bluuebunny/crossref_metadata_2025 huggingface dataset:
Phenotypic and functional properties of murine thymocytes. III. Kinetic analysis of the recovery of intrathymic cytolytic T lymphocyte precursors after in vivo administration of hydrocortisone acetate.
Authors: Ceredig R , Cummings D E
Published: December 2022

The phenotypic and functional properties of cells in the C57BL/6 mouse thymus regenerating after a single dose of 100 mg/kg hydrocortisone acetate (H/C) are described. Functionally, the frequency of anti-H-2d cytolytic T lymphocyte precursors (CTL-P) in thymuses from individual mice was determined by limit dilution analysis of mixed leukocyte microcultures. The initial increase in CTL-P frequency, seen 48 hr post-H/C, was followed at 6 to 8 days by a phase of rapid decrease. The CTL-P frequency returned to a normal level by 28 days post-H/C. Analysis of the results from individual mice suggested that changes in total thymic CTL-P content were independent of the kinetics of thymus regeneration. Phenotypically, whereas the thymus 48 hr after H/C was considerably depleted of Lyt-2+ cells, there followed a rapid increase in the proportion of such cells to normal levels by 14 days post-H/C. In addition, as measured by FLS, a subpopulation of larger, predominantly Lyt-2+ cells was found during the phase of rapid thymic regeneration. With the use of a monoclonal anti-Thy-1.2 antibody, the weakly Thy-1-staining subpopulation of cells was absent from the thymus at 14 days post-H/C. These changes in the phenotypic properties of the post-H/C regenerating thymus were correlated with changes in their functional properties.
Histamine-induced suppressor factor (HSF): inhibition of helper T cell generation and function.
Authors: Garovoy M R , Reddish M A , Rocklin R E
Published: December 2022

The effect of histamine-induced suppressor factor (HSF) on the humoral immune response was examined with the model of polyclonal B cell activation induced during a primary mixed lymphocyte culture (MLC). The number of plaque-forming cells (PFC) generated during MLC was measured by a protein A plaque assay. HSF was produced by incubating lymphocytes from normal subjects with 10(-4) M histamine. The addition of HSF on day 0 to MLC-induced plaques reduced the mean number of IgG, IgM, and IgA PFC by 60 to 80%. HSF supernatants were active at a titer of 1/1000 and suppressed IgG, IgM, and IgA isotypes equally. To study the effect of HSF on the T helper cell component of this reaction, purified T lymphocytes were activated in undirectional MLC and subsequently combined with unprimed B cells to induce a polyclonal PFC response. HSF present during the generation phase of activated T cells or only at the time of co-culture inhibited the total PFC response by 83 +/- 13% and 76 +/- 23%, respectively. The expression of "Ia" and autologous DR antigens normally detected on 50 to 62% of activated T cells generated during MLC were reproducibly reduced to 20% in the presence of HSF. Similarly, the polyclonal B cell response inducible by MLC-derived helper factors (1500 IgG PFC/10(6)) was markedly inhibited. Thus, HSF inhibits MLC-induced polyclonal B cell activation by interfering with the generation and effector function of T helper cells as well as the B cell response to preformed helper factors.
Changes in intracellular glutathione levels in stimulated and unstimulated lymphocytes in the presence of 2-mercaptoethanol or cysteine.
Authors: Zmuda J , Friedenson B
Published: December 2022

Certain low m.w. thiol compounds can enhance various in vitro lymphocyte mitogenic responses, whereas reagents that react with sulfhydryl groups are potent cell poisons. The possible targets for thiols and thiol reagents could include membrane and cytoplasmic proteins, some of which may regulate lymphocyte proliferation. As a step toward understanding how lymphocyte proliferation is enhanced in the presence of thiols, we studied the appearance of both protein and non-protein sulfhydryl groups in stimulated lymphocytes. To help clarify the role of the cellular targets for thiols, we studied the time course of appearance of both protein and non-protein sulfhydryl groups as lymphocytes proliferate under various conditions. In the presence of an appropriate concentration of the thiol 2-mercaptoethanol (2-ME), there is a substantial rise in the level of intracellular glutathione for lymphocytes stimulated with the mitogen concanavalin A (Con A). Otherwise, the level of intracellular glutathione declines, even for Con A-stimulated cells, but the presence of 2-ME partially prevents this decline. Increasing the amount of cysteine in the medium to a level that enhances cell proliferation leads to effects similar to those obtained with 2-ME. Thus, apparently one effect of various thiol reducing agents is to enhance the production of glutathione in lymphocyte mitogenesis and to protect against the loss of glutathione that occurs in resting or proliferating lymphocytes.
Common idiotypic determinant associated with human antibodies to hepatitis B surface antigen.
Authors: Kennedy R C , Dreesman G R
Published: December 2022

Anti-Id antisera generated to purified anti-HBs isolated from two different individuals recognized a common idiotype. This idiotype was detected in purified anti-Hbs preparations from three individuals and also in anti-HBs-positive sera obtained from six hemophilic patients. The ability of both HBsAg and a virus-derived HBsAg native polypeptide to inhibit the idiotype-anti-idiotype reaction suggested that the anti-HBs idiotype was associated with the antibody-combining site.
Demonstration of multiple antigenic sites of the SV40 transplantation rejection antigen by using cytotoxic T lymphocyte clones.
Authors: Campbell A E , Foley F L , Tevethia S S
Published: December 2022

Multiple antigenic sites on the simian virus 40 (SV40) tumor-specific transplantation antigen (TSTA) were detected by the use of cytotoxic T lymphocyte (CTL) clones isolated from continuous cultures of SV40-specific CTL (H-2b). Two independently derived clones, K11 and K19, specific for the SV40 TSTA in association with H-2Db, each recognized a different antigenic determinant of the SV40 TSTA. This conclusion was based on the observation that a human papovavirus BK virus (BKV) transformed cell line, which possesses a T antigen serologically cross-reactive with that of SV40, was lysed by a heterogeneous population of SV40-immune lymphocytes and by clone K19 but not by K11. Therefore, these CTL clones must recognize two different antigenic determinants of the SV40 TSTA:K19 recognizes a cross-reactive determinant of the SV40 and BKV TSTA, whereas K11 is reactive against an SV40-specific determinant.
The characteristics of a monoclonal antibody that binds specifically to rat T lymphoblasts and inhibits IL 2 receptor functions.
Authors: Osawa H , Diamantstein T
Published: December 2022

In order to obtain an anti-interleukin 2 (IL 2) receptor antibody, we immunized mice with phorbol myristate acetate-pulsed rat T lymphoblasts. The spleen cells of the mice were fused with myeloma cells. Several stable clones of hybridoma cells were obtained that produced monoclonal antibodies (mAb) reacting specifically with rat T lymphoblasts. Only one of these mAb, the mAb ART18, showed characteristics of a putative anti-IL 2 receptor antibody. This mAb was produced on a large scale, purified, and characterized. As tested by a binding assay, 125I-labeled mAb ART18 bound to rat T lymphoblasts (7.5 X 10(4) binding sites per cell), but not to thymocytes or spleen cells of rat origin, or to lymphoblasts, thymocytes, or spleen cells of murine origin. Only marginal binding to lipopolysaccharide-stimulated rat lymphoblasts was detected. The mAb ART18 inhibited in a species-specific and dose-dependent manner i) the capacity of rat T lymphoblasts to absorb IL 2 activity and ii) the capacity of rat T lymphoblasts to proliferate in response in IL 2. The function of T lymphoblasts of murine origin was not affected by the mAb ART18. The time course of the acquisition by mitogen-stimulated spleen cells of the capacity to absorb IL 2 activity was paralleled by that of their capacity to bind 125I-labeled mAb ART18. According to these data, the mAb ART18 seems to be directed against an antigenic determinant of the IL 2 receptor molecule.
Characteristics of tolerance induction among adult hapten-specific T lymphocyte precursors revealed by clonal analysis.
Authors: Good M F , Nossal G J
Published: December 2022

Thaptens 2,4,6-trinitrobenzene sulfonic acid (TNBS) and 3-iodo-4-hydroxy-5-nitrophenyl-acetic acid succinamide ester (NIP) on murine splenic and thymic cytotoxic T lymphocyte precursors (CTLp) was studied. Both BALB/c (H-2d) and CBA (H-2k) CTLp can be rendered unresponsive to subsequent in vitro stimulation with either hapten, provided the mice were pretreated with the correct dose of reactive hapten. The unresponsiveness was shown not to be due to toxicity, because treated animals can mount an alloreactive response equivalent to untreated animals. A degree of cross-reactivity in tolerance induction, however, was demonstrated between the two different haptens. Limit dilution analysis of tolerant lymphoid populations reveals that such cells obey single hit (zero order) kinetics and that there is effective deletion of the CTLp. After correction for "anti-self" responses, the anti-TNP CTLp frequency in TNBS-treated BALB/c mice was 10.7 X 10(-6) in contrast to 101.1 X 10(-6) in control animals (89.4% tolerance). Similarly, after correction for "anti-self" responses, the anti-NIP CTLp frequency in NIP-treated animals was 4.7 X 10(-6), in comparison to 42.8 X 10(-6) in control animals (89.0% tolerance). Kinetic studies show that tolerance occurs within 24 hr. This, together with the limit dilution data, argues against the role of suppressor cells, over the cellular range studied.
A monoclonal antibody reactive with Marek's disease tumor-associated surface antigen.
Authors: Lee L F , Liu X , Sharma J M , Nazerian K , Bacon L D
Published: December 2022

An antibody-secreting hybridoma, RPH-6, to Marek's disease tumor-associated surface antigen (MATSA) was produced by somatic-cell hybridization between the mouse myeloma SP2/O-Ag/14 and spleen cells from MSB1 immunized mice. The antibody reacted with 95 to 100% of the cells from eight of 10 chicken MD cell lines and one of two turkey MD cell lines. It did not react with chicken MD cell lines RP1 and SK3, turkey MD cell line RP19, lymphoid leukosis (LL) cell lines, RP9 and RP12, and reticuloendotheliosis virus (REV) cell line RP14. A weak reaction was observed with REV cell line RP13 (10 to 20%), and a very slight reaction with normal chicken spleen cells (1 to 5%). RPH-6 produces immunoglobulin of the IgM class. Cell culture and mouse ascitic fluids have titers of 10(2) and 10(6), respectively, by fluorescent antibody (FA) test against MD tumor cell lines. The antibody did not react with Marek's disease virus (MDV) internal antigen or membrane antigen as detected by indirect FA test on chicken embryo fibroblast cultures grown on coverslips. The 51Cr release assay showed RPH-6 is highly cytotoxic (60% release) only against MD lymphoblastoid cell lines, with antibody prepared from mouse ascites having a cytotoxic titer approaching 10(6). Results from using RPH-6 for differential diagnosis of chicken lymphoid tumors of unknown origin were in complete agreement with those obtained with rabbit anti-MATSA reference serum and pathologic diagnosis.
Level of mIa expression on mitogen-stimulated murine B lymphocytes is dependent on position in cell cycle.
Authors: Monroe J G , Cambier J C
Published: December 2022

Using correlated flow cytometric analysis of cell surface Ia antigen expression (immunofluorescence) and cell cycle phase (pulse-width of axial light extinction), we have quantitated changes in expression of mIa antigen on murine B cells during progression through cell cycle. Our results indicate that density of mIa expressed on mitogen-stimulated B cells increases fourfold to fivefold during the transition from G0 to G1. By early S phase, mIa density has decreased by fourfold to fivefold relative to peak expression. This decrease becomes more evident by late S, G2, and M phases, when an eightfold decrease in mIa antigen density is observed relative to peak levels. This decrease results in mIa antigen expression lower than that of resting, unstimulated B cells. Therefore, maximum mIa antigen expression occurs during G0 to G1 transition and in early G1, when a requirement for I region-restricted, antigen-driven T cell help for thymus-dependent, antigen-driven B cell activation has been demonstrated.
Bone marrow-derived macrophage as accessory cells in antigen-induced T cell proliferation. H-2I region requirements for L-glutamic60-L-alanine30-L-tyrosine10 response.
Authors: Schook L B , Allen P M , Niederhuber J E
Published: December 2022

Marrow stem cells cultured with L cell-conditioned medium were used to produce large numbers of macrophages free of contaminating lymphoid cells or granulocytes. Experiments were performed by using these bone marrow-derived macrophages (BMDM) as antigen-presenting cells (APC) for the terpolymer antigen L-glutamic60-L-alanine30-L-tyrosine10 (GAT). By using antigen-specific T cell proliferation, it was demonstrated that BMDM were equal to splenic macrophages in their capacity to present GAT. Furthermore, when BMDM were pretreated with alloantibodies specific for Ia antigens of the I-A subregion, the T cell proliferative response to GAT was inhibited. The I-A subregion is known to be the site of Ir gene regulation of the GAT response, and antibodies to other subregions had no inhibitory effect. Monoclonal antibodies recognizing a beta-chain product of the I-A subregion (Ia. 17) also inhibited the BMDM from effective antigen presentation. These results were similar to those obtained with purified splenic macrophages.
The effect of alpha 1 antitrypsin on the proliferative response of human peripheral blood lymphocytes.
Authors: Breit S N , Luckhurst E , Penny R
Published: December 2022

In evaluating immune aberrations in patients with alpha 1 antitrypsin (alpha 1 AT) deficiency, we have previously shown that they exhibit enhanced lymphocyte responsiveness to PHA that is serum mediated. In this study, we demonstrate suppression of the PHA response by using purified alpha 1 AT, and also similar but less marked suppression of the Con A response. alpha 1 AT, however, has no effect whatsoever on PWM-induced proliferation. This effect is demonstrable provided alpha 1 AT is added within 4 hr of mitogen activation and is mediated by its action on adherent cells rather than on proliferating lymphocytes. Adherent cells still exhibit this effect if pulsed with alpha 1 AT, then thoroughly washed before their activation. This suggests that it may be inhibiting a membrane serine esterase already activated before the addition of PHA. Thus alpha 1 AT may modulate the activation of T cells through its effect on monocytes, leading to abnormalities in immunoregulation, and hence a predisposition to the development of a variety of immunologic disorders in alpha 1 AT-deficient subjects.
Autoimmune effector cells. IV. Induction of experimental allergic encephalomyelitis in Lewis rats without adjuvant.
Authors: Holda J H , Silberg D , Swanborg R H
Published: December 2022

Experimental autoimmune encephalomyelitis (EAE) was induced in Lewis rats without the aid of adjuvant. Radioresistant cells were activated from spleens of nonimmune donor rats by in vitro culture with myelin basic protein (BP). After adoptive transfer, these cells recruited EAE effector cells in normal syngeneic recipients, as demonstrated after transfer to secondary hosts. The induction of EAE without adjuvant may lead to a better understanding of the mechanisms of autoimmune disease induction.
Identification of a membrane antigen that is distinct from the interleukin 2 receptor and that may be required for interleukin 2-driven proliferative responses.
Authors: Malek T R , Robb R J , Shevach E M
Published: December 2022

During the course of studies designed to raise murine monoclonal antibodies to a guinea pig alloreactive T cell line, an IgM monoclonal antibody (5C3) was identified that completely inhibited the alloantigen and IL 2-dependent proliferative response of the immunizing colony and other alloreactive T cell colonies with different antigen specificities. Tissue distribution on the FACS demonstrated that the surface antigen defined by 5C3 was preferentially expressed on T cells activated by mitogen, alloantigen, or antigen. 5C3 was also a potent inhibitor of alloantigen- and antigen-induced T cell proliferation. In addition, 5C3 substantially inhibited the proliferative response of guinea pig Con A-induced blast cells to guinea pig IL 2-containing culture fluids and to a preparation of partially purified human IL 2 but was a poor inhibitor of mitogen-induced T cell proliferation. Blast cells obtained from the co-culture of mesenteric lymph node cells with Con A in the presence of 5C3 lacked the surface molecule defined by 5C3. The 5C3- and 5C3+ blast populations, nevertheless, absorbed equivalent amounts of IL 2, which suggested that 5C3- blasts were not deficient in the expression of IL 2 receptors. However, 5C3- blast cells failed to proliferate in response to IL 2-containing culture fluids. Furthermore, 5C3 had no inhibitory activity on either the binding or subsequent degradation of radiolabeled human IL 2 by guinea pig IL 2 receptors. These data suggest that 5C3 recognizes a component of a putative IL 2 receptor-effector complex distinct from the IL 2 receptor and that this molecule plays a critical role in the processing or generation of the growth signal IL 2 transmits to T lymphocytes.
A requirement for nonspecific T cell factors in antibody responses to "T cell independent" antigens.
Authors: Endres R O , Kushnir E , Kappler J W , Marrack P , Kinsky S C
Published: December 2022

Using murine splenic B cell preparations depleted of macrophages and rigorously depleted of T cells, we studied the role of nonspecific helper factors in in vitro antibody responses to T cell-independent (TI) type 1 and type 2 antigens. TNP-lipopolysaccharide, TNP-Brucella abortus, and DNP-liposomes containing lipid A were chosen as examples of TI type 1 antigens. DNP-Ficoll and DNP-liposomes without lipid A were chosen as TI type 2 antigens. Only the type 1 antigens were able to elicit significant, albeit very weak, responses without added helper factors. Both type 1 and 2 antigens required factors present in supernatants from concanavalin A-stimulated spleen cells (Con A SN) to stimulate optimum antibody responses. Interleukin 2- (IL 2) containing supernatant from the T cell hybridoma FS6-14.13 supported suboptimal responses to varying degrees with each TI antigen, in contrast to its lack of effect on responses to sheep red blood cells in the absence of additional factors. This activity of the FS6-14.13 supernatant was removed by absorption with the IL 2-dependent T cell line HT-2, suggesting that IL 2 was the active component. Another factor, IL-X, which is distinct from both IL 1 and IL 2 and is also found in Con A SN, was required in addition to IL 2 to achieve optimal responses with both types of TI antigens. These results clearly establish a role for factors derived from T cells in the activation of B cells by both type 1 and type 2 TI antigens.
Nonspecific inhibitor of DNA synthesis elaborated by T acceptor cells. I. Specific hapten- and I-J-driven liberation of an inhibitor of cell proliferation by Lyt-1-2+ cyclophosphamide-sensitive T acceptor cells armed with a product of Lyt-1+2+-specific suppressor cells.
Authors: Malkovský M , Asherson G L , Chandler P , Colizzi V , Watkins M C , Zembala M
Published: December 2022

Lyt-1+2+ hapten-specific T suppressor cells (Ts) from mice injected and then painted with picryl or oxazolone derivatives produce hapten-specific T suppressor factors (TsF) in vitro. Stimulation by painting with contact sensitizer (which need not be specific) gives rise to Lyt-1-2+, I-J+, cyclophosphamide-sensitive T acceptor cells (Tacc). When the Tacc population is armed with TsF and then is exposed to specific antigen in the context of I-J-controlled determinants (antigen-presenting, haptenized spleen cells and Ts sharing the same I-J subregion), a nonspecific inhibitor of DNA synthesis (nsINH) appears in the supernatant. This inhibitor suppresses the primary DNA synthetic response to concanavalin A, lipopolysaccharide, and alloantigens in both syngeneic and allogeneic lymphocytes. The nsINH is only effective when added to lymphocyte cultures less than 8 hr after the stimulation with concanavalin A. The nsINH, however, affects neither primary nor secondary cytotoxicity in vitro. These data suggest the mouse immune system is capable of selective regulation of the response to specific antigen by the production of nonspecific soluble suppressor factor(s).
The synthesis and secretion of granulocyte-monocyte colony-stimulating activity (CSA) by isolated human monocytes: kinetics of the response to bacterial endotoxin.
Authors: Sullivan R , Gans P J , McCarroll L A
Published: December 2022

We studied the kinetics of the synthesis and secretion of granulocyte-monocyte colony-stimulating activity (CSA) by human monocytes stimulated by S. typhi endotoxin. We found that these cells initially secrete copious quantities of CSA when exposed to endotoxin but rapidly become refractory to its stimulatory effect. When monocytes were incubated in liquid suspension cultures, large amounts of CSA were generated during the first 24 hr of culture after the addition of as little as 10 ng/ml of endotoxin to previously unstimulated monocytes. After the addition of endotoxin, CSA secretion abruptly took place after an initial 1 to 2 hr lag phase, and occurred primarily within the first 6 to 12 hr of culture. The addition of puromycin or cycloheximide to the cultures significantly inhibited CSA secretion in response to endotoxin, suggesting that CSA production by stimulated monocytes requires de novo protein synthesis and does not solely result from the release of preformed active CSA. After initial exposure to endotoxin, CSA production by monocytes steadily decreased after 24 hr and ceased after 72 hr. Subsequent reexposure to the same concentration of endotoxin resulted in little CSA production, whereas the refractory state could be overcome by increasing the quantity of endotoxin added. Additional studies demonstrated that refractoriness of monocytes to further CSA production after initial response to endotoxin was not due to a loss of cell viability, degradation of endotoxin, or inhibition of CSA synthesis by soluble metabolites generated by stimulated monocytes. Our data suggest that the capacity of human monocytes to synthesize CSA in response to endotoxin rapidly becomes blunted after initial exposure of the cells to the lipopolysaccharide, but that the refractory state of these cells can be overcome by increasing the concentration of endotoxin. Acquired hyporesponsiveness of cells that produce CSA may in part account for the phenomenon of immediate endotoxin tolerance observed in vivo.
Isoelectric focusing of human anti-diphtheria toxoid antibodies: identical spectrotypes of anti-fragment A antibodies with the same IgG subclass and light chain constant regions are expressed in multiple individuals.
Authors: Morrow C D , Dorey F , Stevens R H
Published: December 2022

The serum antibodies from humans booster-immunized with diphtheria toxoid were analyzed by isoelectric focusing. To restrict the antibody response, we visualized those antibodies that reacted to the Fragment A moiety of diphtheria toxin. Of the 100 normal human donors examined, 20 were estimated to be nonresponders to Fragment A. Sixty-three of the 80 donors who responded to Fragment A had restricted IgG-anti-Fragment A spectrotypes that consisted of three to eight distinct bands. Six series of repeat or shared spectrotypes were delineated among our donor population with repeat frequencies ranging from 0.025 to 0.312. We determined that the repeat spectrotypes were IgG1-kappa anti-Fragment A antibodies. Three percent of our donors had complex IgG-anti Fragment A spectrotypes consisting of greater than 10 bands. In some instances, the complex spectrotypes were composed of simple spectrotypes found among other donors. The complex IgG-anti Fragment A spectrotypes of these donors consisted of both IgG1 and IgG4-anti-Fragment A antibodies. From a statistical analysis, we calculate that only a limited number of IgG-anti-Fragment A spectrotypes are expressed in vivo after booster immunization. Our analysis suggests two groups of IgG-anti-Fragment A spectrotypes exist in our donor population. One IgG-anti-Fragment A spectrotype group is expressed randomly, and a second group is preferentially expressed among individuals in our donor population.
Cell volume and osmotic properties of erythrocytes after complement lysis measured by flow cytometry.
Authors: Bauer J , Valet G
Published: December 2022

The changes of volume distribution curves of erythrocytes during and after lysis by complement or nystatin or in hypotonic buffers were measured by flow cytometry. Biconcave and spheroidal ghosts were observed after complement lysis and spheroidal ghosts were seen only after nystatin and hypotonic lysis. The spheroidal ghosts derived from red cells lysed by complement or nystatin were permeable to sucrose; those from hypotonic lysis were sucrose-impermeable. Spheroidal ghosts after complement lysis remained permeable for sucrose whereas spheroidal ghosts after nystatin lysis resealed after removal of the drug by washing. Biconcave ghosts produced by complement lysis were almost impermeable to sucrose initially and therefore responded to osmotic changes, but they became sucrose-permeable upon prolonged incubation at 37 degrees C. The rate of sucrose equilibration increased as the stability of the biconcave shape diminished with increasing numbers of C5b-9 complexes. At 850 C5b-9 complexes/ghost, the biconcave shape and impermeability for sucrose were completely lost. The results support the hypothesis that complement C5b-9 complexes, in addition to the interaction with the lipid bilayer, may interact with the cytoskeleton of the erythrocyte membrane.
Studies on the mechanism of bacterial resistance to complement-mediated killing. III. C5b-9 deposits stably on rough and type 7 S. pneumoniae without causing bacterial killing.
Authors: Joiner K , Brown E , Hammer C , Warren K , Frank M
Published: December 2022

Gram-positive cocci resist direct killing by serum. The mechanism of resistance was studied by measuring consumption of terminal complement components from serum and uptake of purified, radiolabeled C7 and C9 on rough and encapsulated type 7 Streptococcus pneumoniae. Extensive consumption of C5, C7, and C9 occurred when 5 X 10(8) rough or type 7 pneumococci were incubated for 1 hr in 10% pooled normal human serum (PNHS). Approximately 10,000 molecules of C7 and C9 bound per organism during the same period of incubation. Twenty to 30% of C7 and C9 was released from rough organisms. Release was not due to autolysis since it occurred with glutaraldehyde-fixed organisms as well as in S. pneumoniae that were rendered resistant to autolysis by growth in ethanolamine. Between 10 and 30% of bound 125IC9 counts were eluted from the rough and type 7 organisms by incubation in 1 M NaCl or 0.01 M EDTA, which suggests that bound C5b-9 was not attached by predominantly ionic interactions. Elution of 44 to 74% of 125IC9 from live and glutaraldehyde-fixed organisms by 1% sodium deoxycholate suggests that hydrophobic bonds are involved in C5b-9 attachment. Trypsin cleaved 67 and 55% of 125IC9 counts from live rough and type 7 S. pneumoniae, respectively which indicates that the bound complex is not protected by the cell wall from proteolytic attack. Serum resistance in S. pneumoniae does not represent a failure to form C5b-9 on the bacterial cell wall but apparently reflects a failure of the bound complex to penetrate the thick peptidoglycan layer.
Granulocyte activation by endotoxin. II. Role of granulocyte adherence, aggregation, and effect of cytochalasin B, and comparison with formylated chemotactic peptide-induced stimulation.
Authors: Dahinden C , Fehr J
Published: December 2022

PMN stimulation by endotoxin is heavily dependent on incubation conditions: although marked increase of PMN adhesiveness, enzyme release, and hexose monophosphate shunt activity, as well as superoxide production, occur when endotoxin-challenged cells are incubated in stationary petri dishes, absolutely no such response is observed with cells kept in suspension. Preincubation with cytochalasin B does not alter this reaction pattern. Hyperadhesiveness of endotoxin-challenged PMN is retained after thorough washing, eliminating the possibility that adhesion is due to absorption of endotoxin to petri dishes. In contrast to formylated chemotactic peptides that stimulate PMN adherence as well as aggregation, endotoxin elicits absolutely no aggregatory response (+/- cytochalasin B). Preexposure to endotoxin does not block the aggregatory response to subsequent addition of chemotactic peptides, whereas effects of endotoxin and formylated peptides on PMN in the stationary system are additive. These findings indicate that contact to surfaces is an essential primary event in PMN stimulation.
Studies of allogeneic tumor transplants: induced rejection of advanced tumors by immune alteration of recipients.
Authors: Russell P S , Chase C M , Burton R C
Published: December 2022

In the present experiments, a methylcholanthrene-induced sarcoma (S-702) of B10.D2 origin was found to grow rapidly in B6AF1 mice leading to the death of all recipients in 5 to 9 wk. Nevertheless, immunity to MHC antigens presented by the tumor was readily demonstrable in tumor-bearing mice by their responses to donor strain skin grafts until late in the course of tumor growth, when a nonspecific form of immune suppression developed. In addition, B6AF1 mice preimmunized by exposure to B10.D2 donor strain antigens did not permit tumor growth. Treatment of tumor-bearing B6AF1 mice with CY at 18 days, when the tumors measured over 12-mm in diameter, followed by the i.p. injection of B10.D2 lymphoid cells (at a dosage of from 1.2 to 2.5 X 10(8) cells) resulted in the complete regression of 100% of these large tumors. CY treatment combined with localized immune stimuli in the form of donor strain skin grafts or secondary tumor implants was incapable of producing a sufficiently heightened immune response to cause tumor rejection. A dose of CY temporarily retarded tumor growth in most mice, and in a minority of animals so treated (less than 25%) tumors regressed completely. In syngeneic (B10.D2) animals, CY also temporarily slowed tumor growth, but total regression was never observed. An effective B10.D2 cell inoculum could consist not only of living lymphoid cells but of irradiated (1000 rad) cells as well. Tumor cell suspensions (after irradiation, 10,000 rad) were also effective. These observations suggest local immune factors at the host-tumor interface may have been of importance in the survival of these allogeneic tumor transplants and that CY influenced this state, perhaps through an influence on suppressor cells, allowing subsequent administration of donor strain cellular antigens to induce an effective tumor rejection response.
Enhancement of tumor metastasis and suppression of natural killer cell activity by beta-estradiol treatment.
Authors: Hanna N , Schneider M
Published: December 2022

Treatment of mice with 17 beta-estradiol leads to a selective inhibition of natural killer (NK) cell activity as measured by in vitro cytotoxicity against YAC-1 lymphoma and UV-2237 fibrosarcoma target cells. Activation of NK cells by polyinosinic-polycytidylic acid (poly I.C) and Corynebacterium parvum also was impaired in beta-estradiol-treated mice, whereas activation of tumoricidal macrophages was uncompromised. The beta-estradiol-induced suppression of NK cell activity was associated with a higher incidence of experimental and spontaneous pulmonary metastasis of the syngeneic UV-2237 fibrosarcoma and K-1735 and B16 melanoma tumors. Injection of poly I.C or C. parvum before i.v. tumor challenge markedly inhibited the development of lung tumor colonies in normal recipients, but it was less effective in beta-estradiol-treated mice. Adoptive transfer of normal spleen cells enhanced the NK cell activity and increased the resistance of beta-estradiol-treated mice against hematogenous tumor metastasis. Nude mice treated with beta-estradiol also exhibited a low level of NK cell activity and an enhanced susceptibility to metastasis of allogeneic tumor cells. The potential use of this in vivo model for studies of metastasis of human malignant neoplasms is discussed.
Natural killer (NK) cells as a responder to interleukin 2 (IL 2). II. IL 2-induced interferon gamma production.
Authors: Handa K , Suzuki R , Matsui H , Shimizu Y , Kumagai K
Published: December 2022

In the accompanying paper, we showed that natural killer (NK) cells were a major population in the naive spleens of normal mice that responded directly to a T cell growth factor, interleukin 2 (IL 2), and clonally replicated without other stimulating agents. The cloned cells growing in IL 2 showed a potent NK activity against several NK targets without addition of an NK-activating agent, interferon (IFN). In the present study, therefore, we examined whether these cloned NK cells on their own produced IFN. It was found that all NK clones growing in IL 2 produced IFN in the culture fluids. The titers of IFN produced in the IL 2-containing media correlated well with the number of growing cells. With the culture in the absence of IL 2, neither cell growth nor IFN production could be detected. Addition of Con A into the culture in the IL 2-free media showed no IFN production. The antiserum neutralizing IFN alpha and IFN beta failed to significantly neutralize IFN produced by NK clones. Treatment with either a pH of 2.0 or antiserum neutralizing mouse IFN gamma resulted in a marked reduction of IL 2-induced NK IFN, indicating that a major part of IFN produced was IFN gamma. These results indicate that IL 2 stimulates NK clones to proliferate, accompanied by IFN gamma production. The results also show that an NK clone, when stimulated with Sendai virus, produced a type 1 IFN (IFN alpha and/or IFN beta), suggesting that murine NK cells can produce both type 1 (alpha and/or beta) and type 2 (gamma) IFN, depending on inducers.
Differences in the MHC-restricted self-recognition repertoire of intra-thymic and extra-thymic cytotoxic T lymphocyte precursors.
Authors: Kruisbeek A M , Sharrow S O , Singer A
Published: December 2022

The MHC specificity of TNP-specific pCTL from the thymus and spleen of F1 leads to parent chimeras was evaluated. It was found that in the presence of exogenously added helper factor IL 2, thymic pCTL were restricted to recognizing TNP only in association with host MHC determinants, whereas splenic pCTL recognized TNP in association with either host or donor MHC determinants. Thus, the spleens of F1 leads to parent chimeras contain a pCTL repertoire that is not present intrathymically. Data are presented which suggest that such pCTL did nevertheless differentiate into functional competence in the chimeric host. These results are consistent with extra-thymic differentiation as the mechanism by which such nonthymically restricted pCTL may have developed.
Differences in in vivo distribution and homing of T cell subsets to mucosal vs nonmucosal lymphoid organs.
Authors: Kraal G , Weissman I L , Butcher E C
Published: December 2022

The migratory properties of Lyt-2- and Lyt-2+ T cells in the mouse have been investigated. In short-term in vivo homing studies, Lyt-2- T cells localized consistently more efficiently than Lyt-2+ T cells in Peyer's patches (about 1.5 times as well), whereas both populations localized roughly equivalently in peripheral lymph nodes. These homing characteristics of Lyt-2- and Lyt-2+ subsets are largely independent of their organ source. The specificity of migration appears to be determined by selective recognition of organ-specific determinants on the endothelial cells of high endothelial venules (HEV), specialized venules that mediate the exit of migrating lymphocytes from the blood: In an in vitro assay of lymphocyte binding to HEV in lymphoid organ frozen sections, Lyt-2- cells constituted a significantly and consistently greater proportion of T cells binding to Peyer's patch HEV than of those binding to peripheral node HEV. The homing and HEV recognition preferences of the Lyt subsets are reflected in differences in their in situ representation in mucosal vs nonmucosal lymphoid organs, which suggests that the selective migration of these populations may be an important factor in determining the character of local immune responses.
Demonstration of abnormalities in expression of thymic epithelial surface antigens in severe cellular immunodeficiency diseases.
Authors: Haynes B F , Warren R W , Buckley R H , McClure J E , Goldstein A L , Henderson F W , Hensley L L , Eisenbarth G S
Published: December 2022

Thymic epithelium from three patients with severe cellular immunodeficiency diseases were compared with age-matched normal thymic epithelium using three markers of human thymic epithelium and antibodies against thymosin alpha 1, thymopoietin, and thymosin beta 4. We have previously shown that normal thymic epithelium reacts with antibodies against GQ gangliosides (antibody A2B5) and binds tetanus toxin (TT). In addition, some areas of normal thymic epithelium express human Thy-1 antigen. We found thymic epithelium in patients with severe cellular immunodeficiency diseases to be different from normal subjects. Two children with severe combined immunodeficiency disease (SCID) had thymic epithelium that bound anti-GQ ganglioside antibody but, unlike in normals, did not bind TT. The patient with severe cellular immunodeficiency and normal serum immunoglobulins (Nezelof syndrome) had thymic epithelium that bound TT but, unlike normal thymic epithelium, did not react with anti-GQ ganglioside antibody. Thymic epithelium from both SCID and Nezelof syndrome patients contained thymosin alpha 1, thymopoietin, and thymosin beta 4 and expressed human Thy-1 antigen. In contrast to SCID thymus rudiments, Nezelof thymus contained numerous (though fewer than normal) lymphocytes with mature T cell surface antigens. Thus, using these probes of human thymic epithelium, we have demonstrated heterogeneous defects in thymic epithelial surface marker expression in severe primary cellular immunodeficiency diseases. These defects presumably reflect abnormalities of in vivo thymic epithelial maturation.
Thymic dysfunction in the mutant diabetic (db/db) mouse.
Authors: Dardenne M , Savino W , Gastinel L N , Nabarra B , Bach J F
Published: December 2022

Thymic function has been explored in genetically diabetic homozygous C57BL/KsJ (db/db) mice by evaluating their serum thymic factor (FTS) levels with a rosette assay. As previously reported for other autoimmune mice (NZB or MRL/I mice), the age-dependent decline of FTS levels was significantly accelerated in diabetic mice when compared to heterozygous littermates. Furthermore, FTS inhibitory molecules were detected in db/db mouse sera (as early as 10 wk of age) as evaluated by their ability to absorb in vitro the activity of synthetic FTS in the rosette assay, and in vivo for their capacity to induce the disappearance of endogenous FTS when injected into normal mice. These inhibitors were shown to be immunoglobulins. Histologically, the thymus presented an accelerated involution starting with a cortical lymphocytic depletion and an increased number of Hassall's corpuscles. Ultrastructural studies showed alterations in thymic epithelial cells, mainly represented by an increasing number of cytoplasmic vacuoles. By means of indirect immunofluorescence with anti-FTS monoclonal antibodies, it was shown that the number of FTS+ cells was reduced in db/db mouse thymuses: at the age of 22 wk, diabetic mice had 10 times fewer FTS+ cells than heterozygotes of the same age. Taken together, these results indicate important abnormalities in the thymus of diabetic mice. It is possible that the associated lymphocyte dysfunction plays a role in the pathogenesis of the autoimmune disease presented by db/db mice.
Modulation of fibroblast growth by a lymphokine of human T cell continuous T cell line origin.
Authors: Wahl S M , Gately C L
Published: December 2022

Human peripheral blood T lymphocytes activated by mitogens or specific antigen produce a mediator that stimulates proliferation in a quiescent population of dermal fibroblasts. This lymphocyte product, which may have implications in the fibrosis associated with chronic cell-mediated inflammatory lesions, is not elaborated by unstimulated lymphocytes. Characterization of the fibroblast mitogenic factor differentiated it from a monocyte-derived factor with similar activity. The T cell factor appears to be a protein with an apparent m.w. of 40,000 and an isoelectric point between 5.0 and 5.5. This fibroblast growth factor can be produced in the relative absence of monocytes, and further evidence that the factor is a T cell product stems from the ability of a human T cell line to generate such a factor. Comparison of the biochemical characteristics of the normal T cell product with those of the T cell line revealed them to be indistinguishable. Thus, the T cell line can be used as a source of fibroblast-activating factor (FAF) for more detailed analysis of its structure and function. Further characterization of this lymphokine may contribute to understanding the mechanisms that mediate fibrosis in normal tissue repair as well as in the pathophysiologic fibrotic response associated with certain chronic inflammatory diseases.
Evidence for minor structural variations of class II genes in wild and inbred mice.
Authors: Wakeland E K , Klein J
Published: December 2022

We organized 29 B10.W lines into eight antigenic groups on the basis of similarities in the antigenic phenotypes of their class II antigens. Tryptic peptide fingerprint comparisons of class II antigens in different antigenic groups detected structural variations in 30 to 60% of the tryptic peptides. In contrast, less than 10% of the tryptic peptides were distinguishable in comparisons of class II antigens within the same antigenic group. These results suggest structurally discrete allelic families of class II genes exist in wild mouse populations.
The Qa-1 alloantigens. II. Evidence for the expression of two Qa-1 molecules by the Qa-1d genotype and for cross-reactivity between Qa-1 and H-2K.
Authors: Cook R G , Jenkins R N , Flaherty L , Rich R R
Published: December 2022

The nature of cell surface determinants detected by Qa-1-specific alloantisera and cytotoxic T lymphocytes (CTL) in mice of the H-2f, Qa-1d genotype was investigated. The H-2f, Qa-1d strains A.CA and B10.M express both Qa-1a and Qa-1b encoded alloantigenic determinants (Qa-1.1 and Qa-1.2, respectively), as defined in the prototypic A (or B6-Tlaa) and C57BL/6 (or A-Tlab) strains, respectively. Both anti-Qa-1.1 and -Qa-1.2 sera immunoprecipitate 46K m.w. glycoproteins from H-2f, Qa-1d strains. In addition to 46K m.w. proteins, anti-Qa-1.1 sera, but not anti-Qa-1.2 sera, precipitate 55 to 75K m.w. proteins; the nature of these proteins and their relationship to Qa-1 is unclear at present. Sequential immunoprecipitation experiments and the analysis of several recombinant strains revealed that anti-Qa-1.1 sera also cross-react with a 46K m.w. H-2f-encoded alloantigen, probably H-2Kf. Both Qa-1.1 and the non-Qa-1.1 cross-reacting determinants were detected by polyclonal anti-Qa-1a CTL on the Qa-1d strains. The Qa-1a encoded but not the cross-reacting determinants were detected by a cloned anti-Qa-1a CTL line. Sequential immunoprecipitation experiments on the recombinant strains B6.AC2 and B10.M(17R), which are Qa-1d but not H-2Kf, revealed that the Qa-1.1 and Qa-1.2 determinants do not reside on the same molecule. Furthermore, although Qa-1b-encoded determinants were detected on these strains with anti-sera and with bulk CTL cultures, cloned anti-Qa-1b CTL lines thus far analyzed have failed to react with Qa-1d targets, indicating that some but not all of the prototypic Qa-1b-encoded determinants are expressed by the Qa-1d strains.
The equilibrium constant for the interaction between a monoclonal Fab fragment and an influenza virus neuraminidase.
Authors: Jackson D C , Howlett G J , Nestorowicz A , Webster R G
Published: December 2022

The affinity or equilibrium constant between an Fab fragment derived from monoclonal IgG directed against influenza virus neuraminidase was measured as 4.1 X 10(7) M-1. The method, which makes use of an air-driven ultracentrifuge, is simple and uses extremely small amounts (10(-11) mol) of material. Furthermore, interpretation of the data is based on sound theoretical considerations. The technique also allows m.w. of the interacting species to be measured and the stoichiometry of the reaction to be determined.
Neutralization of influenza virus by normal human sera: mechanisms involving antibody and complement.
Authors: Beebe D P , Schreiber R D , Cooper N R
Published: December 2022

All normal human sera examined neutralized WS/33 H1N1 influenza virus efficiently by one of two antibody-dependent mechanisms. A minority of the sera contained moderate levels of IgG antibody directed against the viral hemagglutinin that had the ability to directly neutralize the virus. The majority of sera tested contained very low levels of IgG anti-hemagglutinin antibody, which was detectable with a specific ELISA but not by conventional HAI assays. Such IgG antibody was unable to directly neutralize the virus. Studies with agammaglobulinemic serum and with sera depleted of and reconstituted with complement components established essential roles for IgG and the components of the classical complement pathway through C3 for neutralization. The components of the alternative and membrane attack pathways were not needed for neutralization. As anticipated from the requirement for IgG and exclusive mediation of neutralization by the classical pathway, the virus-IgG immune complex activated purified C1. Binding of C3 and C4 to the virus was demonstrated, as was classical pathway-mediated triggering of the alternative pathway, with recruitment of properdin. In addition, the H1N1 influenza virus also directly activated the alternative complement pathway in human serum, leading to C3 and properdin deposition on the viral envelope. Such direct alternative pathway activation also required immunoglobulin. However, the alternative pathway alone was unable to neutralize the virus. Thus, most normal sera examined contain low levels of IgG anti-hemagglutinin antibody, which activate the classical pathway of the complement system and neutralize WS/33 influenza virus by deposition of C3 and C4 on the viral envelope.
Induction of thromboxane release from macrophages by anaphylatoxic peptide C3a of complement and synthetic hexapeptide C3a 72-77.
Authors: Hartung H P , Bitter-Suermann D , Hadding U
Published: December 2022

Guinea pig peritoneal macrophages were examined for their response to homologous anaphylatoxic peptide C3a of complement. C3a caused the release of considerable amounts of thromboxane A2 from macrophages maintained in serum-free culture for up to 12 hr. Preincubation of C3a with the Fab portion of a monoclonal anti-C3a IgG abrogated its stimulatory capacity. C3a desarg was found to be rather ineffective in eliciting thromboxane release. Hexapeptide C3a 72-77, comprising the C-terminal amino acids of human C3a, proved also to stimulate the cyclooxygenation pathway of macrophage arachidonic acid metabolism, following the same routes of receptor-ligand interactions (rapid onset of activation, low dose, and cumulative desensitization) as proven for other anaphylatoxin-cell interactions. This novel effect of anaphylatoxins C3a and C3a 72-77 could contribute to inflammatory reactions in which macrophages play a role.