December 2022

Seven-day-old C57BL/10 mice injected with 3 X 10(6) normal adult lymph node cells (NAL) from syngeneic donors were compared with uninjected littermates when 5 to 7 wk old. The direct PFC response to sheep erythrocytes was suppressed by 60%. Responses to PHA and Con A but not LPS were suppressed by 60 to 90%. Primary and secondary (after skin graft rejection) MLR responses were suppressed by 30 to 40%. Active suppressor cells were demonstrable in these mice. Skin grafts incompatible for H-2 were rejected in normal time. NAL derived from adult mice conferred the effect, but not cells from 7-day-old donors. Mice inoculated with Rous sarcoma virus within the 1st day of life and 3 X 10(6) NAL at 7 days of age displayed up to 5-fold increased incidence of primary sarcomas.
The generation of cytolytic activity in mixed leukocyte cultures: cortisone-resistant thymocytes are deficient in helper T lymphocytes.
Authors: Lutz C T
Published: December 2022

Cortisone-resistant (CR) thymocytes did not generate cytolytic activity toward H-2 K or D alloantigen unless they were also stimulated by H-2 I or non-H-2 alloantigens, even though spleen cells generated brisk cytolytic activity toward H-2 K or D alone. Backstimulation by stimulating strain T lymphocytes accounted for neither the response of spleen cells toward H-2 K or D alloantigen nor the response of CR thymocytes to a full set of alloantigens. In addition, lack of non-T accessory cells did not account for the CR thymocyte pattern of reactivity. Rather, CR thymocytes appeared to be relatively deficient in helper T lymphocytes (HTL). CR thymocytes generated specific cytolytic activity toward H-2 D alloantigen when T cell growth factors (TCGF) or cloned alloreactive helper T lymphocytes were added to culture. CR thymocytes contained fewer HTL precursors detected at limit dilution than spleen cells did. Thus spleen cells generated cytolytic activity toward class I alloantigens alone, but under the same culture conditions CR thymocytes had to be stimulated by both class I and class II alloantigens. Class II alloantigens may be required to stimulate cytolytic activity only under culture conditions in which class I-reactive HTL are not sufficient to provide a minimal threshold of help.
Human gamma interferon production by leukocytes induced with monoclonal antibodies recognizing T cells.
Authors: von Wussow P , Platsoucas C D , Wiranowska-Stewart M , Stewart W E
Published: December 2022

The murine monoclonal antibody OKT3 (IgG2), which recognizes the majority of human E-rosette-forming cells, was found to induce interferon (IFN) production in human peripheral lymphocytes in vitro. This IFN was classified as IFN-gamma by virtue of its acid stability, nonneutralization by antisera recognizing human IFN-alpha and beta, and non-cross-reactivity on bovine kidney cells. OKT3 elicited considerable IFN-gamma yields at a concentration of 1 ng/ml, at which only very low mitogenesis of the lymphocytes measured by 3H-thymidine incorporation was seen. The IFN-gamma activity appeared beginning 3 to 6 hr after stimulation and reached its maximum after 36 hr. Anti-human Lyt3, anti-human Lyt2, or anti-human Lyt1 monoclonal antibodies, recognizing all or the majority of the E-rosette-forming cells, did not elicit IFN-gamma production. The OKT4 monoclonal antibody, which defines a subpopulation of T cells containing cells with helper activity, induced marginal levels of IFN-gamma. In contrast, OKT8 monoclonal antibody (defining cytotoxic/suppressor cells) as well as monoclonal or heterologous antibodies recognizing histocompatibility antigens or surface immunoglobulin failed to induce IFN-gamma production. Heterologous anti-thymocyte globulin or anti-lymphocytic serum were effective IFN-gamma inducers. Cell separation studies of the leukocytes demonstrated that E-rosette-forming cells are responsible for production of IFN-gamma induced by OKT3. Treatment of the leukocyte suspension with OKt3 monoclonal antibody plus complement abolished IFN-gamma production induced by this antibody. These results demonstrate that the OKT3 antibody recognizing an antigen present on the majority of the E-rosette-forming cells alone is sufficient to induce IFN-gamma production. OKT3-positive cells are apparently the producers of this IFN-gamma.
Antigen-dependent proliferation of cloned continuous lines of H-2-restricted influenza virus-specific cytotoxic T lymphocytes.
Authors: Andrew M E , Braciale T J
Published: December 2022

The antigenic requirements for in vitro proliferation of several cloned continuous lines of H-2-restricted influenza virus-specific cytotoxic T lymphocytes (CTL) has been examined. The cloned CTL lines were established from individual splenic CTL precursors obtained from A/JAPAN/305/57 (H2N2)-immune F1 (C57BL/6 X BALB/c) donors. The lines were isolated (by limiting dilution in liquid culture) and expanded in the presence of A/JAPAN/305/57-infected irradiated syngeneic (F1) spleen cells and T cell growth factor (TCGF) of rat spleen origin. Optimal proliferation (and long-term in vitro cultivation) of these H-2-restricted CTL lines required both specific antigenic stimulation in the form of virus-infected syngeneic spleen cells and an exogenous source of TCGF. In addition, the antigenic requirements for proliferation of these lines directly reflected the pattern of H-2-restricted influenza virus-specific recognition at the level of target cell recognition and lysis.
Characterization of an antiserum to guinea pig antithrombin III (AT III). I. Reactivity with guinea pig T lymphocytes.
Authors: Clement L T , Yamashita U , Shevach E M
Published: December 2022

In the course of studies investigating the effects of antisera prepared against a variety of guinea pig proteins on lymphocyte function, a goat antiserum prepared against a guinea pig gamma-globulin preparation was found to react with guinea pig T lymphocytes. This antiserum, serum 592, contained a significant titer of antibodies that were cytotoxic for a subpopulation of lymph node cells and thymocytes, and mitogenic for lymph node T cells. Immunoelectrophoretic analysis and selective absorptions of the antiserum demonstrated that the antigen recognized on thymocytes was also present on an alpha 2 globulin in guinea pig plasma, which, on the basis of physiochemical characteristics and heparin-binding affinity, appeared to be guinea pig antithrombin III (AT III). Although the antiserum was shown to contain antibodies to both protein and carbohydrate determinants on the AT III molecule, studies comparing the effects of 7 M guanidine and periodate oxidation on the antigenicity of the AT III determinant also recognized on the thymocytes indicated that this shared antigenic determinant was carbohydrate in nature. The thymocyte membrane molecule bearing this determinant was also isolated and was found to be a 210,000-dalton macromolecule that was very sensitive to proteolytic and/or autolytic degradation. These data raise the interesting possibility that guinea pig lymphoid cells may have a membrane-associated protease inhibitor related to plasma AT III.
Characterization of an antiserum to guinea pig antithrombin III (AT III). II. Stimulation of T lymphocyte proliferation.
Authors: Yamashita U , Clement L T , Shevach E M
Published: December 2022

A goat antibody specific for an antigenic determinant shared between guinea pig antithrombin III (AT III) and thymocytes was shown to be mitogenic for lymph node T lymphocytes in the presence of macrophages. Although the antiserum was not mitogenic for purified populations of B lymphocytes, B lymphocytes were as efficient as T lymphocytes in absorbing the mitogenic activity of the serum. The shared antigenic determinant appeared to be carbohydrate in nature in that native and guanidine-treated AT III, but not periodate oxidized AT III, were capable of inhibiting the mitogenic activity of the serum when added continuously to the cultures. The possibility that the plasma protease inhibitor AT III or an antigenically related membrane protein are involved in the regulation of T cell activation is discussed.
Evidence for interaction between T cell populations of tumor-bearing and normal mice in immune suppression.
Authors: Ingenito G G , Calkins C E
Published: December 2022

Spleen cells of DBA/2 mice bearing subcutaneous implants of the syngeneic tumor L5178Y induce suppression of the in vitro antibody response of normal spleen cells to sheep erythrocytes (SRBC). Cells mediating suppression are detected in the spleens of tumor-bearing mice as early as 24 hr post-implantation but are no longer detected there 15 days post-implantation. These spleen cells are nylon wool nonadherent, sensitive to anti-Thy 1.2 + C and anti-Lyt 1.1 + C, and insensitive to anti-Lyt 2.1 + C treatment. The anti-SRBC response of the unfractionated spleen cells from the tumor-bearing mice is not itself suppressed at the cell numbers used. This along with the finding that suppression occurs in the presence of spleen cells from normal mice suggest that a cell population from the normal mouse spleen is also involved in the suppression. Spleen cells from mice inoculated with irradiated (nonproliferating) L5178Y cells are similarly capable of mediating nonspecific suppression for the same limited period of time after the inoculation. In addition, spleen cells from mice stimulated with several nontumorigenic cellular antigens interact with normal spleen cells to produce suppression. These findings suggest that suppression observed in vitro with spleen cells from these tumor-bearing mice may be the result of antigen-activated cells triggering normal immunoregulatory cells.
Restriction in IgM expression. III. Affinity analysis of monoclonal anti-lactose antibodies.
Authors: Mandal C , Karush F
Published: December 2022

A clonal analysis has been made of the murine BALB/C response to lactose-containing immunogens with respect to the affinity restriction in IgM expression. Monoclonal IgM and IgG antibodies were prepared from antilactosyl hybridomas generated from mice immunized with p-aminophenyl-beta-lactoside (PAPL) coupled to BGG or with a vaccine of Streptococcus faecalis (Strain N). Association constants for the binding of monovalent derivatives of PAPL were measured by quenching fluorescence. These derivatives carried a probe (2,4-dinitrophenyl or 1-dimethylaminonaphthalene-5-sulfonyl) that served to quench the protein fluorescence when the ligand was complexed with the protein. The central finding was that with both immunogens a restriction in the affinity of IgM was demonstrated since the highest values exhibited by IgG antibody exceeded by at least a factor of 50 the highest comparable values of the association constants for IgM antibody. It is suggested that the hypothesis of germ-line restriction, previously proposed as the basis for the affinity restriction of IgM, may also be applicable to the T cell receptor since its distinctive properties parallel those of IgM antibody.
The adjuvant activity of nonionic block polymer surfactants. I. The role of hydrophile-lipophile balance.
Authors: Hunter R , Strickland F , Kézdy F
Published: December 2022

We developed an experimental model for studying the physicochemical basis of the adjuvant activity of surface-active agents. Pluronic polyols are chemically similar but physicochemically diverse surface-active agents composed of polymers of hydrophilic polyoxyethylene and lipophilic polyoxypropylene. One of them, L121, was found to be a powerful adjuvant for increasing antibody formation to BSA in mice when injected in an oil-in-water emulsion. Another, L101, was less effective in increasing antibody formation, but was more effective in inducing granulomatous inflammation. Three others had little effect on either response. The ability of these and other surface active agents to serve as adjuvants correlated with a physicochemical parameter, the hydrophile-lipophile balance (HLB). All of the surfactants with strong adjuvant activity have HLB values of less than 2. They are strongly lipophilic and promote the retention of macromolecules by oil drops in oil-in-water emulsions. We propose that the adjuvant activity of these compounds is dependent on their ability to concentrate the adjuvant, immunogen, and host proteins on hydrophobic surfaces, where they are more effectively presented to cells of the immune system.
Activation of human neutrophils with 1-O-hexadecyl/octadecyl-2-acetyl-sn-glycerol-3-phosphorylcholine (platelet activating factor).
Authors: Shaw J O , Pinckard R N , Ferrigni K S , McManus L M , Hanahan D J
Published: December 2022

1-O-Hexadecyl/octadecyl-2-acetyl-sn-glyceryl-3-phosphorylcholine (AGEPC), the acetylated alkyl phosphoglyceride known as platelet-activating factor, stimulated human neutrophil (PMN) exocytosis, migration, superoxide production and aggregation over a concentration range of 10(-10) to 10(-5) M. AGEPC-induced PMN exocytosis of azurophilic (myeloperoxidase and beta-glucuronidase) and specific (lactoferrin and lysozyme) lysosomal granules was rapid (T 1/2 = 20 sec), dependent on the presence of cytochalasin B, but was not associated with release of cytoplasmic LDH. As seen with the complement-derived peptide stimulus, C5a, AGEPC-initiated PMN enzyme release was dependent on temperature and cellular glycolysis but not on the presence of extracellular Ca++. When analyzed by gradient analysis, PMN migration caused by AGEPC was primarily chemotactic in nature. An unusual feature for both enzyme secretion and migration was a decrease in response between 10(-6) M and 10(-5) M AGEPC. This decreased responsiveness could be explained by rapid PMN desensitization occurring at high AGEPC concentrations, limiting the overall cellular response. Rapid desensitization for exocytosis was demonstrated in PMN stimulated with AGEPC in the absence of cytochalasin B. When cytochalasin B was added subsequently and PMN challenged with AGEPC or C5a, stimulus-specific desensitization to AGEPC but not C5a-induced lysosomal enzyme release occurred. PMN desensitized to C5a responded normally to a subsequent AGEPC challenge. Stimulation of all the PMN functions examined was markedly attenuated with removal of the 2-acetyl group from AGEPC (lyso GEPC). These results suggest that AGEPC stimulates a wide variety of human PMN responses by a receptor-like mechanism, dependent on the short chain fatty acid ester in the 2-position of the alkyl phosphoglyceride.
Glucocorticoid receptors and in vitro corticosensitivity of peanut-positive and peanut-negative human thymocyte subpopulations.
Authors: Ranelletti F O , Piantelli M , Iacobelli S , Musiani P , Longo P , Lauriola L , Marchetti P
Published: December 2022

In 6 human thymus glands, the immature subset of thymocytes was separated from the more mature one, by differential peanut lectin agglutination. These 2 cell subpopulations were analyzed for glucocorticoid receptor content by using a whole cell assay, with (3H)-triamcinolone acetonide as tracer. The unagglutinated thymocytes (peanut negative) contained about 2 times more receptor sites per cell than agglutinated (peanut positive) ones (7650 +/- 1550 S.D. verus 3195 +/- 896 S.D.). The affinity for steroid was similar in both cell subsets, as was the stereospecificity for glucocorticoids, the time-course of steroid-receptor association, and cytoplasmic to nuclear translocation. Despite the greater number of glucocorticoid receptor sites, the peanut-negative thymocyte subpopulation did not differ from the peanut-positive one in its sensitivity to the inhibitory effects of triamcinolone acetonide, as determined by measurements of the incorporation of radiolabeled precursors of protein and DNA. Moreover, the peanut-negative subset appeared more resistant in vitro to the steroid-induced cell lysis as compared to the peanut-positive one. Thus, our data suggest that glucocorticoid receptor density and corticosensitivity are not directly correlated and that the number of glucocorticoid receptor sites may be dependent on the degree of immunologic maturation.
Experimental allergic encephalomyelitis in Lewis rats: immunoregulation of disease by a single amino acid substitution in the disease-inducing determinant.
Authors: Kardys E , Hashim G A
Published: December 2022

A single amino acid substitution in the sequence of the encephalitogenic determinant for Lewis rats destroyed its ability to induce experimental allergic encephalomyelitis (EAE) but generated a potent immunoregulatory sequence capable of suppressing the development of both clinical and histologic signs of EAE. The EAE-inducing determinant (synthetic peptide S6) H-Ala-Gln-Gly-His-Arg-Pro-Gln-Asp-Glu-Asn-OH (residues 75 to 84) of the bovine MBP induced clinical and histologic signs of EAE when it was administered at doses of 0.5 micrograms or higher. Gly substituted for the C-terminal Asn during the synthesis of peptide S6 generated the homologous sequence designated by peptide S79. Peptide S79 failed to induce either clinical or histologic signs of EAE even when it was administered at dosages up to 1000 times higher than those of S6. Similarly, rats pretreated with a single dose of S79 were not only unresponsive to an encephalitogenic challenge but also were capable of transferring unresponsiveness to syngeneic recipients with viable donor lymphocytes. The induction of unresponsiveness that was abrogated by pretreatment with cyclophosphamide suggests the development of an S79-sensitive lymphocyte subset that regulates MBP-induced EAE in Lewis rats.
Increased expression of I-region-associated antigen (Ia) on B cells after cross-linking of surface immunoglobulin.
Authors: Mond J J , Seghal E , Kung J , Finkelman F D
Published: December 2022

Both surface Ig (sIg) surface Ia (sIa) have been shown to have important roles in B lymphocyte activation. In order to investigate a possible relationship between these molecules, we studied the effects of cross-linking of sIg on the expression of sIa, as measured by fluorescence-activated cell sorter analysis of lymphoid cells stained with conventional anti-Ia anti-serum or with fluorescein-labeled anti-Ia antibodies. Exposure of cells for 24 hr in vitro to anti-delta, anti-mu, anti-kappa antibodies, or their F(ab')2 fragments induced a very dramatic increase in expression of sIa. Similarly, i.v. injection of anti-delta antibodies into adult mice induced a 2- to 3-fold increase in expression of B cell sIa on spleen, lymph node, and Peyer's patch lymphocytes. There was no increase under these conditions in expression of other B lymphocyte surface antigens, including H-2, 4B9, and 17C9. Furthermore, exposure of B lymphocytes to antibodies directed to B lymphocyte surface antigens other than sIg did not result in an increase in expression of sIa. The anti-Ig-induced increase in sIa expression appeared to be T independent, required cellular protein synthesis, and required more time to occur than did the cross-linking and removal of sIg. This increase in expression of sIa did not occur on B lymphocytes obtained from mice younger than 3 wk old. This increase in expression of sIa may reflect a proximal event in B lymphocyte activation that occurs after cross-linking of sIg by antigen and that may enhance subsequent cellular interactions involving B lymphocytes.
Activation of the fifth and sixth component of the complement system: similarities between C5b6 and C(56)a with respect to lytic enhancement by cell-bound C3b or A2C, and species preferences of target cell.
Authors: Hänsch G M , Hammer C H , Mayer M M , Shin M L
Published: December 2022

Brief shift of purified C5 and C6 at 0 degrees C to pH 6.4, followed by immediate neutralization, results in the generation of a factor, designated C(56)a, that lyses erythrocytes together with C7, C8, and C9. We compared C(56)a and C5b6 generated by an alternative-pathway convertase, with regard to their action on different target cells. We found tht C(56)a is similar to C5b6 in the following properties: 1) Together with C7, C(56)a forms a stable intermediate on either sheep or guinea pig erythrocytes. 2) Membrane-bound C3b, or A2C incorporated in the membrane, enhances lysis by C(56)a-9, as well as lysis by C5b6-9. We also found that the lysis of EC(56)a7 or EC5b67 intermediates by C8 and C9 depends on the species of the erythrocytes and the species of C8 and C9. Thus, lysis of sheep erythrocytes is more efficient with guinea pig C8 and C9 than with human C8 and C9. In the case of guinea pig erythrocytes, this relationship is reversed, i.e., these cells lyse more efficiently when human C8 and C9 are used. Enhancement of lysis by membrane-bound C3b or A2C does not abrogate this species incompatibility pattern.
Human large granular lymphocytes: spontaneous and interferon-boosted NK activity against adherent and nonadherent tumor cell lines.
Authors: de Landazuri M O , López-Botet M , Timonen T , Ortaldo J R , Herberman R B
Published: December 2022

Human natural killer (NK) cells against K-562 tumor cells have been morphologically identified as large granular lymphocytes (LGL). We have investigated whether LGL are also responsible for NK activity against a variety of adherent and nonadherent tumor cell lines. LGL-enriched fractions, obtained by centrifugation of nonadherent mononuclear cells on discontinuous density gradients of Percoll, contained high levels of spontaneous and interferon- (IFN) boosted NK activity against each of the target cells tested. In contrast, the LGL-depleted functions, which contain most of the typical small lymphocytes, were devoid of NK activity and did not develop appreciable activity after treatment with IFN. The ability of the different fractions to exert NK activity correlated with their capacity to form conjugates with nonadherent tumor cells. Reconstitution of the different fractions with macrophages did not increase the NK activity of already cytotoxic fractions and did not induce appreciable activity in the LGL-depleted fractions. On the other hand, mixtures of highly cytotoxic and nonreactive cell fractions did not reveal the presence of suppressor cells for NK in the nonreactive fractions. Taken together, our data demonstrate that both spontaneous and IFN-boosted NK activity is confined to the LGL-enriched fractions and suggest that LGL are the main effector cells exerting NK cytotoxicity against a variety of adherent as well as nonadherent tumor cell lines.
Analysis of HLA-DR antigens by using monoclonal antibodies: recognition of conformational differences in biosynthetic intermediates.
Authors: Shackelford D A , Lampson L A , Strominger J L
Published: December 2022

The monoclonal antibodies L203 and L227 were used to characterize Ia-like antigens on human B cell lines. The antibodies appear to recognize subsets of HLA-DR molecules on the cell line Raji, which is heterozygous for HLA-DR. However, by using L203 and L227, more than 1 DR-like molecule could not be clearly identified on the DRw8 homozygous cell line MADURA. The 2 antibodies did display different affinities for different forms of the same DR molecule. On DRw8 and cell lines of other DR types, L227 had a greater affinity than L203 for the biosynthetic intermediates of the DR heavy and light chains. L203 displayed a greater affinity than L227 for the mature forms of the DR subunits. The antibody L227 immunoprecipitated the denatured DR light chain, but not the heavy chain, suggesting that the antigenic determinant is on the light chain. The L203 and L227 determinants are not on the N-linked oligosaccharides, since both antibodies recognized nonglycosylated DR antigens from cells treated with tunicamycin. In contrast to the DRw8 homozygous cell line, it was found that DRw6 homozygous B cell lines express at least 2 DR light chains. A xeno anti-HLA-DR (anti-p23,30) serum and L203 recognized both light chains, whereas L227 precipitated only 1 of the 2. A model depicting the expression of the L203 and L227 antigenic determinants on the DRw8 molecule is discussed.