bluuebunny/crossref_metadata_2025 huggingface dataset:
Radioimmunoassay for the Detection of Hepatitis e Antigen (HBeAG) and Antibody (Anti-HBe)
Authors: Fields Howard A. , Bradley Daniel W. , Davis Candace L. , Murphy Bert L. , Schable Charles A. , Maynard James E.
Published: December 2022

A solid phase micro-immunoradiometric assay (micro-SPIRA) for the detection of hepatitis e antigen (HBeAg) and antibody has been developed. Chimpanzee anti-HBe/2 was developed by repeated immunizations with purified antigen containing HBeAg/1 and HBeAg/2. An anti-HBe/2 titer of 1:4 was determined by immunodiffusion (ID) analysis. Anti-HBe/1 was not detected. The anti-HBe IgG used in the assay was purified from plasma by a combination of DEAE-cellulose and affinity chromatography. The sensitivity of the micro-SPIRA for antigen and antibody was 193 ng/ml and 65 ng/ml, respectively. By comparing relative endpoint titers obtained by ID to micro-SPIRA, it was determined that micro-SPIRA for antigen and antibody is 320 and greater than 1300 times more sensitive, respectively, than ID. The specificity of the assay was ascertained by the examination of various non-B specimens. The application of the assay to a panel of 50 hepatitis B surface antigen (HBsAg)-positive specimens resulted in an increase in positivity of 18% for antigen and 22% for antibody.
Distinct Subpopulations of IgG Memory B Cells Respond to Different Molecular Forms of the Same Hapten
Authors: Tittle Thomas V. , Rittenberg Marvin B.
Published: December 2022

Spleen cells from mice primed with the thymus-dependent antigen trinitrophenyl keyhole limpet hemocyanin several months earlier can be cultured in vitro to give vigorous IgG antihapten PFC responses to thymus-dependent (TD) and thymus-independent (TI) forms of the same hapten. Here we show that the IgG memory precursors that respond to these two forms of the hapten constitute functionally distinct subpopulations. We have designated these subpopulations as B1γ and B2γ to represent secondary precursor cells responding to TI and TD antigens, respectively. Three types of evidence for these subpopulations are presented: 1) In vitro secondary IgG responses to TD and TI forms of the TNP hapten are additive when both forms are added to the same culture. 2) The precursor frequencies for the TD and TI antigens are additive, but addition is not observed between two TD or two TI antigens. 3) Each population can be selectively eliminated by BUdR and light treatment without affecting the other population. The ontogenetic relationships between these subpopulations are discussed in relation to all presently proposed subpopulations B1µ, B2µ, B1γ, and B2γ.
Structural Studies on Induced Antibodies with Defined Idiotypic Specificities
Authors: Capra J. Donald , Ju Shyr-Te , Nisonoff Alfred
Published: December 2022

Previous studies in this series have been directed toward the elucidation of the heavy and light chain variable region structures of antibodies raised in A/J mice to the p-azophenylarsonate (Ar) hapten, certain of which bear a cross-reacting idiotype. The present study concerns an analysis of anti-Ar antibodies that arise in A/J mice suppressed for a cross-reacting idiotype. The results indicate that when an idiotype is suppressed and the animal subsequently hyperimmunized, the resultant antibodies are “deviated” into different V-region subgroups, both in the heavy and light polypeptide chains. The study presents the first primary structural analysis of a humoral immune response that has been manipulated by an idiotypic reagent.
Characterization of a B Cell Defect in the NZB Mouse Manifested by an Increased Ratio of Surface IgM to IgD
Authors: Cohen P. , Ziff M. , Vitetta E. S.
Published: December 2022

In an effort to define the cellular basis of abnormalities in polyclonal B cell activation previously noted in NZB mice, the surface immunoglobulin (sIg) isotypes of spleen cells from NZB mice were examined. After lactoperoxidase-catalyzed radioiodination, the cell surface immunoglobulins were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Spleen cells from 8- to 10-week-old NZB mice were found to have an increased ratio of cell surface IgM/IgD compared to cells from 11 control strains. The altered ratio of sIg isotypes was not a consequence of increased proteolytic activity present in NZB cell suspensions or of the presence of cytophilic antibody or autoantibody.

Ontogenetic studies of the sIgM/sIgD (µ/δ) ratio on splenocytes from NZB and BALB/c mice revealed that the former cells had higher µ/δ ratios as early as 2 weeks after birth. By 4 weeks of age the µ/δ ratios were equivalent. Between 4 weeks and 1 year of age, the µ/δ ratios on NZB splenocytes remained constant whereas those on BALB/c splenocytes decreased and reached adult levels at 6 weeks.
Effect of Tunicamycin on IgM, IgA, and IgG Secretion by Mouse Plasmacytoma Cells
Authors: Hickman Scot , Kornfeld Stuart
Published: December 2022

Tunicamycin, an antibiotic that prevents glycosylation of glycoproteins by blocking the formation of N-acetylglucosamine-lipid intermediates, was used to study the importance of glycosylation for the secretion of immunoglobulins by mouse plasmacytoma lines that produce immunoglobulins of different classes. Biosynthetically labeled secreted and intracellular immunoglobulins were measured by immunoprecipitation assays. Tunicamycin, at a concentration of 0.5 µg/ml produced an 81% inhibition of IgM secretion by MOPC 104E plasma cells without significantly affecting the initial rate of synthesis of intracellular IgM. No increase in the intracellular degradation of nonglycosylated IgM could be demonstrated. Tunicamycin also produced a 64% average inhibition of IgA secretion by several mouse IgA-secreting plasmacytoma lines. In contrast, despite inhibiting the incorporation of D-[14C] glucosamine into newly synthesized IgG, tunicamycin only produced a 28% average inhibition of IgG secretion, which was only slightly more than the nonspecific inhibition of secretion of the normally nonglycosylated λ2 light chains by variant MOPC 315 plasmacytomas. These data indicate that the extent of inhibition of immunoglobulin secretion produced by tunicamycin depends on the immunoglobulin class produced by the plasma cell.
Relationship Between Trinitrophenol and H-2 Antigens on Trinitrophenyl-Modified Spleen Cells
Authors: Vitetta Ellen S. , Hart David A. , Forman James
Published: December 2022

Splenic lymphoblasts or normal spleen cells were treated with varying concentrations of TNBS in order to assess whether cell membrane H-2 molecules were derivatized with TNP. Cells treated with high concentrations of TNBS had their cell membrane H-2 molecules derivatized and functioned antigenically as inhibitors in a cold target TNP-CML competition assay. In contrast, cells derivatized with lower concentrations of TNBS had a significant proportion of their membrane proteins derivatized with TNP but did not have their H-2 molecules derivatized. These latter cells were unable to block anti-TNP cytotoxic effector cells in the competition assay. When cells were treated with 3H-TNBS, it was observed that TNP couples to cell membrane H-2, Ia and Ig molecules, and an estimate of the number of TNP molecules bound per cell at varying concentrations of TNBS was determined.

The data obtained are consistent with there being a requirement for TNP to directly derivatize H-2 molecules on the cell membrane in order to create antigenic determinants that can be recognized by cytotoxic anti-TNP effector cells. As an alternative, there may be a requirement for the presence of a high density of TNP molecules per cell rather than direct H-2 derivatization by TNP in order to account for activity.
Age-Related Changes in the Subsets and Functions of Human T Lymphocytes
Authors: Kishimoto Susumu , Tomino Shinhachiro , Inomata Koichiro , Kotegawa Shoji , Saito Taro , Kuroki Masahide , Mitsuya Hiroaki , Hisamitsu Shotaro
Published: December 2022

The purpose of this study was to investigate changes in immunocompetence associated with aging in humans by the use of various immunologic parameters. Volunteers examined in this study were healthy or apparently healthy without diseases affecting the immune system and ranged from 14 to 89 years of age. The results of this study are summarized in Table I.

The absolute number of peripheral blood lymphocytes (PBL) decreased with aging. The mean count was 2043 in the adult group (less than 60 years of age) and 1659 in the aged group (over 60 years of age). When T lymphocytes were assayed by rosette formation with neuraminidase-treated sheep erythrocytes in the presence of fetal calf serum after 1 hr incubation at 4°C, there was no difference in the proportions of T lymphocytes between the groups, whereas the proportions of T cells as assayed by rosette formation with sheep erythrocytes in the absence of fetal calf serum were significantly depressed in the aged group (p < 0.01). On the other hand, the proportion of T lymphocytes assayed by rosette formation with sheep erythrocytes after centrifugation at 1500 RPM for 5 min at room temperature was higher in the aged group than in the adult group (p < 0.02). The proportion of IgG-Fc receptor-bearing T lymphocytes detected by double rosette formation with sheep erythrocytes and chicken erythrocytes sensitized with rabbit IgG antibody was also higher in the aged than in the adult group (p < 0.05). The proportion of complement receptor-bearing lymphocytes did not change with aging. In addition, the proportion of surface Ig-bearing lymphocytes examined after being treated in pH 4.0 acetate buffer for 1 min to remove nonspecifically bound Ig from the cell surface showed no differences between the two groups.

Response to PHA and concanavalin A was impaired in the aged (aged 70 to 83 years) compared to the young adult group (aged 20 to 40 years) (p < 0.01), whereas the response to pokeweed mitogen was of the same magnitude. PHA response was negatively correlated with the proportion of IgG-Fc receptor-bearing T cells. The mixed lymphocyte culture reaction with mitomycin-treated AL1 cells derived from Burkitt's lymphoma was also impaired in the aged group (p < 0.01) as was cell-mediated cytotoxicity to AL1 cells as target cells (p < 0.05).

Levels of IgG and IgA in plasma were significantly higher in the aged than in the adult group (p < 0.001), whereas those of IgM and IgE were not different. Radioimmunoassay determination of Ig classes revealed that the geometric means of IgG, but not IgM or IgA, synthesized in vitro was significantly increased in the aged compared to the young adult group (p < 0.05) when PBL were cultured in the presence of PWM for 7 days. This increase in IgG synthesis in the aged group was attributed to enhanced T helper activity.
Specificity of Antisera from Lewis Rats Immunized with Encephalitogenic Fragment 43–88 of Guinea Pig Myelin Basic Protein
Authors: Fritz Robert B. , Jen Chou C.-H. , Randolph Devery H. , Desjardins Anne E. , Kibler Robert F.
Published: December 2022

Inbred Lewis rats were immunized with encephalitogenic fragment 43–88 of guinea pig myelin basic protein emulsified in complete Freund's adjuvant. After recovery from experimental allergic encephalomyelitis (EAE) the animals were given a booster immunization, bled, and the specificity of the individual anti-fragment anti-sera was examined by direct binding assays by using radioiodinated fragment 43–88 and peptides 43–67, 68–88, and 79–88. Competitive-binding experiments with these peptides in competition with labeled fragment 43–88 were also done. The results of these experiments indicated that the immunodominant region of fragment 43–88 was the carboxy-terminal half of the molecule. Individual antisera recognized different antigenic determinants within this region.
Natural Killing and Antibody-Dependent Cellular Cytotoxicity Are Mediated by Different Mechanisms and by Different Cells
Authors: Koren Hillel S. , Williams Margaret S.
Published: December 2022

Natural killing (NK) in humans, as well as in other species, has been shown to be specific for antigenic determinants present on the surfaces of a variety of tumor cells. Physical separation of NK cells from K cells, which mediate antibody-dependent cellular cytotoxicity (ADCC), has not been successful; however, there is indirect evidence suggesting that these activities are distinct. To further study the relationship between NK and K cells, competitive inhibition techniques were employed.

NK cells can be blocked via two mechanisms: 1) by direct inhibition with NK-sensitive tumor cells binding to NK receptor sites present on the effector cells and 2) by steric inhibition resulting from the binding of antibody-coated cells to the FcR on the effector cells. K cells, however, lack the NK receptor site(s) but are FcR+, and can therefore be blocked only by antibody-coated cells. We therefore postulate that NK and K cells are two separate lymphoid populations. NK cells bear receptor site(s) for NK determinants and FcR, whereas K cells bear only FcR.
Antigen-Initiated B Lymphocyte Differentiation
Authors: Howard Maureen C. , Baker Julie A. , Shortman Ken
Published: December 2022

The effects of specific and nonspecific stimuli on the cell cycle status of secondary B cells responding in either an adoptive immune assay or in tissue culture were investigated with a “cell suicide” procedure. Spleen cells from unstimulated or recently stimulated long-term hapten-primed “memory” mice were exposed to high specific activity 3H-TdR before AFC-progenitor assay; in all experiments, AFC-progenitor activity was assessed as a response to NIP-POL antigen.

Secondary adoptive transfer or cell culture AFC-progenitors from animals not receiving immediate prestimulation were not in a state of rapid cell cycle. Secondary AFC-progenitors detected in adoptive immune assays were activated into rapid cell cycle by nonspecific stimuli. Secondary AFC-progenitors responding in cell cultures were not activated by the same nonspecific stimuli, but showed a proliferative response to specific antigen. These results are analogous to those presented in the preceding paper for primary B cells. They suggest that, like primary B cells, secondary B cells contain distinct adoptive transfer AFC-progenitor (termed “preprogenitor”) and culture AFC-progenitor (termed “direct progenitor”) subsets. These various primary and secondary B cell subsets are incorporated into a model of B cell development.
Suppressor Cells Present in the Spleens of Trypanosoma Cruzi-Infected Mice
Authors: Ramos Celso , Schädtler-Siwon Ilse , Ortiz-Ortiz Librado
Published: December 2022

Infection with Trypanosoma cruzi decreases the ability of spleen cells from mice to respond to either T cell, concanavalin A (Con A), or B cell, lipopolysaccharide (LPS), mitogens. The effect of infection on the mitogenic response depends on the elapsed time between the day of infection and the time of mitogen presentation. Responses early in infection are normal, whereas later responses to either mitogen are depressed. Spleen cells from late trypanosome-infected mice inhibit the ability of normal spleen cells to respond to Con A or LPS. The cell in the T. cruzi-infected spleen cells responsible for this effect is nonadherent, sensitive to treatment with anti-mouse thymus serum plus complement, but insensitive to treatment with anti-immunoglobulin plus complement. These data indicate that infection with T> cruzi elicits over time the generation of T cells suppressive to T and B cell mitogenic responses.
Immunoregulatory Properties of Human Esophageal Tumor Extract
Authors: Mohagheghpour Nahid , Parhami Behnaz , Dowlatshahi Kambiz , Kadjehnouri Dara , Elder John H. , Chisari Francis V.
Published: December 2022

Noncytotoxic aqueous extracts of esophageal tumors (TEx) inhibit the spontaneous uptake of 3H-thymidine by human peripheral blood mononuclear cells (PBM). TEx also inhibits mitogen- and antigen-induced PBM blastogenesis, mitogenesis, 3H-thymidine uptake, and 3H-leucine incorporation. Inhibition is reversible and is mediated by a heat-labile protein with a m.w. of approximately 70 to 80,000 daltons. Inhibitory activity is not due to trivial binding of mitogen or nucleotide by the inhibitor, nor is it entirely tumor specific since similar but quantitatively less activity is extractable from adjacent normal esophageal mucosa. Assorted cell surface membrane phenomena such as contact inhibition of in vitro endothelial cell proliferation, lymphocyte E rosette formation, and cap formation are unaffected by TEx. In contrast, immunoglobulin synthesis by pokeweed mitogen-stimulated PBM is enhanced by TEx. The potential importance of the local production of immunoregulatory agents like TEx in the immunologic control of tumor cell growth is discussed.
Immune Responses to Weakly Immunogenic Virally Induced Tumors
Authors: Devens Bruce , Naor David
Published: December 2022

Splenocytes from A mice injected with YAC-1 or RBL5 could generate, after in vitro culture with or without stimulation, a genetically nonrestricted cytotoxic response against the allogenic tumor RBL5. YAC-1 tumor is an in vitro carried tumor induced in A mice (H-2a) by Moloney virus. RBL5 tumor is a Rauscher virus-induced tumor of C57BL/6 mice (H-2b). These tumors cross-react serologically.

The effector cells that were generated after the in vitro cultivation recognized tumor-associated antigens on the target cells. H-2 alloantigens were not recognized by the effector cells. The effector cells that killed RBL5 tumor in a genetically nonrestricted manner were identified as T cells. The in vivo carried tumor YAC, in contrast to the in vitro carried tumor YAC-1, could not induce anti-RBL5 reactive cells in A mice. Instead, YAC tumor induced suppressor cells in A mice, which could abrogate the anti-RBL5 cytotoxic response of RBL5-primed splenocytes, but not that of YAC-1 primed splenocytes.
Characterization of the Immunosuppressive State during Schistosoma Mansoni Infection
Authors: Attallah A. M. , Smith A. H. , Murrell K. D. , Fleischer T. , Woody J. , Vannier W. E. , Scher I. , Ahmed A. , Sell K. W.
Published: December 2022

Analysis of a murine model of schistosomiasis revealed that both the thymus (T)- and bursa (B)-derived compartments of the immune system are modified during acute infection. The functional capacity of T and B lymphocytes to respond to mitogenic stimuli and the humoral response to thymus-dependent (SRBC) and thymus-independent (DNP-Ficoll) antigens are severely depressed. In addition, it was found that suppressor cells capable of inhibiting the response of normal lymphocytes to SRBC arise during acute infection. Although the splenic frequency of T (ϑ) and B (Ig+) cells remained constant during chronic infection, quantitative changes were detected in each population. In the T cell pool there was a decrease in the percentage of Ly-1+ cells and a concomitant increase in Ly-1+, 2+, 3+ cells, whereas the B cell pool showed a progressive loss of complement receptor-bearing lymphocytes, which apparently was the result of inactivation of surface complement receptor by a serum factor specifically found in infected mice. Characterization of the serum factor strongly suggests it is an immune complex. Thus, it appears that both suppressor cells and immune complexes contribute to changes noted in the immune system during acute schistosomiasis. Additional studies carried out in mice after unisexual infection revealed that egg production is not a necessary prerequisite for several of the immunologic phenomena associated with acute schistosomiasis.
Association between H-2 and Vaccinia Virus-Induced Antigens on the Surface of Infected Cells
Authors: Senik Anna , Neauport-Sautes Catherine
Published: December 2022

The relationship between H-2 molecules and vaccinia virus-induced antigens on the surface of H-2d infected cells was investigated by the differential redistribution method and by the blocking capacity of monospecific anti-H-2 sera on an anti-vaccinia cell-mediated cytotoxicity (CMC). Capping of either H-2K or H-2D molecules upon addition of monospecific anti-H-2 sera was followed by the complete redistribution of viral antigens, suggesting the formation, on the cell membrane, of complexes composed of H-2K, H-2D molecules and vaccinia virus-induced antigens. However, not all H-2 molecules were involved in this association since i) free H-2K and H-2D molecules still moved independently on the cell surface, and ii) capping of vaccinia virus-induced antigens failed to induce the redistribution of all the H-2K and H-2D molecules. In addition, either monospecific anti-H-2K or anti-H-2D antiserum was found to exert potent blocking activity on anti-vaccinia CMC, indicating also a close topographical relationship between H-2K, H-2D molecules and vaccinia virus-induced antigens.
Regulation of Cytotoxic Lymphocyte Responses in New Zealand Mice by Alloantigen-Activated Spleen Cells
Authors: Huston David P. , Rich Robert R. , Steinberg Alfred D. , Truitt Gary A.
Published: December 2022

The ability of autoimmune New Zealand mice and control mice to suppress the generation of cytotoxic lymphocytes was studied. These experiments demonstrated that allosensitized spleen cells from both 1- and 6-month-old female NZB, NZW, and NZB/NZW F1 mice, or culture supernatants of such cells, could suppress the in vitro generation of cytotoxic lymphocytes. In addition, the cytotoxic lymphocyte responses of NZW and NZB/NZW F1 mice were suppressed by allosensitized cells and their culture supernatants. In contrast, cytotoxic lymphocyte responses of NZB, as well as CBA/J and CBA/N mice, although readily suppressed by allosensitized spleen cells, were resistant to suppression by the culture supernatants of these allosensitized cells.

To investigate further NZB and CBA resistance to cytotoxic lymphocyte suppressor factor(s), F1 hybrids of resistant × susceptible and resistant × resistant strains were studied. F1 hybrids of strains suppressed by culture supernatants and strains resistant to supernatant suppression were all suppressed, demonstrating suppression to be a dominant phenotypic trait. F1 hybrids of two strains resistant to supernatant suppression were not suppressed; thus, resistance was recessive without gene complementation. Furthermore, all strains resistant to suppression by culture supernatants were suppressed by the allosensitized cells. This cell-supernatant dichotomy suggests that allosensitized spleen cells can suppress several different steps in a cytotoxic immune response.
Defective Tumoricidal Capacity of Macrophages from A/J Mice
Authors: Boraschi Diana , Meltzer Monte S.
Published: December 2022

Although macrophages from Mycobacterium bovis, strain BCG-infected C3H/HeN mice were highly cytotoxic to tumor cells in vitro, cells from BCG-infected A/J mice were not tumoricidal. Varying the dose of BCG or the time of infection did not evoke cytotoxic activity with A/J macrophages. Inflammatory responses to BCG infection in A/J mice were relatively normal: the yield of macrophages was about ¼ that from C3H/HeN mice; however, no differences between strains were detected for inflammation-induced increases in macrophage phagocytic capacity or in percentage of peroxidase-positive cells. Similarly, production of macrophage activation factor activity by tuberculin-stimulated BCG-immune spleen cell cultures was also intact in A/J mice. Thus, despite adequate inflammatory responses and production of active lymphokines, macrophages from BCG-infected A/J mice were not activated for tumor cytotoxicity. Under certain conditions, however, these macrophages could develop cytotoxic activity. Macrophages from BCG-infected but not control A/J mice developed tumoricidal activity after further in vitro treatment with lymphokines, bacterial lipopolysaccharides (LPS), or certain plant lectins. Peritoneal exudate macrophages first treated with lymphokines also developed cytotoxic activity but only after further treatment with LPS or lectins. The phenotypic expression of the A/J genetic defect in macrophage cytotoxic activity was very similar to the defect of lipid A-unresponsive C3H/HeJ mice except for the concentration of LPS required for cytotoxic activity by lymphokine-treated cells. Other responses of A/J mice to LPS were also different from those of the unresponsive C3H/HeJ mice. Responses of A/J mice to the lethal toxicity of LPS or of A/J macrophages to direct toxicity of LPS in vitro, responses influenced by the Lps gene, were normal. On the other hand, macrophage cytotoxic activity and spleen cell proliferative responses to LPS in vitro, responses also controlled by the Lps gene, were abnormal. We conclude that the genetic basis for the macrophage tumoricidal defect of A/J mice represents either a previously undescribed allelic change at the Lps locus or, what is more likely, gene mutation(s) at entirely different loci.
Specific Binding of T Lymphocytes to Macrophages
Authors: Lyons Clifford R. , Tucker Thomas F. , Uhr Jonathan W.
Published: December 2022

The effect of anti-Ia alloantiserum on the capacity of selected peritoneal exudate lymphocytes (selected PEL) to bind to antigen-pulsed F1 (responder × nonresponder) macrophages was investigated. With the use of selected PEL for antigens under Ir gene control, it was shown that anti-Ia serum to the responder haplotype blocked adherence of selected PEL to antigen-pulsed macrophages whereas anti-Ia serum to the nonresponder haplotype did not. The target cell of the anti-Ia alloantiserum appeared to be the macrophage because anti-13 Ia in contrast to anti-2 Ia did not inhibit binding of F1 (2 × 13) DNP-GL selected PEL to DNP-GL pulsed strain-2 Mφ (responder strain). Taken together with previous experiments that indicate that an antibody to the native protein antigen employed is unable to block specific binding, the present results suggest that T cells may recognize fragments of exogenous antigen in association with Ia molecules.
Immune Response to Levan
Authors: Bona Constantin , Lieberman Rose , House Shirley , Green Ira , Paul William E.
Published: December 2022

Pretreatment of BALB/c mice with antisera to a cross-reactive idiotype (E109IdX) expressed on many anti-bacterial levan (BL) and anti-inulin (Inu) antibodies leads to a prolonged suppression in production of IdX-bearing molecules in response to BL immunization. There is a comparable suppression in numbers of plaque-forming cells secreting IdX-bearing anti-BL and anti-Inu molecules. Furthermore, spleen cells from anti-E109IdX pretreated mice are unable to transfer to irradiated recipients the ability to produce IdX-bearing anti-BL and anti-Inu antibodies. These results indicate that the suppressive effect is at the precursor level and not simply a clearance of antibodies bearing the IdX.

Suppression of IdX production can be achieved by pretreating nu/nu BALB/c mice with anti-E109IdX antibodies. Furthermore, spleen cells from pretreated mice do not inhibit the capacity of spleen cells from normal mice transferred to irradiated recipients to produce E109IdX in response to BL. This indicates that the suppression of IdX production in the anti-BL system is T independent and probably represents direct inhibition of precursors by anti-IdX.
IgE Mediated Triggering of Rat Basophil Leukemia Cells: Lack of Evidence for Serine Esterase Activation
Authors: Taurog Joel D. , Fewtrell Clare , Becker Elmer L.
Published: December 2022

Inhibition of mediator release from mast cells and basophils by diisopropylfluorophosphate (DFP) and other organophosphorus compounds known to inhibit serine esterases has in the past led to the hypothesis that immunologic triggering of these cells involves an activatable serine esterase. In this study we have shown that two nonphosphorylating or poorly phosphorylating structural analogs of two potent phosphorylators inhibit release of incorporated serotonin from cultured rat basophil leukemia cells. We conclude that, by itself, inhibition of immunologic mast cell triggering by phosphorylating organophosphorus compounds can no longer be considered evidence for involvement of an activatable serine esterase in mast cell triggering.
Physicochemical Characterization of Lymphocyte-Activating Factor (LAF)
Authors: Mizel Steven B.
Published: December 2022

The LAF produced by the mouse macrophage cell line, P388D1, is a single polypeptide chain of m.w. 12,000 to 16,000 daltons. Native LAF was destroyed by Streptomyces griseus protease, but not by trypsin, chymotrypsin, and papain, although in the presence of 8 M urea, papain completely destroyed LAF activity. LAF did not bind to concanavalin A-Sepharose, suggesting that LAF does not contain significant amounts of mannosyl or glycosyl residues. Since LAF activity was not inactivated by a treatment of reduction and alkylation the active conformation of LAF does not appear to be dependent on disulfide linkages. LAF was not irreversibly denatured by 8 M urea or 0.1 to 0.5% SDS. On SDS-polyacrylamide gels, the m.w. of LAF was 12,000 daltons, as compared to a value of 16,000 daltons, as determined by gel filtration. The isoelectric point of LAF was 5.0 to 5.4 as determined on 7.5% acrylamide gels (pH 3 to 10). On the basis of these results it appears that the P388D1 cell line-derived LAF is a relatively stable molecule that shares several physicochemical properties with normal human and mouse macrophage-derived LAF.
Immunoaffinity Purification of Schistosoma Mansoni Soluble Egg Antigens
Authors: Harrison D. Janiece , Carter Clint E. , Colley Daniel G.
Published: December 2022

Schistosoma mansoni egg antigens were purified from a heterogeneous mixture of soluble egg antigens (crude SEA) with an immunoaffinity column that consisted of the specific anti-SEA antibodies contained in 16-week S. mansoni-infected mouse serum bound to Sepharose 4B. On sodium dodecyl sulfate (SDS) gel electrophoresis, the purified antigen fraction yielded at least eight bands staining with Coomassie blue and at least five bands reacting with periodic acid-Schiff (PAS). All of the proteins in the antigenic fraction appear to contain carbohydrate residues. Upon immunoelectrophoresis the antigen yielded four precipitin arcs. The antigenic fraction isolated by means of the immunoaffinity column was then compared to various fractions obtained from concanavalin A (Con A) chromatography of SEA. The results of Ouchterlony immunodiffusion and immunoelectrophoresis indicate that the antigenic fraction isolated by immunoaffinity purification of SEA contains the major antigens found in the fractions obtained from Con A chromatography of SEA. The results of SDS gel electrophoresis indicate that the major PAS-reacting bands of the antigenic fraction isolated by immunoaffinity purification are found in the 3rd peak (bound fraction) resulting from Con A chromatography of SEA, whereas the major Coomassie blue-staining band in the isolated antigenic fraction is found in the 2nd peak (unbound fraction) from Con A chromatography of SEA.
Structural Studies on the Murine Ia Alloantigens
Authors: Cook Richard G. , Siegelman Mark H. , Capra J. Donald , Uhr Jonathan W. , Vitetta Ellen S.
Published: December 2022

Murine Ia alloantigens encoded by the I-A and I-E/C subregions were isolated from radiolabeled splenic lysates and examined by NH2-terminal sequence analysis. Haplotype-associated sequence variation was detected in the β, but not α, subunits of both the A and E/C alloantigens. The A β-polypeptides from k and b haplotypes show four differences in the 12 positions compared, whereas the E/C β-polypeptides from k and r haplotypes show two differences in the 13 positions compared. No sequence variation was detected between the Ak and Ab α-chains (six positions compared) or between the E/Ck and E/Cr α-chains (11 positions compared). Homology relationships between these murine Ia alloantigens and the human Ia (DR) alloantigens are also presented.
Natural Thymocytotoxic Autoantibodies in Autoimmune and Normal Mice
Authors: Eisenberg Robert A. , Theofilopoulos Argyrios N. , Andrews Brian S. , Peters C. J. , Thor Lee , Dixon Frank J.
Published: December 2022

Natural thymocytotoxic autoantibodies (NTA) were found in all mouse strains. Among those strains that show autoimmune syndromes resembling human systemic lupus erythematosus (SLE), the NZB and NZB×NZW had high levels of NTA, the BXSB had moderate levels, and the MRL/1 and MRL/n had very low levels. In addition, some normal strains had high levels, sometimes even higher than the autoimmune strains. The NTA were mostly IgM and were present, but not concentrated, in the cryoprecipitates of the autoimmune mouse strains. In most strains, they were directed toward an antigen shared by thymocytes and brain. The failure to find high levels of NTA in all autoimmune mouse strains, as well as the finding of very high levels in some normal strains, make it unlikely that such autoantibodies are a fundamental etiologic factor in all murine SLE.
Suppressor Cells for the Afferent Phase of Contact Sensitivity to Picryl Chloride: Inhibition of DNA Synthesis Induced by T Cells from Mice Injected with Picryl Sulfonic Acid
Authors: Thomas Wayne R. , Watkins Madeleine C. , Asherson Geoffrey L.
Published: December 2022

Previous reports have shown that picryl sulfonic acid (PSA) induces suppressor T cells that inhibit the effector phase of contact sensitivity, whereas its DNP counterpart, dinitrobenzenesulfonate (DNBS) induces cells that inhibit the afferent phase of sensitization. Accordingly, cells from mice injected with DNBS, but not PSA, could be shown to inhibit the DNA synthesis in the lymph nodes that occurs during sensitization. It is now shown that PSA does induce T cells that suppress DNA synthesis but this can only be detected with enriched T cells or by using a regimen of PSA injection different from previously used to induce suppressor cells for the effector phase. The T cells did not affect responses to oxazolone or dinitrofluorobenzene (DNFB) and were distinguishable from suppressors of the efferent phase in that they could be produced in adult thymectomized but not cyclophosphamide-treated mice. T cells from mice injected with DNBS that inhibited DNA synthesis to DNFB had the same properties.
Characteristics of in Vitro Production of Antibodies to DNA in Normal and Autoimmune Mice
Authors: Sawada Shigemasa , Talal Norman
Published: December 2022

The in vitro production of antibodies to dsDNA was studied with spleen cells from normal and autoimmune mice. After culture for 4 days, the binding of dsDNA in the culture supernatant was measured by a radioimmunoprecipitation assay. The production of antibodies to dsDNA by spleen cells appeared at 15 hr after culture and reached a plateau at 24 hr. No antibodies were produced by thymus cells or splenic T cells. The specificity for dsDNA was shown by competitive inhibition with nonradioactive nucleic acids. Autoimmune strains of mice (NZB/NZW, BXSB, MRL/1) produced more antibodies to dsDNA than did several control strains. Young B/W mice and control strain mice produced mainly IgM antibodies, whereas older B/W mice produced predominantly IgG antibodies to dsDNA. The in vitro production of antibodies to dsDNA by aged B/W spleen cells was macrophage and T cell dependent.
Hanganutziu-Deicher Antigen and Antibody in Pathologic Sera and Tissues
Authors: Nishimaki Tomoe , Kano Kyoichi , Milgrom Felix
Published: December 2022

Heterophile, Hanganutziu-Deicher (H-D) antigen was studied in pathologic sera by means of inhibition of agglutination of bovine erythrocytes by H-D antibodies. H-D antigen was demonstrated in 38% of random cancer sera, 25% of lymphoma or leukemia sera, 25% of leprosy sera, 8% of infectious mononucleosis sera, 6% of rheumatoid arthritis sera, and 27% of synovial fluids of rheumatoid arthritis patients. None of 134 normal human sera gave positive results. Some of the inhibition-positive cancer sera formed precipitation lines with H-D antibody-containing sera. Over 50% of various extracts of cancer tissues as well as spleens of lymphoma or leukemia patients were shown to contain H-D antigen by means of the inhibition test.
Natural Cytotoxicity of Mouse Monocytes and Macrophages
Authors: Tagliabue Aldo , Mantovani Alberto , Kilgallen Marianne , Herberman Ronald B. , McCoy James L.
Published: December 2022

Adherent-phagocytic cells were obtained from the peripheral blood and the unstimulated peritoneal cavity of normal C3H/HeN mice and tested for their cytotoxicity in an in vitro 3H-TdR release assay and in a growth inhibition assay. These cells had appreciable cytolytic and cytostatic activity, which varied with the attacker: target (A:T) cell ratio, against TU5, an SV40 virus-transformed BALB/c tumor cell line. Cytolytic activity was first detected after 48 hr of incubation and was considerably higher at 72 hr. In contrast, this line was resistant to cytotoxicity by nonadherent lymphoid cells at the A:T ratios employed, and adherent cells from the peritoneal cavity were not cytotoxic against YAC-1 target cells in a 4-hr 51Cr release assay. The natural cytotoxicity of adherent cells from the peritoneal cavity was largely inhibited by treatment with silica. These results indicated that the natural reactivity observed here was mediated by macrophages and not by natural killer cells. The cytotoxic reactivity of the macrophages from normal mice could not be attributed to activation by a variety of possible in vivo or in vitro stimuli. Cells from germ-free nude mice had cytolytic activity, as did cells cultured in medium containing endotoxin-free fetal bovine serum or normal mouse serum. Activation by interferon produced in vitro was also unlikely since marcophages cultured in the presence of anti-interferon were also cytotoxic. As previously described for activated macrophages, peritoneal macrophages from normal C3H/HeJ mice were significantly less cytotoxic than macrophages from syngeneic C3H/HeN mice. The monocytes and macrophages seemed to have somewhat different properties since the addition of lymphokine-containing medium to the adherent peripheral monocytes increased their cytotoxicity, whereas no enhanced activity was observed by adding this supernatant to resident peritoneal macrophages. The normal macrophages and monocytes reacted against both syngeneic (1023) and allogeneic (TU5) tumor cells as well as mouse fibroblast cells transformed by SV40 virus (SV40-3T3). Similar activity was also observed against a human cell line derived from a patient with breast cancer (G11) and against human fibroblast cells transformed by SV40 (SV40-2931 and SV40-2303), but not against nontransformed fibroblasts from the same donors. Thus, normal murine peripheral monocytes and peritoneal macrophages possess natural cytotoxic activity that is selective for tumor and virus-transformed cells.
Induction of Cytocidal Macrophages after in Vitro Interactions between Listeria-Immune T Cells and Macrophages—Role of H-2
Authors: Farr Andrew G. , Wechter William J. , Kiely Jeanne-Marie , Unanue Emil R.
Published: December 2022

Twenty-four-hour cultures of macrophages from non-immune mice with T cells from mice immune to Listeria resulted in the acquisition of cytocidal activity by the macrophages. The macrophages killed P815 tumor cells in a chromium-release assay. Macrophages from non-immune mice given peptone or thioglycollate injections were much better effector cells than resident macrophages. Experiments established the conditions for generation of cytolytic macrophages: the specificity of T cells to Listeria and the need for macrophages to specifically interact with heat-killed Listeria organisms. Cells from the immune T cell inoculum did not participate in the killing process by the activated macrophages.

The interactions leading to the generation of cytocidal macrophages required homology at the I region between macrophages and T cells, as well as the presence of macrophages bearing Ia molecules. Cultures of T cells and macrophages with heat-killed Listeria contained a factor that, when added to peptone macrophages, resulted in their activation for cytolytic activity. The factor could activate syngeneic or allogeneic macrophages, or macrophages lacking detectable Ia molecules. The immunogenic moiety associated with Listeria was short-lived.
Induction of Secretion of IgM from Cells of the B Cell Line 38C-13 by Somatic Cell Hybridization
Authors: Eshhar Zelig , Blatt Cila , Bergman Yehudit , Haimovich Joseph
Published: December 2022

Cells of the murine B cell line 38C-13 possess immunoglobulins of the IgM class on their surface but do not secrete them. Upon hybridization of 38C-13 cells with murine myeloma cells, hybridoma clones were obtained that secreted both pentameric IgM of 38C-13 origin and the myeloma protein. All hybridoma clones synthesized and secreted large amounts of homogeneous IgM with a half disappearance time of about 2 hr, typical of mature plasma cells. Concomitantly with the induction of IgM secretion, the hybridoma cells lost their surface IgM. The possibility of separate pathways for the synthesis of membrane and secreted IgM is discussed.
Properties and Applications of Monoclonal Antibodies Directed against Determinants of the Thy-1 Locus
Authors: Marshak-Rothstein Ann , Fink Pamela , Gridley Thomas , Raulet David H. , Bevan Michael J. , Gefter Malcolm L.
Published: December 2022

Fusion of cells of the mouse myeloma line, P3/X63-Ag8 with spleen cells from AKR/J mice immunized against C3H thymocytes or from (BALB/c × BALB.K)F1 mice immunized against AKR/J thymocytes gave rise to hybrid cell lines that continuously secrete antibodies specific for the Thy-1.2 and Thy-1.1 antigens, respectively. Monoclonal antibodies from four such cell lines were analyzed in detail. All were 19S IgM, and, in the presence of complement (C), had high lytic titers on T cells of the appropriate antigenicity. Their specificity was shown by lysis of thymocytes from Thy-1 congenic mouse strains, A/J(Thy-1.2) and A.Thy 1.1. Furthermore, they lyse only 60 to 70% of lymph node cells, suggesting cytotoxicity for mature T cells and not B cells. Treatment of peripheral lymphocyte populations with monoclonal antibody plus C eliminated effector cytotoxic T lymphocytes, their precursors, and the mitogenic response to Con A, but did not affect the response to LPS. Purified, fluorescein-labeled monoclonal anti-Thy-1 antibody could be used to distinguish T and B cells. Purified antibody coupled to Sepharose 6MB was used to separate viable T and B cells. Two independently isolated anti-Thy-1.2 hybridomas are indistinguishable and bind the same determinant whereas a third is unique and may bind a separate site.
Respective Influence of Extrinsic and Intrinsic Factors on the Age-Related Decrease of Thymic Secretion
Authors: Bach Marie-Anne , Beaurain Geneviève
Published: December 2022

The serum level of a circulating thymic factor (FTS) described in our laboratory diminishes in mice after adult thymectomy and with age. However, thymuses from old mice, when grafted into young adult thymectomized recipients lacking circulating FTS, can still partially restore the circulating FTS level of the recipients, whereas newborn thymuses are less efficient in restoring the serum level of FTS in old recipients than in young adult thymectomized recipients. Taken together, our results suggest that in addition to an intrinsic deficiency of the thymic secretion, “environmental” factors play a role in the disappearance of circulating FTS with age, the more so since we observed in old mouse sera factors inhibiting the in vitro biologic activity of FTS, factors that are absent from young mouse sera.
Antibody-Dependent Cell-Mediated Cytotoxicity against Cells Infected with Respiratory Syncytial Virus: Characterization of in Vitro and in Vivo Properties
Authors: Meguro Hidenori , Kervina Marija , Wright Peter F.
Published: December 2022

An in vitro 51Cr release assay for human antibody-dependent cell-mediated cytotoxicity (ADCC) against HeLa cells infected with respiratory syncytial virus (RSV) has been characterized by using leukophoresed and adherent cell-depleted adult lymphocytes. Lymphocytes from RSV seronegative children were also competent as effector cells.

Sera from children with: 1) primary and recurrent natural RSV infections, or 2) live attenuated RSV vaccine infection were examined to characterize the behavior of ADCC antibody in vivo. After natural RSV infection ADCC antibody rose and fell more rapidly than neutralizing antibody. In two children undergoing primary RSV infection with attenuated vaccine, neutralizing antibody was formed in the absence of detectable ADCC antibody. The nonparallel behavior of ADCC and neutralizing antibodies suggests the heterogeneity of either the antigen involved or the mechanism of antibody production in the two antibody systems.
Differentiation Antigens on Normal and Abelson Virus Transformed Lymphocytes
Authors: North John R. , Raschke William C.
Published: December 2022

Rabbit antisera to Abelson leukemia virus (A-MuLV)-induced murine lymphomas have been analyzed by absorption with a variety of murine lymphoma lines. Antibody binding to a panel of cell lines and normal lymphocytes was visualized by using hapten-sandwich indirect membrane immunofluorescence. Novel membrane antigens thereby detected are shared between lymphosarcomas, B lymphomas, normal B lymphocytes, and normal membrane immunoglobulin negative (sIg-) bone marrow cells, but are not found on T cells, thymic lymphomas, plasmacytoid lymphomas, or myelomas. The existence of such shared differentiation antigens suggests that sIg- A-MuLV-induced lymphosarcomas may be transformed B cell precursors. Since differences in the expression of these antigens on individual plasmacytoid lymphoma lines were found, this category of lymphomas may include cells at a variety of differentiation states.
Sex Hormone and Glucocorticoid Receptors in the Bursa of Fabricius of Immature Chicks
Authors: Sullivan David A. , Wira Charles R.
Published: December 2022

Estrogen, androgen, progestin, and glucocorticoid binding was measured in the immature chick bursa of Fabricius. Studies with whole bursal cytosol indicated the presence of distinct and noninteracting estradiol, dihydrotestosterone, progesterone, and dexamethasone receptors with mean dissociation constants of 0.1, 0.3, 7.8, and 12 nM and average binding site concentrations of 1.9, 13.6, 42, and 520 fmoles/mg protein, respectively. Binding to saturable sites was specific for steroids with estrogen, androgen, progestin, or glucocorticoid activity. For example, estrone and diethylstilbestrol competed with estradiol, whereas progesterone, dihydrotestosterone, and dexamethasone did not. Similarly, testosterone competed for dihydrotestosterone binding, whereas estradiol, progesterone, and dexamethasone competed very little. In the presence of 5 µM cortisol, progestosterone binding was competed for by the anti-progestin R-5020, but not by dexamethasone, estradiol, and dihydrotestosterone. Competition for dexamethasone binding was observed with cortisol and corticosterone but not with estradiol, dihydrotestosterone, and progesterone.

When cytosol was prepared from bursal cell pellets that were predominantly lymphoid in nature, glucocorticoid but not estrogen, androgen, or progestin binding was identified. Since only glucocorticoid binding was found, it suggests that estrogen, androgen, and progestin receptors are present in epithelial cells of the bursa. Binding of dexamethasone in intact cells to cytosol and nuclear forms of the receptor complex and the temperature-dependent translocation of receptor complex from the cytosol to the nucleus were also shown. The results presented indicate that estrogens, androgens, progestins, and glucocorticoids interact with cells in the bursa of Fabricius at physiologic concentrations.