bluuebunny/crossref_metadata_2025 huggingface dataset:
The V region sequence of lambda Bence-Jones protein Wh: evidence for separate germ-line sets within lambda-subgroups.
Authors: Kiefer C R , Patton H M , McGuire B S , Garver F A
Published: December 2022

The complete variable region sequence of the glycosylated Bence-Jones protein Wh has revealed that the most hypervariable segments of the first and third complementarity-determing regions (CDR) are identical with those of another glycosylated Bence-Jones protein (Nei) of the same lambda-subgroup. The carbohydrate is attached in both proteins at the identical sequences in the third CDR, and the sequences of the first CDR differ at only one position in fourteen. Because CDR sequence comparisons more sensitively reflect overall germ-line V gene similarities, proteins Wh and Nei could possibly reflect a "set" relationship analogous to the mouse V kappa isotype. Cluster analyses of currently available V lambda sequences supported this suggested relationship.
Production of Con A-induced helper T cell replacing factor requires a T cell and an Ia-positive non-T cells.
Authors: Swain S L , Dutton R W
Published: December 2022

Evidence is presented that indicates that two cells are required for the production of Con A-induced, helper T cell replacing factor. The factor production involves both a T cell and an IA,IE/C-positive non-T cell. The T cell and the Ia-positive cell need not be H-2 compatible. Limiting dilution analyses suggested that the Ia-positive cell is limiting in normal spleen. A hypothesis is put forward that the activation of T cells by Con A may involve "gluing" normally specific, functional T cells to the Ia expressing cells with which they would normally interact in antigen-specific activation.
Differentiation of Thy 1-bearing cells from progenitors in long-term bone marrow cultures.
Authors: Jones-Villeneuve E V , Rusthoven J J , Miller R G , Phillips R A
Published: December 2022

Lone-term marrow cultures, established according to the technique described by Dexter, contain a colony-forming cell (CFU-T) that produces Thy 1-positive progeny after stimulation with PHA-induced conditioned medium from human leukocytes. When 10(-7) M hydrocortisone is added to the medium for the long-term cultures CFU-T can be maintained in vitro for at least 16 weeks. Cells from individual colonies lack precursors for cytotoxic lymphocytes but can provide nonspecific help required for the generation of cytotoxic lymphocytes.
Characterization of the stimulator cells in the murine primary mixed leukocyte response.
Authors: Minami M , Shreffler D C , Cowing C
Published: December 2022

Macrophage-enriched and depleted spleen cell populations were examined for their ability to stimulate a murine 1 degree mixed leukocyte response. A quantitative comparison of unfractionated spleen cells, Sephadex g-10 passed spleen cells, 4-hr glass adherent and 24-hr glass adherent spleen cells revealed that a potent 1 degree MLR stimulator cell was phagocytic and that T and B lymphocytes failed to stimulate a response. By using several inb;ed and recombinant mouse strains congenic for different subregions of the MHC, it was found that macrophage-enriched cells were capable of inducing an MLR against K, D, I-A, and I-E differences and against K and D mutants. Both macrophage-enriched and depleted spleen cells failed to stimulate responses when the stimulator and responder differed at I-J, I-C, S, or G. Moreover, MLR stimulation induced by K or D region differences alone was dependent upon stimulator cells that expressed Ia antigens. Therefore, it would appear that only a minor subpopulation of the cells that express the relevant MHC alloantigens is immunogenic in the murine 1 degree MLR.
Lymphocytes bearing Fc receptors for IgE. IV. Formation of IgE-binding factor by rat T lymphocytes.
Authors: Yodoi J , Ishizaka K
Published: December 2022

Culture of mesenteric lymph node cells from rats infected with Nippostrongylus brasiliensis resulted in the release of a soluble factor (IgE-binding factor) that can inhibit rosette formation of Fc epsilon R(+) lymphocytes with IgE-coated red cells. The factor is specifically absorbed with IgE-coated Sepharose. It has a m.w. of between 10,000 and 20,000. The major source of IgE-binding factor appears to be Fc epsilon R(+) T cells. The formation of IgE-binding factor by the lymphocytes was enhanced by IgE added to the culture medium. Evidence was obtained that Fc epsilon R(+) cells are involved in the induction of IgE-induced factor formation. Normal rat lymphocytes cultured alone failed to release IgE-binding factor, but incubation of normal lymphocytes with rat IgE resulted in the formation of IgE-binding factor and an increase in the proportion of Fc epsilon R(+) cells. It was also found that normal T cells formed the soluble factor upon incubation with IgE. In the induction of factor formation by normal lymphocytes, Fc gamma R(+) cells are essential; an Fc gamma R-depleted fraction failed to form IgE-binding factor upon incubation with IgE. The results suggest that interaction of IgE with Fc epsilon R(+) T cells and Fc gamma R(+) T cells induces the formation of IgE-binding factor(s).
Active site radioimmunoassay for human urokallikrein and demonstration by radioimmunoassay of a latent form of the enzyme.
Authors: Silver M R , ole-MoiYoi O , Austen K F , Spragg J
Published: December 2022

A radioimmunoassay specific for the active site in urokallikrein has been developed with a monospecific antibody that neutralizes the enzymatic activities of urokallikrein and a radioligand purified so as to maintain the active site. In order to favor the involvement of the antibody with high affinity for the active site in the competition between urokallikrein in biological fluids and radiolabeled urokallikrein, the radioligand was separated from denatured radiolabeled urokallikrein by affinity chromatography for the active site. The concentration of specific IgG used in the assay bound approximately 80% of the radioligand, which was displaced in a dose-related fashion by 0.2 to 2.5 ng of the unlabeled urokallikrein. When random urine samples from 21 healthy volunteers were assessed for endogenous urokallikrein by both active site radioimmunoassay and kinin generation, there was a linear relationship between the results of the two assays with a correlation coefficient of 0.89. A combined correlation plot of the results of the bioassay and the active site radioimmunoassay for the 21 urine samples before and after trypsin activation gave a linear regression line with a correlation coefficient of 0.91. The finding that trypsin activation increased the urokallikrein concentration of urine to a similar extent in both the radioimmunoassay and the bioassay means that latent urokallikrein was not detected until its active site was uncovered.
Experimental allergic encephalomyelitis supernatant transfer activity (EAE-STA) in Lewis rats:immunobiologic and initial biochemical properties.
Authors: Whitacre C C , Paterson P Y
Published: December 2022

Supernatants derived from incubated lymph node cells (LNC) of Lewis rats sensitized to neuroantigen-adjuvant have the capacity to transfer experimental allergic encephalomyelitis (EAE) to syngeneic recipients. Sensitization of donors with guinea pig or rat spinal cord adjuvant by either of two routes of injection was effective in generating EAE supernatant transfer activity (EAE-STA). Despite transfer of typical EAE histopathologic lesions, which were intense and disseminated throughout the neuraxis in some animals, recipient animals invariably remained clinically well. Donor sensitization with purified myelin basic protein of guinea pig or rat origin was conspicuously ineffective in generating EAE-STA. Absorption studies revealed that EAE-STA was diminished after exposure to guinea pig or rat spinal cord or guinea pig or rat kidney, but it was not demonstrably reduced after absorption with neonatal rat spinal cord lacking encephalitogenic activity or with guinea pig or rat myelin basic protein or lysozyme. EAE-STA is stable at -20 degrees C/4 days, -70 degrees C/2 days, 4 degrees C/18 hr, and 56 degrees C/1 hr, and has a m.w. in excess of 100,000 daltons.
The cellular nature of concanavalin A-stimulated antibody-dependent cell-mediated cytotoxicity.
Authors: Connolly J M , Wunderlich J R
Published: December 2022

Enhanced antibody-dependent cytotoxic cell (ADCC) activity, generated in vitro by concanavalin A (Con A) stimulation of human peripheral blood lymphocytes, was analyzed to identify the nature of the killer (K) cell. Stimulated and unstimulated 2-day cultured cells were separated into sheep erythrocyte rosetting (E+) and non-rosetting (E-) subpopulations, and ADCC activity was determined by using the slopes of linear dose-response plots as a measure of K cell activity. Quantitative comparisons were made by examining activity on a per cell basis ("specific" activity) and total K cell activity recovered per subpopulation. This analysis revealed that although the E- cells were enriched for specific ADCC activity, the E+ subpopulation contained the major proportion of the total recovered activity and nearly all of the Con A-induced increase in K cell activity.
Role of sialic acid in the macrophage glycolipid receptor for MIF.
Authors: Liu D Y , Petschek K D , Remold H G , David J R
Published: December 2022

Acidic glycolipids from guinea pig macrophages enhance the response of macrophages to migration inhibitory factor (MIF), suggesting a role of glycolipid receptors for this lymphocyte mediator. Neuraminidase treatment of these glycolipids results in the loss of their biologic activity. This activity remains intact after incubation of the glycolipids with beta-galactosidase. In order to investigate whether sialic acid is essential for the macrophage's response to MIF, macrophages were incubated with neuraminidase. Neuraminidase treatment of peritoneal exudate cells results in the abrogation of macrophage responsiveness to MIF. Other exoglycosidases such as beta-galactosidase and beta-glucosidase had no effect upon the macrophage response. The effect of neuraminidase was found to be reversible within 18 hr. These experiments suggest that macrophage glycolipids containing sialic acid are components of the macrophage receptor for MIF.
Newly transformed schistosomula spontaneously lose surface antigens and C3 acceptor sites during culture.
Authors: Samuelson J C , Sher A , Caulfield J P
Published: December 2022

The binding of rat anti-schistosome antibody and rat C3 to the surface of newly transformed schistosomula of S. mansoni was measured by quantitative immunofluorescence during the first 48 hr of their development in vitro. Schistosomula, cultured in media either with or without serum, lost antibody or C3 from their surface exponentially with a halftime of 5 hr for both labels. The loss of the surface molecules is seen in parasites that are labeled and then cultured as well as in parasites cultured and then labeled, indicating that the labeling procedure itself is not inducing the observed change. This immunochemical modification in the schistosomulum surface appears to be independent of host molecule adsorption and intrinsic to the development of the parasite.
Interferon activation of "pre-spontaneous killer" (pre-SK) cells and alteration in kinetics of lysis of both "pre-SK" and active SK cells.
Authors: Targan S , Dorey F
Published: December 2022

Interferon augments spontaneous killer cellular cytotoxicity (SKCC). This modulation may be vital in both tumor resistance and host viral defenses. To date, whether increased numbers of effector cells or activation of individual effector cells is responsible for this augmentation has not been established. Using a single cell assay where SK killing is directly visualized by the reading of trypan blue uptake at various time points by effector-target cell conjugates plated in agarose gel, we measured in vitro alterations in percentage of conjugate formation, number of active SK cells, and kinetics of SK cell lysis at the single cell level after 1 hr of interferon incubation. After interferon, there is no difference in number of cells that bind K-562 targets. This augmentation of cytotoxicity is due to both recruitment of new effector cells and an activation of the lytic process of both the new and already active SK cells.
Failure of lymphokine-producing lymphocytes from aged humans to undergo activation by recall antigens.
Authors: Sohnle P G , Larson S E , Collins-Lech C , Guansing A R
Published: December 2022

In order to understand better the causes of diminished delayed hypersensitivity in the elderly, we studied lymphocyte function in 10 healthy subjects over 80 years of age. Markedly decreased antigen-stimulated lymphokine production (3 of 30 assays positive versus 24 of 31 in young control subjects) was the most significant finding in the elderly subjects and appeared to be the best explanation for their reduced cutaneous responses to recall antigens. Although mitogen-stimulated lymphocyte transformation in the elderly group was more sensitive to suppression by prostaglandin E2 than that of the young group, direct prostaglandin action did not appear to explain the diminished antigen-stimulated lymphokine responses. The nonspecific mitogen concanavalin A elicited normal lymphokine responses in 9 of 10 elderly subjects, indicating that lymphocytes from the elderly have a normal capacity for lymphokine production when activated by a sufficient stimulus. Therefore, diminished delayed hypersensitivity in aged humans may be related to deficient lymphokine production, which in turn appears to involve a decreased capacity of the lymphocytes producing these mediators to undergo activation by recall antigens.
Purification, immunochemical, and biologic characterization of the Schistosoma circulating M antigen.
Authors: Carlier Y , Bout D , Strecker G , Debray H , Capron A
Published: December 2022

Circulating M antigen, specific for genus Schistosoma, was previously described in serum, urine, patients' milk, and in serum and urine of animals infected by S. mansoni. The M antigen was thermostable and soluble in trichloroacetic acid. It was not hydrolyzed by protease, ribonuclease, amylase, or neuraminidase but destroyed by sodium metaperiodate. In the present study, we have purified the M ag by using trichloroacetic acid solubility, DEAE Sephadex, and immunoadsorption. The M ag showed a neutral electric charge, a m.w. heterogeneity, and was only stained by periodic acid-Schiff. The composition study revealed M ag was a glycoprotein with a polysaccharide moiety (63% of the molecules) particularly rich in galactose, fucose, glucosamine, and mannose, and with a high molecular ratio of serine and threonine. The presence of O-glycosidic linkage allowed M ag to be considered as a mucin or a mucus glycoprotein-like component. It was localized in the cell wall of the gut of adult worms.
The specificity of uptake of model immune complexes and other protein aggregates by the murine reticuloendothelial system.
Authors: Finbloom D S , Abeles D , Rifai A , Plotz P H
Published: December 2022

We have studied the selectivity of the murine reticuloendothelial system in recognizing and removing from the blood model immune complexes and several aggregated proteins. Although both IgG anti-DNP model immune complexes and micro-aggregated albumin exhibited rapid hepatic uptake, which is saturable, they did not cross-inhibit each other. Aggregates of ovalbumin, a glycoprotein rich in mannose (a sugar recognized by Kupffer cells), had no effect on the uptake by the liver of either IgG immune complexes or micro-aggregated albumin. The finding that transferrin aggregates were not taken up by the liver at all suggests that aggregation alone is not sufficient to overcome the lack of a specific hepatic receptor for that protein. Thus, several mechanisms exist whereby aggregated proteins can be cleared from the blood by the reticuloendothelial system, an observation consistent with the selective failure of reticuloendothelial function found in several human diseases.
Modulation of tumor cell susceptibility to humoral immune killing through chemical and physical manipulation of cellular lipid and fatty acid composition.
Authors: Schlager S I , Ohanian S H
Published: December 2022

Line-10 tumor cells cultured for 24 hr in lecithin-rich normal human plasma or with synthetic lecithin showed a 5- to 8-fold increase in their lecithin:sphingomyelin mole ratio without being affected in their total lipid content or cholesterol:phospholipid mole ratio. These cells were more sensitive to killing by antibody plus complement (C) than untreated controls. Line-10 cells that underwent a homogeneously catalyzed hydrogenation reaction were reduced 6-fold in their content of unsaturated fatty acid compared to controls; the lipid content of these cells was largely unaffected. These cells were more resistant to antibody-C mediated killing than controls. These modifications in cellular lipid and fatty acid composition could be reversed when the cells were recultured for 24 hr in serum-containing tissue culture medium; the cells regained control levels of susceptibility to antibody-C killing at this time. These results suggest that by manipulating the lipid or fatty acid composition of a tumor cell, either indirectly by changing the lipid composition of the environment in which the cell resides or by directly altering the chemical nature of a cellular lipid constituent, the susceptibility of the cell to humoral immune killing can be modulated.
Idiotypic analysis of anti-GAT antibodies. VII. Common idiotype on hybridoma antibodies to poly(Glu60 Ala40)
Authors: Ju S T , Pierres M , Germain R N , Benacerraf B , Dorf M E
Published: December 2022

A single DBA/2 mouse, immunized with L-glutamic acid60-L-alanine40 (GA), was used to produce hybridoma cell lines. Seven hybridoma anti-GA antibodies were obtained for idiotypic analyses. Two hybridoma anti-L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT) antibodies, preferentially reactive to GA, were studied in parallel. Anti-idiotypic antisera to purified anti-GAT and anti-GA serum antibodies and to hybridoma anti-GA antibodies were analyzed by idiotype binding and inhibition of idiotype binding assays. Five of the nine hybridoma antibodies exhibited common GA-1 idiotypic specificities previously demonstrated on the majority of anti-GA antibodies of inbred mouse strains of differing immunoglobulin heavy chain linkage groups; these hybridoma antibodies also possessed private idiotypic determinants. Two GA-1 negative hybridoma anti-GA antibodies appeared identical by immunochemical criteria, arguing that somatic hybridization does not artifactually generate private idiotypic determinants. The results demonstrate that the common GA-1 idiotype system is associated with a family of nonidentical but idiotypically related antibody molecules present in a single DBA/2 mouse, and these antibodies are part of the "GA-1 idiotypic family".
Mechanisms of immune lysis. II. CTL-induced nuclear disintegration of the target begins within minutes of cell contact.
Authors: Russell J H , Dobos C B
Published: December 2022

Within minutes of CTL-target interaction, the intracellular disintegration of the target cell nucleus begins. This nuclear event satisfies the ionic and temperature requirements that have been described by others as important in CTL-mediated lysis. Such nuclear disintegration does not occur in Ab+C-lysed targets. Thus the target cell nucleus appears to signal a primary event in CTL-mediated lysis that distinguishes the CTL lytic mechanism from that of Ab+C.
The role of immunoglobulin receptors and T cell mediators in B lymphocyte activation. I. B cell activation by anti-immunoglobulin and anti-idiotype reagents.
Authors: Primi D , Lewis G K , Goodman J W
Published: December 2022

The possibility that the failure of anti-mouse immunoglobulin (Ig) antibody to induce antibody synthesis by B cells might be due to reversible receptor blockade was investigated. Murine spleen cells were cultured for 3 days in the presence of minute quantities of intact of (Fab') fragments of rabbit anti-mouse Ig antibody. Thereafter, the cells were washed and either trypsin treated or not before reculturing for 18 hr. Only cells that had been trypsinized after culturing with either intact or fragments of anti-Ig gave a vigorous polyclonal antibody response. This response was extremely T dependent, since T cells or culture supernatants from Con A-activated T cells were required for the B cell response. Moreover, anti-delta was much more effective than anti-mu in inducing antibody synthesis. Finally, the use of three different anti-idiotypic antisera rather than anti-Ig reagents selectively activated the specific idiotype in each instance. The findings demonstrate that anti-Ig reagents can potentiate the response of B cells to signals delivered by T cells.
Isolation of biosynthetically-labeled Fc-binding proteins from detergent lysates and spent culture fluid of a macrophage-like cell line (P388D1).
Authors: Loube S R , Dorrington K J
Published: December 2022

Lactoperoxidase-catalyzed surface iodination of P388D1 mouse macrophage cells has led to the identification of a 57,000 m.w. protein as a candidate for the cell's receptor for the Fc region of IgG. THe present work extends these studies and shows, by biosynthetic incorporation of radiolabeled amino acids and sugars, that the protein in question is a glycoprotein, which appears to be shed or released from the cells in a slightly altered form during the labeling period. Use of labeled amino acids as probes permitted the detection of a large number of contaminating proteins in these preparations; some of these may represent degradation products of the putative FcR. One of the contaminants was identified as actin; the binding of the putative receptor to IgG appeared to be quite independent of any actin-IgG interactions.
Phagocytosis of immune complexes mediated by IgM and C3 receptors by macrophages from mice treated with glycogen.
Authors: Mantovani B
Published: December 2022

Phagocytosis mediated by IgM and C3 receptors was studied by in vitro experiments with peritoneal macrophages from mice treated with repeated i.p. injections of glycogen. The immune complexes were sheep red cells sensitized with mouse IgM antibody or antibody and complement. Macrophages from the inflammatory exudate induced by glycogen were able to phagocytize immune complexes of red cells and IgM antibodies in contrast to normal macrophages, which can only bind to these complexes. Evidence was also found for phagocytosis mediated by interaction with C3 receptors in the macrophages from the glycogen-treated animals. The implications of these findings in the mechanism of phagocytosis and the possible relevance of the observed changes in the function of macrophage receptors for the afferent and efferent branches of immune response are discussed.
Negatively selected H-2bml and H-2b cells stimulated with vaccinia virus completely discriminate between mutant and wild-type H-2K alleles.
Authors: Doherty P C , Bennink J R , Wettstein P J
Published: December 2022

Lymphocyte populations from B6, C-H-2bml (KbmlDb) mutant mice cannot, after both in vivo and in vitro negative selection for alloreactivity, be induced to recognize vaccinia virus presented in the context of H-2Kb. This finding may mean that the T cell receptor(s) expresses a component that is very specific for a particular "active site" on the self-H-2 molecule. Alternatively, (if recognition is directed at a virus-H-2 complex) the more similar 2 H-2 molecules are, the more likely it may be that precursor thymocytes in the mutant with the capacity to bind H-2Kb + vaccinia virus may be deleted during ontogeny as a result of cross-reaction with H-2Kbml + endogenous antigen.
Fc (IgG) receptors on rat basophilic leukemia cells.
Authors: Segal D M , Sharrow S O , Jones J F , Siraganian R P
Published: December 2022

Receptors for IgG and IgE on rat basophilic leukemia cells (the 2H3 subline of RBL-IV) were detected by their ability bind IgE and IgG in fluorescence or radioactive assays. The 2H3 cells have approximately two-thirds as many receptors for IgG as for IgE. The IgE receptors bind monomeric IgE with high affinity and IgE binding is uninhibited by high concentrations of monomeric or oligomeric IgG. In contrast, the IgG receptors bind both (rabbit and rat) IgG and (rat) IgE. The affinity of IgG receptors for monomeric IgG and IgE is much lower than that of the IgE receptor for its ligand; binding of radiolabeled IgG or IgE to IgG receptors can be detected only with cross-linked oligomers. Although both receptors are sensitive to tryptic digestion than IgE receptors. Dual laser flow microfluorometric studies revealed that the numbers of IgG and IgE receptors per cell varied considerably among 2H3 cells and that the distributions of the 2 types of receptors were independent of each other. 2H3 cells released histamine through an IgE-mediated system, but cross-linked IgG, even in saturating amounts, did not elicit histamine release, nor have any effect upon IgE-induced release. Moreover, 2H3 cells would not mediate ADCC of IgG-coated chicken red blood cells.
T lymphocyte tolerance and early appearance of virus-induced cell surface antigens in Moloney-murine leukemia virus neonatally injected mice.
Authors: Collavo D , Zanovello P , Biasi G , Chieco-Bianchi L
Published: December 2022

Regression of Moloney-murine sarcoma virus- (M-MSV) induced sarcomas in normal adult mice is accompanied by generation of virus-specific cytotoxic T lymphocytes (CTL). However, when neonatal mice that were injected with Moloney-murine leukemia virus (M-MuLV carrier) were subsequently challenged as adults with M-MSV, the sarcomas did not regress nor did they generate CTL. This failure to produce CTL cannot be ascribed to nonspecific immunodepressive effects or to suppressor cell generation since M-MuLV carrier mice exhibit normal reactivity after allogeneic cell stimulation. Moreover, addition of M-MuLV-infected cells as the third party to cultures does not reduce activity of CTL from M-MSV immune mice. Since M-MSV and M-MuLV possess common antigens, the observed unresponsiveness was considered in relationship to induction of a T lymphocyte tolerance, which may follow introduction of foreign antigens at an early stage of development. In fact, it was observed that as early as 10 days after injection, thymus, lymph node, and spleen from M-MuLV carrier mice express virus-induced cell-surface antigens that not only are targets for M-MSV-immune CTL, but also induce in vitro a strong specific cytotoxic response. In addition, a cold target inhibition assay disclosed that the same antigens are shared by both M-MuLV infected and leukemia cells, even though they are less expressed on the surface of the former. The finding that the cytotoxicity of alloreactive lymphocytes from M-MuLV carrier mice is reduced after preincubation with M-MSV immune CTL confirms that virus infection does not bring about functional inactivation of lymphocytes. Finally, it was observed that virus antigen presence on lymphocytes from M-MuLV neonatally injected mice is closely related to subsequent leukemia development.
The effects of electrophoretically "slow" and "fast" alpha-2 macroglobulin on mixed lymphocyte cultures.
Authors: Hubbard W J , Hess A D , Hsia S , Amos D B
Published: December 2022

It has been determined that alpha-2 macroglobulin (alpha 2M) is more suppressive of a mixed lymphocyte response (MLR) when complexed with proteinase than in its "native" state. Other alpha 2M preparations showed a moderate level of MLR suppression, but it is unlikely that this is a result of interaction with cellular proteinases. A panel of other proteinase inhibitors (alpha 1 PI, SBTI, BPTI, TLCK) did not suppress the MLR to the same extent as alpha 2M either when bound with or free from trypsin. A dose-responsive pattern of MLR suppression similar to that observed with purified proteinase-complexed alpha 2M was seen with serum containing proteinase-complexed alpha 2M. The population of cells that apparently conveys the suppressive property is the adherent cells (putative monocytes), which can reduce the MLR almost as well as unfractionated cells when exposed to alpha 2M. Most of these properties of alpha 2M were demonstrable in "serumless" medium with qualitative similarity to the MLR obtained in cultures performed with conventional serum supplemented medium. It was found that alpha 2M-trypsin complexes must be presented at or near culture initiation and remain in contact with the cells for a minimum of approximately 4 hr to have its optimum effect.
Monoclonal antibodies to guinea pig Ia antigens. I. Production, serologic, and immunochemical characterization.
Authors: Burger R , Clement L , Schroer J , Chiba J , Shevach E M
Published: December 2022

Xenogeneic antibodies to guinea pig Ia antigens were produced by application of the hybridoma technique. BALB/c mice were immunized with the Ia-positive B cell leukemia line EN-L2C of strain 2 guinea pigs, and the spleen cells were fused to the mouse myeloma line NS-1. Culture supernatants of hybrids growing in the selective HAT medium were screened in a binding assay, and hybrids selectively reactive with EN-L2C but not with its Ia-negative variant BZ-L2C were cloned. Ascitic fluid was produced and used as a source of antibody. Four independently derived antibodies proved to be specific for Ia antigens. Two of the antibodies (25E3 and 25E11) seemed to recognize alloantigenic determinants on strain 2 Ia antigens in that they bound only to the cells of strain 2 but not to the cells of strain 13 animals. Two other antibodies (22C4 and 27E7) were found to react with both strain 2 and strain 13 cells, and immunoprecipitation experiments revealed that these antibodies were directed to common determinants shared between certain strain 2 and strain 13 Ia antigens. All antibodies were cytotoxic in the presence of complement for EN-L2C (but not for BZ-L2C) leukemia cells and for lymph node, spleen, peritoneal exudate cells, thymocytes, and purified T cells of strain 2 guinea pigs. In addition, 22C4 was cytolytic for cells of both strains. These results indicate that antibodies can be obtained through the hybridoma technique that are able to distinguish different determinants on Ia antigen molecules of te guinea pig. These antibodies should provide a useful tool for further study of the role of Ia antigens in cellular interaction and Ia-Ir gene function.
The potential importance of soluble deoxynucleotidase activity in mediating deoxyadenosine toxicity in human lymphoblasts.
Authors: Carson D A , Kaye J , Wasson D B
Published: December 2022

Deoxyadenosine and its nucleotides have been implicated in the pathogenesis of the immune dysfunction associated with a genetic deficiency of adenosine deaminase (ADA). We have previously shown that when ADA is blocked with a synthetic inhibitor, human T lymphoblastoid cell lines are more sensitive to deoxyadenosine toxicity, dephosphorylate deoxyadenosine nucleotides at a slower rate, and have much lower levels of ecto-5'-nucleotidase than most B cell lines. It seemed unlikely, however, that an enzyme on the outer surface of the lymphocyte plasma membrane could regulate intracellular deoxynucleotide catabolism. We now report that human lymphoblasts also contain a soluble deoxynucleotidase activity that is distinguishable from the plasma membrane enzyme by several criteria. In multiple human lymphoblastoid cell lines of varying origin and phenotype. soluble deoxynucleotidase correlated significantly (rs = 0.80, p < 0.001) with sensitivity to deoxyadenosine toxicity.
Functional properties of lymphocyte subpopulations in hepatitis B virus infection. I. Suppressor cell control of T lymphocyte responsiveness.
Authors: Chisari F V , Castle K L , Xavier C , Anderson D S
Published: December 2022

Hepatocellular injury in hepatitis B virus infection may be produced by an autoaggressive hepatocytotoxic immune response. To test the hypothesis that acquired suppressor cell defects may participate in such a response, we assessed the functional integrity of 2 suppressor cell populations in patients with type B viral hepatitis. Spontaneous suppression of the 1-way mixed lymphocyte response by radiation-resistant, adherent peripheral blood mononuclear cells decreases during the acute phase of disease, returns towards normal with clinical recovery, but remains depressed in patients with chronic hepatitis. The degree of spontaneous suppressor cell dysfunction correlates inversely with at least 1 biochemical parameter of hepatocellular injury (SGPT). The functional integrity of this suppressor cell fluctuates during chronic hepatitis and may reflect currently undefined biologic variables in this disease. Mitogen-induced suppression on lymphocyte activation by radiation resistant, nonadherent suppressor cells is also depressed in acute and chronic hepatitis, but it does not correlate with biochemical evidence of hepatocellular injury on an individual-patient basis. Documentation of these generalized defects of nonspecific suppressor cell function establishes a basis for the possible existence of specific anomalies of immuno-regulation that may permit the expression of normally suppressed auoaggressive hepatocytotoxic immune mechanisms in viral hepatitis.
A monoclonal antibody (BA-1) reactive with cells of human B lymphocyte lineage.
Authors: Abramson C S , Kersey J H , LeBien T W
Published: December 2022

NALM-6-M1 is an acute lymphoblastic leukemia cell line previously shown in our laboratory to express the pre-B lymphocyte phenotype, i.e., cytoplasmic IgM+, surface immunoglobulin-. Hybridomas were produced against this cell line by fusing spleen cells from hyperimmunized mice with NS-1 mouse myeloma cells. One monoclonal antibody derived from this fusion, designated BA-1, reacted with peripheral blood B lymphocytes, chronic lymphocytic leukemias, pre-B-ALL, most non-Hodgkin's lymphomas, and most non-T, non-B-ALL. BA-1 showed weak reactivity with B lymphoblastoid cell lines and failed to react with multiple myeloma and pokeweed mitogen-induced plasma cells. BA-1 also reacted with peripheral blood granulocytes and the erythroleukemia cell line K-562. No reactivity was seen with cells of T lymphocyte origin, platelets, red cells, monocytes, or acute myelocytic leukemias. Evidence is presented indicating that the determinant recognized by BA-1 is not surface immunoglobulin, HLA-DR, or receptors for C3 or Fc. We conclude that monoclonal antibody BA-1 may be useful in the study of early stages of human B lymphocyte development.
An immunomorphologic study of adenosine deaminase distribution in human thymus tissue, normal lymphocytes, and hematopoietic cell lines.
Authors: Chechik B E , Schrader W P , Minowada J
Published: December 2022

Adenosine deaminase (ADA) has been detected immunohistochemically in human thymus. The enzyme was localized predominantly in cortical thymocytes. Occasional lymphocytes in the medulla were also positive for ADA. Blood vessels, connective tissue, and Hassall's corpuscles were not stained for the enzyme. Using single-cell immunofluorescence and immunoperoxidase assays, we found that thymocytes and lymphoid cells of peripheral blood (PBL) and tonsils were heterogeneous with respect to ADA expression. About 70% of thymocytes were strongly stained for the enzyme whereas weak staining was seen in 20% of cells. About 10% of thymocytes were ADA negative. Twenty percent of PBL and tonsil cells were strongly positive for ADA, 10% of cells were negative for the enzyme, and weak staining was seen in the remainder. Bone marrow mononuclear cells were not stained for ADA. One hundred percent of lymphoblasts of 3 T cells leukemia lines were strongly stained for the enzyme whereas weak staining was seen in a pre-B cell leukemia line, 4 B cell lymphoma/normal lines, 2 non-T, non-B cell leukemia lines and 2 myeloid cell leukemia lines. There was a good correlation between intensity of cellular staining and quantity and activity of ADA detected in cell extracts by radioimmunoassay and enzymatically. The development of immunomorphologic methods for the detection of ADA provides a tool to study the role of the enzyme in function(s) and differentiation of normal and leukemic cells.
The ADCC capacity of macrophages from C3H/HeJ and A/J mice can be augmented by BCG.
Authors: Koren H S , Meltzer M S , Adams D O
Published: December 2022

Macrophages elicited from the peritoneal cavity of C3H/HejJ and A/J mice by bacillus Calmette-Guérin (BCG), although ineffective in mediating tumor cytotoxicity, had augmented capacity to mediate antibody-dependent cellular cytotoxicity (ADCC) against both erythroid and neoplastic target cells. The level of augmented ADCC was comparable to that expressed by BCG-activated macrophages from control C3H/HeN mice. The data suggest that the defective potential for cytolytic activation in 2 genetically deficient strains of inbred mice does not extend to augmentation of capacity for mediating ADCC and that full expression of cytolytic activation is not necessary for expression of augmented cytolysis of antibody-coated targets.
Recovery from a viral respiratory infection. I. Influenza pneumonia in normal and T-deficient mice.
Authors: Wells M A , Albrecht P , Ennis F A
Published: December 2022

Experiments were performed to serve as a basis for analyzing the immune responses of the host to a viral respiratory tract infection. In a comparative study, inbred BALB/c and nude (athymic mice bred on a BALB/c background) mice were infected intranasally with mouse adapted A/Port Chalmers/1/73 influenza virus. Survival was prolonged in the nude mice (16 days vs 10 days for BALB/c mice, t = 3.5, p less than 0.01), but significantly fewer of the nude mice survived 21 days (16% vs 42% of BALB/c mice). In addition, virus persisted longer in the lungs of nude mice (5.8 log10 of infectious virus on day 21 vs no detectable virus in BALB/c mice) and lung pathology progressed more slowly but lasted longer in nude mice, as measured by immunofluorescent and histologic examination of pulmonary tissue. Antibody production was significantly lower and there was no cytotoxic T cell response detected in lymphoid cells prepared from the spleens, peripheral blood, or the lung after the infection of nude mice. A significant increase in the number of lymphocytes isolated from the lung was observed by day 7 (t = 5.8, p less than 0.001) in the BALB/c mice but not until day 14 (t = 4.9, p less than 0.001) in the nude mice. Lymphocytes from nude mice did not respond to influenza virus antigens or Concanavalin A (Con A) but did respond to lipopolysaccharide (LPS), whereas lymphocytes from BALB/c mice responded to all 3 preparations.
An adherent, radiosensitive accessory cell necessary for mouse cytolytic T lymphocyte response to viral antigen.
Authors: Fast L D , Fan D P
Published: December 2022

A cell type in addition to T lymphocytes and macrophages was found to be necessary for the in vitro stimulation of mouse anti-Sendai viral T lymphocytes in response to subviral antigens. This cell type was found to be glass and nylon wool adherent, to bear no surface immunoglobulin or Thy 1.2 antigen, and to be radiosensitive. This type of cell was not needed when the stimulatory antigen was virus-infected cells.
Pyridine nucleotide-dependent generation of hydrogen peroxide by a particulate fraction from human neutrophils.
Authors: DeChatelet L R , Shirley P S
Published: December 2022

The catalytic oxidation of [14C]-formate to 14CO2 was adapted to measure H2O2 formation in cellfree system. Standard curves employing glucose-glucose oxidase and xanthine-xanthine oxidase demonstrated linearity between 14CO2 evolution and enzyme concentration. A particulate fraction from human neutrophils was capable of oxidizing [14C]-formate; this reaction was dependent upon the presence of catalase, reduced pyridine nucleotide, and cellular material. Reaction increased with time of incubation and protein concentration, although not in a strictly linear fashion. The pH optimum was approximately 5.5 NADPH was a significantly better substrate than NADH, although both were capable of generating H2O2. The particulate fraction derived from phagocytizing cells was more active than a corresponding fraction from resting cells with either substrate. H2O2 production was abnormal in particulate fractions derived from 2 patients with chronic granulomatous disease. H2O2 production was markedly inhibited by superoxide dismutase or cytochrome c (scavengers of superoxide anion) but not by scavengers of singlet oxygen or hydroxyl radical. Reaction was greatly stimulated by the addition of manganous ion. These results are consistent with the hypothesis that the respiratory burst in human neutrophils is initiated by an oxidase that can utilize either NADPH or NADH but exhibits a marked preference for the former. Further, the inhibitor studies strongly support a mechanism involving an initial enzymatic reaction followed by a self-sustaining free radical reaction involving superoxide anion.
Regulatory role of IgE-binding factors from rat T lymphocytes. V. formation of IgE-potentiating factor by T lymphocytes from rats treated with Bordetella pertussis vaccine.
Authors: Hirashima M , Yodoi J , Ishizaka K
Published: December 2022

An i.p. injection of Bordetella pertussis vaccine (BP) into rats induced the formation of soluble factors that had affinity for IgE (IgE-binding factors). The factor was detected in the serum of BP-treated animals 5 to 7 days after the treatment. Their circulating lymphocytes as well as spleen cells spontaneously released IgE-binding factors in the serum of BP-treated rats and those released from their circulating lymphocytes had affinity for lentil lectin, and the ability to selectively potentiate an in vitro IgE response of DNP-OA primed cells to homologous antigen. The molecular size of IgE-potentiating factor was between 10,000 and 20,000, and was comparable to that formed by lymphocytes of rats infected with Nippostrongylus brasiliensis. Evidence was obtained that IgE-potentiating factor was derived from Fc epsilon R(+) T cells, with a T cell marker identified by monoclonal antibody W 3/25. Their production of IgE-potentiating factor may be the basis of the adjuvant effect of BP on the IgE response.
Inhibition of murine plaque-forming cell responses in vivo by ribavirin.
Authors: Peavy D L , Powers C N , Knight V
Published: December 2022

The effects of ribavirin, a potent inhibitor of RNA and DNA virus replication, on the generation of primary plaque-forming cell (PFC) responses in vivo has been investigated. Intraperitoneal administration of ribavirin 1 day after immunization of C3H/HeJ mice with sheep erythrocytes (SRBC) suppressed splenic IgM and IgG PFC responses by 60 to 90%. Primary IgM PFC responses of C3HeB/FeJ mice to bacterial lipopolysaccharides (LPS), a T-independent antigen, were also inhibited to a similar extent. Inhibition of splenic PFC responses, without significant reduction in nucleated cell recoveries, was dose dependent between 0.5 and 4 mg ribavirin. Varying the time of treatment determined that optimal inhibitory activity occurred when ribavirin was administered simultaneously or 1 day after antigen. Kinetic analysis of PFC responses in ribavirin-treated mice revealed that suppression did not result from a delay in development since reduced numbers of PFC were found at all times after immunization. Ribavirin-treatment after primary sensitization in vivo also impaired the capacity of spleen cells to develop secondary PFC responses in vitro, indicating that ribavirin also inhibited memory cell generation.
Flow cytofluorometric analysis of the binding of Vicia villosa lectin to T lymphoblasts: lack of correlation with cytolytic function.
Authors: MacDonald H R , Mach J P , Schreyer M , Zaech P , Cerottini J C
Published: December 2022

The relationship between the binding of Vicia villosa (VV) lectin and the expression of cytolytic function in T lymphoblasts has been investigated using flow cytofluorometric techniques. Spleen cells activated in vitro in 5-day mixed leukocyte cultures (MLC) were incubated sequentially with VV, rabbit anti-V antiserum, and fluoresceinated sheep anti-rabbit IgG. When these stained MLC cells were passed on a flow cytometer gated to exclude nonviable cells and small lymphocytes, a single heterogeneous peak of fluorescence was seen, as compared to control MLC cells that had not been incubated with VV. Fluorescence of lymphoblasts was dependent upon lectin dose and was eliminated when staining was performed in the presence of N-acetyl-D-galactosamine, the appropriate competitive sugar for VV. T cell blast populations activated against H-2, Mls, or parasite antigens all had comparable levels of fluorescence after staining with VV, although the cytolytic activity of these cells varied widely. Furthermore, when MLC lymphoblasts binding large or small amounts of VV were sorted on the basis of their relative fluorescence intensity and tested for cytolytic function, no appreciable difference in activity between the 2 populations was observed. These results are inconsistent with the hypothesis that VV binds selectively to cytolytic T lymphocytes.
Age-associated changes in Epstein-Barr virus-induced human lymphocyte autoantibody responses.
Authors: Fong S , Tsoukas C D , Frincke L A , Lawrance S K , Holbrook T L , Vaughan J H , Carson D A
Published: December 2022

In the present investigation we have studied the capacity of the Epstein-Barr virus (EBV), a ubiquitous human B lymphocyte polyclonal activator, to induce cultured peripheral blood lymphocytes from umbilical cords, young adults (20 to 40 yr old), and elderly adults (75 to 90 yr old) to form IgM antibodies to human IgG or human thyroglobulin. The EBV preparation used was shown to exert its B cell stimulatory effect independently of T cell-suppressor effects. The cultures were studied in limiting dilution analyses, and the data were taken to represent relative numbers of B cell precursors of autoantibody-forming cells in the 3 age groups. The results showed: 1) the EBV-inducible IgM anti-IgG and anti-thyroglobulin-producing cells increased in number from birth to old age; 2) the rise occurred at different times of life for the 2 autoantibodies, anti-IgG reactive B cells increasing between birth and young adulthood, and anti-thyroglobulin reactive B cells between young adulthood and old age; 3) the apparent relative avidity of the anti-IgG for antigen was higher in the elderly adults than in the younger adults. We believe these findings are determined by differences in the frequencies of the respective self-reactive B cells in the circulation. What physiologic or environmental factors determine the differential expansions of the human autoreactive B lymphocytes for IgG and thyroglobulin are not known. It seems possible that individual variations in the sizes of these pools may be a factor in determining susceptibility to autoimmune disease.