bluuebunny/crossref_metadata_2025 huggingface dataset:
Inhibition of antibody-dependent eosinophil-mediated cytotoxicity by heparin.
Authors: Kierszenbaum F , Ackerman S J , Gleich G J
Published: December 2022

Killing of bloodstream forms of Trypanosoma cruzi, the unicellular parasite that causes Chagas' disease in humans, by human eosinophils in the presence of specific antibody to the parasite was inhibited by the polyanion heparin in a dose-dependent manner. The concentration of heparin required to completely abolish eosinophil-effected killing of the flagellate increased as the eosinophil to parasite ratio increased. These results suggest that antibody-dependent eosinophil-mediated cytotoxicity is mediated by basic constituents of the eosinophil granule.
Immunosuppressive effects of mouse seminal plasma components in vivo and in vitro.
Authors: Anderson D J , Tarter T H
Published: December 2022

Mouse seminal plasma (SP), a mixture of aqueous extracts of prostate, seminal vesicle, and epididymis, exerts potent immunosuppressive effects in vivo. The SP mixture completely suppressed both primary and secondary humoral immune responses in mice to low immunizing doses of antigen (bovine serum albumin or washed epididymal sperm), and also significantly suppressed the antibody response to high doses of immunizing antigen. The humoral immune response to epididymal sperm was also suppressed when sperm were incubated in SP and then washed before immunization and when SP was administered at a secondary site (i. p.) at the time of sperm immunization (subcutaneous). When SP components were tested individually in in vitro assays, all of them (prostate, seminal vesicle, and epididymis) suppressed mitogen-induced lymphocyte transformation. Prostatic fluid also inhibited complement-mediated hemolysis in a standard immune hemolytic assay. These data indicate that potent immunosuppressive factors are present at several locations within the male reproductive tract; these factors may serve to protect sperm from immunologic damage and prevent sensitization of females to sperm antigens after insemination.
Membrane-bound and secreted IgA contain structurally different alpha-chains.
Authors: Sitia R , Kikutani H , Rubartelli A , Bushkin Y , Stavnezer J , Hammerling U
Published: December 2022

Three different forms of alpha-chains are synthesized by BF0.3 and 615.2, two cloned cell lines derived from the murine B lymphoma 1.29. The three forms of alpha-chains differ in size, pI, cellular location, and rate of turnover. They were identified by means of lactoperoxidase-catalyzed radioiodination, internal 14C or 35S labeling, and immunofluorescence techniques as membrane-bound(alpha m), secreted (alpha s), and intracellular (alpha ic) proteins. Comparison of immunoglobulin products of the two lymphoma lines with those of a hybridoma cell line, Id 150, which secretes IgA of the 1.29 idiotype but lacks membrane IgA, confirmed the assignments of alpha m, alpha s, and alpha ic. Results of biosynthetic labeling of BF0.3, 615.2, and Id 150 in the presence and absence of tunicamycin suggest that the difference in m.w. and charge observed between alpha m and alpha s can be attributed to differences in primary amino acid structure rather than different degrees of glycosylation.
Effect of interferons on protein synthesis in human lymphocytes: enhanced synthesis of eight specific peptides in T cells and activation-dependent inhibition of overall protein synthesis.
Authors: Cooper H L , Fagnani R , London J , Trepel J , Lester E P
Published: December 2022

Lymphoblastoid and fibroblast IFN inhibited PHA stimulation of overall protein synthesis in human lymphocytes by ca. 30%. Inhibition occurred within the first 6 hr of PHA treatment and was not progressive. DNA synthesis at 48 hr was inhibited to the same extent. Overall protein synthesis in resting lymphocytes was not detectably inhibited by IFN concentrations up to 1000 U/ml. Thus, inhibition of protein synthesis and subsequent reduction of cell proliferation by IFN require certain early events in mitogen activation. Resting lymphocytes were not unresponsive to IFN treatment, however. Two-dimensional electrophoretic analysis of newly synthesized proteins after IFN treatment showed enhanced synthesis of a specific set of eight peptides (I-peptides), which were shown to be synthesized in T lymphocytes. This enhancement was produced by both IFN-alpha and IFN-beta after 4 to 6 hr of exposure and was identical for all lymphocyte donors studied. After growth stimulation, IFN treatment produced no enhancements of additional peptides, although the original eight I-peptides were enhanced as usual. It is concluded that the biochemical activities of the I-peptides, which remain to be determined, cannot inhibit protein synthesis in resting lymphocytes, but may do so after mitogen activation, when the major physiologic restriction of lymphocyte protein synthesis is released. Alternatively, the I-peptides may be unrelated to regulation of protein synthesis but may be involved in viral protection or enhancement of NK activity.
Immunochemical studies of infectious mononucleosis. VIII. A glycoprotein from sheep erythrocytes with sialic acid-dependent receptor properties.
Authors: Fletcher M A , Caldwell K E , Latif Z A , Cayer M , Claflin A
Published: December 2022

A glycoprotein was solubilized from sheep erythrocyte membranes with hot aqueous ethanol. The glycoprotein was purified by phosphocellulose chromatography, ethanol precipitation, lipid solvent extraction, and DEAE chromatography. In water solution the glycoprotein existed as globular aggregates with a diameter of 7.1 +/- 2.2 nm. In the presence of sodium dodecyl sulfate, 80% of the material exhibited a subunit m.w.app of 27,000. Approximately 10% of the material had a m.w.app of only 9000 and another 10% had a m.w.app of 35,000. All three fractions were reactive with Paul-Bunnell heterophile antibody from the sera of patients with infectious mononucleosis and with Limulus polyphemus lectin. These activities were destroyed by neuraminidase treatment. Complete inhibition of the rosetting of sheep red blood cells by 4 X 10(5) human peripheral blood lymphocytes was seen at 100 to 200 micrograms glycoprotein/ml. Neuraminidase-treated glycoprotein was not inhibitory. Pronase-derived sialoglycopeptide was inhibitory. Most likely, the receptor for lymphocytes resides in the carbohydrate portion of the glycoprotein. By using 125I-glycoprotein, binding studies were carried out that yielded an estimate of approximately 2 X 10(5) binding sites for sheep erythrocyte glycoprotein per lymphocyte. Purified glycoprotein contained 44.4% amino acid. Carbohydrate components and their molar ratios were sialic acid (1.0): galactose (1.0):N-acetylglucosamine (1.3): N-acetylgalactosamine (1.2).
Tissue origins of human polymeric and monomeric IgA.
Authors: Kutteh W H , Prince S J , Mestecky J
Published: December 2022

Human tissues were examined for their ability to produce monomeric and polymeric forms of IgA in vitro. Mononuclear cells were examined for the presence of cytoplasmic immunoglobulins and J chain (by immunofluorescence) and for the amounts and molecular forms of intracellular and secreted IgA (by radioimmunoassay). Cultured bone marrow cells were the largest producers of total IgA, and approximately 90% of the secreted IgA was monomeric. Spleen cells also produced predominantly monomeric IgA. Intestinal lamina propria cells secreted the greatest amounts of polymeric IgA, but also produced significant amounts of monomeric IgA. Tonsillar, lymph node, and peripheral blood cells produced approximately equal proportions of monomeric and polymeric IgA. These results suggested that bone marrow may be a major source of serum monomeric IgA, and that lymphoid tissues associated with secretory surfaces may provide a greater proportion of polymeric IgA than other lymphoid tissues. There was a positive correlation between the production of polymeric IgA and the presence of cytoplasmic J chain; tissues that produced predominantly monomeric IgA generally displayed fewer cells containing cytoplasmic J chain.
Modulation of granulomatous hypersensitivity. IV. Immunoglobulin and antibody production by vigorous and immunomodulated liver granulomas of Schistosoma mansoni-infected mice.
Authors: Boros D L , Amsden A F , Hood A T
Published: December 2022

The interlesional production of immunoglobulins and SEA-specific antibodies was examined in vitro in cultured hepatic granulomas isolated from Schistosoma mansoni-infected mice. Vigorous lesions of 8-wk and immunomodulated lesions of 20-wk infected mice were cultured in serum-free medium for 48 hr; the supernatant fluid was concentrated, dialyzed, and tested for immunoglobulins by immunodiffusion. Whereas cultures of vigorous granulomas contained only IgG1, those of immunomodulated lesions yielded IgG1, IgG2a, IgG2b, IgG3, and IgA immunoglobulins. Both types of lesions incorporated 14C-labeled leucine into IgM and IgG class immunoglobulins thus proving intralesional synthesis. The immunoglobulins also had specific anti-SEA activity proven by passive hemagglutination and in vivo PCA test. The kinetics of SEA-specific IgM and IgG antibody-forming granuloma lymphocytes was examined by the plaque assay after the dispersal of the lesions. At 8 wk of the infection the number of IgM antibody-producing lymphocytes was low and that of IgG was negligible. In subsequent weeks both IgM and IgG antibody-forming cells increased in numbers. The IgM producer cells peaked at 12 to 16 wk and by 32 wk they dropped to barely detectable levels. The IgG antibody-producing lymphocytes peaked in numbers in the immunomodulated lesions at 20 wk and also disappeared by 32 wk. The kinetics of the granuloma lymphocytes as well as the magnitude of their response differed from those of splenic cells. Intralesional antibody production may promote antigen sequestration, complex formation, and occasional tissue injury. The participation of locally produced antibodies in the modulation of the granulomatous inflammatory response remains to be established.
Monoclonal antibodies to Chlamydia trachomatis: antibody specificities and antigen characterization.
Authors: Stephens R S , Tam M R , Kuo C C , Nowinski R C
Published: December 2022

Nineteen independent hybrid cell lines that produce monoclonal antibodies to Chlamydia trachomatis surface antigens were prepared by the fusion of mouse myeloma cells with lymphocytes of mice that were immunized with C. trachomatis immunotypes B, C, and L2. Seven serologically distinct reaction patterns were detected by microimmunofluorescence (micro-IF) of elementary body (EB) preparations when culture fluids were tested against a panel of 18 chlamydial serotyping reference strains. These reaction patterns demonstrated genus-, species-, subspecies-, and type-specific distributions. Additionally, these antibodies were tested in parallel against reticulate body (RB) preparations of several chlamydial strains. Monoclonal antibodies that reacted with genus-specific antigens reacted preferentially with RB, whereas antibodies that reacted to species-, subspecies-, or type-specific antigens reacted equivalently to both RB and EB. Physiochemical characterization of antigens recognized by the different monoclonal antibodies was assessed by heat treatment, pronase digestion, periodate oxidation, and immuno-blot techniques. The genus-specific antigen was a heat-stable, pronase-resistant, and relatively periodate-sensitive component of less than 10,000 m.w. The species-, subspecies-, and type-specific antigens were heat stable, pronase sensitive, and periodate resistant. The antibodies that detected species- and subspecies-specific antigens predominantly reacted in immuno-blots with the 40,000 m.w. major outer membrane protein. These monoclonal antibodies now provide a new approach for the precise serologic classification and detection of different C. trachomatis strains.
Lysis of hapten-labeled cells by anti-hapten IgG and complement: effect of cell surface hapten density.
Authors: Circolo A , Borsos T
Published: December 2022

Rabbit anti-methotrexate (MTX) IgG antibody was used to study the effect of MTX density on sheep red cells on the lysis of cells by complement (C). Under conditions in which the density of IgG did not vary, the number of lytic IgG complexes remained approximately the same when the average distance between MTX molecules was between 5 and 7 nm, but decreased rapidly with an average distance less than 10 nm. The results were interpreted to mean that in addition to aggregation for IgG to acquire maximal C in fixing and activating properties, the angle of the Fab arms of the molecule must not be larger than about 50 to 60 degrees. The data also suggested IgG molecules with Fab angles of less than about 30 degrees might also not be able to induce C-mediated cell lysis.
Structure of Ia antigens: identification of dimeric complexes formed by the invariant chain.
Authors: Koch N , Hämmerling G J
Published: December 2022

A 2-dimensional gel technique was used for the identification of disulfide-linked polypeptide complexes formed by Ia chains and their corresponding subunits. Several dimers containing the invariant (Ii) chain were found. The most prominent one consisted of 2 Ii chains. In addition, Ii molecules were found to be covalently linked to 3 unknown components of 41,000, 27,000, and 10,000 to 15,000 m.w. The dimeric form of Ii exists also after alkylation of free SH-groups with iodoacetamide, which suggests that formation of the Ii dimer is not the result of experimental conditions. Furthermore, noncovalently associated dimers formed by processed alpha- and beta-chains as well as by their precursors alpha p and beta p could be detected. No mixed associated chains composed of precursors and processor molecules were observed, which indicates that processing and insertion into the membrane take place only after dimerization.
The area of attachment of cytotoxic T lymphocytes to their target cells shows high motility and polarization of actin, but not myosin.
Authors: Ryser J E , Rungger-Brändle E , Chaponnier C , Gabbiani G , Vassalli P
Published: December 2022

Conjugates of cytotoxic T lymphocytes attached to their target cells were studied by double immunofluorescence on fixed smears to detect simultaneously the localization of actin and myosin within the cells. Actin was found to be polarized in the area of attachment of the lymphocytes (but not of the target cells), whereas myosin remained evenly distributed in the cytoplasm. When living conjugates were brought to 37 degrees C to induce cytotoxicity, this pattern remained unchanged, but observation by interference reflexion microcinematography revealed a high motility of the lymphocytes in the contact area. This localized motility in the area of attachment associated with a peculiar actin polarization, which has no equivalent in any type of cell contact presently known, could represent a necessary step in the sequence of events leading to target cell killing by cytotoxic T lymphocytes.
Ultrastructural events in the translocation of polymeric IgA by rat hepatocytes.
Authors: Takahashi I , Nakane P K , Brown W R
Published: December 2022

It has been proposed that polymeric IgA is translocated from plasma to bile across hepatocytes of the rat liver by a secretory component-mediated, vesicular transport. To define the ultrastructural details of the proposed transport mechanism, we employed peroxidase-labeled antibody immunocytochemistry to localize secretory component in the rat liver and monitor the hepatic translocation of homologous myeloma polymeric IgA infused i.v. Secretory component was found associated with the endoplasmic reticulum, Golgi complexes, cytoplasmic vesicles, and plasma membranes of the sinusoidal and canalicular surfaces of hepatocytes; secretory component at the sinusoidal surface was most prominent in micropinocytic invaginations or pits. Livers were examined for the sites of polymeric IgA 5, 15, and 30 min after infusion. Evidence was obtained that polymeric IgA is translocated across hepatocytes by a series of events: 1) polymeric IgA binds selectively to secretory component on the external surface of the sinusoidal plasma membrane; 2) secretory component-IgA complexes are internalized in endocytic vesicles; 3) the vesicles migrate through the cytoplasm without association with lysosomes or Golgi complexes; 4) the vesicles fuse with the cytoplasmic surface of the bile canalicular membrane, where secretory component-IgA complexes are released into bile by exocytosis.
Phagocytosis of bacteria by macrophages: changing the carbohydrate of lipopolysaccharide alters interaction with complement and macrophages.
Authors: Liang-Takasaki C J , Mäkelä P H , Leive L
Published: December 2022

Salmonella transductants and recombinants differing the O-antigenic side chain of their lipopolysaccharide are taken up at different rates by the murine macrophage-like cell line J774. Bacteria containing abequose, mannose, rhamnose, and galactose in O-antigenic side chain were taken up at the slowest rate; the one containing tyvelose instead of abequose was taken up at an intermediate rate; and the one containing mannose, N-acetylglucosamine, and glucose, instead of the above sequence, was taken up at the highest rate. These rates correlate well with the known virulence of these strains; the most virulent is the one taken up slowest, the one taken up at an intermediate rate is less virulent, and the one taken up fastest is the least virulent. The differences in ingestion rates reflect differences in affinity of the bacteria for the macrophages and not in the rate of ingestion once interaction has occurred, suggesting a receptor-mediated process. The majority of uptake is probably dependent on complement, as shown by the requirement for a serum component(s) destroyed by heating at 56 degrees C or by incubation with zymosan. Specific antibody is not required. We therefore postulate that relative virulence in vivo may reflect the relative ability of the polysaccharide of bacterial lipopolysaccharide to activate complement, thus determining the susceptibility of the bacteria to ingestion via the complement receptor of phagocytic cells.
Cross-reactivity with Escherichia coli K100 in the human serum anticapsular antibody response to Haemophilus influenzae type B.
Authors: Insel R A , Anderson P W
Published: December 2022

Escherichia coli K100 is known to induce antibodies cross-reactive with the capsular polysaccharide (CP) of Haemophilus influenzae b (Hib); the cross-immunogenicity is found consistently in a number of mammalian species including man. We have studied the reciprocal cross-immunogenicity of Hib or its purified CP in man. Of 25 adults and 13 children responding to systemic immunization with Hib CP, only four and four, respectively, made K100 CP cross-reactive antibody, determined by radioantigen-binding inhibition. The detection of preimmunization cross-reactive antibody was not statistically predictive of induction of postimmunization cross-reactive antibody. Cross-reactive antibody was detected in only one of 11 children making an anticapsular response to Hib systemic infection and two of four children responding to asymptomatic nasopharyngeal colonization. In contrast, cross-reactivity was detected in the naturally occurring anti-Hib-CP antibody of 27 of 42 children. Thus, humans responding to Hib or its CP often do not make antibody to the K100-cross-reactive determinant(s), implying that the cross-reactive natural antibodies frequently seen in children must have a stimulus other than Hib.
Induction of proliferation and Ig production in human B leukemic cells by anti-immunoglobulins and T cell factors.
Authors: Yoshizaki K , Nakagawa T , Kaieda T , Muraguchi A , Yamamura Y , Kishimoto T
Published: December 2022

The proliferation and differentiation of human leukemic B cells (B-CLL cells) with anti-Ig and T cell-derived helper factors are described. Stimulation of B-CLL cells with anti-Ig and T helper factors could induce proliferation as well as differentiation into IgM- and IgG-producing cells. Neither anti-Ig nor T helper factors alone could induce any proliferation and/or differentiation of B-CLL cells. Not only whole molecules of anti-Ig but also F(ab')2 fragments could induce proliferation and differentiation of B-CLL cells in the presence of T helper factors, but monovalent Fab' fragments were not effective. Induction of both IgM and IgG with the same idiotype was confirmed by immunofluorescent and SDS-PAGE analysis. By employing an IL 2-dependent cytotoxic T cell line and a TRF-responsive B cell line, T cell factors were separated into a fraction with IL2 activity but no TRF activity and a fraction with TRF activity but no IL 2 activity by chromatofocusing. Anti-Ig and IL 2 fraction could induce proliferation of B-CLL cells, but TRF fraction was not effective for the induction of proliferation in anti-IG-stimulated cells. For IgM and IgG production, anti-Ig and both IL 2 and TRF fractions were required. Depletion of IL 2 fraction in the first 2 days' culture inhibited Ig production, whereas the absence of TRF fraction in the first 2 days did not show any inhibitory effect on Ig production.
Constitutive and mitogen-induced production of T cell growth factor by stable T cell hybridoma lines.
Authors: Altman A , Sferruzza A , Weiner R G , Katz D H
Published: December 2022

Stable T cell growth factor- (TCGF; IL 2) producing cloned T cell hybridoma lines were constructed by fusing murine alloantigen-activated T cells with the 8-azaguanine-resistant lymphoma line, BW5147. Many, but not all, clones of one of these hybridomas, i.e., hybridoma 24, secreted TCGF constitutively, but production was markedly enhanced by stimulation with T cell mitogens. Large numbers of TCGF-secreting hybridoma cells in a stable functional state could be obtained from histocompatible mice inoculated with cloned T cell hybridomas. Moreover, such in vivo-derived hybridoma cells could be stimulated sequentially with mitogen at least twice to secrete their biologically-active product, resulting in larger TCGF yields from the same cells. The secreted product of these T cell hybridoma lines resembled TCGF isolated from other cellular sources in that it: a) supported the growth of a TCGF-dependent T cell line; b) provided help for the induction of alloantigen-reactive cytotoxic T lymphocytes from thymocyte precursors; c) facilitated concanavalin A-induced mitogenic responses of low thymocyte numbers; d) had an apparent m.w. of 30,000 to 40,000 by gel filtration chromatography; and e) was eluted from DEAE-Sephacel ion-exchange chromatography columns by salt concentrations of 30 to 150 mM NaCl. The ability of these T cell hybridomas to grow in vivo and retain their functional characteristics in a stable form should prove useful in terms of providing large numbers of TCGF-secreting cells and studying in vivo aspects of the production of TCGF as well as other immunoregulatory mediators.
Hapten-specific B cell repertoire probed by hybridoma technology: selection and characterization of representative clonotypes from the antibody-forming cell pool.
Authors: Bloor A G , Jou Y , Hoeplinger C , Gartner J E , Mayers G L , Bankert R B
Published: December 2022

In the course of this study, more than 3000 phthalate-specific antibody-forming cell hybrids were identified using the hybridoma technology. With the aid of a rapid screening assay and an extensive library of phthalate analogs, it was possible to assign selected hapten-specific clones to one of 11 distinct fine-specificity sets. This compartmentalization of the phthalate-specific hybridomas has made it possible to focus attention upon a single manageable portion of the phthalate-specific repertoire. Fourteen clones from a single fine-specificity set were selected for further immunochemical characterization. Five of these clones were found to secrete an antibody that was indistinguishable in isoelectric focusing. Affinity-purified, high-resolution anti-idiotype antibodies were prepared with specificity for the antibodies produced by one of these clones (i.e., 4C7). A major portion of the serologically defined private idiotype (4C7 IdI) was shown to be associated with the ligand-combining site. Our results indicate that the five clones that share a common spectrotype also express the 4C7 IdI. Two other independently derived clones from two distinct fusions also share this idiotype. The 4C7 IdI was also identified in affinity-purified anti-phthalate antibodies derived from a pool of phthalate-immune serum (conventional antibody) and from affinity-purified antibodies derived from a pool of serum from unimmunized BALB/c mice (natural antibody). The 4C7 IdI is thus considered to represent a repeating clonotype in the phthalate-specific repertoire of BALB/c mice, and will serve as one of several useful clonal markers that are being developed for studies of the mechanism regulating idiotype expression.
The capacity and mechanism of bone marrow antibody formation by thymus-independent antigens.
Authors: Koch G , Lok B D , van Oudenaren A , Benner R
Published: December 2022

Primary immunization of mice with certain thymus-independent (TI) antigens (i.e., TNP-LPS and DNP-Ficoll) leads to antibody formation in the bone marrow (BM). TNP-Brucella abortus, Pneumococcus pneumoniae organisms, and alpha-(1,6) dextran, on the other hand, do not induce a BM antibody-producing plaque-forming cell (PFC) response. This paper deals with the mechanism underlying antibody formation in the BM to TNP-LPS and DNP-Ficoll. The majority of the BM-localizing PFC induced by TNP-LPS are formed within the BM from the proliferating lymphocyte pool, because this response was found to be resistant to splenectomy and sensitive to treatment with hydroxyurea (HU) before immunization. This local activation of newly formed B cells requires in addition to the antigenic signal of TNP-LPS the mitogenic signal from the lipid A component of LPS. In contrast, the BM PFC response to DNP-ficoll was reduced in splenectomized mice and resistant to HU treatment before the primary immunization. Thus, antibody formation in the BM to DNP-Ficoll is mainly dependent on long-lived B cells that migrate from the peripheral lymphoid organs into the BM.
Immune recognition in the streptococcal carditis of mice: the role of macrophages in the generation of heart-reactive lymphocytes.
Authors: Dos Reis G A , Gaspar M I , Barcinski M A
Published: December 2022

The role of immune T cells and of streptococcus-pulsed M phi in the production of cardiac lesions, as well as the ability of M phi to present streptococcal antigens to mouse lymphocytes, was investigated. Cells of mice infected with extracts of group A streptococcus were able to induce the appearance of heart lesions when transferred to syngeneic receptors as well as to transfer DTH reactions to syngeneic heart extracts. Streptococcus-pulsed M phi were also able to induce heart lesions and an increase in the serum CPK activity when injected into syngeneic receptors. This last phenomenon was only observed in mice aged 5 mo or more. Furthermore, it was shown in an in vitro model of T cell proliferation that peritoneal M phi pulsed with group A streptococci are able to induce a specific response to syngeneic cardiac extracts. M phi pulsed with group G streptococcus failed to induce such a response. Those findings are discussed as part of a model for the induction of rheumatic cardiac lesions in which M phi display a central role by selecting antigenic determinants from the pathogenic organism for presentation to immunocompetent cells.
Human serum-free mixed lymphocytes response: the stereospecific effect of insulin and its potentiation by transferrin.
Authors: Strom T B , Bangs J D
Published: December 2022

We have established a serum-free human mixed lymphocyte response (MLR). Proliferative unidirectional responses are discerned using peripheral blood leukocytes obtained from unrelated individuals and family members mismatched for HLA while MLR conducted between HLA identical siblings are negative. Resting lymphocytes lack insulin and transferrin receptors; activated lymphocytes are known to express insulin and transferrin receptors. Interestingly, physiologic concentrations of insulin and transferrin synergistically potentiate DNA synthesis in serum-free allogeneic cultures. To verify that the insulin effect was due to a stereospecific interaction between insulin and classical, high-affinity insulin receptors, insulin and a series of insulin-like peptides were compared in their ability to bind to alloactivated T cells and amplify the MLR. The results demonstrate the stereospecificity of the insulin effect in the MLR and underscore the importance of lymphocytic insulin receptors in the process of lymphocyte activation.
Thymus cell migration: cells migrating from thymus to peripheral lymphoid organs have a "mature" phenotype.
Authors: Scollay R
Published: December 2022

To gain information on the lineage relationship of cells leaving the thymus, we studied the phenotype of thymus emigrants within hours of their exit. The migrants were identified in the peripheral lymphoid organs by their fluorescence, 3 to 4 hr after intrathymic injection of a solution of fluorescein isothiocyanate, a technique that initially only labels thymocytes. Migrants identified in this way were analyzed with rhodamine-anti-Thy-1 or rhodamine peanut agglutinin (PNA). They were found to express Thy-1 antigen and PNA binding sites at levels very similar to those found on the majority of peripheral T cells or medullary thymocytes and quite different from cortical thymocytes. Taken together with our previous experiments on Lyt-1, Lyt-2, and H-2 levels, the data show that cells leaving the thymus are quite mature in phenotype and are indistinguishable from peripheral T cells by all the criteria examined.
The specificity of in vivo tolerance to haptens in NZB and normal mice after exposure to hapten-modified syngeneic spleen cells.
Authors: Cowdery J S , Laskin C A , Steinberg A D
Published: December 2022

We have evaluated hapten-specific hyporesponsiveness induced by in vivo administration of TNP-modified syngeneic spleen cells (TNP-SC). Pretreatment of non-autoimmune mice led to hyporesponsiveness to challenge with either TNP or the closely related hapten DNP coupled to Ficoll. There was also a significant reduction of the direct PFC response after challenge with TNP-HGG. In tolerized mice challenged with TNP-HGG, the IgM portion of the serum response was similarly suppressed; however, the total serum antibody as well as the indirect PFC response was not suppressed. There was no tolerance at all when the mice were challenged with DNP-HGG. Thus, exposure to TNP-SC results in an incomplete form of hapten-specific B cell tolerance. This tolerance is selective for the IgM isotype and does not extend to the cross-reactive hapten DNP on a thymic-dependent carrier, although it does extend to a DNP on a thymic-independent carrier. Autoimmune NZB mice were defective with regard to tolerance after injection of hapten-modified syngeneic spleen cells. They did manifest a reduced direct PFC response to the challenge with TNP-Ficoll, but failed to demonstrate cross-tolerance to DNP-Ficoll challenge. Moreover, they did not have suppression of the hapten-specific IgM response after challenge with TNP on the thymic-dependent carrier. These abnormalities in tolerance induction in NZB mice to modified self may help to explain the loss of self-tolerance that occurs spontaneously and is expressed as autoimmune disease.
Expression of histoincompatibility antigens during spermatogenesis in the rat.
Authors: Söderström K O , Sege K , Andersson L C
Published: December 2022

The expression of the AgB-SB and -LD (Ia-like) antigens on spermatogenic cells has been investigated by using a protein A-containing Staphylococcus Cowan I rosetting assay. The results show that the Ia-like antigens are present on all spermatogenic cells although the relative expression seems to decrease slightly concomitant with the differentiation. The AgB-SD antigens are detected on spermatogenia, leptotene, and zygotene spermatocytes but not on later developmental stages.
A phorbol ester (TPA) can replace macrophages in human lymphocyte cultures stimulated with a mitogen but not with an antigen.
Authors: Koretzky G A , Daniele R P , Nowell P C
Published: December 2022

A culture system was developed in which human peripheral blood mononuclear cells (PBM) depleted of macrophages did not proliferate in response to the lectin mitogen PHA or to the soluble antigen of tetanus toxoid. These cells were able to respond to both mitogen and antigen if purified autologous macrophages were added back to the culture. The response to PHA was partially restored by supplementing the cultures with supernatants from LPS-stimulated macrophages or with partially purified human interleukin 1 (IL 1). The response to tetanus was not restored by reconstitution with these materials. The phorbol ester, 12-O-tetradecanoyl-phorbol-13-acetate (TPA), has been shown to have IL 1-like effects in other species and is a polyclonal activator of human T and B lymphocytes. In this study, we tested the ability of TPA to replace macrophages in human lymphocyte cultures stimulated with mitogen or with antigen. Small doses of TPA (50 ng/ml) completely replaced macrophages in the PHA-stimulated cultures; however, in doses of up to 400 ng/ml, TPA was not able to replace macrophages in cultures stimulated with tetanus. Thus, TPA appears to mimic the macrophage-replacing ability of soluble factors (IL 1, macrophage supernatants) in the triggering of human lymphocytes.
Macrophages as effector cells of protective immunity in murine schistosomiasis. III. Loss of susceptibility to macrophage-mediated killing during maturation of S. mansoni schistosomula from the skin to the lung stage.
Authors: Sher A , James S L , Simpson A J , Lazdins J K , Meltzer M S
Published: December 2022

Newly transformed skin-stage and lung-stage schistosomula were compared in terms of their susceptibility to killing mediated by activated mouse macrophages in vitro. Although skin-stage schistosomula were readily killed by macrophages activated as a consequence of either BCG or Schistosoma mansoni infection and used either as cell monolayers or in suspension, lung-stage larvae appeared to be totally resistant to this effector mechanism and survived normally when reinjected into mice. Resistance of schistosomula to in vitro damage by macrophages was evident as early as 18 hr after host infection and was complete in worms recovered at 42 hr. The insusceptibility of lung-stage larvae is apparently not due to a defect in effector cell-target contact, because the induction of extensive macrophage adherence to the worms by the addition of anti-mouse red blood cell antisera to the cultures had no effect on parasite viability. These findings provide additional support for the concept that schistosomula during their development to the lung stage undergo a generalized change affecting their susceptibility to a variety of different immunologic effector mechanisms.
Inhibition of autologous rosette formation by monoclonal antibody to the sheep erythrocyte receptor.
Authors: Scheffel J W , Swartz S J
Published: December 2022

The commercial monoclonal HuLyT-3, and which recognizes a 55,000-dalton polypeptide associated with the receptor for SRBC on human T lymphocytes, was capable of blocking rosette formation between human T cells and autologous human RBC. Lysostripping of the T cell surface with HuLyT-3 abolished rosette formation with both sheep and autologous RBC. The data suggest that a common structure is involved in the spontaneous rosetting of both sheep and human RBC by human T cells.
Murine I-Ak alpha-chain subspecies with glycosylation differences.
Authors: Cowan E P , Schwartz B D , Cullen S E
Published: December 2022

In isolating homogeneous populations of murine Ia antigens by SDS-PAGE we have found that immunoprecipitated I-Ak molecules from whole spleen possess an electrophoretically heterogeneous population of alpha-chains. SDS-PAGE of 3H-mannose-labeled I-Ak immunoprecipitates revealed that the alpha-chain peak was divided into three regions (alpha 1, alpha 2, and alpha 3), each of which migrated as a distinct individual band when isolated and reelectrophoresed. Only alpha 1 and alpha 2 labeled with 3H-fucose. Nonequilibrium pH gradient electrophoresis excluded the possibility that any of the species was invariant chain, but rather suggested that the subspecies were structurally related and that alpha 3 has fewer sialic acid residues than alpha 1 and alpha 2. Sialic acid differences between alpha 1, alpha 2, and alpha 3 were further suggested by SDS-PAGE and by peptide mapping of neuraminidase-treated immunoprecipitates. Tryptic peptide maps of 3H-leucine-labeled alpha 1, alpha 2, and alpha 3 were identical, indicating that the polypeptide backbone of the molecules is the same for all subspecies. Our results suggest that alpha 1 and alpha 2 have fucose and sialic acid and thus possess complex type oligosaccharides, whereas alpha 3 lacks these sugars and possess high mannose-type saccharides. The potential relationships of the alpha-chain subspecies is discussed. In addition, we demonstrate the usefulness of high-pressure liquid chromatography in the isolation of glycopeptides for their subsequent analysis.U
Studies on the behavior of H-Y incompatible skin grafts in rats.
Authors: Chen H D , Silvers W K
Published: December 2022

Studies on the behavior of H-Y incompatible skin grafts in rats indicate that 1) there is considerable variation in the ability of females of different strains to reject male skin isografts; 2) small, male trunk skin isografts are more likely to be rejected than similarly sized ear or tail skin grafts; 3) other factors besides the MHC plan an important role in determining the ability of females to reject male skin isografts; 4) the ability of adult male skin grafts to induce unresponsiveness to H-Y depends upon their size (large grafts are more effective than small) and the genotype of the host; 5) even in those strains in which females usually accept small, male trunk skin isografts, such grafts are acutely rejected by sensitized females; and 6) the total amount of H-Y incompatible trunk skin the host is exposed to, rather than the size of each graft, determines their fate.
Biochemical comparison of MHC antigens isolated from mutant C3HfB/HeN and parent C3H/HeN mice.
Authors: Callahan G N , Martin W J , Pardi D
Published: December 2022

H-2K antigen heavy chains were isolated from C3HfB/HeN and C3H/HeN spleen cells by immunoprecipitation and high pressure liquid chromatography and were compared by tryptic peptide mapping. Glycopeptides obtained from 3H-glucosamine-labeled H-2Kk (C3H) and H-2Kkv1 (C3Hf) heavy chains were found to be identical, indicating that previously observed differences between the two molecules are due to alterations in amino acid sequence. I-A subregion antigen alpha- and beta-chains were also isolated from C3H and C3Hf spleen cells by immunoprecipitation and SDS-polyacrylamide gel electrophoresis, and were compared by tryptic peptide mapping analyses. Material isolated from 3H-glucosamine-labeled cells revealed that I-A alpha- and beta-chains were glycosylated, and antigens from C3H and C3Hf mice contained identical glycopeptides. Similar analyses of I-A antigen alpha- and beta-chains isolated from C3H and C3Hf spleen cells labeled with 3H-arginine or 3H-lysine revealed no differences in the amino acid sequences of the I-A antigens of these two strains. These findings indicate that all of the unique immunologic properties of mutant C3Hf cells can be attributed to modifications of the amino acid sequence of the MHC K locus gene product.
Receptors for IgA on rabbit lymphocytes. II. Characterization of their binding parameters for IgA.
Authors: Stafford H A , Knight K L , Fanger M W
Published: December 2022

The specificity of rabbit receptors for IgA (RFc alpha) was investigated on lymphocytes isolated from the Peyer's patches and spleen. Specifically was determined by the inhibition of IgA rosette formation with intact and proteolytic fragments of rabbit and human secretory IgA (SIgA), human myeloma proteins, and secretory component. RFc alpha bound human SIgA as efficiently as rabbit SIgA. Of the human IgA tested, RFc alpha preferentially bound to the IgA2 subclass and most avidity to a multimeric IgA molecule. In addition., the site of the RFc alpha-IgA interactions was localized to the C alpha 2 domain in the Fc portion of IgA. RFc alpha on spleen lymphocytes showed the same pattern of binding specificity toward the human IgA subclasses as the RFc alpha on lymphocytes from Peyer's patches. However, differences between the spleen and Peyer's patches were observed in the interaction of RFc alpha and the rabbit IgA subclasses. RFc alpha on lymphocytes from the Peyer's patches bound the rabbit IgA-g subclass more efficiently than the rabbit IgA-f subclass, whereas the converse was true for the spleen RFc alpha. Human secretory component inhibited IgA but not IgG or IgM rosette formation as a result of its interaction with lymphocytes. These studies suggest: 1) that there may be two types of RFc alpha expressed by lymphocytes in certain tissues, and 2) that there are two closely associated receptors on the lymphocyte--one specific for secretory component and the other for structures within the C alpha 2 domain of IgA.
Mouse lymph node germinal centers contain a selected subset of T cells--the helper phenotype.
Authors: Rouse R V , Ledbetter J A , Weissman I L
Published: December 2022

Cells staining for Lyt-1 are more frequent than cells staining for Lyt-2 in both primary follicles and the cuffs of secondary follicles; there is an even more striking predominance of cells bearing only Lyt-1 in germinal centers. In addition, there is an increase in the total percentage of cells bearing T cell antigens in germinal centers compared to primary follicles. These differences in phenotype and distribution of T cell populations indicate the T cells in B cell areas, and especially in germinal centers, are not randomly selected, but rather represent a specific subpopulation of T cells enriched for the helper phenotype (Lyt-1+2-), perhaps involved in the development and/or function of germinal centers.
Ipr gene control of the anti-DNA antibody response.
Authors: Pisetsky D S , Caster S A , Roths J B , Murphy E D
Published: December 2022

The influence of the Ipr gene on the anti-DNA antibody response was investigated in MRL and B6 Ipr/Ipr inbred mice, MRL +/+ mice less than a yr of age produced low levels of anti-DNA antibody, whereas older animals of this strain demonstrated levels in some instances comparable to those of the more severely affected MRL Ipr/Ipr mice. This result indicates a tendency to autoreactivity in MRL mice independent of the Ipr gene. To determine whether other mice bearing the Ipr gene would also express autoantibodies, the anti-DNA antibody responses of B6 Ipr/Ipr mice were studied. This strain was development by matings to transfer the Ipr gene into another inbred background and allow evaluation of the action independent of other disturbances of the MRL mice. Mice of this strain produced antibodies to DNA, with female animals displaying significantly higher levels than males. This result demonstrates that the Ipr gene can stimulate autoantibody production in mice other than the MRL strain and does not require abnormalities unique to this background to potentiate autoreactivity.
Successful murine tumor allotransplantation after total lymphoid irradiation.
Authors: Hoppe R T , Dorie M J
Published: December 2022

C3H/Km (H-2k) mice were treated with fractionated total lymphoid irradiation (TLI) (17 x 200 rad) and then were inoculated with 3 x 10(6) tumor cells from the EMT6 tumor cell line derived from the BALB/c (H-2d) mouse. One group of mice also received an i.v. infusion of BALB/c bone marrow (3 x 10(7) cells). Tumors in control animals (no TLI) became palpable within 10 days after inoculation, but subsequent tumor regression was rapid, with complete disappearance in all animals by 23 days. Initial tumor growth was exponential in both groups of mice treated with TLI, but tumors regressed after 30 days in the animals treated with TLI alone. In the group treated with TLI and bone marrow infusion exponential tumor growth continued until pulmonary metastases developed and all animals died. The TLI-treated mouse provides a model for tumor allografting. Further development of this model may demonstrate its efficacy in human tumor xenotransplantation.
Constitutive production of a unique lymphokine (IL 3) by the WEHI-3 cell line.
Authors: Lee J C , Hapel A J , Ihle J N
Published: December 2022

It has been recently demonstrated that conditioned media from Con A-stimulated splenocyte cultures contain a novel lymphokine termed IL 3. IL 3 is characterized by its ability to induce 20-alpha-hydroxysteroid dehydrogenase in spleen cells from young nu/nu mice. In search of a convenient source for biochemical and in vivo studies of this lymphokine, a number of established murine cell lines were screened for the constitutive and induced production of Il 3. It was found that WEHI-3 cells, originally designated as a myelomonocyte cell line, produced high levels of IL 3 constitutively. Added mitogen and/or phorbol acetate did not enhance the production of IL 3. Production of IL 3 varied among various sublines of WEHI-3. IL 3 purified from WEHI-3-conditioned media has biochemical and biologic characteristics identical to those obtained from Con A-conditioned media. WEHI-3 conditioned media, however, generally contained 100-fold higher levels of IL 3 than conditioned media from Con A-stimulated splenic lymphocytes.
Demonstration of primary cytotoxic T cells in venous blood and cerebrospinal fluid of children with mumps meningitis.
Authors: Kreth H W , Kress L , Kress H G , Ott H F , Eckert G
Published: December 2022

Cryopreserved lymphocytes from venous blood and cerebrospinal fluid (CSF) of 10 children with mumps meningitis were tested in 5-hr 51Cr-release assays against uninfected and mumps virus-infected PHA-blasts. Lymphocytes from all patients were cytotoxic to autologous mumps virus-infected target cells, but completely failed to lyse histoincompatible virus-infected PHA-blasts. Cytotoxicity was specific for the infecting virus, and was mediated by E rosette-forming lymphocytes. The effector cells were present over 2 to 3 wk after onset of meningitis. Mumps viral antigens appeared to be preferentially recognized in association with HLA B determinants. The results show that specifically sensitized cytotoxic T cells (CTL) are induced in patients with mumps meningitis. These cells circulate in venous blood and are locally enriched in CSF. Based on clinical observations, it is proposed that mumps-specific CTL play an immunopathologic role.