bluuebunny/crossref_metadata_2025 huggingface dataset:
Analysis of recombinant inbred lines derived from "autoimmune" (NZB) and "high leukemia" (C58) strains: independent multigenic systems control B cell hyperactivity, retrovirus expression, and autoimmunity.
Authors: Datta S K , Owen F L , Womack J E , Riblet R J
Published: December 2022

The relationship of B cell hyperactivity and retrovirus expression to other autoimmune traits were examined in recombinant inbred (N X 8 RI) lines derived from NZB and C58 progenitor strains. Although both NZB and C58 mice expressed high levels of xenotropic virus, the RI lines segregated in virologic phenotype, as high or low expressors of the endogenous virus. The expression of the C58-derived ecotropic virus occurred in only one-half of the RI lines, and its expression in the remaining lines of mice appeared to be suppressed by the NZB-derived allele at the Fv-1 locus. The inheritance of B lymphocyte abnormalities of the NZB progenitor strain was investigated by studying spontaneous and SRBC-induced production of IgM by the spleen cells of the RI lines. These two phenotypes of B cell hyperactivity were found to be determined by independently segregating genes and they were not linked to immunoglobulin structural gene loci. The strain distribution patterns of virus expression and B cell hyperactivity in the RI lines did not match with each other or with the inheritance patterns of other immunologic abnormalities, such as defective AMLR and production of autoantibodies.
Idiotypes of anti-Ia antibodies. II. Effects of in vivo treatment with xenogeneic anti-idiotype.
Authors: Epstein S L , Masakowski V R , Sharrow S O , Bluestone J A , Ozato K , Sachs D H
Published: December 2022

The effects of in vivo treatment with xenogeneic anti-idiotypic antibodies were examined in an anti-Ia idiotypic system. Monoclonal antibody 14-4-4S, specific for Ia.7, has been shown to bear idiotopes that are expressed at readily detectable levels in conventional alloantibody responses. Sera from mice treated with purified anti-idiotypic antibodies (anti-Id) were found to contain inhibitory activity in an ELISA specific for the 14-4-4S Id, whereas sera from control mice treated with heterologous normal Ig did not. In addition, sera of anti-Id-treated C3H.SW mice contained specific anti-I-E activity, shown by binding to B10.A(2R) but not B10.A(4R) LPS blasts in flow microfluorometry. The anti-I-E induced by anti-Id included more IgG1 than IgG2. Even though a significant amount of anti-I-E activity was present in the serum, absorption analysis showed that most of the idiotope-positive antibody was not I-Ek-specific. Penetrance of induction of anti-I-E by anti-Id was 100% in the C3H.SW mice tested, and activity persisted in the serum for at least 8 to 9 mo in some cases. B10 mice produced only marginal anti-I-E activity after treatment, suggesting that induction is due to specific triggering rather than due entirely to a resemblance of anti-Id to the I-E antigen. The results thus indicate long-lasting alterations in an anti-Ia idiotypic system in the absence of exposure to conventional antigen, and represent specific manipulation of anti-Ia immunity.
Prepubertal orchidectomy induces thymic abnormalities in aging (NZB X SJL)F1 male mice.
Authors: Dumont F , Barrois R , Habbersett R C
Published: December 2022

Female but not male (NZB X SJL)F1 (NS) mice develop abnormalities of their intrathymic lymphocyte population in the course of aging. To determine the role played by androgens in this sex-related difference, we monitored the evolution of the cellular composition of the thymus in NS males deprived of androgens by prepubertal orchidectomy. Although in young mice this operation resulted in a twofold enlargement of the thymus, there was no histologic alteration or major change in the surface phenotype and mitogenic reactivities of the thymocytes, which suggests that all thymocyte subsets were increased to the same extent. In 12-mo-old control (BALB/c X SJL)F1 mice, prepubertal orchidectomy also produced an equal expansion (1.4-fold increase) of all thymocyte subsets. In contrast, in 12-mo-old orchidectomized NS males, there was a marked depletion of the thymic cortex and a hyperplasia of the medullary lymphoid tissue reflecting the selective expansion of a subset of phenotypically mature T cells (dull Thy-1+, Lyt-1+2+/-, dull PNA+) together with the emergence of intrathymic surface immunoglobulin-bearing cells. These latter cells probably represented B cells because there was a concomitant augmentation of the mitogenic responsiveness in vitro of thymic cell suspensions to lipopolysaccharide. Such thymic abnormalities induced by prepubertal orchidectomy in old NS males resemble those occurring spontaneously in the NS females. This suggests that the absence of thymic disease in intact NS males is primarily due to a suppressive effect of androgens.
Monoclonal antibodies for immunochemical analysis of methanogenic bacteria.
Authors: Conway de Macario E , Macario A J , Kandler O
Published: December 2022

Sixty-nine hybridomas were generated to produce monoclonal antibodies to species of methanogens representing three of the four families accepted at the present time: Methanobacteriaceae, Methanococcaceae, and Methanomicrobiaceae. The antibody of each of 29 hybridomas cross-reacted with a methanogen of the same species or genus as the immunizing (homologous) strain, whereas the antibody of the other 40 cell lines reacted only with the homologous strain. Inhibition-blocking experiments with compounds of known composition and structure were used to define the fine specificity of antibodies of four hybridomas representing the four genera of the methanogens used for immunization. The combining site of the monoclonal antibody against M. thermoautotrophicum delta H examined is specific for a structure in the pseudomurein involving N-acetyl-glucosamine but not the (1-3) linkage or the C-terminus gamma-Glu-Ala of the peptide, both of which are characteristic of pseudomurein, the cell-wall peptidoglycan distinctive of the Methanobacteriaceae. In contrast, the monoclonal antibody to the other methanogen of this family examined, M. arboriphilus DH1, recognizes a determinant involving gamma-Glu-Ala. Thus pseudomurein expresses at least two dissimilar antigenic determinants in different portions of the molecule. The monoclonal antibodies against M. vannielii SB and M. hungatei JF1, whose families do not possess pseudomurein, did not display specificity for analogues of pseudomurein.
Fibroblast stimulation in schistosomiasis. II. Functional and biochemical characteristics of egg granuloma-derived fibroblast-stimulating factor.
Authors: Wyler D J , Rosenwasser L J
Published: December 2022

The pathogenetic mechanisms that underlie hepatic fibrosis in schistosomiasis are unknown, but may be under the regulation of molecules secreted by the hepatic granulomas that encase the helminth eggs. Previous studies in mice demonstrated that isolated Schistosoma mansoni egg granulomas can elaborate in vitro substances that stimulate fibroblast proliferation and collagen synthesis. The present study provides the initial characterization of the granuloma-derived factor(s). Serum-free cell culture supernatants from isolated granulomas contained activity that stimulated the uptake of (3H)-thymidine by quiescent human dermal fibroblasts. This activity was present in fractions (Sephadex G-200 chromatography) with estimated m.w. of 30,000 to 40,000. Activity eluted with linear salt gradients from ion exchange columns (DEAE Sephadex) with 0.4 to 0.7 and 1.5 to 1.8 M NaCl (pH 8.4). Activity was present in fractions with approximate pI of 6 to 6.5 and 4, prepared by flat-bed isoelectrofocusing of crude supernatant in granular gel. Activity that stimulated thymocyte proliferation in an interleukin 1 (IL 1) assay was present in crude granuloma supernatants as well as in the partially purified fractions that contained fibroblast-stimulating activity. We conclude that the granuloma-derived fibroblast-stimulating factor has biologic activity similar to IL 1 but on the basis of m.w. and pI determination is distinct from IL 1.
Analysis and preliminary purification of glycoproteins isolated from eggs in the urine of patients with Schistosoma haematobium infection.
Authors: Hamburger J , Lustigman S , Siongok T K , Ouma J H , Mahmoud A A
Published: December 2022

Preliminary analysis and purification of glycoproteins from Schistosoma haematobium eggs were carried out with a small quantity of antigenic material obtained from the urine of infected human patients. A soluble egg extract was 125I-labeled and was fractionated by lectin affinity chromatography for separating egg glycoproteins. The crude glycoprotein fraction was run on SDS-PAGE to yield three polydisperse peaks with Rf values of 0.31, 0.57, and 0.84. 125I-labeled egg glycoproteins were further fractionated by ion exchange chromatography to yield four peaks or shoulders. One of these peaks constituted the major labeled egg glycoprotein of S. haematobium (MEGL-H) in a relatively pure form as determined by SDS-PAGE, and its estimated m.w. was 70,000. This glycoprotein was consistently and highly reactive serologically with a serum pool from schistosomiasis haematobia patients by a Farr-type radioimmunoassay (RIA). A limited cross-specificity study of inhibition RIA indicated that S. mansoni eggs contain components that cross-react only partially with MEGL-H. These results focus attention on MEGL-H as a potential serodiagnostic probe.
Characterization of HNK-1+ (Leu-7) human lymphocytes. I. Two distinct phenotypes of human NK cells with different cytotoxic capability.
Authors: Abo T , Cooper M D , Balch C M
Published: December 2022

Human lymphocytes with NK and K cell activities can be identified by the HNK-1 (Leu-7) monoclonal antibody. In these experiments, subsets of HNK-1+ cells from blood and bone marrow were distinguished by their expression of other cell surface antigens, their morphology, and their NK functional capability. Two-color immunofluorescence analysis revealed that subpopulations of HNK-1+ cells in blood expressed antigens found on mature T cells (e.g., T1, T3, T4, T8), but none expressed antigens characteristic of immature T cells (T6, T9). The majority of HNK-1+ cells (greater than 60%) also expressed a myeloid antigen (M1), whereas a minority (less than 25%) expressed HLA-DR. HNK-1+ cells were separated into T3- and T3+ subsets with the fluorescence-activated cell sorter and analyzed for their morphology and NK cell function. HNK+T3- cells exhibited a high level of NK activity against K562 target cells and contained many cytoplasmic granules. On the other hand, HNK+T3+ cells had low NK activity and a paucity of cytoplasmic granules. The cell sizes of HNK+T3- and HNK+T3+ cells were indistinguishable by light-scatter analysis. When these cell fractions were analyzed further, a reciprocal relationship between T3 and M1 antigen expression was observed. These results thus delineate two distinct subsets of human HNK-1+ cells in blood with different cytotoxic capability: HNK+T3-M1+ and HNK+T3+M1- cells. Analysis of bone marrow cells demonstrated that only 0.7% of the nucleated cells expressed the HNK-1 antigen; virtually all of these cells expressed both the T3 and T8 antigens but lacked the M1 antigen. Thus, a majority of HNK-1+ cell population in blood were T3-M1+, whereas almost all bone marrow HNK-1+ cells were T3+M1-. We propose that these subsets of cells represent different stages in NK cells differentiation.
Studies of the induction of anti-DNA in normal mice.
Authors: Smith H R , Green D R , Raveche E S , Smathers P A , Gershon R K , Steinberg A D
Published: December 2022

Injection of PBA has previously been demonstrated to induce anti-DNA. In the present study, we found that the combination of neonatal thymectomy and chronic administration of PBA (LPS + poly rI . rC) led to significantly higher anti-DNA levels than either PBA or thymectomy separately. These results suggested that a thymic regulatory process normally serves to suppress anti-DNA after chronic PBA exposure. Indeed, antigen-nonspecific suppressor function was found to be deficient in such thymectomy + PBA-treated mice. In addition, the cells of such mice in vitro interfered with the development of normal suppressor function by control cells.
Alloreactive cloned T cell lines. VI. Multiple lymphokine activities secreted by helper and cytolytic cloned T lymphocytes.
Authors: Prystowsky M B , Ely J M , Beller D I , Eisenberg L , Goldman J , Goldman M , Goldwasser E , Ihle J , Quintans J , Remold H , Vogel S N , Fitch F W
Published: December 2022

Culture supernatants generated by alloantigenic or lectin stimulation of a cloned helper T lymphocyte, designated L2, contain interleukin 2 (IL 2), granulocyte/macrophage colony-stimulating factor (CSF), B cell stimulating factor (BCSF), macrophage (Ia+)-recruiting factor (MIRF), (Ia+)-inducing activity, gamma-interferon, Fc receptor-enhancing activity, macrophage migration inhibitory factor (MIF), macrophage activation factor (MAF), interleukin 3 (IL 3), and a factor responsible for prolonging the synthesis and secretion of the fourth and second components of complement by guinea pig peritoneal macrophages. Erythropoietin was not detected. A spontaneously arising variant of L2, designated L2V, produces much lower quantities of macrophage-stimulating activities, IL 2, and interferon. However, when compared to L2, L2V produces much higher levels of BCSF, equivalent amounts of IL 3, and slightly smaller amounts of CSF. Unlike L2V, a cytolytic clone, designated L3, secretes lymphokines that primarily affect macrophage function. The time course of lymphokine production by L2 cells indicates that for the six lymphokine activities studied there are three different times at which maximal or near maximal levels are reached, as follows: 1) IL 2, 12 to 24 hr; 2) IL 3 and CSF, 24 to 48 hr; and 3) (Ia+)-inducing activity, MAF, and interferon, 48 hr or later. Only IL 2 activity disappears during the 8-day culture cycle. The time course data and the differential production of activities by the three types of lymphocyte clones suggest that at least four terminal effector lymphokine molecules account for the ten biologic activities tested.
Replicative properties of lipopolysaccharide-induced murine B lymphocytes: detection of three different responses to mitogenic restimulation.
Authors: Norin A J , Short M K
Published: December 2022

A number of investigators have reached different conclusions regarding the proliferative behavior of murine B lymphocytes stimulated by lipopolysaccharide (LPS). We examined the replicative properties of LPS-stimulated splenic B cells in primary culture using a density transfer technique to detect DNA replication. The method selectively monitors those lymphocytes that synthesized DNA in a first division and determines whether their progeny undergo subsequent replication, providing information about the growth characteristics of the first generation of LPS-responsive cels. Three different proliferative responses were observed. In one type, the initial signal that stimulated parental cells to divide was perpetuated in their progeny in that daughter cell proliferation occurred in the absence of mitogen (19 to 37% of the parental cells, mode I). In another type of response, the proliferation of daughter cells depended on further exposure to mitogen (10 to 17% of the parental cells, mode II). In a third response, proliferation of daughter cells did not occur even in the presence of mitogen (54 to 64% of the parental cells, mode III). Three types of growth responses were also observed for Con A-stimulated T cells. A larger proportion of T cells, however, were restimulated to an additional round of DNA replication (40 to 55% of the cells, mode II) when compared to LPS-stimulated B cells. These in vitro growth responses suggest alternate pathways by which antigens may regulate immune reactions.
Interleukin-2 does not induce murine B cells to secrete Ig.
Authors: Pure E , Isakson P C , Paetkau V , Caplan B , Vitetta E S , Krammer P H
Published: December 2022

We previously defined two T cell-derived lymphokines, BDCF mu and BCDF gama, which induce B cell differentiation in the absence of IL 2. Although these studies indicated that IL 2 was not required to induce Ig secretion, the effect of IL 2 on B cell differentiation was not established. In the present studies we have assessed the ability of purified IL 2 to induce IgM or IgG secretion. Our results demonstrate that IL 2 does not induce IgM secretion by BCL1 tumor cells, nor does it enhance BCDF mu-induced IgM secretion by these cells. Similarly, IL 2 had no effect on LPS or BCDF gamma-induced IgG secretion by normal B cells. Thus, we conclude that IL 2 does not act directly on B cells to induce differentiation.
Role of T cells and adherent cells in age-related decline in murine interleukin 2 production.
Authors: Chang M P , Makinodan T , Peterson W J , Strehler B L
Published: December 2022

Spleen and lymph node cells from young (3 to 4 months) and old (24 to 36 months) mice of two inbred lines and one hybrid stock were assessed for the effect of age on IL 2 production. The results revealed that the production of IL 2 by old spleen and lymph node cells is reduced to 42% that of young cells. Cell counting, IL 2 inhibition and absorption, and cell mixture studies were then performed to determine the basis for the reduced IL 2 production. Cell counting studies revealed that the reduced IL 2 production by old cells cannot be caused by a decrease with age in the total number of Lyt 1+ T helper cells or by an increase with age in Lyt 2+ T suppressor cells. IL 2 inhibition, IL 2 absorption, and young-old spleen cells mixture studies revealed that the decrease in the level of IL 2 activity with age is caused neither by the formation of excessive amounts of soluble IL 2 inhibitors, by the generation of excessive numbers of IL 2 responsive blast cells, nor by an increase in suppressor cells. Further cell mixture studies revealed a decrease in IL 2 production by young T cells in presence of old adherent cells, which did not approach that of the old T cell-old adherent cell mixture, and an increase in IL 2 production by old T cells in the presence of young adherent cells, which did not approach that of the young T cell-young adherent cell mixture. These results would indicate that the decline in activity of both the T cells and adherent cells involved in IL 2 production are responsible for the reduction. Finally, it was found that the reduced proliferative activity of Con A-stimulated old spleen cells can be enhanced to the same extent as that of young spleen cells by exposing them to exogenous IL 2.
Procedures for the purification of interleukin 3 to homogeneity.
Authors: Ihle J N , Keller J , Henderson L , Klein F , Palaszynski E
Published: December 2022

A procedure is described for the routine purification of IL 3 to homogeneity from WEHI-3-conditioned media. The techniques employed include ammonium sulfate fractionation, DEAE-cellulose, hydroxylapatite, and G-75 Sephadex column chromatography. The last step in purification involves chromatography on C18 hydrophobic supports in RP-HPLC systems, which results in the coelution of a protein peak and IL 3 activity. This purification sequence results in approximately a 1,000,000-fold purification from the initial starting material with yields of 5 to 10% of the initial activity. typically, 150 liters of conditioned media yields 2 to 10 micrograms of IL 3. The purified material was homogeneous by SDS-PAGE analysis and had an apparent m.w. of 28,000. Purified IL 3 had a specific activity of approximately 0.05 ng/unit of activity. Additional criteria used to establish the relationship of the 28,000-dalton protein to IL 3 include the ability of an antiserum against IL 3 to concomitantly immunoprecipitate the iodinated protein and to inhibit its biologic activity as well as the ability of the iodinated protein to bind specifically to cell lines known to require IL 3 for growth.
Allospecific human T cell lines and clones which mediate HLA-DR restricted helper activity.
Authors: Friedman S M , Thompson G S , Principato M A
Published: December 2022

In order to analyze the immunoregulatory activity of allospecific human T cells, we have combined the techniques of limiting dilution culture and IL-2 dependent T cell growth to generate cloned alloreactive T cell lines (TCL). These TCL have been expanded in continuous culture for greater than 8 mo with retention of stable phenotypic and functional characteristics. For example, phenotypic analysis of alloproliferative TCL, utilizing a panel of monoclonal antibodies, demonstrate that these cultures are comprised of T cells belonging exclusively to the T3+ T4+ T8- "helper" or "inducer" T cell subset. Coculture of cloned alloproliferative TCL cells with a panel of allogeneic stimulators reveals that these clones are specifically reactive against the HLA-DRw1 determinant. Of greater interest, coculture of selected alloproliferative TCL cells with DRw1+ but not DRw1- B cells results in a vigorous polyclonal response as measured by the reverse hemolytic plaque assay. Although major histocompatibility complex restriction of this T-B interaction operates at the inductive level, help is at least partially unrestricted at the effector level as alloantigen-activated TCL cells provide demonstrable, although not maximal, help for DRw1- B cells.
Human B cell differentiation. III. Enhancing effect of monoclonal anti-immunoglobulin D antibody on pokeweed mitogen-induced plasma cell differentiation.
Authors: Kuritani T , Cooper M D
Published: December 2022

The effects of monoclonal anti-delta antibodies on pokeweed mitogen (PWM) responses of blood mononuclear cells (MNC) were studied. Treatment with anti-delta antibody enhanced both B cell proliferation and plasma cell differentiation, which are T cell-dependent responses. The anti-delta enhancement of plasma cell differentiation, predominantly of IgM plasma cells, was surprising because PWM-responsive subpopulations of B cells have been shown to lack IgD and their plasma cell differentiation is easily and selectively suppressed by anti-mu, -gamma and -alpha antibodies. Treatment of MNC with monoclonal anti-delta antibody enhanced the number of IgM plasma cells induced by PWM stimulation by approximately threefold. The degree of enhancement was dependent upon the concentration of anti-delta antibody, and the F(ab')2 fragments were effective. Maximal enhancement was obtained either when MNC were preincubated with anti-delta antibody for 1 day before PWM stimulation or when anti-delta antibody was added with PWM at the beginning of 7-day cultures. Anti-delta antibody had little or no effect when added 1 to 3 days after the initiation of PWM stimulated cultures. Anti-delta treatment overnight induced a population of small IgM+IgD+ B cells to enlarge and converted them from poor to good PWM responders. The results are discussed in the context of a model which proposed that differentiation of both immature and preactivated mature IgD- cells can be inhibited by signals generated via surface immunoglobulin cross-linkage, whereas this stimulus enhances differentiation of the intermediate IgD+IgM+ B cells.
In vitro inhibition of pancreatic B cell function by lymphocytes from diabetics with associated autoimmune diseases: a T cell phenomenon.
Authors: Boitard C , Chatenoud L M , Debray-Sachs M
Published: December 2022

Lymphocytes from insulin-dependent diabetic patients were previously shown to suppress insulin release from mouse islet cells in vitro. Glucagon release was not suppressed. In order to further analyze this phenomenon, lymphocytes from three insulin-dependent diabetic patients with associated autoimmune diseases were treated by using the panning method for cell separation before testing on islet cell suspensions. The OKT3+, OKT3-, and OKT4- cell subsets were obtained. Insulin release was suppressed by the OKT3+ (T lymphocyte-enriched) subset, but not by the OKT3- (T lymphocyte-depleted) subset. These results suggest that T lymphocytes are directly involved in the suppression of insulin release in this model. Furthermore, the OKT4- (T helper-depleted) subset also suppressed insulin release.
Immune deficiency in the X-linked lymphoproliferative syndrome. I. Epstein-Barr virus-specific defects.
Authors: Harada S , Sakamoto K , Seeley J K , Lindsten T , Bechtold T , Yetz J , Rogers G , Pearson G , Purtilo D T
Published: December 2022

Eleven males with XLP were evaluated for EBV-specific antibodies during periods of 2 to 7 yr. Variable responses to EBV-specific antigens were found. All 11 patients had subnormal anti-EBNA titers, which probably reflected a T cell deficiency. The patients showed four different patterns in their anti-VCA response: 1) two boys who had experienced malignant lymphoma mounted no antibodies at all; 2) two patients showed intermittent anti-VCA titers; 3) four males had persistently elevated anti-VCA titers; and 4) three patients showed normal anti-VCA titers. ADCC against EBV-infected cells was abnormally low in six patients and was elevated in two patients given gamma-globulin. ADCC titers did not correlate with anti-VCA titers. However, most patients with XLP failed to effect regression of autologous EBV-infected lymphoblastoid cell lines, indicating a deficiency in long-lived T cell-mediated immunity to EBV.
Mouse monoclonal antibodies to chicken VH idiotypic determinants reactivity with B and T cells.
Authors: Chanh T C , Chen C L , Cooper M D
Published: December 2022

We have prepared mouse monoclonal antibodies against idiotypic (Id) determinants on chicken antibodies to N-acetylglucosamine (NAGA) and p-aminobenzoic acid (PABA) made by inbred line EL 6(3) birds. The monoclonal anti-NAGA Id antibody, termed CId-1, reacted with affinity purified antibodies to NAGA, but not with antibodies specific for PABA, arsanilic acid (Ars), phosphorylcholine (PC), or with normal chicken IgG and IgM. The monoclonal anti-PABA ID antibody, termed CId-2, reacted with anti-PABA antibodies and to a lesser extent with anti-Ars antibodies, but not with anti-NAGA, anti-PC, and normal IgG and IgM. The Id determinants were found among antibodies to NAGA and PABA made by outbred and inbred lines of White Leghorn chickens. The binding of the CId-1 and CId-2 antibodies to intact homologous anti-NAGA and anti-PABA antibodies, respectively, was not hapten-inhibitable in either case. Both anti-Id antibodies reacted specifically with isolated homologous heavy chains, suggesting VH Id specificities. The monoclonal CId-1 and CId-2 antibodies were reactive by immunofluorescence with approximately 0.9 and 0.2%, respectively, of the circulating lymphocytes and with approximately 0.4 and 0.15 of plasma cells. CId-1+ and CId-2+ bursal cells were first detected on the 16th and 14th days of incubation, respectively; both reached maximal frequencies by the 17th day of incubation. The CId-2 antibody reacted exclusively with immunoglobulin-positive cells. The CId-1 antibody also reacted with a subpopulation (0.4%) of immunoglobulin-negative lymphocytes from normal and agammaglobulinemic chickens, and thus would appear to recognize an idiotypic determinant expressed by certain clones of B and T cells.
Immunoelectron microscopic localization of idiotypes and allotypes on immunoglobulin molecules.
Authors: Roux K H , Metzger D W
Published: December 2022

We have developed a novel approach to the analysis of antigenic (allotypic and idiotypic) determinants on intact immunoglobulin molecules. Immune complexes composed of IgG in combination with anti-idiotype or anti-allotype antibody were "visualized" by transmission electron microscopy. Individual Fab fragments of anti-idiotype or anti-allotype antibody, when bound to the IgG, altered the "Y" configuration in a reproducible and interpretable manner. Anti-idiotype antibody (either as Fab or IgG) bound to the terminus of the presumed V region of the IgG molecule, thus extending the apparent length of the Fab arms. Analysis of a rabbit VH framework allotype (a1) revealed that the determinant(s) is (are) located on the lateral portion of the V region of IgG. Binding of the anti-a1 Fab fragments was always at approximately right angles to the axis of the Fab arms of IgG. Fab antibody to the rabbit kappa light chain (b4) allotype bound to the lateral portion of the terminal half of the IgG Fab arms. This technique should be of value in localizing less well defined immunoglobulin determinants.
Structure of the murine plasma cell alloantigen PC-1: comparison with the receptor for transferrin.
Authors: Goding J W , Shen F W
Published: December 2022

The plasma cell alloantigen PC-1 was isolated from C1.18 myeloma cells by immunoprecipitation and was analyzed by polyacrylamide gel electrophoresis. It was found to consist of two similar or identical disulfide-bonded polypeptide chains, each of Mr 115,000. The mobility of PC-1 in nonequilibrium pH gradient electrophoresis was similar to that of bovine serum albumin (pI 4.9). The PC-1 antigen is therefore similar to the transferrin receptor in Mr, charge, subunit composition, disulfide bonding, and developmental regulation. Similarities can also be detected by peptide mapping with subtilisin, but not with staphylococcal V8 protease. It is suggested that the PC-1 protein and the transferrin receptor may have had a common evolutionary origin, and may have similar functions.
Human lung mast cells: purification and characterization.
Authors: Schulman E S , MacGlashan D W , Peters S P , Schleimer R P , Newball H H , Lichtenstein L M
Published: December 2022

Detailed studies of the biochemistry and pharmacology of mast cell-mediated inflammatory disorders have been hampered by the inability to purify human mast cells. We now report techniques to purify human lung mast cells to apparent homogeneity. The major purification steps are: 1) dispersion of lung fragments into a single-cell suspension with enzyme combinations (pronase-chymopapain, collagenase-elastase); 2) partial purification by countercurrent centrifugation elutriation (CCE); and 3) affinity column chromatography. Enzymatic dispersion yielded suspensions with congruent to 10(6) mast cells per gram of lung parenchyma in purities of 1.2 to 9.7%. Dispersed mast cells responded comparably to those in parent lung fragments to challenge with anti-human IgG and pharmacologic agonists. Elutriation of lung cell suspensions yielded mast cell-enriched fractions with purities up to 70%. High purity mast cell fractions were combined, passively sensitized with purified human penicillin (BPO)-specific IgE, and purified by a BPO-affinity column chromatography procedure. Post elutriation mast cell purities of 29 +/- 3.5% were increased to 84 +/- 3% (range 65 to 98%) by the affinity column. Short-term (24 hr) culture of column-purified mast cells allowed adherence of non-mast cell contaminants to tissue culture plates, further increasing purity (up to 100%). Purified mast cells were intact and functional as assessed by dye exclusion, survival in short-term culture, IgE-mediated histamine release, and modulation of release by the pharmacologic agonists adenosine, IBMX, prostaglandin E2, and fenoterol.
The effect of hypophysectomy on thymic aging in mice.
Authors: Harrison D E , Archer J R , Astle C M
Published: December 2022

Surgical removal of the pituitary (hypophysectomy) followed by endocrine supplementation in middle-aged rats has been reported to reverse immunologic decline with age. We attempted to confirm and extend these reports by using a well-defined and readily available mouse model system. Hypophysectomy and endocrine supplementation in 8- to 9-mo-old C57BL/6J (B6) male retired breeder mice improved some, but not all, T dependent immune functions tested at 15 mo of age. In hypoxed mice, spleen cell proliferation in response to phytohemagglutinin (PHA) in vitro, and delayed type hypersensitivity (DTH) responses to sheep red blood cells (SRBC) measured by footpad swelling improved to levels shown by young controls. Direct anti-SRBC plaque-forming cell (PFC) responses by spleen cells, and serum agglutination responses against SRBC were not improved. Hypoxed mice had larger thymuses and much higher ratios of cortex-medulla areas than did age-matched controls. Hair regrowth after shaving was much faster in hypoxed mice. Nevertheless, hypophysectomy reduced mean and maximum longevities. These results conflict in several ways with the previously reported studies in rats, in which direct PFC responses and maximum longevities were improved by this treatment. There have been no previous studies of the effect of hypophysectomy and endocrine supplementation on thymic aging in mice, nor has it previously been reported that this treatment causes improvements in PHA and DTH responses and in thymic morphology. These effects show that at least some aging processes are reversible in aging individuals. They also suggest that hypophysectomy of middle-aged mice will be useful for studying neuroendocrine and thymic interactions that occur during the aging process.
Wheat germ agglutinin specifically inhibits formyl peptide-induced polymorphonuclear leukocyte chemotaxis.
Authors: Perez H D , Ong R , Khanna K , Banda D , Goldstein I M
Published: December 2022

Evidence that surface membrane glycoproteins of polymorphonuclear leukocytes (PMN) are involved in stimulus-response coupling prompted us to examine effects on these cells of various plant lectins. We have found that wheat germ agglutinin (WGA) (1.0 microgram/ml) completely, specifically, and irreversibly inhibits directed migration (chemotaxis) of human PMN toward the synthetic peptide, N-formylmethionyl-leucyl-phenylalanine (FMLP) (0.1 to 100 nM). This effect of WGA was not shared by subagglutinating concentrations of either concanavalin A or Bandeirea simplicifolia lectin. In contrast to its effects on FMLP-induced chemotaxis, WGA did not influence other FMLP-induced PMN responses (i.e., selective discharge of lysosomal enzymes from cytochalasin B-treated cells, generation of superoxide anion radical(s). WGA also did not influence PMN chemotactic responses to either the complement-derived peptide, C5a, or the lipoxygenase product, leukotriene B4. Inhibition of FMLP-induced chemotaxis by WGA was not reversed by washing WGA-treated cells, but was reversed (and prevented) by N-acetyl-D-glucosamine (not by N-acetyl-D-galactosamine or mannosamine). WGA did not affect either orientation or stimulated random motility of PMN, and did not interfere with specific binding to PMN of (3H)-FMLP. A derivative of WGA with 10-fold less agglutinating activity for human erythrocytes was prepared by treating the native lectin with cyanogen bromide and formic acid. The derivative also inhibited FMLP-induced PMN chemotaxis specifically and selectively. These data suggest that WGA specifically inhibits FMLP-induced PMN chemotaxis by attaching to N-acetyl-D-glucosamine residues at a locus on the PMN plasma membrane that is distinct from the binding site of the FMLP receptor.
Chemotactic factors trigger their own oxidative inactivation by human neutrophils.
Authors: Clark R A
Published: December 2022

Human neutrophils exposed to phagocytosable particles or soluble activating agents are able to inactivate chemotactic factors. The present experiments demonstrate that the peptide chemoattractant, f-met-leu-phe, serves as a sufficient secretory stimulus to trigger its own functional inactivation by neutrophils. This phenomenon is dependent on the concentration of the chemoattractant and the duration of its exposure to neutrophils. It requires low numbers of viable neutrophils and physiologic concentrations of chloride and is blocked by addition of azide, cyanide, catalase, or methionine, but not methionine sulfoxide, superoxide dismutase, or heated catalase. Neutrophils from patients with myeloperoxidase deficiency of chronic granulomatous disease are not able to inactivate the chemotactic factor unless purified myeloperoxidase or hydrogen peroxide, respectively, is added. Thus, chemotactic peptides can trigger neutrophils to secrete myeloperoxidase and hydrogen peroxide, which together with a halide destroy the biologic activity of the peptide, probably by oxidizing a methionine residue. These observations support the pathophysiologic significance of oxidative inactivation of humoral mediators as a negative feedback inflammatory control mechanism.
The Influence of Age on the Skin Sensitiveness of Tuberculous Guinea Pigs
Authors: Freund J.
Published: December 2022

Young experimentally infected guinea pigs show slight if any reaction to the injection into the dermis of a watery extract of B. tuberculosis despite the presence of gross lesions of tuberculosis in the regional lymph nodes and in the spleen. The adult mother pigs infected under the same conditions when injected with the same extract show at site of injection marked redness, edema and necrosis.
Prothymocytes in postirradiation regenerating rat thymuses: a model for studying early stages in T cell differentiation.
Authors: Castellanos R C , Leizerovitz R , Kaiser N , Galili N , Polliack A , Korkesh A , Galili U
Published: December 2022

Prothymocytes were obtained from regenerating thymuses of intrathymic-irradiated, bone marrow-shielded rats. In contrast to cortical thymocytes, which are small nondividing cells containing nuclear TdT, prothymocytes are characterized by their large size, high mitotic activity, lack of natural attachment, absence of PNA-binding capacity, nonexpression of membranal thymic specific antigens, and absence of nuclear TdT. In addition, these cells are capable of responding to the mitogens Con-A and PHA, and are sensitive to in vitro lysis by physiologic concentrations of corticosterone and cortisol. Prothymocytes incubated for 3 days on thymic monolayers differentiated into small lymphocytes expressing cortical thymocyte characteristics. Light and electron microscopy studies demonstrated the infiltration of prothymocytes from the circulation via the thymic blood vessel wall into the perivascular sinuses. Prothymocytes isolated from the thymuses, however, did not exhibit specific "homing" to the thymus when transfused back into the animals. In view of the observed accelerated thymic repopulation in adrenalectomized rats, and the high in vitro glucocorticoid sensitivity of the prothymocytes, it is suggested that thymic homeostasis is regulated by specific effect of adrenocortical hormones on the prothymocyte subset.
Direct demonstration of the human suppressor inducer subset by anti-T cell antibodies.
Authors: Morimoto C , Reinherz E L , Borel Y , Schlossman S F
Published: December 2022

Prior studies indicated that sera of patients with active juvenile rheumatoid arthritis (JRA) contain anti-T cell antibodies reactive with the T4+ inducer population. More important, depletion of this T cell subset with JRA anti-T cell antibodies (JRA+ T cells) and C abrogated T5/T8+ suppressor T cell function. In the present study, we utilized Ig-coated plate techniques and JRA anti-T cell antibodies to fractionate the T4+ population into T4+JRA+ and T4+JRA- subsets and characterize the individual T4+ inducer subset. It was shown that whereas only the T4+JRA- population responded maximally to the soluble antigens, TT and mumps, both T4+JRA+ and T4+JRA- subsets proliferated equally well to mitogens and alloantigens. Furthermore, B cell immunoglobulin production induced by T4+JRA- T cells was approximately twice that induced by the reciprocal T4+JRA+ subset. In contrast, the T4+JRA+ subset alone activated T8+ T cells to become suppressor effector cells. These results suggest that the T4+JRA+ subset is the inducer of suppressor subpopulation whereas the T4+JRA- subset functions maximally as the inducer of B cells. It is believed that the suppressor inducer population may have a central role in the immunoregulatory network in man.
Immunohistologic analysis of lymphoid infiltrates in primary Sjogren's syndrome using monoclonal antibodies.
Authors: Adamson T C , Fox R I , Frisman D M , Howell F V
Published: December 2022

The characterization of lymphocytes infiltrating salivary glands in patients with primary Sjogren's syndrome (1 degree SS) yields insights to disease pathogenesis that are not revealed by studies of the corresponding peripheral blood lymphocytes (PBL) alone. We analyzed salivary gland lymphocytes (SGL) and PBL in 14 patients with untreated 1 degree SS using monoclonal antibodies that detect T cells, T cell subsets, B cells, and antigens associated with lymphocyte activation. A four-step biotin-avidin immunoperoxidase technique was used for salivary gland frozen sections; cell suspensions and PBL were stained cytofluorographically. A predominance of T cells (Leu 1 = L17F12; Leu 4 = OKT3) was found in SGL (greater than 75%) and PBL (76 +/- 9%) with the majority belonging to the Leu 3a (OKT4) subset. A minority of B cells (anti-delta, -kappa, -lambda) was present in both SGL and PBL; however, a subset of B cells defined by monoclonal antibody B532 was present in SGL (5 to 20%) but was absent from PBL. An increased prevalence of activation antigens (Ia; OKT10) was found on SGL T cells (greater than 50% positive) compared to PBL T cells (less than 15% positive). These studies demonstrate that specific antigenic markers on lymphocytes at the site of inflammation in 1 degree SS differ significantly from those of the corresponding PBL. These differences emphasize that theories of disease pathogenesis of 1 degree SS must include studies on SGL.
In vivo interleukin 2 administration augments the generation of alloreactive cytolytic T lymphocytes and resident natural killer cells.
Authors: Hefeneider S H , Conlon P J , Henney C S , Gillis S
Published: December 2022

Interleukin 2 (IL 2) is a T cell growth factor that has been shown to modulate several in vitro immune responses. Produced by T cells, the lymphokine has been highly purified and used in a series of in vivo studies to examine the effect of IL 2 on murine cytotoxic T cell (CTL) and natural killer (NK) cell reactivity. By employing an immunization protocol known to generate CTL activity against allogeneic tumors, in vivo administration of highly purified IL 2, either in concert, with or 2 days after, tumor administration, resulted in an augmented CTL response as compared to effector cells harvested from untreated, alloimmunized control animals. Characterization of the effector cells responsible for the augmented cytolytic activity showed them to be of the T cell lineage and to be specific for the appropriate immunizing tumor cell. Furthermore, we were able to demonstrate that administration of purified IL 2 to naive, non-antigen-challenged recipients resulted in substantial potentiation of NK cell activity. The augmented cytolytic reactivity was mediated by NK cells as evidenced by effector cell lysis of NK-susceptible but not NK-insusceptible tumor targets. The results of these studies suggest IL 2 functions in vivo as an immune response regulator and may have a beneficial effect as an in vivo immunopotentiator.
Structural heterogeneity of the human Ia molecular pool as detected by cross-reacting mouse monoclonal antibodies.
Authors: Accolla R S , Pierres M
Published: December 2022

Three monoclonal antibodies (Mab), H39.49.5, H40.242.3, and H40.164.3, produced in an A.TH anti-A.TL system and reacting with determinants expressed either on I-Ak, on I-Ek, or on I-Ak and I-Ek molecules, respectively, have been used to analyze, at the structural level, the molecules of the human Ia pool against which they cross-react. The results indicate that the human Ia pool can be dissected out by the use of the A.TH anti-A.TL Mab in distinct families, which display structural variations not only among each other but also with respect to the previously defined NG1 and NG2 Ia subsets. In two distinct Ia pools analyzed (from Raji cells: HLA-DR 3,W6, and from LG-2 cells: HLA DR 1,1), the three A.TH anti-A.TL cross-reacting antibodies recognized molecules in which the allelic polymorphism was confined to the beta subunits. Moreover, within the same Ia pool, the alpha subunits as well as the beta subunits were shown to be different from each other. These results are discussed in the context of the present knowledge of the heterogeneity of the human Ia pool and of the possible similarity between gene products of the mouse H-2-I region and the human HLA-D counterpart.
Immunoregulatory factors derived from human tumors. II. Partial purification and further immunobiochemical characterization of a human sarcoma-derived immunosuppressive factor expressing HLA-DR and immunoglobulin-related determinants.
Authors: Roth J A , Osborne B A , Ames R S
Published: December 2022

An immunoregulatory factor (IRF) that suppresses Con A-mediated peripheral blood mononuclear cell (PBM) proliferative responses was partially purified by DEAE anion exchange chromatography and affinity chromatography from a 3 M KCl extract of a human liposarcoma. The factor (m.w. = 70K) co-purified with albumin, monitored by two-dimensional gel electrophoresis, and demonstrated a heterogeneous isoelectric point (pI 7.6-7.8). Xenoantisera produced against the DEAE-purified fraction and coupled to Affigel 10 removed suppressive activity that could subsequently be eluted by glycine-HCl, pH 3.5. An anti-albumin column partially removed activity, but the unbound 70K factor could still be detected in the column effluent. A xenoantiserum to this 70K effluent coupled to acrylamide beads completely removed the immunosuppressive activity in immunodepletion experiments. Further direct binding enzyme-linked immunoassays (ELISA) and solid-phase immunoabsorption experiments with monoclonal antibodies to human anti-HLA-DR framework determinants and a constant region of the IgM mu-chain demonstrated determinants on the 70K factor recognized by these antibodies.
Purification and biochemical characterization of human liver-derived inhibitory protein (LIP).
Authors: Grol M , Schumacher K
Published: December 2022

The liver-derived inhibitory protein (LIP), described by us in 1974 (1), has now been purified and biochemically characterized. LIP was purified 166-fold from liver extract (LE) by a combination of gel filtration and ion exchange chromatography, using AcA 44, DEAE-cellulose, hydroxylapatite and Sephadex G-150 superfine. The m.w. of purified LIP was determined by gel filtration to be 67,000 daltons and by SDS-PAGE to be 37,000 daltons. The pI was in the range of 8.83 to 8.96. The protein is temperature labile (60 to 70 degrees C) and acid labile (pH 4). The specific lymphocyte inhibitory activity of the purified protein is 0.15 microgram/culture. LIP is distinct from other liver-derived inhibitory proteins.
The interaction of human and rodent IgE with the human basophil IgE receptor.
Authors: Conrad D H , Wingard J R , Ishizaka T
Published: December 2022

The cross-reactivity of the human IgE receptor with mouse and rat IgE was studied. Using leukocytes from a patient with chronic myelogenous leukemia, in which the mononuclear fraction contained up to 75% basophils, both rat and mouse IgE were found to inhibit the binding of 125I-human IgE to the human basophilic leukemia (HBL cells). About 15-fold more rodent IgE was required for 50% inhibition of binding than unlabeled human IgE (hIgE). Dose-response studies using increasing amounts of rodent vs human 125I-IgE indicated that the HBL cells had about 8000 receptors per cell for hIgE and 5500 receptors per cell for rodent IgE. When the HBL cells were surface labeled with 125I and subsequently solubilized with non-ionic detergent, the labeled hIgE receptor could be isolated by either affinity chromatography on IgE-Sepharose (either human or rodent) or by immunoprecipitation with hIgE and anti-IgE. By SDS-PAGE on 10% gels, the receptor had a m.w. of 58,000 daltons. The solubilized receptors exhibited some rebinding to hIgE-Sepharose, and this rebinding could be inhibited by either human or rodent IgE but not by human IgG. Both the HBL cells and normal human basophils could be passively sensitized with murine IgE anti-DNP for antigen-induced histamine release. The minimum concentration of the mouse IgE antibody for sensitizing normal basophils was 20 to 200 ng/ml. Pretreatment of basophils with human IgE, but not human IgG, abrogated the capacity of the murine IgE antibody to sensitize the cells for histamine release, which indicated that the human and rodent IgE were interacting with the same receptor.