bluuebunny/crossref_metadata_2025 huggingface dataset:
Natural and antibody-dependent cell-mediated activity against Salmonella typhimurium by peripheral and intestinal lymphoid cells in mice.
Authors: Nencioni L , Villa L , Boraschi D , Berti B , Tagliabue A
Published: December 2022

Cell-mediated immune responses were assessed employing a 2-hr in vitro cytotoxicity assay against S. typhimurium. It was observed that lymphocytes from GALT as well as from peripheral lymphoid organs possessed natural antibacterial activity, whereas macrophages were devoid of this spontaneous activity. The distribution of this newly described natural activity was PPL greater than MnL greater than IEL = SpL = PBL greater than PoL; this did not correlate with the organ distribution of NK activity against YAC-1 tumor cells, which was PBL greater than SpL = IEL greater than MnL = PoL = PPL. Moreover, the phenotype of the splenic effector cell of the natural activity against S. typhimurium showed some differences from that of NK activity. In fact, both these cells were asialo GM1+, Fc-receptor+, nonadherent, and nonphagocytic, but the former was Thy-1.2- and the latter Thy-1.2+. The effector cell of the natural antibacterial activity in the Peyer's patches had the same phenotype as the splenic one. It was then observed that the antibacterial activity could be augmented by the addition of immune antibodies against S. typhimurium. This was particularly evident employing IEL, SpL, and PBL as effector cells, whereas PPL and MnL did not show any antibody-dependent antibacterial activity. Furthermore, these last two populations could not mediate ADCC against CRBC. Employing selective methods to deplete cell populations, we observed that, at least at the splenic level, there is also a cell that differs in its phenotypic characteristics from that mediating natural antibacterial activity but that plays a role in the antibody-dependent reactions. In conclusion, these results suggest that natural and antibody-dependent antibacterial mechanisms might be important in defense against S. typhimurium, particularly at the gastrointestinal level, where many bacterial infections first take place and begin to interact with the host immune system.
Pichinde virus-specific cell-associated suppression of primary footpad swelling in an inbred strain of Syrian hamsters.
Authors: Chan M , Clark D , Rawls W E
Published: December 2022

Pichinde virus causes a lethal disease after i.p. inoculation of adult MHA hamsters; other strains of Syrian hamsters are resistant to this lethal infection. During studies of cell-mediated immune responses to Pichinde virus, it was noted that MHA hamsters survived infection when the virus was given in the footpad. However, unlike the resistant LSH and LVG strains of hamsters, the MHA hamsters did not manifest a footpad swelling response. Failure of the MHA hamster to respond to a footpad inoculation of Pichinde virus was shown to be virus-specific and appeared to be mediated by a cell-associated suppressor mechanism.
Immunization with idiotypic immunoglobulin protects against development of B lymphocytic leukemia, but emerging tumor cells can evade antibody attack by modulation.
Authors: Stevenson F K , Gordon J
Published: December 2022

Immunization of strain 2 guinea pigs with idiotypic IgM obtained from serum of animals in the terminal phase of the L2C leukemia produced high levels of circulating anti-idiotypic antibody that was cytotoxic for the tumor cells in the presence of syngeneic C. Such animals showed good protection against injected tumor cells, but after a long delay, leukemia did develop. As tumor cells appeared in the blood, anti-idiotype was consumed, but the cells displayed little or no surface Ig and could not be killed in vitro by anti-idiotype and C. However, as serum antibody levels fell, the cells displayed increasing amounts of surface IgM lambda of the same idiotype and regained their susceptibility to the antibody and C in vitro. It appears, therefore, that the ability of tumor cells to modulate their target idiotype can present a considerable problem for antibody-mediated attack.
Regulation of the immune response to tumor antigens. IX. In vitro Lyt-1+2- cell proliferative responses to cellbound or subcellular tumor antigen.
Authors: Carter R H , Drebin J A , Schatten S , Perry L L , Greene M I
Published: December 2022

We investigated the cellular basis of immune reactivity to the S1509a fibrosarcoma in tumor-immune A/J mice. In a Winn assay, immune Lyt-1+2- T cells are capable of retarding S1509a tumor growth in naive A/J mice. In vitro proliferation to S1509a is also mediated by tumor-immune Lyt-1+2- T cells. This response is specific to the immunizing tumor and appears 5 to 7 days after reexposure to the tumor in vivo. Proliferation also requires the presence of a population of adherent cells. In fact, adherent peritoneal exudate cells pulsed with tumor membrane fragments derived from S1509a cells can stimulate proliferation. Proliferation is blocked by the addition of anti-I-Ak monoclonal antibody to the culture medium without complement or by treatment of the responder population with anti-I-Ak and complement. In vitro responsiveness is also inhibited by the presence of tumor-specific suppressor T cells in vivo. These observations suggest in vitro proliferation may provide a potential means of defining tumor antigens and cell-surface structures involved in tumor immunity.
Evidence for thymic regulation of autoimmunity in BXSB mice: acceleration of disease by neonatal thymectomy.
Authors: Smith H R , Chused T M , Smathers P A , Steinberg A D
Published: December 2022

BXSB mice spontaneously develop an autoimmune syndrome characterized by autoantibody production, immune complex renal disease, and lymphadenopathy containing B cells. The male BSXB mouse has a rapid onset of disease that is determined by defective stem cells. Although the B cell hyperactivity has been traced to the stem cell, previous studies did not evaluate the T cells of BXSB mice with regard to a role in modifying their natural history. The present study was carried out in an attempt to determine whether or not thymic regulation played a role in modulating the stem-B cell abnormalities in BXSB mice. It was found that neonatal thymectomy led to a marked increase in autoantibody production (anti-RBC, anti-DNA), a dramatic increase in lymphadenopathy, and worsening of renal disease. Flow cytometric analysis of the lymph node cells from intact and thymectomized mice demonstrated a loss of Lyt-2+ cells from the thymectomized mice and an increased proportion of Ly-1+, Thy-1.2- cells. These results indicate that although the B cell hyperactivity of BXSB mice is determined by a marrow stem cell defect, thymic regulation normally serves to dampen that B cell hyperactivity; neonatal thymectomy reduces the thymic regulation of autoimmunity in the BXSB males and leads to excessive B cell numbers and function.
Constitutive production of a factor supporting B lymphocyte differentiation by a T cell hybridoma.
Authors: Elkins K , Cambier J C
Published: December 2022

To investigate the role of soluble T cell products during B cell differentiation more fully, we have produced T cell hybridomas by the fusion of normal helper T cells with the T cell lymphoma BW5147. In this report we describe the production of one such hybrid, 14G3, the subclone 14G3.1F2, and the functional activity of the constitutive product. The hybrid supernatant acts exclusively in antigen-nonspecific, but antigen-dependent, promotion of B cell differentiation. It is optimally effective in the presence of small amounts of EL4 supernatant. It does not itself contain any detectable IL 2 or BCGF or interferon activity, however. 14G3.1F2 activity is probably an important component of the conventional TRF preparations produced by mixed lymphocyte populations, and will be useful in further dissection of the contributions of different soluble T cell products to B cell differentiation.
Identification of human natural killer soluble cytotoxic factors (NKCF) derived from NK-enriched lymphocyte populations: specificity of generation and killing.
Authors: Farram E , Targan S R
Published: December 2022

Activated natural killer (NK) cells were induced by stimulation of normal peripheral blood lymphocytes with a B cell line for 5 days or by interferon activation for 1 hr. They were enriched for large granular lymphocytes (LGL) by separation on Percoll density gradients. Supernatants were obtained from incubation mixtures of LGL and NK-sensitive target cells (MOLT-4 or K562). Although some lysis could be detected at 16 hr, optimal NK-sensitive target cell lysis by this supernatant was evident after 40 hr incubation by using a microcytotoxicity assay and trypan blue exclusion. Supernatants derived by co-culture of NK targets with either NK-depleted cell populations or NK cells alone were not cytolytically active. Target cell requirements for both the generation and cytotoxic activity of NKCF were specific. That is, only the NK-sensitive target cells MOLT-4 and K562, and not the NK-resistant cells YAC-1, EL4, IM9, and Raji, could induce release of, or be lysed by, NKCF. Furthermore, the requirement for both Ca2+ and Mg2+ in the generation of NKCF suggests that both target cell binding and initiation of programming for lysis are required for its release. NKCF lytic potential was stable for 2 mo at -20 degrees C and partially stable at 60 degrees C. NKCF activity could be substantially removed by prior absorption on the NK-sensitive target cells, K562 or MOLT-4, but not Raji or IM9 (resistant) cells. Therefore, it appears that the specificity of this cellfree lytic factor depends on its recognition of, or its being recognized by, appropriate target cells. The properties of NKCF will enable the study of the mechanism involved in the lethal hit of NK cell-mediated cytotoxicity.
Activation of the classical pathway of human complement by a human monoclonal IgE, IgE(DES).
Authors: Saint-Remy J M , Magnusson C G , Masson P L
Published: December 2022

Activation of the classical pathway of human complement by monoclonal IgE from patient DES was demonstrated by using IgE(DES) coupled to latex particles. This material depleted human serum of C1 and C4 hemolytic activities. In addition, C3bi was deposited in a calcium-dependent way onto the insolubilized IgE as shown by the agglutination of latex by conglutinin. The alternative pathway was also activated. These anticomplementary activities were dose and time dependent. Moreover, we confirmed that another monoclonal IgE, IgE(PS), activated the alternative pathway exclusively. Particular attention was paid to exclude contamination by other immunoglobulins or C-reactive protein, generation of artifacts due to the chemical coupling, and the presence of proteolytic enzymes in the IgE(DES) preparation. Moreover, evidence is also presented against the involvement of IgG or IgM anti-IgE autoantibodies that could activate the classical pathway after their binding to insolubilized IgE(DES). Although one cannot exclude the possibility that IgE(DES) or IgE(PS) are abnormal proteins, these findings suggest the existence of an isotypic or allotypic variation of IgE.
Arachidonic acid metabolism in the murine eosinophil. III. Effect of nonsteroid anti-inflammatory drugs on lymphokine-directed eosinophil migration in vivo.
Authors: Rand T H , Clanton J A , Colley D G
Published: December 2022

In vitro studies of murine eosinophils indicated that lymphokine-stimulated metabolism of arachidonic acid by a lipoxygenase pathway was required for a migration response. In this study we tested the effects of drugs that affect arachidonic acid metabolism on lymphokine-dependent eosinophil accumulation in vivo by the use of 111In-labeled eosinophils. Indomethacin at different dosages either stimulated (1 mg/kg) or partially inhibited (5 to 25 mg/kg) eosinophil accumulation. Aspirin had no significant effects in doses up to 50 mg/kg. BW755C, a drug that inhibits both cyclooxygenase and lipoxygenase, dramatically inhibited eosinophil accumulation at 25 to 125 mg/kg. Pretreatment in vitro of 111In-labeled eosinophils with some drugs known to inhibit lipoxygenase reduced their subsequent accumulation at a lymphokine-injected site in vivo, but the high concentrations required for inhibition suggested that the effects of the drugs were at least partially reversible. Pretreatment with indomethacin did not inhibit the ability of 111In-labeled eosinophils to accumulate at the site of lymphokine injection in vivo. It may be anticipated from these results that drugs inhibiting lipoxygenase will have critical effects on manifestations of immunologic reactions that recruit eosinophils.
Antibodies reactive with liposomal phospholipids are produced during experimental Trypanosoma rhodesiense infections in rabbits.
Authors: Richards R L , Aronson J , Schoenbechler M , Diggs C L , Alving C R
Published: December 2022

Antibodies against phospholipids appeared spontaneously during the course of experimental Trypanosoma rhodesiense infections in rabbits. These antibodies were observed in rabbits infected either with a lethal strain or with a strain newly discovered to give a spontaneous self-cure. Serum antibodies reacting with liposomes containing dimyristoyl phosphatidylcholine (DMPC), phosphatidylinositol (Pl), phosphatidylinositol phosphate (PlP), or cardiolipin were detected at 3 to 4 wk by complement-mediated release of trapped marker from liposomes. Antibodies were also detected against a trypanosomal lipid fraction (TrF2) that contained Pl as a major constituent. The antibody activities against DMPC, Pl, or TrF2 all reacted (or cross-reacted) with DMPC, and were removed from the serum by adsorbing with liposomes containing DMPC as the only phospholipid. Phosphocholine inhibited the antibodies reactive with liposomes containing either DMPC or DMPC and Pl as phospholipids. Antibodies against PlP, however, reacted only with liposomes containing PlP and were not removed by adsorbing with liposomes lacking PlP. We conclude that anti-phospholipid antibodies appear during the course of trypanosomal infections that either undergo apparent self-cure or are lethal, and at least two anti-phospholipid antibody specificities can be detected.
Transformation of murine LMTK- cells with purified HLA class I genes. I. Modification of conformation of murine beta 2-microglobulin upon its association with HLA heavy chains.
Authors: Lemonnier F A , Le Bouteiller P P , Malissen B , Golstein P , Malissen M , Mishal Z , Caillol D H , Jordan B R , Kourilsky F M
Published: December 2022

Murine LMTK- cells were unexpectedly found to cross-react with a murine anti-human beta 2-microglobulin (beta 2-m)monoclonal antibody (m.Ab) after transformation with cosmid clones containing different purified HLA class I genes. The same cross-reactivity was observed with CTP 34 B4 (murine x human) somatic hybrid cells, which express class I molecules constituted of human HLA heavy chains and murine beta 2-m. Inhibition studies of the complement-dependent cytolysis mediated by the cross-reacting m.Ab indicated that isolated murine beta 2-m does not express the cross-reacting determinant, suggesting that its expression by the transformed cells reflects conformational modification of murine beta 2-m upon its association with HLA heavy chains. These results illustrate one of the possible post-translational mechanisms through which the antigenicity of a polypeptide chain can be modified. They might provide a serologic marker of the third domain of HLA class I heavy chains. Finally, because quantitative differences of reactivity with the anti-human beta 2-m m.Ab were observed, depending on the HLA class I genes used for transformation, these results individualize two families of HLA class I heavy chains responsible for different conformational modifications of murine beta 2-m.
Recombinant interferon-gamma increases HLA-DR synthesis and expression.
Authors: Basham T Y , Merigan T C
Published: December 2022

It has been shown that all three classes of interferons enhance the expression of the major histocompatibility class I antigens (HLA-A,B,C;H-2) on a wide variety of cell types (1-10). However, their effect on the expression of the class II antigens (HLA-DR, Ia), which play a major part in cellular interactions that initiate an immune response, is more controversial. The predominate findings have been that the interferons specifically increase the synthesis and expression of only the class I antigens (3, 4, 6, 8, 10, 11). We report here that recombinant interferon-gamma (IFN-gamma) increases the synthesis and expression of the HLA-DR (la-like) antigens as well as beta 2-microglobulin (beta 2-m), a low m.w. subunit of HLA, on human melanoma cells. No increase in HLA-DR was detected on these melanoma cells with leukocyte interferon (IFN-alpha) at doses 400 times higher than the maximum dose of IFN-gamma. These findings were extended to show that pure IFN-gamma also increases the expression of the HLA-DR antigens on normal peripheral blood monocytes, whereas recombinant IFN-alpha at a similar dose had little effect on the expression of this surface antigen. These findings suggest a specialized role for IFN-gamma in immune regulation in comparison with IFN-alpha.
Depletion of human NK cells with monoclonal antibodies allows the generation of cytotoxic T lymphocytes without NK-like cells in mixed cultures.
Authors: Strassmann G , Bach F H , Zarling J M
Published: December 2022

In vitro stimulation of human mononuclear cells with x-irradiated autologous lymphoblastoid cell line (LCL) or allogeneic normal cells in mixed leukocyte cultures (MLC) was previously shown to result in the generation of OKT3+ OKT8+ cytotoxic T lymphocytes (CTL) lytic for allogeneic and autologous LCLs and also of natural killer- (NK) like cells that are OKT3- and primarily OKT8- and are lytic for HLA- NK-sensitive K562 cells. The origin of the NK-like cells was not previously known because, although the majority of fresh human NK cells react with monoclonal antibodies OKM1 and B73.1, lymphocytes bearing these markers are not detected several days after the onset of MLC, when NK-like cells are present. In this study, experiments were undertaken to determine whether NK-like cells generated after stimulation with x-irradiated pooled allogeneic normal cells (poolx) or with autologous LCL are derived from cells expressing antigens reactive with monoclonal antibodies OKM1 or B73.1, which react with fresh NK cells. Mononuclear cells, depleted of monocytes, were stained with OKM1 or B73.1 and fluorescein-labeled goat anti-mouse IgG. Lymphocytes depleted of OKM1+ or B73.1+ cells, by fluorescence-activated cell sorting, and lymphocytes that were stained but not sorted were stimulated for 7 days with either poolx or autologous LCL. The generation of NK-like activity was decreased at least 90% after depletion of cells reactive with OKM1 or B73.1, whereas the generation of CTL against autologous and allogeneic LCL was minimally affected. These findings show that NK-like cells generated in MLC are derived from cells that express the phenotype of fresh NK cells (OKM1+ or B73.1+) and that CTL can be generated in cultures in which relatively little NK-like activity is concomitantly detected, by depleting NK cells with monoclonal antibodies before stimulation.
Inhibition of ongoing myeloma IgE synthesis in vitro by activated human T cells.
Authors: Hassner A , Saxon A
Published: December 2022

The ability of activated T cells to suppress ongoing IgE synthesis in vitro was assessed using U266--a human myeloma cell line spontaneously producing IgE. T cells were able to inhibit U266 IgE synthesis in the presence of 10 micrograms/ml of Con A by 41.8% (p less than 0.01). T cells preincubated with 10 or 50 micrograms/ml of Con A and washed extensively were still able to inhibit U266 IgE synthesis in the absence of Con A by 41 and 46% (p less than 0.05 and p less than 0.02, respectively). The decrease in IgE measured was due to inhibition of newly formed IgE by U266, as shown by control experiments with cycloheximide. The inhibition was not due to the simple depletion of nutrient growth factors by the activated T cells, as it did not occur with MOLT-4, T cells that are very active metabolically; nor could it be reversed with medium containing IL 2 and B cell growth factors. Culture supernatants of Con A-activated T cells were also able to suppress IgE synthesis by U266 (21%; p less than 0.01), which suggests that upon appropriate activation, T cells secrete material(s) with inhibitory properties for IgE synthesis. Activation of T cells by mixed lymphocyte culture using puromycin-treated lymphoblastoid cell lines as stimulators also generated T cells that had suppressive activity for IgE synthesis. T cells activated with Con A and subsequently incubated with IgE demonstrated a diminished ability to suppress IgE synthesis. This observation is in agreement with the finding that patients with high levels of IgE may lack isotype-specific suppressor T cells for spontaneous IgE secretion. However, T cells from such patients have so far shown variable loss of IgE suppressive function. These results suggest that human IgE synthesis is susceptible to inhibition at a very differentiated stage, and this may be important in expression of allergic diseases.
Changes in B lineage cell population in liver and spleen of normal neonatal mice.
Authors: Jyonouchi H , Kincade P W
Published: December 2022

The frequency and characteristics of B lymphocyte lineage cells in neonatal murine liver and spleen were studied during the first 10 days after birth. These were distinguished as B cells with surface IgM (slgM), immediate precursors of B cells (pre-B cells) lacking slgM but containing micron-heavy chains of IgM, and earlier precursors that did not synthesize immunoglobulin but could be detected with monoclonal 14.8 antibody. Experiments were also done to relate these to cells capable of clonal proliferation in mitogen-containing semisolid agar cultures and cells that acquire this function only after preculture in liquid medium. Newborn liver contained large numbers of early precursors as well as pre-B cells, and culture studies revealed that a majority of the colony-forming B cells present at that time were slg-. Adherent accessory cells in newborn liver suspensions facilitated the maturation of these into functional B cells in vitro. At most ages, however, numbers of slg+ B cells detected in that tissue were surprisingly low. Possible explanations for this include a rapid exit of newly formed B cells and their immediate precursors from liver and/or a high rate of abortive lg gene rearrangements during the neonatal period. In contrast, whereas the spleen contained early precursors and pre-B cells at birth, these cells steadily declined in number with age as the numbers of slgM+ B cells increased. Adherent cells in liver but not spleen of immunodeficient CBA/N mice suppressed B lymphocyte formation in semisolid or liquid cultures. These observations document population dynamics in B lineage cells during a critical period of development.
Interactions of lectins and monoclonal antibodies with human mononuclear cells. I. Specific inhibition of OKT4 and OKT8 binding by Ricinus communis agglutinin and wheat germ agglutinin.
Authors: Boldt D H , Dorsey S A
Published: December 2022

We used flow cytometry to examine effects of lectins on interactions between human lymphocytes and the anti-T cell monoclonal reagents OKT4 (T helper-specific) and OKT8 (T suppressor-specific). Wheat germ agglutinin (WGA) inhibited OKT8 binding to lymphocytes by a mean 77% and Ricinus communis agglutinin (RCA-I) inhibited OKT4 binding by 66%. Inhibition was abolished in each case by appropriate carbohydrate hapten inhibitors of lectin binding, indicating it was mediated by the lectin saccharide combining sites. Neither WGA nor RCA-I inhibited binding of OKT3, a pan-T cell monoclonal reagent. In addition, a group of other lectins with a variety of nominal carbohydrate specificities did not inhibit OKT4 or OKT8 binding. Preincubation experiments and gel filtration indicated that inhibition in each case was due to competition between lectin and monoclonal for binding to cell surfaces, not to direct lectin-monoclonal antibody interactions. Treatment of lymphoid cells with OKT8 and complement reduced OKT8- and WGA-binding cells concurrently, whereas treatment with OKT4 and complement did not reduce percentages of either type of cell. Similarly, specific depletion of OKT8-binding cells abolished the mitogenic response to WGA but not that to PHA. Cell populations enriched for WGA-binding cells prepared by flow cytometry and cell sorting demonstrated parallel enrichment for OKT8-binding and depletion of OKT4-binding cells. Therefore, these data demonstrate specific inhibition of OKT4 and OKT8 binding by the lectins, RCA-I and WGA, respectively. Inhibition was mediated by lectin binding to lymphoid cell surfaces, perhaps directly to the T4 or T8 antigens. The observations indicate that lectins may prove useful for investigating structural features of some immunologic cell surface markers. Furthermore, they provide the possibility that certain in vitro effects of lectins on immune function may result from their interactions with molecules such as the T4 and T8 antigens.
Secretion of immune interferon and generation of cytotoxic T cell activity in nude mice are dependent on interleukin 2: age-associated endogenous production of interleukin 2 in nude mice.
Authors: Klein J R , Bevan M J
Published: December 2022

We have examined the relationship between the Con A-mediated induction of CTL effector function and immune IFN secretion in congenitally athymic nude mice. Spleen cells from young adult nude mice (2 mo of age) were found to require an exogenous source of IL 2 for the activation of a CTL effector response and for the release of immune IFN. However, the response of spleen cells from nude mice 9 mo of age differed from that of younger animals in that exogenous IL 2 was not required for the activation of CTL effector function and immune IFN secretion. Spleen cells from old nude mice were found to be capable of making IL 2 upon Con A stimulation similar to normal thymus-bearing animals. The data demonstrate that the maturation of IL 2-secreting cells can occur in athymic mice, that IL 2 is a requisite signal not only for the development of CTL effector function but also for the production of immune IFN, and that CTL effector function and immune IFN release are closely associated immunologic events.
Modulation of the biologic activities of IgE-binding factor. II. Physicochemical properties and cell sources of glycosylation-enhancing factor.
Authors: Iwata M , Huff T F , Uede T , Munoz J J , Ishizaka K
Published: December 2022

T lymphocytes of rats treated with Bordetella pertussis vaccine (BP) formed a soluble factor that enhanced the glycosylation of IgE-binding factors during their biosynthesis, and provided the latter factors with the biologic activity to potentiate the IgE response. The present experiments demonstrated that pertussigen (leukocytosis-promoting factor) from BP induced normal rat spleen cells to form the glycosylation-enhancing factor. The same factor was obtained by incubation of normal spleen cells with 5 micrograms/ml, but not 2 micrograms/ml, concanavalin A. When normal rat mesenteric lymph node cells were incubated with the glycosylation-enhancing factor together with IgE, IgE-binding factors formed by the cells selectively potentiated the IgE response. The IgE-binding factors formed by the same cells upon incubation with IgE alone neither enhanced nor suppressed the IgE response. The glycosylation-enhancing factor changed the nature of IgE-binding factors formed by the rat-mouse T cell hybridoma, 23A4. IgE-binding factors induced by IgE alone lacked affinity for lentil lectin, whereas those induced by IgE in the presence of the glycosylation-enhancing factor had affinity for the lectin. The cell source of the glycosylation-enhancing factor appeared to be W 3/25+ Fc gamma R+ T cells. The glycosylation-enhancing factor was protein in nature and had a m.w. of about 25,000. The factor had affinity for acid-treated Sepharose and could be recovered from the beads by elution with lactose. The factor was different from interleukin 2 with respect to both its affinity for galactose and its isoelectric point.
Preliminary structural characterization of the leukocyte cell surface molecule recognized by monoclonal antibody TA-1.
Authors: LeBien T W , Bradley J G , Koller B
Published: December 2022

TA-1 is a monoclonal antibody identifying a cell surface molecule with a broad distribution on normal and malignant human leukocytes. Preliminary structural studies performed by using radioimmunoprecipitation and sodium dodecyl sulfate polyacrylamide gel electrophoresis indicated that TA-1 recognizes a two-chain polypeptide of approximately 170 kilodaltons (KD) and 95 KD under reducing conditions. The same experiment conducted under nonreducing conditions yielded bands of approximately 155 KD and 110 KD, suggesting the existence of intrachain disulfide bonds in both subunits. Both polypeptide chains were labeled with tritiated sodium borohydride after treatment of cells with neuraminidase and galactose oxidase, thereby demonstrating that both were glycosylated. Tryptic peptide mapping indicated that the 170-KD and 95-KD subunits did not have significant homology in peptide composition. We are designating this newly defined human leukocyte bimolecular complex gp 170/95.
Selective defect of natural killer and killer cell activity against lymphomas in SJL mice: low responsiveness to interferon inducers.
Authors: Kaminsky S G , Nakamura I , Cudkowicz G
Published: December 2022

Three unrelated inbred strains of H-2s mice, SJL/JN, A.SW, and B10.S, were classified according to the levels of splenic NK activity against susceptible lymphoma targets, with or without prior administration of the interferon (IFN) inducer p-I:C to the prospective donors of splenocytes: 1) SJL/JN had low endogenous activity that responded poorly to p-I:C-induced augmentation; 2) A.SW exhibited low levels of spontaneous activity that were inducible to higher levels by p-I:C; and 3) B10.S showed high endogenous levels of activity that were augmented to a still higher level by p-I:C treatment. In vitro incubation of spleen cells with partially purified IFN-beta for up to 12 hr augmented NK activity of A.SW and B10.S but not of SJL mice. The relative levels of K cell-mediated ADCC in the three strains were similar to those of NK activity, including augmentability by p-I:C; however, the capacity to perform three additional cell-mediated immune functions, i.e., natural cytotoxicity against a fibrosarcoma target, adherent cell-mediated ADCC against antibody-coated chicken RBC, and alloimmune T cell-mediated cytotoxicity against appropriate targets, did not distinguish one strain from another. The low NK phenotype in SJL/JN remained distinctive throughout postnatal life, but there was a tendency for the highest levels of NK activity to occur in 4 to 8-wk-old mice.
Direct effect of complement factor C5a on the contractile state of isolated smooth muscle cells.
Authors: Scheid C R , Webster R O , Henson P M , Findlay S R
Published: December 2022

The complement-(C) derived factor C5a has long been recognized as a potent contractile agonist in smooth muscle (1,2); however, controversy remains as to whether the effects of this anaphylatoxin are direct or secondary to the release of histamine (3) and/or other mediators (4-8) from nonmuscle cells within the tissue. To resolve this controversy, we have assessed the contractile effects of purified human C5a and C5a des Arg in a homogeneous preparation of enzymatically dispersed smooth muscle cells derived from the stomach of the toad, Bufo marinus. This preparation, which is insensitive to histamine at concentrations as high as 10(-4) M, responds normally to a variety of electrical (9), mechanical (10), and pharmacologic (11, 12) stimuli. These smooth muscle cells also respond to purified human anaphylatoxin; exposure to the cells to purified human C5a or C5a des Arg produce contractions of the smooth muscle cells that are accompanied by increased Ca2+ influx. The contractile response was unaffected by antagonists to histamine or acetylcholine but was reduced by 30% by pretreatment with the leukotriene antagonist FPL55712. A direct contractile effect of C5a on amphibian smooth muscle cells is suggested.
Induction of B lymphocyte proliferation by monoclonal anti-Lyb 2 antibody.
Authors: Subbarao B , Mosier D E
Published: December 2022

Monoclonal antibody to Lyb 2, a differentiation antigen present on all B cells, has been used to study the role of Lyb 2 molecules in B cell activation. Monoclonal anti-Lyb 2 antibody (m-anti-Lyb 2) transforms resting B cells into blast cells and induces proliferation in these activated B cells. The proliferative response to anti-Lyb 2 is a property of the Lyb 5+ subset of B cells, since the antibody fails to stimulate B cells from mice expressing the CBA/N immune defect. B cells activated by anti-Lyb 2 mature into antibody-secreting cells in the added presence of T cell-replacing factors contained in the supernatants from concanavalin A-activated T cells. Thus, Lyb 2 molecules may participate in the delivery of stimulatory signals during the early phases of B lymphocyte activation.
The involvement of antigen-presenting cells and suppressor cells in the ultraviolet radiation-induced inhibition of secondary cytotoxic T cell sensitization.
Authors: Jensen P J
Published: December 2022

When mice were treated with ultraviolet (UV) radiation before immunization with the skin sensitizers, trinitro-chlorobenzene or dinitrofluorobenzene, they showed greatly depressed levels of priming for a secondary in vitro cytotoxic response against haptenated cells. Because the irradiation and skin painting were done on separate sites, these results suggest that the UV radiation had a systemic effect. The priming response of irradiated mice to allogeneic stimulation was normal, indicating some antigen selectivity to the inhibitory effect of UV radiation. A defect in antigen-presenting cells, previously demonstrated in UV-irradiated mice, was found to be largely responsible for the depressed priming response observed in these animals. In addition, the UV-irradiated, immunized mice possessed suppressor cells that were capable of blocking priming for cytotoxic responses against haptenated cells in normal mice.
Functional lymphocytes in the chicken pineal gland.
Authors: Cogburn L A , Glick B
Published: December 2022

The primary antibody response of lymphoid tissue occupying the pineal gland of 6-wk-old chickens was studied subsequent to injection of the carotid artery with sheep red blood cells (SRBC) or bovine serum albumin (BSA). Injection of SRBC did not produce plaque-forming cells (PFC) among pineal lymphocytes whereas BSA stimulated synthesis of anti-BSA immunoglobulin in pineal lymphoid tissue. A cytotoxic assay using appropriate anti-lymphocyte sera indicated that single-cell suspensions of pineal lymphocytes were composed of 42% B lymphocytes and 51% T lymphocytes. Bursal and thymic lymphocytes labeled with tritiated thymidine migrated into pineal lymphoid tissue when injected into 4- and 5-wk-old naive chicks. These observations indicate that the bursa and thymus make equivalent contributions to the lymphoid mass in the chicken pineal gland. Challenge of pineal-established lymphocytes by antigen introduced via the blood vascular system suggests that soluble antigens--rather than particulate ones--stimulate antibody production in the pineal gland. Collectively, these studies indicate that the pineal gland should be considered as a functional component of the chicken's lymphomyeloid system.
Analysis of cellular immune response to EBV by using cloned T cell lines.
Authors: Slovin S F , Schooley R T , Thorley-Lawson D A
Published: December 2022

Eight cloned T cell lines specific for Epstein Barr virus-transformed B lymphocytes were derived. In the presence of the autologous virus-infected B cells, the T cell lines show HLA-restricted cytotoxic activity and also secrete alpha-interferon in sufficient amounts to inhibit infection and transformation. Four of these clones showed restriction to a single HLA locus (two for A3, and two for B7) and three showed exquisite self-restriction lysing only autologous targets. These seven clones expressed the classical cell surface phenotype of cytotoxic T cells being T3, 8, 11, and la-positive and T4-negative. An eighth clone that lacked the T8 surface marker appeared to recognize both B7 and BW51. HLA restriction was confirmed: 1) by the ability of a monoclonal antibody against an HLA-A,B,C framework antigen (W6-32) to block the cytotoxicity; 2) the failure of the clones to lyse Daudi, an EBV-positive, HLA-A,B, C-negative cell line; and 3) successful competition of the cytotoxicity by autologous but not allogeneic cold targets. The cloned T cells do not kill EBV-negative targets such as autologous pokeweed mitogen blasts and cell lines including CEM and the natural killer cell target K562. The results suggest T cell clones may be generated against an EBV-associated membrane antigen on transformed B cells, perhaps equivalent to the lymphocyte-determined membrane antigen, and that the recognition is restricted by a single HLA determinant. We propose that single T cells can play multiple roles in controlling EBV infection in vitro and in vivo including the elimination of transformed cells by cytotoxicity and the prevention by secreted interferon of further re-infection and transformation.
Characterization of anti-tubular basement membrane antibodies in rats.
Authors: Zanetti M , Wilson C B
Published: December 2022

Autoimmune tubulointerstitial nephritis was induced in Brown-Norway (BN) rats by immunization with bovine (Bov) tubular basement membrane (TBM) in complete Freund's adjuvant. Serum antibodies thus produced reacted to a greater extent with Bov than BN TBM antigens by indirect immunofluorescence and by radioimmunoassay with particulate (P) and collagenase-solubilized (CS) TBM. The quantities of antibodies reactive with CS TBM correlated with the intensity of tubulointerstitial pathologic changes. Antibodies eluted from kidneys reactive with BN TBM by indirect immunofluorescence were 508 times more concentrated in the kidney than in the serum, compared with 15 times for Bov TBM-reactive antibodies. The reactivity of eluted antibodies to P BN TBM was inhibited by 70% after absorption with BN CS TBM. A major CS TBM antigen of 42,000 m.w. was identified by polyacrylamide gel electrophoresis. This antigen was present in both Bov and BN TBM, and may be important in triggering autoantibody formation in this model. Lewis rats immunized under the same conditions produced antibodies reactive with BN TBM by immunofluorescence but failed to develop immune deposits in TBM of their own kidneys. Analysis of serum anti-TBM antibodies in Lewis rats revealed a selective lack of reactivity with either homologous or autologous CS TBM. These results suggest that the ability to make an immune response to one or more elements of CS TBM plays a major role in the development of autoimmune tubulointerstitial nephritis in rats.
Clonal analysis of HLA-restricted virus-specific cytotoxic T lymphocytes from cerebrospinal fluid in mumps meningitis.
Authors: Fleischer B , Kreth H W
Published: December 2022

T lymphocytes were cloned directly from the cerebrospinal fluid of a patient with mumps meningitis by limiting dilution in the presence of irradiated feeder cells and T cell growth factor. Of 84 colonies analyzed, 41 were cytotoxic, as shown by their ability to exert phytohemagglutinin-dependent killing. Of these, 39 showed specificity for the autologous mumps-virus infected target cells. The cytotoxic T cell colonies showed the same pattern of HLA restriction as bulk cultures of CSF lymphocytes. This study shows that it is possible to perform functional assays on inflammatory exudate cells at the clonal level. The data also suggest that recruitment of effector cells from the peripheral compartment into the meningeal spaces in mumps meningitis is highly antigen-specific.
Suppression of T cell cytotoxicity by nude mouse spleen cells: reversal by monosaccharides and interleukin 2.
Authors: Palladino M A , Ranges G E , Scheid M P , Oettgen H F
Published: December 2022

The effects of monosaccharides on the suppression of cytotoxic T cell generation by spleen cells from nu/nu mice were examined. Suppression of the B6 anti-BALB/c response and the B6 anti-C3H response was reversed by alpha-methyl-D-galactoside (alpha MG) but not other sugars, including beta MG. Suppression was associated with a decrease in the level of IL 2, which suggests competition; this decrease was also reversed by alpha MG.
In vivo analysis of allogeneic lymphocyte interaction: activation of suppressor T cells by an I-J-restricted allogeneic effect factor (AEF).
Authors: Bromberg J S , Delovitch T L , Kaufman K , Greene M I
Published: December 2022

The preceding paper detailed the production and fractionation of a T cell-derived I-J-specific allogeneic effect factor (AEF) and analyzed its ability to provide help in a T cell-depleted, in vitro primary anti-sheep erythrocyte response. The identical AEF fractions were examined in this study for their ability to elicit suppression in a delayed-type hypersensitivity assay. Previous reports showed that low or suboptimal doses of antigen, presented i.v. on a cell surface, induce a precursor or primed set of suppressor T cells (pre Ts). These cells manifested antigen-specific suppression only in the presence of a T cell-mediated I-J-specific allogeneic effect induced in vivo against the pre Ts. The experiments reported here examined the ability of alloactivated T cell-derived I-J-specific AEF components to replace the in vivo I-J allogeneic effect. The results show that certain AEF components can indeed provide the signal(s) necessary for activation of suppression. Size and charge separation of the crude AEF preparation revealed several components, some of which could independently serve as appropriate inductive signals. One of these components proved to be biochemically identical to interleukin 2 (IL 2) and accounted for some of the genetically unrestricted AEF activity observed; other higher m.w. molecules also possessed unrestricted activity. Another component provided the requisite activational signals and this 68,000-dalton, pI 5.6 molecule(s) was I-J restricted. These findings are discussed in terms of models of lymphocyte subset interactions and activation.
The origin of chemiluminescence produced by neutrophils stimulated by opsonized zymosan.
Authors: Cheung K , Archibald A C , Robinson M F
Published: December 2022

The luminol-dependent chemiluminescence (CL) of neutrophils phagocytosing zymosan is inhibited by superoxide dismutase (SOD), catalase, sodium benzoate, and 2,5-dimethyl furan. In the present report it is shown that inhibition by SOD and 2,5-dimethyl furan is diminished and removed, respectively, by the omission of glucose from the incubation medium. Zymosan-induced CL is also inhibited by inhibitors of arachidonic acid (AA) metabolism, including 5,8,11,14-eicosatetraynoic acid, nordihydroguaiaretic acid, quinacrine, indomethacin, and aspirin, by prostaglandins E1 and E2, theophylline, and dibutyryl cyclic AMP (cAMP), and by the addition of AA, sodium fluoride, and xanthine oxidase plus xanthine to the cell suspension. These findings lead us to postulate that the metabolism of AA via the lipoxygenase (and cyclooxygenase) pathway(s) is the source of CL observed in neutrophils after phagocytosis. Reactive oxygen species produced as a result of activation of NAD(P)H oxidase provide oxidizing agents for the oxidation of AA along these pathways. It is also suggested that elevated levels of cAMP induced by prostaglandins synthesized via the cyclooxygenase pathway may play a role in the regulation of the zymosan-induced CL response.
Skin allografts generate an enhanced production of histamine and histamine-producing cell-stimulating factor (HCSF) by spleen cells in response to T cell mitogens.
Authors: Dy M , Lebel B
Published: December 2022

In response to T cell mitogens, spleen cells produce a large amount of histamine, whereas no or a slight increase is observed after B cell mitogen stimulation. This increased histamine production results from the effect of a factor having all the characteristics of HCSF (histamine-producing cell-stimulating factor) already described in secondary MLC supernatant. This factor is produced by Thy-1, 2, Lyt-1, 2-positive cells. Spleen cell cultures derived from skin-allografted mice during rejection produce more histamine in response to T cell mitogens than do spleen cells from normal or syngeneic grafted mice. Such a phenomenon is not observed in response to B cell mitogens. A striking association is found between enhanced histamine synthesis and skin allograft rejection. This phenomenon results from a) a five to 10-fold increase in HCSF production by allograft recipient spleen cells in response to T cell mitogens, and b) an increase in HCSF sensitivity of these spleen cells.
An antigen cross-reactive with gp52 of mammary tumor virus is expressed on a B cell subpopulation of mice.
Authors: Tax A , Ewert D , Manson L A
Published: December 2022

The reactivity of murine lymphoid tissue with biotinylated F(ab')2 fragments of monoclonal antibody VE7 (BIOT VE7), which reacts with gp52 of murine mammary tumor virus (MuMTV), was tested with fluoresceinated avidin in an indirect fluorescent antibody assay on live splenocytes. A small percentage of splenic lymphocytes in C3H/He mice infected exogenously with MuMTV, and in C57BL/6 mice that, like C3H/He, harbor several endogenous MuMTV proviruses in their genomes, were reactive with the monoclonal antibody. The antigen-positive splenocytes were shown to represent a subpopulation of B cells. The possible nature of the B cell-associated antigen recognized by monoclonal antibody VE7 is discussed.
Viral inhibition of lymphocyte mitogenesis: interference with the synthesis of functionally active T cell growth factor (TCGF) activity and reversal of inhibition by the addition of same.
Authors: Wainberg M A , Vydelingum S , Margolese R G
Published: December 2022

We have investigated the mechanisms whereby co-incubation of several types of virus particles with human lymphoid cells in the presence of T cell lectins leads to inhibition of the proliferative response that otherwise ensues. The data indicate that, in the absence of infection, such inhibition can be reversed by the addition to cultures of relatively high concentrations of fluids rich in T cell growth factor (TCGF) activity. The ability of these fluids to achieve such reversal of inhibition is both concentration- and time-dependent. Addition of the factor to virus co-incubated cells more than 26 hr after culture initiation does not restore responsiveness. We have also shown that virus co-incubated cultures are deficient with respect to their ability to synthesize detectable levels of TCGF activity in the presence of phytohemagglutinin. In contrast, the use of relatively dilute virus preparations (less than 10 particles per cell) permits partial responsiveness to lectin as well as the synthesis of moderate levels of TCGF. These finding suggest that viral inhibition of lymphocyte mitogenesis is mediated directly or indirectly by interference with the synthesis of functionally active TCGF activity.