bluuebunny/crossref_metadata_2025 huggingface dataset:
Strain variation in BCG-induced chronic pulmonary inflammation in mice: control by a cyclophosphamide-sensitive thymus-derived suppressor cell.
Authors: Moore V L , Mondloch V M , Pedersen G M , Schrier D J , Allen E M
Published: December 2022

We previously reported that only certain strains of inbred mice develop intense chronic granulomatous inflammation (CGI) in the lungs and spleen in response to an i.v. injection of killed BCG in an oil-in-saline emulsion (BCG-E). The capacity to respond did not appear to be controlled by genes within the H-2 complex; subsequent studies have shown that genes linked to the Igh allotype complex influence the development of CGI. In other systems, unresponsiveness to certain antigens has been shown to be because of cyclophosphamide- (Cy) sensitive suppressor cells. We therefore used Cy as a probe to study mechanisms of unresponsiveness in low-responder (LR) CBA mice. The results indicate that LR mice could be converted into high responders (HR) by treatment with 100 mg of Cy per kilogram of body weight 2 days before injection with BCG-E. In addition, the effects of Cy were inhibited by the provision of syngeneic whole or purified spleen T cells from mice injected 7 days previously with BCG-E. Cells responsible for abolition of the Cy effect were sensitive to anti-Thy-1 serum + C. Thus, the intensity of BCG-E-induced CGI in mouse lungs is controlled by a population of Cy-sensitive T lymphocytes.
Immunologic consequences of antibiotic-induced abridgement of bacterial infection: effect on generation and loss of protective T cells and level of immunologic memory.
Authors: North R J , Berche P A , Newborg M F
Published: December 2022

Ampicillin was employed in mice to determine the effect of rapidly abridging bacterial infection on the generation of T cell-mediated antibacterial immunity. It was found that antibiotic-induced abridgement of infection with Listeria monocytogenes had a pronounced effect on the generation of splenic T cells capable of adoptively immunizing normal recipients against a Listeria challenge infection. Ampicillin given at the time of maximum bacterial growth at 24 hr caused a striking reduction in the number of protective T cells produced at the time of peak response on day 6. Although the greatest effect was caused by giving ampicillin early in infection, a significant reduction in peak T cell production occurred even when Ampicillin was given as late as day 5. On the other hand, the effect of abridging infection at the onset of decay of the anti-Listeria response was to cause protective T cells to be lost from the spleen at a much faster rate. These results clearly show that regardless of any immunoregulatory mechanism involved, the duration of generation, and the onset and duration of decay of the anti-Listeria response are determined by the number of replicating Listeria in the tissues. Moreover, Ampicillin-induced abridgement of infection, either before or at the time of peak primary response, resulted in the expression of greatly reduced levels of immunologic memory at a later time. This indicates that memory cells are generated throughout the entire course of the primary anti-Listeria response, including the period of its decay.
Mechanism of action of macrophage-derived suppressor factor produced by soluble immune response suppressor-treated macrophages.
Authors: Aune T M , Pierce C W
Published: December 2022

After a 2-hr incubation with soluble immune response suppressor (SIRS), a product of concanavalin A-activated murine T cells, macrophages release a factor, M phi-derived suppressor factor (M phi-SF), which nonspecifically suppresses immune responses in vitro. The mechanism(s) of action of M phi-SF and range of cell types affected by M phi-SF have been investigated. M phi-SF suppressed antibody responses to background levels if added at culture initiation and by 80 to 90% if added as late as 2 hr before assay. Primary and secondary IgM and IgG antibody responses, proliferative responses to T cell and B cell mitogens, antibody and protein secretion, and the division of several tumor cell lines in culture were inhibited by M phi-SF. Division of synchronized tumor cells was inhibited when M phi-SF was added at any point prior to and during mitosis; this inhibition could be reversed with 2-mercaptoethanol. In the presence of M phi-SF, asynchronous tumor cells accumulated in the cell cycle just prior to cell division and could be released into mitosis by 2-mercaptoethanol. These data indicate that M phi-SF inhibits cell division by causing a block at or in mitosis and suggest that M phi-SF may be a general inhibitor of cellular proliferation and possibly of protein secretion.
Activation of murine macrophages. I. Different pattern of activation by poly I:C than by lymphokine or LPS.
Authors: Taramelli D , Varesio L
Published: December 2022

The ability of poly I:C to activate mouse macrophages (M phi) to become tumoricidal was evaluated and compared with the ability of 2 other agents, lipopolysaccharide (LPS) and M phi-activating factor (MAF), to induce a tumoricidal state. All these agents were able to stimulate proteose-peptone-elicited M phi to kill RL male 1 tumor cells in an 18-hr 51Cr release cytotoxicity assay. High levels of cytotoxicity were obtained with concentrations as low as 1 microgram/ml of LPS or poly I:C and with 1/81 dilution of MAF. However, in the presence of reagents shown to contain less than 0.01 ng/ml of LPS by the LAL assay (LPS free), we found that poly I:C induced strong reactivity, whereas MAF was ineffective. The addition of 10 ng/ml of LPS during the stimulation period did not enhance the cytotoxicity induced by poly I:C, but it did restore MAF-induced, M phi-mediated cytotoxicity. In addition, poly I:C induced strong tumoricidal activity in resident M phi and in peritoneal exudate cells from the genetically defective C3H/HeJ mice that normally do not respond to LPS and MAF treatment. Therefore, it seems that although LPS is required as a second signal for MAF-induced cytotoxicity, such a second signal is not required for poly I:C-induced cytotoxicity. From the above results, it appears that poly I:C is a more powerful activating agent than LPS and MAF and either activates M phi via a different pathway or is effective on subpopulations of M phi that are not activated by the other agents.
"Mature" thymocytes are not glucocorticoid-resistant in vitro.
Authors: Triglia D , Rothenberg E
Published: December 2022

Dexamethasone, administered in vitro either continuously or for a 30-min pulse period followed by a chase of 18 to 24 hr, is shown to decrease protein synthetic rates as well as cellular viabilities in a dose-dependent manner in murine thymocytes. Differential effects on cortical and medullary thymocyte subpopulations should be detectable by alterations in the rates of synthesis of specific molecules which are stable in vitro, relative to total protein synthesis, in the absence of steroids. However, terminal deoxynucleotidyl transferase (TdT), a marker for cortical cells, and Qa-1 and H-2, preferentially expressed in medullary cells, continue to be produced as constant fractions of total synthesis even after treatment with up to 10(-6) M dexamethasone. Furthermore, thymocytes obtained from normal and in vivo cortisone-treated mice show little, if any, difference in their intrinsic sensitivities to dexamethasone in vitro. The results reported here suggest that a corticoresistant thymocyte per se does not exist in vitro and that the thymus may produce factor(S) in vivo that protect the medullary subpopulation from in vivo glucocorticoid-induced lysis.
Suppressive effects of cyclosporin A on the induction of alloreactivity in vitro and in vivo.
Authors: Wang B S , Heacock E H , Collins K H , Hutchinson I F , Tilney N L , Mannick J A
Published: December 2022

The purpose of the present study was the investigation of the effect of cyclosporin A (CsA) on the induction of alloreactivity in vitro and in vivo. Addition of CsA to mouse mixed lymphocyte cultures (MLC) not only inhibited lymphocyte proliferation but also prevented the generation of alloreactive cytolytic lymphocytes (CL). It was necessary to add CsA within the first 3 days of a 5-day MLC in order to achieve a significant suppressive effect. Lymphocytes, after being cultured in MLC with CsA for 4 days or longer, were incapable of being activated upon re-exposure to the same alloantigens although their responses to unrelated antigens remained intact, indicating antigen specificity of the suppression induced by CsA and its long-lasting effect. Furthermore, lymphocytes from mice treated with CsA after allosensitization failed to manifest primary cytotoxicity and could not be reactivated in a secondary MLC. Finally, CsA had no effect on those CL already generated, suggesting that CsA acts upon the induction of CL rather than the effector phase.
Mycoplasma contamination in human tumor cell lines: effect on interferon induction and susceptibility to natural killing.
Authors: Birke C , Peter H H , Langenberg U , Müller-Hermes W J , Peters J H , Heitmann J , Leibold W , Dallügge H , Krapf E , Kirchner H
Published: December 2022

Cleaning of human tumor cell lines from arginine-dependent nonfermentative Mycoplasma orale 1 (MO1) by a recently developed technique profoundly altered several in vitro properties of the cell lines. Four melanoma lines (Mel I, Mel St, Mel K, IGR3) and 1 ovarian carcinoma line (Ro) induced human leukocyte interferon (IFN-alpha) only in the mycoplasma-infected state and not in the mycoplasma-free state. MO1-infected tumor lines were generally more susceptible to natural killer (NK) cell-mediated lysis than their mycoplasma-free counterparts. Reinfection of cleaned tumor lines with MO1 restored their interferonogenicity and the increased susceptibility to NK lysis. Thus, the amplifying role of MO1 infection on NK target lysis occurred in connection with an increased production IFN-alpha during the assay period. The human erythroleukemia cell line K562 was exceptional in that it also induced high levels of IFN in an apparently mycoplasma-free state and was unaffected in its susceptibility ot NK lysis by infection with MO1. Possible implications of these findings for the biologic significance of the NK reaction are discussed.
Genetic linkage of resistance to Listeria monocytogenes with macrophage inflammatory responses.
Authors: Stevenson M M , Kongshavn P A , Skamene E
Published: December 2022

The mobilization of adequate numbers of mononuclear phagocytes to inflammatory foci was measured in Listeria-resistant and Listeria-sensitive mice. Resistant strains, such as B10.A, were found to have a 2- to 3-fold greater accumulation of peritoneal macrophages after i.p. treatment with a variety of nonspecific inflammatory stimuli in comparison to sensitive strains, such s A/J. In addition to low macrophage inflammatory responses, A/J mice had fewer resident peritoneal macrophages. Moreover, measurement of chemotaxis in vitro of equal numbers of thioglycollate-induced macrophages showed cells from A/J mice to be less responsive to complement-derived chemotactic factors. Thus, the mononuclear phagocyte systems of resistant B10.A and sensitive A/J mice were quantitatively and qualitatively different. A survey of inbred mouse strains revealed that these differences were not peculiar to A strain mice and that, in general, the level of the in vivo macrophage inflammatory response correlated with the level of resistance to Listeria in a given strain. Like resistance to Listeria, the macrophage inflammatory response was found to be genetically controlled by an autosomal, non-H-2-linked gene(s) expressed as incompletely dominant. Backcross analysis showed genetic linkage of the macrophage inflammatory response with resistance to Listeria. Thus, the results of this study provide formal evidence that the cellular basis of the genetically determined, enhanced resistance to Listeria is an augmented pool size of mononuclear phagocytes, and the prompt mobilization of these cells to foci of infection.
I-Kk and H-2Kk antigens are shed as supramolecular particles in association with membrane lipids.
Authors: Emerson S G , Cone R E
Published: December 2022

The form of 125I-labeled membrane alloantigens (I-Ak and H-2Kk) and immunoglobulin M shed by murine splenic lymphocytes labeled by lactoperoxidase-catalyzed radioiodination was determined by gel chromatography and ultracentrifugation. I-Ak and H-2Kk antigens are shed as particles ranging in size from 3 X 10(5) to 2 X 10(6), whereas membrane IgM is shed as an individual unit consonant with its native size of 180,000. The effects of detergents on the size of shed I-Ak and H-2Kk and the coprecipitation of (3H) lipids with these proteins suggest that I-Ak and H-2Kk are shed in association with local membrane lipids as small membrane-derived vesicles, whereas membrane IgM is shed without detectable associated lipid.
Primary in vitro anti-KLH antibody formation by peripheral blood lymphocytes in man: detection with a radioimmunoassay.
Authors: Morimoto C , Reinherz E L , Schlossman S F
Published: December 2022

In the present report, a primary in vitro human antibody response to KLH was investigated. Peripheral blood lymphocytes were incubated with antigen for 5 days and then cultured in the absence of KLH for 4 additional days. Maximal anti-KLH antibody production, as measured by radioimmunoassay, occurred at a cell density of 1 X 10(6) and at an antigen concentration of 5 micrograms per culture. The antibody produced was shown to be predominantly of the IgM isotype and specific for KLH antigen in several binding assays. Moreover, no antibody was generated in the absence of T lymphocytes. This in vitro antibody-forming system should be of considerable use in the analysis of the cellular requirements for antibody production and the genetic control of the immune response in man.
IgE antibody production in guinea pigs treated with cyclophosphamide.
Authors: Graziano F , Haley C , Gundersen L , Askenase P W
Published: December 2022

Guinea pig anaphylactic responses usually involve IgG1 antibodies. Although IgE antibody production has been accomplished, a reliable and sustained method for production has not been described. We have investigated a method of enhancing IgE antibody production in guinea pigs. The criteria for IgE production were homologous passive cutaneous anaphylaxis (PCA) antibody activity that passed an affinity column of rabbit anti-guinea pig IgG1, heat lability (56 degrees C for 3 hr), and a long sensitization period in skin (7 days). Hartley guinea pigs were primed and boosted monthly with 1 microgram Picryl-Ascaris plus 1 mg of alum i.p. Some Hartley guinea pigs also received cyclophosphamide (250 mg/kg) before the first boost. English short-hair guinea pigs (previously shown to be good IgE producers) were immunized similarly but received no cyclophosphamide. Hartley animals receiving no cyclophosphamide inconsistently made low titered IgE anti-hapten antibody (1:100). Fifteen of 15 Hartley animals that received cyclophosphamide had high titers (1:2000) of IgE anti-hapten antibody. English short-hair animals had moderate titers (1:400), and not all animals responded. Thus, cyclophosphamide converted IgE nonresponder Hartley guinea pigs to uniformly high responders. This method provides material to study the biologic properties of IgE in this species, and also provides a model to study the apparent suppression of IgE responses in Hartley guinea pigs.
How complement kills E. coli. I. Location of the lethal lesion.
Authors: Wright S D , Levine R P
Published: December 2022

We have studied the action of human complement (C) on E. coli membranes. We find, as have others, that C disrupts the outer membrane (OM), allowing the release of periplasmic proteins. In addition, we have found 1) that in the complete absence of lysozyme, C damages the inner membrane (IM), 2) IM damage is different from OM damage in that only small molecules traverse a damaged IM whereas macromolecules traverse damaged OM, 3) IM damage and OM damage occur with identical kinetics and dose response, suggesting that IM and OM damage are closely coupled events, and 4) upon the addition of purified C8 and C9 to the washed cellular intermediate, E. coli C 1-7, both IM and OM are damaged coordinately. These results, taken together, suggest that C damages E. coli membranes by acting at a site contiguous with both membranes. We speculate that C may simultaneously gain access to both membranes by acting at the junctions between IM and OM.
Serum-mediated suppression of nonspecific B cell activation. II. Relative resistance of B cells from the NZB mouse strain to regulation by a natural inhibitor in normal mouse serum.
Authors: Walker S M , Sturtevant J E , Weigle W O
Published: December 2022

Recent work has shown that normal mouse serum (NMS) and plasma contain inhibitory substance(s), termed NMS-In, that suppress proliferative and polyclonal antibody responses elicited in vitro with various B cell mitogens. In this study, the NZB mouse, which has a high degree of spontaneous polyclonal B cell activity, was examined for possible reduced amounts of NMS-In. Also, the capacity of NZB B cells to be inhibited by NMS-In was determined. It was found that sera from NZB mice had normal amounts of NMS-In, as assessed by inhibition of mitogenesis of spleen cells in culture induced by bacterial endotoxin (ET). However, B cells of the NZB mouse were found to be abnormal, requiring approximately 4 to 7 times more NMS-In to inhibit polyclonal antibody synthesis and mitogenesis elicited by ET than B cells of more immunologically normal mice. Removal of T cells by treatment with anti-thy 1.2 and complement did not change the sensitivity of the NZB B cells to NMS-In. Insensitivity to NMS-In was expressed in B cells from relatively young NZB mice, e.g., 9 to 15 wk of age. The relative resistance of B cells of the NZB mouse to regulation by a natural inhibitor in NMS could partially serve to explain their high level of spontaneous antibody synthesis. Furthermore, the fact that B cells of the NZB mouse were ultimately susceptible to inhibition by relatively large amounts of NMS-In affords the opportunity in the future to suppress polyclonal antibody activity in the NZB mouse with purified NMS-In in an attempt to ameliorate autoimmune disease.
Monoclonal antibody OKT-9 recognizes the receptor for transferrin on human acute lymphocytic leukemia cells.
Authors: Goding J W , Burns G F
Published: December 2022

The monoclonal antibody OKT-9 recognizes a surface protein of human lymphocytes that consists of a disulfide bonded homodimer of m.w. 200,000 intact and 95,000 reduced. A similar protein is precipitated by transferrin-agarose, but not by agarose alone. Peptide mapping by limited proteolysis shows that the proteins precipitated by OKT-9 antibodies and transferrin-agarose are homologous. It is concluded that OKT-9 antibodies recognize the transferrin receptor. Expression of receptors for transferrin may be useful as a marker for activated or dividing cells.
MIs locus recognition by a cloned line of H-2-restricted influenza virus-specific cytotoxic T lymphocytes.
Authors: Braciale V L , Braciale T J
Published: December 2022

This report demonstrates the expression of strong MIs locus MIsd) recognition by a cloned line of H-2-restricted influenza virus-specific CTL. This clone of F1 (H-2b/d; MIsb) origin was found to specifically proliferate in response to uninfected cells of CBA/J (H-2k, MIsd) origin but not to uninfected B10.BR or CBA/CaJ cells (H-2k, MIsb). In addition, proliferation by this cTL line was observed in response to histocompatible cells expressing cross-reactive MIsa determinants (DBA/2, NZB; H-2d, MIsa). This recognition was observed only at the level of CTL proliferation. The CTL line exhibited no cytotoxic activity for target cells of these MIs types. These observations are contrasted with the response of another cloned H-2-restricted influenza-specific CTL line that simultaneously exhibits alloreactivity for H-2k. The significance of these results for T lymphocyte recognition is discussed.
Surface antigens on mouse natural killer cells: use of monoclonal antibodies to inhibit or to enrich cytotoxic activity.
Authors: Seaman W E , Talal N , Herzenberg L A , Herzenberg L A , Ledbetter J A
Published: December 2022

In studies using monoclonal antibodies to cell-surface antigens we have identified 2 new antigens (H11 and 7.2) expressed on mouse NK cells. These are shared with T cells but not B cells. We have also shown that NK cells express T200 but lack detectable ThB or Lyt-1. The T200 and H11 surface molecules were implicated in the mediation or regulation of natural killing; monoclonal antibodies to T200 and H11 inhibited natural killing when added to the cytotoxicity assay but did not interfere with T cell cytotoxicity against the same target. The inhibitory effect of anti-T200 is consistent with recent evidence showing that antibodies to the Ly-5 polymorphic determinant on T200 block natural killing. The H11 determinants is on a different molecule. The absence of Lyt-1 and ThB on NK cells permitted development of a rapid and simple method for separating NK cells from T cells and B cells. Spleen cells incubated with rat monoclonal antibodies to Lyt-1 (on all T cells) and ThB (on all B cells) were 95% removed by adherence to Petri dishes coated with antiserum to rat immunoglobulin. The natural killer activity in the nonadherent population was enriched 16-fold.
A novel human T cell antigen preferentially expressed on mature T cells and shared by both well and poorly differentiated B cell leukemias and lymphomas.
Authors: Kamoun M , Kadin M E , Martin P J , Nettleton J , Hansen J A
Published: December 2022

A new lymphocyte differentiation antigen shared by all normal T cells and some malignant B cells was defined by a monoclonal antibody designated 12.1. This antibody reacted with all peripheral blood T cells but not with normal B cells and B cell lines. Analysis with a fluorescence activated cell sorter showed that the expression of 12.1 antigen changes during T cell maturation. Most thymocytes, blasts of acute T cell leukemia, and cells from established leukemic T cell lines bear a small amount of 12.1 antigen. In contrast the majority of peripheral blood T cells, activated T cells, and leukemic T cells of the Sezary syndrome bear a large amount of 12.1 antigen. Unexpectedly, antibody 12.1 reacted with leukemic cells from most patients with B-type chronic lymphocytic leukemia (CLL) and some patients with lymphosarcoma cell leukemia (LSCL). Among these leukemias, expression of the 12.1 antigen was not correlated with the stage of B cell maturation, with the amount of surface immunoglobulin on the cells, or with the presence or absence of monoclonal gammapathy. In a comparative serologic analysis the antigen defined by antibody 12.1 was distinct from the p67 T cell antigen (defined by monoclonal antibody 10.2) that is also known to be expressed by B-type CLL cells.
Analysis of a fetal calf serum-induced T cell line. I. H-2 restricted growth and expression of membrane-bound and released molecules bearing 5936-idiotypic determinants.
Authors: Suzan M , Bourgois A , Boned A , De Preval C , Golstein P , Rubin B
Published: December 2022

It was previously established that a fetal calf serum-induced C57BL/6 T cell line that induces T and B cell differentiation could be kept proliferating in vitro only if cultured in the presence of irradiated syngeneic spleen cells and FCS. The present experiments were performed in order to investigate a) whether this cell line was a pure T cell line, b) whether the cells in this cell line (called line 12) were homogeneous with regard to Lyt phenotype, c) whether its growth was H-2 restricted, and d) whether line 12 cells reacted with our anti-idiotype (5936) and anti-T cell receptor allotype/isotype (6036) antisera. The results showed that line 12 consisted of T cells of Lyt 1+, 2.3- and phenotype. Its growth and proliferation was restricted to Kb and/or IAb alloantigen, and this phenomenon was observed with isolated Lyt 1+, 2.3- T cells. Line 12 cells reacted with both 5936 and 6036 antisera, and the positive cells were of Lyt 1+, 2.3- phenotype. Thus, our data indicate that Lyt 1+, 2.3- line 12 T cells interact with FCS and Kb/IAb alloantigen via receptors, which may bear 5936 and 6036 antisera-defined determinants. However, because these antisera only react with a subpopulation of Lyt 1+, 2.3- cells, proof that the same T cell has both MHC specificity and B cell idiotypic determinants will require further experimentation. 5936 and 6036 antisera-reactive molecules could be isolated from the supernatants of line 12 cells. Such molecules had characteristics similar to the 50,000 m.w. form of receptor molecules isolated from B6 anti-CBA T cell supernatants: a single chain polypeptide carrying both 5936 and 6036 antisera-defined determinants.
Protection of neonatal mice against herpes simplex viral infection by human antibody and leukocytes from adult, but not neonatal humans.
Authors: Kohl S , Loo L S , Pickering L K
Published: December 2022

Neither human immune globulin nor human mononuclear leukocytes (containing lymphocytes and monocytes) from herpes simplex virus (HSV) seronegative adult donors could protect infant C57BL/6 mice from a lethal HSV infection. In contrast, a combination of 5 X 10(6) leukocytes and a subneutralizing dose of antibody protected the mice. Leukocytes (5 X 10(6) or 1 X 10(7)) from neonatal humans and antibody could not mediate this protection. This is the first correlation between defective human neonatal leukocyte antibody-dependent cellular cytotoxicity (ADCC) in vitro and defective human neonate leukocyte-antibody protection against HSV infection in vivo in mice.
Suppression of the late stages of mitogen-induced human B cell differentiation by FC gamma receptors (FC gamma R) released from polymorphonuclear neutrophils.
Authors: Bich-Thuy L T , Samarut C , Brochier J , Revillard J P
Published: December 2022

During short incubation in serum-free medium, polymorphonuclear neutrophils (PMN) release soluble material that can be characterized as receptors for Fc IgG (Fc gamma R) on the following evidence: it agglutinates erythrocyte-IgG antibody (EAG) complexes, it prevents the binding of EAG to EAG-rosette-forming cells, and it binds to EAG-rosette-forming cells after modulation of their Fc gamma R, allowing the formation of 'passive' rosettes. These Fc gamma R were isolated by affinity chromatography on sepharose 4B-IgG. This material was shown to interfere with the differentiation of peripheral blood B cells into Ig-secreting cells in cultures stimulated by pokeweed (PWM) or Nocardia opaca (NOC) extracts. The number of Ig-secreting cells determined by a reverse hemolytic plaque assay using protein A-coated sheep erythrocytes was decreased by addition of Fc gamma R over a wide range of dilutions. The number of Ig-containing cells was diminished in PWM-stimulated cultures, but not in cultures stimulated with NOC. Fc gamma R did not affect cell viability, nor did they interfere with plaque-forming cell assay. Fc gamma R was not suppressive when added before the 3rd day or after the 6th day of culture. The suppressor factor was shown to remain associated with Fc gamma R after elution at acidic pH; it was removed by absorption on Sepharose 4B-IgG, but not on pepsin-digested F(ab')2 fragments. The suppressive factor as well as the capacity to restore EAG rosette formation by modulated lymphocytes were destroyed by heating (56 degrees C, 30 min) or by freezing and thawing. Properties of Fc gamma R released from PMN in this system are similar to those of Fc gamma R released from unstimulated human peripheral blood lymphocytes (PBL) and to those of mouse Ig-binding factor produced by alloactivated T cells or T cell lines.
Collaboration between histoincompatible T and B lymphocytes: preferential induction in vivo of antibodies to antigens borne by foreign T cells.
Authors: Piguet P F , Vassalli P
Published: December 2022

We have compared, in vivo, with nude mice and, in vitro, with cultures of T depleted spleen cells, several components of the humoral response resulting from the helper effect of allo- or xenogeneic T cells on mouse B lymphocytes; the response directed to the histocompatibility antigens (or other surface antigens) borne by foreign T lymphocytes, the effect on the response to an additional unrelated antigen, namely sheep red blood cells, and finally a possible nonspecific polyclonal activation. In vivo, injection of allo- or xenogeneic T lymphocytes into nude mice (with or without SRBC) induced a strong anti-T cell response, manifested by the presence of high titered IgG alloantibodies, without significant enhancement of the response to SRBC. In vitro, addition of xenogeneic T lymphocytes to cultures of T-depleted spleen cells did enhance an anti-T cell and an anti-SRBC response. The preferential induction of an anti-T cell response, observed in vivo, implies a peculiar form of T-B cell collaboration where the B lymphocytes binding to the foreign and activated T cells are selectively activated, suggesting that there is no absolute restriction for specific T-B cell collaboration across the histocompatibility barrier.
Adoptive transfer of experimental allergic encephalomyelitis in SJL/J mice after in vitro activation of lymph node cells by myelin basic protein: requirement for Lyt 1+ 2- T lymphocytes.
Authors: Pettinelli C B , McFarlin D E
Published: December 2022

Optimal conditions were established for the adoptive transfer of experimental allergic encephalomyelitis (EAE) in SJL/J mice. Lymph node cells from SJL/J mice primed in vivo with myelin basic protein (BP) were incubated in vitro with BP. These cells proliferated specifically to BP and when transferred at the optimal conditions into syngeneic mice induced EAE in 100% of the recipients. The in vitro proliferative response to BP was dependent on the presence of Lyt 1+ 2- T lymphocytes. Furthermore, when the activated LNC were treated before transfer with anti-Thy 1 or anti-Lyt 1 antibody and C, neither clinical nor histologic signs were observed in the recipients, whereas treatment with anti-Lyt 2 antibody and C had no effect. These results indicated that Lyt 1+ 2- T cells are responsible for the transfer of EAE.
Specific unresponsiveness to skin allografts in anti-lymphocyte serum-treated, marrow-injected mice: participation of donor marrow-derived suppressor T cells.
Authors: Maki T , Gottschalk R , Wood M L , Monaco A P
Published: December 2022

Sequential changes of cell-mediated immune reactivities were examined in anti-lymphocyte serum-(ALS) treated, C3H/He (C3H; H-2k) bone marrow-injected (C57BL/6 X A)F1 (B6AF1; H-2b/k.d) mice bearing enhanced C3H skin grafts. Spleen cells of these mice exhibited marked suppression of the proliferative response to phytohemagglutinin and concanavalin A. When the spleen cells were assayed for the direct lymphocyte-mediated cytotoxicity against H-2k targets, their lytic activity remained low until the time of graft rejection, in contrast to the increasingly high cytotoxic activity exhibited by spleen cells of control B6AF1 mice given only ALS and C3H skin grafts. When spleen cells of marrow-injected B6AF1 mice were cultured with mitomycin-C treated C3H spleen cells, the proliferative response was significantly suppressed the throughout the course, despite the early appearance of high "secondary-type" cytotoxic activity. Co-culture experiments demonstrated the presence of C3H antigen-specific suppressor cells in the ALS-treated, marrow-injected mice bearing intact allografts. Treatment of spleen cells with anti-H-2, anti-Thy 1 and anti-I-J sera and C revealed that the suppressor cells present late in the marrow-injected mice were T cells of donor C3H bone marrow cell origin.
Effects of gamma-irradiation on lymphocyte subpopulations participating in the development of the cytotoxic T lymphocyte response.
Authors: Susskind B M , Faanes R B
Published: December 2022

Differences in radiosensitivity among lymphocyte subpopulations involved in generating allogeneic cytotoxic T lymphocyte (CTL) responses were studied. Gamma irradiated splenic lymphocytes from C57BL/6 mice (B6, H-2b) were stimulated in vitro with DBA/2 (H-2d) spleen cells or the DBA/2 mastocytoma, P815. Limiting dilution analysis indicated that the cytotoxic T lymphocyte effector cells (CTLe) for either antigen arose from the same pool of cytotoxic T lymphocyte precursors (CTLp), and that the D37 value of the B6 anti-H-2d CTLp was approximately 165 R. In mixed lymphocyte cultures, however, differential effects of irradiation were seen between the 2 antigens in the ability of irradiated responder B6 spleen cells to generate CTLe. For induction of cytotoxic activity against DBA/2 splenocytes, the CTLp was the most radiosensitive component of the response, whereas the generation of a cytotoxic response against P815 was limited by the radiosensitivity of an Lyt-1 T helper cell (D37 approximately 85 R). The radiosensitive Lyt-1 cell could be replaced by mixed lymphocyte reaction supernatants or semipurified interleukin 2 (IL-2) from Con A-stimulated murine spleen cells. The requiremet of B6 anti-H-2d CTLp for a radiosensitive accessory cell in the CTL response against P815, in contrast with their activation by DBA/2 splenocytes, may center on the fact that the tumor cells do not possess Ia antigens.
Plaque-forming cell responses to trinitrophenyl (TNP)-L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT) in microcultures are not under conventional Ir gene control.
Authors: Letvin N L , Benacerraf B , Germain R N
Published: December 2022

The IgM plaque-forming cell response to trinitrophenyl (TNP)-conjugated L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT) was studied in an in vitro microculture system. Contrary to expectations, this response was not found to be under conventional H-2 linked Ir gene control. Thus, both classical responder BALB/c (H-2d) and non-responder DBA/1 (H-2q) spleen cells gave equivalent anti-TNP PFC responses to TNP-GAT in these cultures. Experiments were performed to determined if haptenation had changed the GAT carrier so as to remove it from GAT-specific Ir gene control. It could be demonstrated that TNP-GAT elicited in vivo anti-GAT PFC responses showing typical Ir control in the BALB/c and DBA/1 strains; that anti-hapten and anti-carrier PFC responses to DNP-GAT in vivo were similarly controlled; and that the TNP-GAT compound remained a T cell- and Ia+ accessory cell-dependent antigen in vitro. Furthermore, the microculture system allowed GAT-specific T helper cells to be detected in the spleens of DBA/1 mice treated with GAT in vivo under conditions eliciting a predominant suppressor T cell response under usual conditions of assay. These findings contrast with the Ir gene regulation of TNP-(T,G)-A--L responses seen under identical culture conditions. The implications of these results for our understanding of the site of Ir gene action and the target of suppression in the GAT model are discussed.
Hapten-specific murine colony-forming B cells. II. Delineation of a tolerogen-sensitive subpopulation of colony-forming B cells.
Authors: Pillai P S , Scott D W
Published: December 2022

B cell subpopulations were studied by using B cell cloning procedures and an in vitro tolerance induction model. Fluorescein- (FL) specific B cells from normal spleens were isolated by using FL gelatin plates and were then cultured in semisolid agar in the presence or absence of tolerogen. Hapten-specific cells grew in soft agar to form discrete colonies. Colony growth is dependent on "mitogens" present in agar, sheep red blood cells (SRBC), and lipopolysaccharide (LPS). For example, SRBC plus LPS potentiate the growth of an increased number of colony-forming B cells (CFU-B) compared to either additive alone. These CFU-B could be triggered by a specific antigen to yield plaque-forming cells (PFC). With tolerogen (FL-sheep gamma-globulin) present in the agar, the number of FL-specific CFU-B was reduced by 25 to 50%. The ability of the remaining colonies to form PFC upon antigenic stimulation was also reduced. This reduction in CFU-B numbers, however, was observed only when the agar contained both SRBC and LPS as mitogenic potentiators of growth; no effect of tolerogen on CFU-B numbers was seen when cells were grown with either additive alone. Interestingly, the effect of tolerogen on CFU-B numbers was abrogated when peritoneal macrophages, in addition to SRBC plus LPS, were present during cloning. It is postulated that unique subpopulations of B cells form colonies under varied cloning conditions and that those CFU-B grown with SRBC plus LPS display an increased sensitivity to growth inhibition by tolerogen.
A comparison of the interactions of the mitogenic and nonmitogenic lima bean lectins with human lymphocytes.
Authors: Munske G R , Pandolfino E R , Magnuson J A
Published: December 2022

Binding and mitogenic characteristics of the 2 Phaseolus lunatus (lima bean) lectins have been examined with human peripheral blood lymphocytes. Chemical cross-linking of the nonmitogenic lima bean lectin produced a species that stimulated human lymphocytes as well as or better than the mitogenic lima bean lectin, which is a T lymphocyte mitogen with requirement for monocyte participation. The maximal stimulation and the dose response to the cross-linked lima bean lectin did not significantly differ from that observed with the mitogenic lectin. We have used fluorescein-labeled lima bean lectins to show that both lectins share mutually exclusive binding sites on lymphocytes. Our results strongly support the concept that multiple valence of lectins is important in inducing mitogenesis. Both the mitogenic and nonmitogenic lectins demonstrated selective binding by labeling only 70% od human peripheral blood lymphocytes. The fraction not binding lectin is a population of T lymphocytes. Competitive binding studies with the lima bean lectins and other N-acetyl-D-galactosamine-specific lectins suggest that the cell surface receptors for these various lectins may be quite different. We have also studied the response of human lymphocytes to the lima bean lectins and soybean agglutinin after neuraminidase treatment. As previously demonstrated (22), neuraminidase treatment of the cells drastically altered the binding and mitogenic response to soybean agglutinin. Lima bean lectin binding to the treated cells was also markedly increased, but the mitogenic response was essentially unaffected.
Immunochemical relations between pneumococcal group 19 and Klebsiella capsular polysaccharides.
Authors: Lee C J , Koizumi K
Published: December 2022

Pneumococcal type 19F polysaccharide, contained in the current U.S. licensed pneumococcal vaccine, is a poor immunogen in young children. Capsular polysaccharides of Klebsiella, reported to be cross-reactive with the pneumococcal type 19F, were isolated. Immunochemical relations between pneumococcal group 19 and Klebsiella capsular polysaccharides were studied by immunodiffusion with hyperimmune typing antisera, immune passive hemolysis, and plaque-forming cell response in mice following injection of polysaccharides. Klebsiella K2, K9, K32, K47, and K48 polysaccharides were reported to cross-react with horse pneumococcal type 19 antiserum by direct precipitation. However, these Klebsiella polysaccharides did not react in immunodiffusion with horse or with rabbit pneumococcal 19F and 19A typing antisera. Furthermore, pneumococcal type 19F and 19A polysaccharides did not precipitate with rabbit K2, K9, K32, K47, and K49 typing antisera. The cross-reactivity of the Klebsiella and pneumococcal polysaccharides was studied by 2 other methods. Rabbit pneumococcal 19F antiserum induced by low passive hemolysis of sheep erythrocytes coated with Klebsiella polysaccharides. In contrast, Klebsiella typing antisera exhibited higher hemolytic activity with erythrocytes coated with type 19F polysaccharides. The cross-reactivity of K2 and pneumococcal 19F polysaccharides was also exhibited by the direct IgM plaque-forming cell response. The cross-reactivity, determined by precipitin reactions using animal antisera, may not predict the cross-immunogenicity of purified polysaccharides in humans.
Altered idiotype response to phosphocholine in mice bearing an x-linked immune defect.
Authors: Kenny J J , Guelde G , Claflin J L , Scher I
Published: December 2022

The x-linked CBA/N defect results in an altered idiotype expression among the anti-phosphocholine (PC) antibodies produced after antigenic challenge with the thymic dependent antigen PC-KLH but does not preclude the response to this hapten as previously suggested. The majority of immune-defective F1 male mice can be divided into 2 groups based on their T15 idiotype profile. Group 1 mice fail to produce anti-PC antibodies bearing the T15 idiotype in either a primary or secondary response, whereas group 2 mice produce low levels of T15 idiotype; however, this idiotype often appears only after secondary immunization. These responses are distinct from the anti-PC response of normal F1 females, which is predominantly of the T15 idiotype. In addition to the altered idiotype expression, F1 male mice exhibited a greatly reduced primary anti-PC response compared to normal mice, and secondary responses were approximately one-third that of normal mice. The delayed expression of anti-PC antibodies in immune defective mice appears to be due to their inability to produce IgM anti-PC antibodies in either a primary or secondary response to PC-KLH.
Quantitative immunofluorescent analysis of surface phenotypes of murine B cell lymphomas and plasmacytomas with monoclonal antibodies.
Authors: Lanier L L , Warner N L , Ledbetter J A , Herzenberg L A
Published: December 2022

In this study, a large series of murine B lymphomas and plasmacytomas were examined by quantitative flow cytometry analysis using a panel of monoclonal antibodies against murine differentiation antigens. These cell lines appear to represent subpopulations of lymphoid cells arrested at distinct stages of differentiation. In general, the pattern of reactions of these monoclonal antibodies with the B cell neoplasms was comparable to the reactions seen with normal cells in the same lineage. The Thy-1.2, Lyt-2, and T-30 differentiation antigens were not detected on any B lymphoma or plasmacytoma. However, certain surface Ig-positive B lymphomas do express the Lyt-1 antigen. With respect to other differentiation markers examined, including E2, F1, ThB, Lgp-100 (Ly 9.1), G2, and T-200 (Ly 5), the reaction of the B cell tumors reflected the expression of these markers on comparable normal cells. This investigation also emphasized the marked intratumor and intertumor heterogeneity that is observed when cloned cell lines are analyzed quantitatively for a large number of surface markers. Thus, this approach may be particularly useful in defining heterogeneity in maturation states within cloned tumor cell lines.
Recognition of MOPC-460 variable region determinants by polyclonally distributed triggering receptors on B lymphocytes.
Authors: Primi D , Juy D , Le Guern C , Sanchez P , Cazenave P A
Published: December 2022

Purified M460 protein-induced untreated and anti-Thy 1.2 and complement-treated murine spleen cells to proliferate and differentiate in vitro. Fc receptor complexing was not responsible for the phenomenon, since M460 Fab fragments retained the B cell triggering activity. Activation was inhibitable by the addition of DNP-glycine to cultures and could also be detected by using spleen cells from various strains of mice, including the C3H/HeJ, which is genetically nonresponding to LPS. Hybrid molecules were constructed between the M460 proteins and the products of 2 myelomas that do not activate B cells, namely, X25 and X24. The only hybrid molecules that retained triggering activity were (L460-Hx25)2. Interestingly, the same molecule was also the only one recognized by an anti-M460 monoclonal antibody (F6(51)) that has been previously shown to induce B cells to proliferate and differentiate. The data suggest the existence on B lymphocytes of a network of polyclonally distributed mitogen receptors that share determinants cross-reacting with immunoglobulin idiotypes.
Adoptive transfer of experimental autoimmune thyroiditis (EAT) with in vitro activated lymph node cells from thyroglobulin-sensitized guinea pigs: characterization of the cell that transfers EAT.
Authors: Braley-Mullen H , Tompson J G , Sharp G C , Kyriakos M
Published: December 2022

Relatively low numbers of lymph node cells (LNC) from Strain 2 or Strain 13 guinea pigs sensitized with guinea pig thyroglobulin (GPTG) could transfer experimental autoimmune thyroiditis (EAT) to normal syngeneic recipients after in vitro culture with GPTG. The EAT induced in recipients of cultured LNC was similar in incidence and severity to EAT induced by active immunization with GPTG in complete Freund's adjuvant. The cells that were effective in transferring EAT were shown to be immunoglobulin-negative, nylon wool nonadherent, and Ia-negative. The effector cells were sensitive to irradiation after in vitro activation, indicating that cell proliferation in the recipient is required for development of EAT. Recipient animals often developed moderate to severe lesions of EAT yet none had detectable delayed hypersensitivity to GPTG and the majority also had no detectable anti-GPTG antibody.
Adenosine-5'-triphosphate-(ATP) mediated stimulation and suppression of DNA synthesis in lymphoid cells. I. Characterization of ATP responsive cells in mouse lymphoid organs.
Authors: Ikehara S , Pahwa R N , Lunzer D G , Good R A , Modak M J
Published: December 2022

The effects of various nucleotides and nucleosides on DNA synthesis of mouse lymphocyte populations were studied. Significant stimulation of DNA synthesis was observed in the cells from bone marrow or the thymus in the presence of ATP. In contrast, the DNA synthesis of the cells from spleen, lymph nodes, and peripheral blood was markedly inhibited by ATP. Guanosine 5'-triphosphate had nonspecific stimulatory effects on the DNA synthesis of various lymphoid cells, whereas cytidine triphosphate had no effect. When thymocytes or bone marrow cells were separated by 1 x g velocity sedimentation, a distinct cell population was identified as being responsible for the ATP-mediated stimulation of DNA synthesis. Further characterization of ATP-responsive cells revealed that the highest concentration of terminal deoxynucleotidyl transferase, a marker enzyme for precursor T cells, was present in these cells. ATP-mediated stimulation of DNA synthesis may, therefore, serve as a specific marker restricted to a certain population of differentiating T cells.
Two distinct murine differentiation antigens determined by genes linked to the Ly-6 locus.
Authors: Auchincloss H , Ozato K , Sachs D H
Published: December 2022

Two separate monoclonal antibodies, 34-11-3S and 34-2-11S, produced by the fusion of C3H anti-BDF1 sensitized spleen cells to the SP2/0 myeloma variant cell line, were found to detect alloantigens on subpopulations of murine lymphocytes. Analysis of the strain specificities of these antibodies showed that both detected antigens determined by genes closely linked to the Ly-6 locus. However, analysis of the tissue distributions of antigen-bearing cells indicated that the 2 antibodies reacted with distinct antigens, only 1 of which was similar in distribution to the classically described Ly-6.2 antigen. Functional studies performed using cells treated with antibody plus C showed that these 2 antibodies defined lymphocyte subpopulations that are different from each other and from those defined by several other reported Ly-6-related antibodies. The data suggest that the Ly-6 locus consists either of multiple tightly linked genes exhibiting marked linkage disequilibrium or of a single gene, the protein product of which is subject to post-translational modifications during cell differentiation leading to the expression of new determinants.