bluuebunny/crossref_metadata_2025 huggingface dataset:
A human myeloma cell line that does not express immunoglobulin but yields a high frequency of antibody-secreting hybridomas.
Authors: Pickering J W , Gelder F B
Published: December 2022

We selected an 8-azaguanine-resistant variant of a human myeloma cell line (RPMI 8226) by cloning the parental cells on a feeder layer of mouse spleen cells in the presence of increasing concentrations of 8-azaguanine. Culture media and cellfree extracts of both the parental and variant (8226 AR/NIP4-1) cell lines were assayed for production of immunoglobulin heavy and light chains by double immunodiffusion and for lambda-chain by radioimmunoassay. Secretion of free lambda-chain by the parental cell line was confirmed. In contrast, no immunoglobulin heavy or light chains were detected in culture medium of the variant cell line by either immunodiffusion or radioimmunoassay. No intracellular lambda-chain could be detected in the variant cells by radioimmunoassay of cellfree extracts or by immunofluorescence of fixed cells. Hybridomas were produced by fusion of 8226AR/NIP4-1 cells with lymphocytes from a mesenteric lymph node recovered at surgery from a hypertransfused renal transplant recipient. Twenty hybrid culture supernatants were assayed for immunoglobulin by double immunodiffusion, and 15 contained either IgG (lambda) or IgG (kappa). None produced IgM or IgA. An IgG (kappa)-producing hybridoma was shown by immunofluorescence not to express lambda-chain. A second fusion between the variant cell line and spleen cells from a renal transplant patient produced a stable hybridoma secreting IgM (lambda) antibody specific for the I antigen.
A human helper T cell clone secreting both killer helper factor(s) and T cell-replacing factor(s).
Authors: Kaieda T , Okada M , Yoshimura N , Kishimoto S , Yamamura Y , Kishimoto T
Published: December 2022

A human helper T cell clone (d4), which showed its helper effect on the differentiation of both T and B cells, was established by MLC reaction of normal T cells against a B lymphoblastoid cell line (CESS) followed by cloning in the presence of IL2 and x-irradiated CESS and autologous non-T cells. d4 cells helped the induction of cytotoxic T cells against UV-treated CESS cells. Antigen-stimulated d4 cells secreted helper factor(s) involved in the induction of cytotoxic T cells (killer helper factor(s), KHF), and KHF activity could be separated into two fractions, one with the m.w. of 15,000 to 20,000 and the other with the m.w. of 45,000 to 50,000. The factor with 15,000 to 20,000 m.w. showed IL 2 activity; the other factor showed gamma-interferon activity without IL 2 activity, suggesting that both IL 2 and gamma-interferon exerted KHF activity. d4 cells or their culture supernatant showed helper activity in the induction of IgG in a B cell line (CESS). The helper activity of the supernatant (TRF) was absorbed with CESS cells but not with IL 2-dependent CTLL, whereas KHF activity was absorbed with IL 2-dependent CTLL but not with CESS cells. The results showed that TRF and KHF were distinct molecules and a single helper T cell clone could secrete helper factors for both B and T cells.
Alkaline phosphatase activity as a membrane marker for activated B cells.
Authors: García-Rozas C , Plaza A , Díaz-Espada F , Kreisler M , Martínez-Alonso C
Published: December 2022

Alkaline phosphatase activity was assayed by a microtiter assay (with p-nitrophenylphosphate used as substrate) in the plasma membrane of mouse spleen cells activated in vitro by the B cell mitogen LPS and the T cell-dependent B cell mitogen, PWM. No activity was detected in spleen cells cultured in the presence of the T cell mitogens PHA and Con A. This alkaline phosphatase activity was detected in the blast-enriched 30 to 40% fraction of discontinuous Percoll gradients in LPS-treated cultures, and the enzymatic activity assayed was susceptible to inhibition by the alkaline phosphatase inhibitors EDTA and L-phenylalanine. These data support the idea that the membrane alkaline phosphatase activity could be used as a marker for activated B cells.
Concanavalin A-induced human lymphocyte mitogenic factor: activity distinct from interleukin 1 and 2.
Authors: Duncan M R , George F W , Hadden J W
Published: December 2022

Fresh human mononuclear cells (MNL) cultured in serum-free medium and stimulated with concanavalin A (Con A) produce a lymphokine functionally similar to the previously described antigen-induced lymphokine human lymphocyte mitogenic factor (LMF). T lymphocytes appear to be the source of Con A-induced LMF and the presence of adherent monocytes is required for maximal production. The proliferative response to LMF, assayed either as supernatant fluids in the presence of alpha-methyl-D-mannoside or as partially purified 27,000 to 35,000 m.w. fractions from Bio-Gel P-100 columns, indicates LMF is distinct from Con A. LMF acts almost exclusively on B lymphocyte-enriched (E rosette-negative) MNL while exhibiting only minimal activity on T lymphocytes and no activity on monocytes. LMF preparations induce both B cell proliferation and the maturation and proliferation of E rosette-negative T cell progenitors into E rosette-positive cells. Assay of interleukin 1 (IL 1) and interleukin 2 (IL 2) preparations for LMF activity implicate IL 2 as the T cell progenitor stimulating factor. Assay of the proliferation of human IL 2-dependent alloreactive T lymphocyte line/ATLL) indicates that LMF preparations contain IL 2, IL 2 activity, however, is completely removed by absorption of LMF preparations by ATLL cells, whereas LMF activity (B cell proliferation) is unabsorbable, thus distinguishing LMF from IL 2. These absorbed LMF preparations contain no IL 1 activity as assayed by murine thymocyte proliferation, nor does IL 1 have LMF activity. These results indicate that Con A-induced LMF contains a functionally distinct lymphokine with the ability to act as a B cell growth factor.
An epidermal cell-derived cytokine triggers the in vivo synthesis of serum amyloid A by hepatocytes.
Authors: Sztein M B , Luger T A , Oppenheim J J
Published: December 2022

Lipopolysaccharide (LPS) stimulates macrophages to produce a number of closely related soluble mediators, including interleukin 1 (IL 1), endogenous pyrogen (EP), and serum amyloid A (SAA) inducer, which are able to induce hepatocyte production of SAA, an acute-phase reactant. Keratinocytes also produce a soluble mediator, epidermal cell-derived thymocyte-activating factor (ETAF), which is very similar to IL 1. The experiments reported here show that unpurified ETAF-containing preparations, as well as ETAF after chromatography by gel filtration (Sephacryl S-200), anion-exchange on DEAE, phenyl-Sepharose, or isoelectrofocusing, were all able to induce the in vivo production of SAA by hepatocytes. Additionally, ETAF and SAA inducer exhibited the same pattern of temperature sensitivity. These observations add further support to the hypothesis that ETAF, IL 1, EP, and SAA inducer comprise a group of closely related mediators and that nonlymphoid cells can produce mediators that can regulate systemic inflammatory reactions.
Membrane phenotype of the rat cytotoxic T lymphocyte.
Authors: Gilman S C , Rosenberg J S , Feldman J D
Published: December 2022

We have examined rat cytotoxic T lymphocytes for expression of W3/25, OX8, Ia, Thy-1 antigens, and Fc gamma receptors using an effector cell-target cell conjugate formation assay in conjunction with immunofluorescence techniques. Lymph node, spleen, and peritoneal exudate T cells from Lewis rats immunized with allogeneic BN tumor cells specifically bound to and lysed BN tumor targets and BN blast cells, but did not bind or lyse syngeneic Lewis sarcoma cells, Lewis blast cells, or Lou/M blast cells. The numbers of binding and cytotoxic T lymphocytes were greatest in peritoneal exudate cells of immunized rats, less in spleens, and least in lymph nodes. Seventy to 80% of the lymphocytes bound to tumor targets were OX8+ T lymphocytes; less than 12% expressed W3/25, Ia, Thy-1, or Fc gamma R. Moreover, only OX8+ T cells efficiently lysed the target cells to which they were bound. The membrane phenotype of rat cytotoxic T lymphocytes was: OX8+, W3/25-, Ia-, Thy-1, and Fc gamma R-. Monoclonal OX8 antibody did not inhibit target cell binding or subsequent lysis by effector T cells, and there was no diminution of target cell binding or cytotoxic activity when the OX8 antigen was shed from the cell surface before interaction with target cells. There was no preferential association of OX8 antigen at the interface between the effector and target cell. Thus, OX8 antigen marks a subset of rat T lymphocytes that are cytotoxic but the molecule appears not to play a functional role in the cytotoxic process.
The human autologous mixed lymphocyte reaction. III. Immune circuits.
Authors: Smolen J S , Chused T M , Novotny E A , Steinberg A D
Published: December 2022

The human autologous mixed lymphocyte reaction represents the proliferative response of T cells to determinants presented on autologous non-T cells. Purified T4+ cells vigorously proliferate in response to stimulation by either (B + null) cells or M phi, whereas purified T8+ cells proliferate very little without a source of help. Such help can be provided by mitomycin C-treated T4+ cells, which indicates that proliferation of T4+ cells is not necessary for the help. M phi suppress the human AMLR, as measured by the response of T cells to stimulation by (B + null) cells. The target of this M phi-induced suppression was found to be the T4+ inducer cell. In contrast to the suppressive effects of M phi on T4+ cells, M phi did not suppress T8+ cells. The available data suggest that the human AMLR is a two-part inducer circuit. One part can be stimulated preferentially by M phi and the other by (B + null) cells; however, M phi prevent activation of the (B + null) cell part of the circuit. These results may provide an explanation for the differential activities of different regulatory circuits without and also with antigenic stimulation.
Acquisition of immune competence by a subset of human cortical thymocytes expressing mature T cell antigens.
Authors: Umiel T , Daley J F , Bhan A K , Levey R H , Schlossman S F , Reinherz E L
Published: December 2022

Previous studies indicated that the human thymus is composed of several discrete compartments. Cortical thymocytes are reactive with a monoclonal antibody, anti-T6 (TL-like) and coexpress antigens defined by antibodies anti-T4 and anti-T8. In contrast, most medullary thymocytes are unreactive with anti-T6, express either T4 or T8 antigens, and more importantly, brightly stain with anti-T3 and anti-T1, which define mature T cell antigens. Only a minor population of cortical thymocytes strongly express T1 and T3 antigens (T1+T3+). In the present study, we characterized this latter subpopulation of brightly staining T1+ cortical thymocytes. Because murine and human cortical thymocytes alone bear receptors for peanut agglutinin (PNA), cortical PNA+ thymocytes were separated from medullary PNA- thymocytes by fluorescence-activated cell sorting and were subsequently fractionated into PNA+T1+ (strongly anti-T1-reactive) and PNA+T1- (unreactive or weakly anti-T1-reactive) populations. The PNA+T1+ cells represented only 10 to 20% of thymic cortical lymphocytes. Whereas the PNA+T1- subset was unresponsive to PHA or alloantigen even in the presence of IL 2, the PNA+T1+ subset gave a brisk response to both stimuli on addition of IL 2. This latter response was similar to that of the PNA-T1+ medullary thymocytes. These results suggest that acquisition of mature T cell antigens is associated with immunocompetence despite a cortical localization.
Size of the transmembrane channels produced by complement proteins C5b-8.
Authors: Ramm L E , Whitlow M B , Mayer M M
Published: December 2022

It has been shown previously that erythrocytes can be lysed by complement proteins C5b-8, albeit at a much lower rate than C5b-9. We have now performed kinetic sieving experiments with resealed erythrocyte ghosts using sucrose (0.9 nm molecular diameter) and inulin (3.0 nm molecular diameter) as markers. We found that treatment of the ghosts with C5b-8 released sucrose, but not inulin. Addition of C9 to ghosts carrying C5b-8 dramatically increased the rate of sucrose flux and, in addition, caused release of inulin. Hence, unlike C5b-9 channels, those formed by C5b-8 measure less than 3 nm in diameter. Formation of C5b-8 channels was very slow compared with that of C5b-9 channels. Also, we found that about two-thirds of the C5b-8 ghosts did not have sucrose-releasing channels, but such channels were formed on reaction with C9.
Cell cycle analysis of lymphocyte activation in normal and autoimmune strains of mice.
Authors: Raveche E S , Steinberg A D , DeFranco A L , Tjio J H
Published: December 2022

The spontaneous spleen cell proliferation and the proliferation induced by in vivo or in vitro stimulation with such polyclonal B cell activators (PBA) as LPS, poly rI.rC, and anti-mu were studied in normal and autoimmune mice. The various murine models of autoimmunity differ in the level of naturally occurring splenic cellular hyperactivity as well as in the ability of their spleen cells to be further stimulated in vitro by polyclonal stimulators. Both the NZB strain and the MRL/Ipr strain had markedly increased numbers and percentages of spontaneously proliferating spleen cells, whereas the BXSB strain did not. Nonautoimmune strains were found to have very small numbers of activated cells in the spleen. However, such normal strains could be induced in vivo to mimic the natural splenic hyperactivity observed in older NZB and MRL/Ipr autoimmune strains by the injection of polyclonal B lymphocyte stimulators. In contrast, old hyperactive NZB mice were not further induced to undergo proliferation by in vivo administration of such stimulators. Density-separated, T depleted, spleen cells of normal and autoimmune mice were stimulated in vitro with PBA in 48-hr cultures. Cells from old MRL/Ipr and NZB mice were abnormal in both the anti-mu response and the LPS response; BXSB mice had normal anti-mu responses. These studies suggest that there is no prerequisite for spontaneous splenic hyperactivity in the development of autoimmunity. In addition, different PBA stimulate separate subsets of B cells that differ in their state of activation in the various autoimmune strains. Finally, different B cell subsets appear to be abnormal in different types of autoimmune mice.
Long-term in vitro culture of murine mast cells. III. Discrimination of mast cells growth factor and granulocyte-CSF.
Authors: Yung Y P , Moore M A
Published: December 2022

Long-term in vitro growth of murine mast cells was dependent on the presence of a mast cell growth factor (MCGF) present in media conditioned by mitogen-activated splenic leukocytes or by various murine leukemic cell lines. MCGF shared a number of properties with granulocyte colony-stimulating factor (G-CSF). Both factors were present in media conditioned by the myelomonocytic leukemic WEHI-3 and the T cell lymphoma, LBRM-33 cell lines. They were relatively sensitive to trypsin treatment, and were resistant to boiling temperature. NZB mice that failed to respond to WEHI-3-derived G-CSF also failed to respond to MCGF. MCGF differed from G-CSF, however, in sensitivity to neuraminidase and lactoferrin, an inhibitor of macrophage CSF production, suppressed G-CSF production by WEHI-3 cells without affecting MCGF production. Furthermore, peritoneal cells produced G-CSF but not MCGF when stimulated with lipopolysaccharide. In vitro production of MCGF by normal spleen cells required the presence of T lymphocytes and is relatively macrophage-independent. The role of T cells in the maturation and growth of mast cells and the physiologic function of MCGF are discussed.
Dissociation between cutaneous basophil hypersensitivity and basophil sensitization.
Authors: Lett-Brown M A , Thueson D O , Grant J A
Published: December 2022

We investigated the antigen-specific sensitization of basophils in the peripheral circulation of guinea pigs using an in vitro histamine release assay. Animals were primed for cutaneous basophil hypersensitivity (CBH) by intradermal injection of either keyhole limpet hemocyanin (KLH) or ovalbumin (OA) in saline. Basophils isolated from these animals 7 days after sensitization did not release histamine to the specific antigen in vitro. On day 9, however, basophils did release histamine when challenged with the antigen. These results were in contrast to 24-hr skin tests that showed a typical CBH profile both on day 7 and day 9. When animals were immunized via the footpad by emulsification of antigen in incomplete Freund's adjuvant, basophil sensitization (i.e., histamine release) either occurred earlier (day 7, KLH) or at lower antigen concentrations (OA). Thus, dissociation of in vivo CBH and in vitro basophil sensitization was demonstrated. The lack of correlation was only seen early after immunization and depended on the route of immunization used. An additional finding was a transient blood basophilia that also was dependent on the route of immunization, the antigen used, and the time post-sensitization. Our results are consistent with the hypothesis that at early times post-immunization, CBH is mediated by T cells and not via homocytotropic antibodies, but that at later times such antibodies may also play a role in the elicitation of CBH reactions.
Altered levels of mononuclear leukocytes in tumor-bearing rats: decrease of helper T lymphocytes and increase of suppressor cells.
Authors: Fernandez-Cruz E , Gilman S C , Feldman J D
Published: December 2022

In the spleen and peripheral blood of BN rats with progressive tumors, W3/25+ T helper cells were significantly reduced and OX8+ T suppressor/cytotoxic cells were significantly increased. The ratio of helper to suppressor elements was decreased to 1.6 from a ratio of 3 in normal BN rats without tumors, and this decreased ratio correlated with tumor growth. When tumors were eliminated in vivo by infusion of effector cells (W3/25+ T lymphocytes), the levels of W3/25+ and OX8+ T cells returned to normal and the ratio of helper to suppressor/cytotoxic cells in the spleen and peripheral blood reverted to 3.0 or higher. Macrophages and null cells, T-sIg-, were also elevated in the spleen and peripheral blood of rats bearing expanding tumors and returned to normal levels after cure. Assays of spleen cells for cell-mediated cytotoxicity in rats with large tumors revealed little or no specific cytotoxicity. Cytotoxic activity was high in spleen of rats cured of their neoplasms by infusion of helper cells.
Inhibition of cytotoxic T lymphocyte activity by concanavalin A.
Authors: Sitkovsky M V , Pasternack M S , Eisen H N
Published: December 2022

Several lectins were tested for the ability to interfere with the recognition or lysis of target cells by cytotoxic T lymphocytes (CTL). Of those tested, concanavalin A (Con A) had the greatest effect: at 0.25 to 10 micrograms/ml, it profoundly blocked antigen-specific lysis of target cells by cloned and uncloned CTL in various allo- and syngeneic systems. Separate pretreatment of CTL and target cells showed that the effect of Con A is exercised predominantly on CTL. Various experimental results ruled out several possible explanations for the inhibitory effect of Con A, such as: CTL-CTL self-killing by nonspecific, lectin-dependent cytotoxicity, impairment of lateral mobility of CTL surface components, and cell agglutination, preventing recycling of CTL among target cells. The findings suggest that subagglutinating concentrations of Con A block CTL activity by binding to and interfering with CTL surface structures that have relatively high affinity for Con A and are critically involved in recognition or lysis of target cells.
Formation of IgE-binding factors by rat T lymphocytes. VI. Cellular mechanisms for the formation of IgE-potentiating factor and IgE-suppressive factor by antigenic stimulation of antigen-primed spleen cells.
Authors: Uede T , Ishizaka K
Published: December 2022

Analysis of cellular mechanisms of formation of IgE-binding factors by KLH-primed spleen cells revealed that the presentation of KLH to KLH-primed T cells by adherent cells resulted in the formation of lymphokines that in turn stimulated unprimed lymphocytes to form IgE-binding factors. Lymphokines released from KLH-alum-primed spleen cells induced normal lymphocytes to form IgE-potentiating factors, whereas those released from KLH-CFA-primed spleen cells induced the formation of IgE-suppressive factors. Fractionation of the lymphokines from KLH-primed spleen cells and analysis of cell sources of the lymphokines revealed that multiple factors are involved in the selective formation of one or the other IgE-binding factors. Thus, KLH-alum primed splenic T cells from "inducers" of IgE-binding factors and glycosylation-enhancing factors upon antigenic stimulation, and these factors in combination stimulate unprimed W 3/25+ Fc gamma R+ T cells to form IgE-potentiating factors. Antigenic stimulation of KLH-CFA-primed T cells results in the formation of the "inducers" and glycosylation-inhibiting factors, and these two lymphokines collectively stimulate unprimed W 3/25+ Fc gamma R+ T cells to form IgE-suppressive factors. Cell sources of "inducers" are W 3/25+ Fc gamma R- T cells, cells different from the source of IgE-binding factors. The glycosylation-enhancing factor is derived from W 3/25+ F gamma R+ T cells in KLH-alum-primed spleen; the glycosylation-inhibiting factor is derived from OX 8+ T cells in KLH-CFA-primed spleen. Evidence was obtained that these lymphokines, which modulate the protein glycosylation of IgE-binding factors during their biosynthesis, are derived from antigen-primed T cells and determine the nature of IgE-binding factors formed.
Abnormalities of immunoregulatory T cell subsets in patients with insulin-dependent diabetes mellitus.
Authors: Horita M , Suzuki H , Onodera T , Ginsberg-Fellner F , Fauci A S , Notkins A L
Published: December 2022

Patients with insulin-dependent diabetes mellitus (IDDM) have autoantibodies that react with cells in the islets of Langerhans. To determine whether these patients suffer from a more generalized immunoregulatory disorder, the ratio of phenotypic helper to suppressor cells was evaluated by specific monoclonal antibodies. Our experiments showed that the helper/suppressor cell ratio was significantly increased in patients with IDDM of less of 2 mo duration and then gradually returned to normal. Despite the alteration in the helper/suppressor cell ratio, there was no evidence for polyclonal activation as measured by the number of immunoglobulin-secreting plaque-forming cells in the peripheral blood. There was, however, a significant increase in the number of spontaneous plaque-forming cells in patients suffering from both IDDM and Hashimoto's thyroiditis (HT). Nonetheless, immunoglobulin production after stimulation with pokeweed mitogen was not different in diabetics with or without HT when compared to normal controls. These findings suggest that subtle changes in the immunoregulatory system occur during the early stages of IDDM.
A monoclonal antibody against a surface antigen shared by human large granular lymphocytes and granulocytes.
Authors: Rumpold H , Kraft D , Obexer G , Böck G , Gebhart W
Published: December 2022

A monoclonal IgM antibody VEP13 was raised against a cell membrane antigen by immunizing BALB/c mice with a cell preparation enriched for human large granular lymphocytes (LGL). These cells are known to include the effector cells responsible for natural killer (NK) and killer (K) cell activity. By indirect immunofluorescence, VEP13 antibody reacted with 21.4 +/- 7.9% of peripheral blood lymphocytes (PBL) and more than 95% granulocytes but not with B lymphocytes, monocytes, erythrocytes, thrombocytes, thymocytes, and immature bone marrow cells. Cell suspensions enriched for T cells by means of E-rosetting using neuraminidase-treated sheep red blood cells contained 14.7 +/- 5.5% of VEP13+ cells. When PBL were treated with VEP13 antibody and complement, NK activity against a variety of target cells was greatly reduced and K cell activity was also strongly affected. Enrichment experiments of VEP13+ cells by means of the fluorescence-activated cell sorter revealed that all the NK activity resides in the VEP13+ fraction. This cell fraction was a homogeneous population of LGL. Studying the coexpression of various other antigens as defined by monoclonal antibodies, 84.6 +/- 4.6% of VEP13+ cells also expressed M1, 81.6 +/- 5.5% T10, 47.9 +/- 14.4% Leu 2a, 51.6 +/- 17.0% human Lyt-3, and 6.0 +/- 4.0% HLA-DR antigens. Leu 1, Leu 3a, and T6 antigens were found to be absent from VEP13+ cells. We conclude that VEP13 antibody should be helpful in determining LGL in peripheral blood as well as in tissue sections of tumor patients.
In vitro influenza virus-specific antibody production in man: antigen-specific and HLA-restricted induction of helper activity mediated by cloned human T lymphocytes.
Authors: Lamb J R , Woody J N , Hartzman R J , Eckels D D
Published: December 2022

Cloned human T lymphocytes induced with influenza A virus (A/Texas/1/77) and maintained in continuous culture with T cell growth factor were assayed for helper function in the in vitro production of anti-influenza antibody. Helper function mediated by both cloned helper T cells and normal peripheral blood lymphocytes was highly antigen dose-dependent, requiring lower concentrations than that necessary to induce blastogenesis. Optimal help was observed with 1 X 10(2) cloned T cells per culture, whereas excess helper cells inhibited the response. After culture with influenza A virus-induced cloned helper T cells, the antibodies formed were directed against influenza A and not B virus. Furthermore, the cloned helper T cells despite being specific for matrix protein collaborated in the production of predominantly anti-hemagglutinin antibody, suggesting associative recognition of the two discrete antigens. Cellular interactions between cloned helper cells from an HLA-Dw1,3 DR1,3 individual and erythrocyte rosette-negative cells required HLA-Dw1; DR1 compatibility for the production of specific antibody. This was confirmed by using subclones. Finally, it was observed that supernatants of the cloned helper T cells contained functional activity capable of replacing the parent cells in the production of anti-influenza A virus antibody.
Two subgroups of HLA Bw44 defined by cell-mediated lympholysis that differ in Bw44 expression on platelets and in patterns of genetic linkage disequilibrium.
Authors: Tekolf W A , Biddison W E , Aster R D , Shaw S
Published: December 2022

Possible immunogenic heterogeneity of the HLA-Bw44 antigen was investigated using cytotoxic T lymphocytes (CTL) generated between donors identical for HLA-A2,3,-B7,w44. Highly discriminatory CTL combinations were identified that defined two subgroups of Bw44, designated 44.1 and 44.2. Out of 47 Bw44-positive donors tested in a population study, 30 were lysed by the CTL defining 44.1, and 19 were lysed by the CTL defining 44.2. All Bw44 cells could be typed as either 44.1 or 44.2, except two Bw44-positive cells that were phenotypically homozygous for the serologically defined Bw44 antigen and were lysed by both CTL. No Bw44-negative donors (zero out of 37) expressed either 44.1 or 44.2, although cold target blocking was required to eliminate a contaminating reactivity of one CTL population on Bw35 and some Bw45 cells. CTL were also raised between responder/stimulator combinations mismatched for Bw44. These CTL lysed all Bw44-positive target cells, indicating a CML antigen shared by all Bw44 cells. But clear discrimination of the 44.1 and 44.2 subgroups was obtained when appropriate cold target blocking cells were added. All donors with 44.2 expressed high levels of serologically detectable Bw44 on their platelets, and all with 44.1 expressed low levels (p less than 0.005). Furthermore, population studies indicate that 44.1 is in positive linkage disequilibrium with HLA-A2 and possibly DR4, whereas 44.2 is in positive linkage disequilibrium with HLA-DR7 and possibly HLA-A23, -A26, and -A29. These data suggest the existence of two genetically and functionally different subgroups of Bw44 antigens.
The murine sex-limited protein (Slp): reassessment of its sex limitation.
Authors: Ferreira A , Eichinger D , Nussenzweig V
Published: December 2022

Previous studies in which an alloantiserum was used to measure Slp levels indicated that in certain inbred strains of mice (Slpa), this protein was sex-limited, that is, present only in males. We raised several monoclonal antibodies directed against different epitopes of Slp and used them to develop a sensitive two-site immunoradiometric assay. Using this assay we detected Slp in serum of all Slpa females previously thought to be phenotypically negative. The levels of Slp in these females are about 0.2 to 4% of that of the males of the same inbred strain. The molecule found in serum of females was isolated by affinity chromatography and was found to have the characteristic three-chain structure of male Slp. These findings establish that the presence of Slp in Slpa females is the rule rather than the exception. Quantitative differences similar in magnitude to those found between males and females were also detected among Slpa males; i.e., Sd males have about 100 times more Slp than Sp males. The mechanisms by which androgens determine the extensive quantitative male-female differences, and by which the S region determines large variations among Slpa males, are unknown.
Three M components IgA lambda, + IgG kappa n + IgG kappa h in one patient. II. Structurally different kappa n and kappa h chains associated with H(gamma) chains sharing idiotypic determinants.
Authors: Bouvet J P , Liacopoulos P , Pillot J , Banda R , Tung E , Wang A C
Published: December 2022

Structural analysis of purified IgG kappa h and IgG kappa n molecules of DA myeloma paraproteins indicated that the two IgG had different electrophoretic mobility and that L-kappa h had an unusually heavy m.w. (30,000). Peptide mapping showed the existence of additional peptides in the L-kappa h map when compared to the L-kappa n map. Total amino acid analysis showed that L-kappa h contained two additional cysteine residues and certain other amino acids than L-kappa n contained. Sequence of the first 25 NH2-terminal amino acids showed differences at positions 4, 5, 15, 18, and 21, but both L-kappa h and L-kappa n belong to the same V kappa IV subgroup. IgG kappa n but not IgG kappa h reacted with anti-gamma 1 antiserum, indicating that H chains of these two paraproteins were also different. Sera from rabbits immunized with IgG kappa n or IgG kappa h, and rendered specific for idiotypic determinants by appropriate absorption, were used for idiotype characterization of these components. Immunodiffusion, immunoelectrophoresis, and direct hemagglutination demonstrated that IgG kappa h and IgG kappa n cross-reacted partially. Inhibition tests disclosed that the main anti-idiotypic antibody was directed against a conformation structure of the complete IgG kappa h molecule, whereas cross-reaction was due to partial idiotypic similarity between H chains of the IgG kappa h and IgG kappa n. In addition, each of these paraproteins seemed also to bear private idiotypes on their H and L chains.
Immune responses to complex protein antigens I. MHC control of immune responses to bovine albumin.
Authors: Riley R L , Wilson L D , Germain R N , Benjamin D C
Published: December 2022

Murine T cell proliferative and antibody responses to the multi-determinant protein bovine serum albumin (BSA) are controlled by Ir genes mapping within the H-2 gene complex. Strains possessing the H-2k, H-2a, and H-2d haplotypes are classified as high responders to BSA. In contrast, H-2b strains are low responders to BSA. Genetic mapping experiments employing strains with recombinant H-2 haplotypes indicate that both T cell proliferative and antibody responses are at least in part regulated by genes within the I-A subregion. Studies on the inhibition of T cell proliferation by monoclonal anti-Ia antibodies are consistent with the assignment of an Ir gene for BSA to the I-A subregion and strongly suggest a role for genes within the I-E/C subregions as well. The MHC-mediated control of antibody responses did not affect the affinity or the isotype of the antibody produced. The relative quantities of antibody specific for each of the three domains of BSA appears to be regulated by H-2-linked BSA Ir genes, and domain III antigenic determinants were found to be dominant in the responses of low-responder mice and in the early response of high-responder mice. This domain III epitope dominance essentially disappears by the tertiary response of high-responder mice.
Inhibition of basophil histamine release by anti-inflammatory steroids. II. Studies on the mechanism of action.
Authors: Schleimer R P , MacGlashan D W , Gillespie E , Lichtenstein L M
Published: December 2022

The glucocorticosteroids inhibit the IgE-dependent release of histamine by human basophils with an order of potency that very closely parallels that found in vivo (i.e., triamcinolone acetonide greater than dexamethasone greater than beta-methasone greater than prednisolone greater than hydrocortisone much greater than progesterone approximately tetrahydrocortisone approximately 0). The effect is seen after a 24-hr preincubation with nanomolar to micromolar concentrations of glucocorticoid. In contrast, release of histamine stimulated by the formyl methionine containing peptide f-met-leu-phe, the calcium ionophore A23187, and the tumor-promoting phorbol diester 12-O-tetradecanoylphorbol-13-acetate was not inhibited by 24-hr incubation with the potent glucocorticoid dexamethasone. Dexamethasone inhibited anti-IgE-induced histamine release without altering its rate, suggesting that the glucocorticoids do not inhibit histamine release by elevating the intracellular level of cAMP. Dexamethasone did not consistently alter either the total or occupied basophil IgE Fc receptor number, and therefore the glucocorticoid effect does not appear to be due to the modulation of cell surface Fc epsilon receptor content. These data indicate that steroid hormones inhibit basophil IgE-dependent activation through a specific glucocorticoid receptor. The mechanism by which they do so appears not to involve an elevation of cAMP or a shedding of cell surface Fc epsilon receptors. Further, because the glucocorticoids did not inhibit release initiated by the PLA2-dependent stimuli f-met-leu-phe, A23187 and TPA, the inactivation of IgE-dependent histamine release by glucocorticoids may not be the result of PLA2 inhibition.
The capsule of cryptococcus neoformans passively inhibits phagocytosis of the yeast by macrophages.
Authors: Kozel T R , Gotschlich E C
Published: December 2022

We examined the mechanism by which cryptococcal capsular polysaccharide inhibits phagocytosis of Cryptococcus neoformans by macrophages. O-Acetyl and carboxyl groups are major structural features of serotype D polysaccharide. Serotype D capsular polysaccharide or encapsulated whole cryptococci were de-O-acetylated by alkaline hydrolysis with 0.1 M NaOH. Carboxyl groups were reduced by treatment with 1-ethyl-3(3-dimethylaminopropyl)carbodiimide followed by reduction with sodium borohydride. De-O-acetylated or carboxyl-reduced polysaccharides had phagocytosis-inhibiting properties that did not differ appreciably from the untreated polysaccharide; thus, neither the O-acetyl nor the carboxyl groups were essential phagocytosis-inhibiting determinants. Antiserum specific for these noninhibitory groups was obtained by adsorption of cryptococcal antiserum with de-O-acetylated or carboxyl-reduced whole cells to produce antiserum enriched respectively with antibody specific for the O-acetyl and carboxyl groups. These adsorbed antisera showed opsonic activity for the untreated yeast that was similar to the unadsorbed antiserum when these antisera were compared at identical levels of precipitating antibody. We also examined the ability of Fab' fragments of opsonic IgG to reduce the amount of anticapsular IgG needed to opsonize the yeast. No synergy was noted between Fab' fragments and undigested IgG in opsonization. These results are consistent with a passive mechanism for inhibition of phagocytosis. The capsule does not directly modulate phagocytic function, but instead, presents a surface that is not recognized by the phagocyte. This absence of recognition is corrected by opsonizing antibody specific for any surface determinant on the capsule.
Generation of long-term human cytolytic cell lines with persistent natural killer activity.
Authors: Krensky A M , Ault K A , Reiss C S , Strominger J L , Burakoff S J
Published: December 2022

Human cell lines maintained by in vitro stimulation with the HLA-A, B-negative, DR-positive, Epstein Barr virus-transformed, lymphoblastoid cell line Daudi in the presence of conditioned medium demonstrated significant NK activity for over 6 wk in continuous culture. These cells lyse K562 and a broad panel of lymphoblastoid cell lines but do not lyse normal peripheral blood lymphocytes or pokeweed mitogen blasts. They possess the sheep red blood cell receptor but lack other T cell markers (Lyt-3+, OKT3-). Natural killer activity correlated with the presence of a Mac 1-positive subpopulation of cells present in these long-term lines.
Characterization of HNK-1+ (Leu-7) human lymphocytes. II. Distinguishing phenotypic and functional properties of natural killer cells from activated NK-like cells.
Authors: Abo T , Balch C M
Published: December 2022

HNK-1+ and HNK-1- lymphocytes from human blood were sorted with a fluorescence-activated cell sorter and compared for their phenotypic and functional properties. The HNK-1+ cells exhibited virtually no response to either mitogens (PHA, Con A, and PWM) or allogeneic cells, because the 3H-thymidine uptake for HNK-1+ cells was only 1 to 3% of that for HNK-1- cells. On the other hand, the HNK-1- cells responded efficiently to these stimuli and acquired a potent killing activity against K562 and other target cells after stimulation. The proliferating lymphoblasts did not acquire the HNK-1 antigen. These activated HNK-1- cells had a wider spectrum of spontaneous cytotoxicity than did HNK-1+ cells against 12 different target cells. Two distinct populations of effector cells for spontaneous killing can thus be distinguished on the basis of HNK-1 antigen expression. The classically defined NK cells are HNK-1+, whereas effector cells activated by either mitogens or allogeneic cells are HNK-1-. The HNK-1+ cells were not generated from the pool of HNK-1- cells under any of the culture conditions tested. The NK cells defined by the HNK-1 antibody are thus a functionally distinct population of cells. Although some HNK-1+ cells may also express T cell-associated antigens and sheep erythrocyte receptors, they lack some important functional properties ascribed to T cells.
Broadly reactive murine cytotoxic cells induced in vitro under syngeneic conditions.
Authors: Dorfman N , Winkler D , Burton R C , Kossayda N , Sabia P , Wunderlich J
Published: December 2022

Mouse spleen cells became cytotoxic in short-term 51Cr-release assays for a wide variety of target cells after 5 days of culture in vitro with polyinosinic acid in a system that was otherwise entirely syngeneic. This study characterizes these effector cells with respect to target specificity, effect of donor age, time course of their appearance, mouse strain differences, and expression of differentiation antigens Thy-1, Lyt-1, Lyt-2, NK-1, and asialo GM1. The combination of properties of this cytotoxic cell response that make it unique are that a) the broadly reactive cytotoxic activity developed from unprimed spleen cells in the absence of either foreign cells or foreign serum; b) the response did not peak until 4 to 5 days of culture in vitro; c) the broad reactivity pattern included freshly dispersed primary syngeneic sarcoma cells and cultured syngeneic fibroblasts but did not include syngeneic lymphoblast target cells; d) the response was largely monoclonal as defined by target cell binding; and e) cytotoxic cell activity was sensitive in complement-mediated treatments to both anti-NK and anti-theta but not to anti-Lyt-2, anti-Lyt-1, or anti-asialo GM1. Both high- and low-responding mouse strains have been identified.
Development of cytotoxic T lymphocyte precursor cells in T cell colonies grown in vitro.
Authors: Ching L M , Miller R G
Published: December 2022

T cell colonies grown in vitro from normal mouse spleen cells contain cytotoxic T lymphocyte precursor cells (CLP) that develop from immature precursors within the colonies. The colony-forming unit for CLP development (CFU-TCLP) has two components. One is Thy-1-, Lyt-1-, Lyt-2- and is found in spleen, nude spleen, and bone marrow. It can by itself produce a colony, but these colonies do not contain CLP. The other component is Thy-1+, Lyt-1-, Lyt-2- and is found in spleen and lymph node. The Thy-1+ component gives rise to the CLP that develop in the colony. A single cell in this component can give rise to multiple progeny CLP. Each colony develops CLP of several different specificities. The full repertoire developed includes specificities specific for allogeneic H-2, TNP-modified-self H-2 and TNP-modified-allogeneic H-2. Although CLP specific for TNP-modified-allogeneic stimulator cells are definitely present, there is a distinct preference for TNP-modified self. The results lead to the tentative conclusion that the T cell repertoire can develop in the absence of the special environment provided by the thymus.
Regulation of murine macrophage Ia antigen expression by an immune interferon-like lymphokine: inhibitory effect of endotoxin.
Authors: Steeg P S , Johnson H M , Oppenheim J J
Published: December 2022

The initiation of antigen-specific, T-dependent immune responses by macrophages requires their expression of Ia antigens. Ia antigen-deficient (Ia-) peritoneal exudate or P388D1 cell line macrophages, when incubated for 2 to 3 days with partially purified immune interferon (IFN-gamma), expressed Ia antigen, as detected by antibody-and-complement-mediated cytotoxicity. This paper reports that bacterial endotoxin (lipopolysaccharide, LPS) inhibited both IFN-gamma induction of macrophage Ia antigen expression and IFN maintenance of the Ia+ state in a dose-dependent manner. In the absence of IFN-gamma, LPS had no significant effect on macrophage Ia antigen expression. The inhibitory effects of LPS were abrogated by the addition of indomethacin into the culture medium. Further, 10(-10) to 10(-6) M exogenous prostaglandin E2 or 10(-6) to 10(-4) M exogenous dibutryl 3'5' cyclic adenosine monophosphate also inhibited IFN-gamma regulation of macrophage Ia antigen expression. The data suggest that LPS inhibits IFN-gamma regulation of macrophage Ia antigen expression by stimulating macrophage prostaglandin E2 production, and consequently enhancing intracellular cAMP levels. The data outline an inhibitory pathway involved in the regulation of macrophage Ia antigen expression, and may explain, in part, reported immunosuppressive effects of LPS.
The limited diversity of the mouse gamma-chains anti-GAT repertoire does not seem to be noticeably amplified upon class switch.
Authors: Rocca-Serra J , Mazie J C , Moinier D , Leclercq L , Somme G , Theze J , Fougereau M
Published: December 2022

NH2-terminal sections of H and L chains isolated from five monoclonal anti-GAT antibodies derived from BALB/c mice have been sequenced upon to residue 43. Four among these five antibodies, sharing similar public idiotypic determinants, possess extremely conserved sequences, both for the H, which is apparented to the VH II type, and the L chains, which belong to the V kappa I subgroup. VH sequences are identical up to residue 43 and contain the common sequences (residues 1 to 32) defined for the H chains derived from the DBA/2 IgM anti-GAT monoclonal antibodies. Light chains are also remarkably conserved, a rather unusual situation for kappa-chains. The fifth antibody that expresses only part of the public idiotypic determinants contains very distinctive H and L chains. Its heavy chains are close to the VH I subgroup, whereas its kappa-chains permit definition of a new V kappa subgroup. The repertoire appears to be highly conserved between BALB/c and DBA/2 mice, and does not seem larger in IgG than in IgM antibodies. This latter observation does not speak in favor of a switch-linked amplification of diversity.
Glycosylation and secretion of an altered immunoglobulin heavy chain in mouse myeloma MOPC 315.
Authors: Sonenshein G E , McCormack J
Published: December 2022

A variant line (LV-1) of mouse myeloma MOPC 315 (IgA, lambda 2) has lost the ability to synthesize L chain. It synthesizes an altered H chain (H' chain) that is turned over intracellularly and is not secreted. Rescue of H' chain secretion can be accomplished by fusion of LV-1 to a variant of another myeloma line, MPC 11 (IgG2b, kappa), which only synthesizes a light chain. The hybrid (X-2) secretes the H' chain in a four chain structure (kappa 2 alpha' 2). In wild-type MOPC 315 cells, it was reported previously that inhibition of core sugar addition blocks the secretion of the H chain polypeptide. We have studied glycosylation in MPOC 315 wild-type, LV-1 variant, and X-2 hybrid cell lines. The ability of all three lines to add the core sugars mannose and glucosamine to heavy chain was demonstrated. Due to the instability of the H' chain in LV-1, it is difficult to assess H' chain fucosylation directly. To study fucose addition in LV-1, the enveloped virus vesicular stomatitis (VSV), which can infect the three lines, was utilized. The fucosylation and secretion of VSV glycoprotein G was discernible in all three lines; however, only LV-1 cannot activate free fucose, and instead fucosylates through conversion of the mannose intermediate. Normal fucose addition to H chain in a wild-type cell occurred immediately before secretion. The fact that degradation of H' chain in LV-1 begins before fucosylation suggests that the rescue of H' chain secretion by formation of the X-2 hybrid is due to the acquired presence of a suitable L chain rather than complementation of a sugar defect. These observations indicate that proper assembly of the polypeptide components of some secretory proteins, e.g., Ig molecules, is required for the secretion of the individual chains.
Analogous antigenic alterations elicited in C3 by physiologic binding and by denaturation in the presence of sodium dodecylsulfate.
Authors: Nilsson U R , Nilsson B
Published: December 2022

The expression of three antigenic subsets of C3--the C3(S), the C3(N), and the C3(D) antigens--by soluble and target-bound forms of C3 was studied. The C3(S) subset is stable and is expressed by native as well as denatured C3 (exposure to sodium dodecyl sulphate (SDS) M greater than or equal to 10(-3)). The C3(N) and C3(D) subsets are labile and are expressed by native and denatured C3, respectively. Antisera to native C3, anti-C3(S-N), react with the C3(S) as well as the C3(N) subset. Antisera to isolated C3 subunits react exclusively with the C3(D) subset. A separation of anti-C3(S) and anti-C3(N) antibodies was accomplished by adsorbing the anti-C3(S-N) antiserum with insolubilized, denatured C3, anti-C3(N) antibodies remained unadsorbed. Anti-C3(S) antibodies were adsorbed and subsequently eluted from the denatured C3. Agglutination studies with EAC1423b cells showed significant agglutination with anti-C3(S) and anti-C3(D) antisera but reduced agglutination with anti-C3(N) antisera. Agglutination by anti-C3(D) antisera was unaffected in the presence of EDTA serum containing converted or unconverted C3. These data suggest an antigenic modification of C3b-b' upon binding that mirrors the antigenic transition associated with SDS denaturation of C3.