bluuebunny/crossref_metadata_2025 huggingface dataset:
Role of lipomodulin, a phospholipase inhibitory protein, in immunoregulation by thymocytes.
Authors: Hirata F , Iwata M
Published: December 2022

The treatment of murine thymocytes with anti-lipomodulin antibody during Con A stimulation causes selective loss of suppressor activity, but not of helper activity on PFC assay, when co-cultured with T cell-depleted spleen cells. Interaction of the antibody with responder cells in thymocyte culture were necessary in the early stage rather than in the later stage of lymphocyte activation by Con A, which suggests that anti-lipomodulin antibody acts in the stage of suppressor T cells generation. When thymocytes were cultured with purified lipomodulin for 48 hr, suppressor activity was induced. Lipomodulin as detected by radioimmunoassay was found to be released from T cells with the phenotype of I-J+, Lyt-1-, Lyt-2+. The immunoprecipitates from the media of Con A-stimulated thymocyte with anti-I-Kk antibody and anti-lipomodulin antibody were analyzed on SDS-gel electrophoresis. I-J products had m.w. 36,000 and 24,000, whereas lipomodulin had m.w. 36,000, 24,000, and 15,000. Because anti-I-Jk antibody could precipitate 125I-labeled lipomodulin purified from rabbit neutrophils, these results suggest that lipomodulin is a product of I-J genes that induces suppressor T cells.
Epitopes associated with a synthetic hepatitis B surface antigen peptide.
Authors: Ionescu-Matiu I , Kennedy R C , Sparrow J T , Culwell A R , Sanchez Y , Melnick J L , Dreesman G R
Published: December 2022

A synthetic peptide (SP1), corresponding to the amino acid residues 122 through 137 of the major polypeptide derived from hepatitis B surface antigen (HBsAg), subtype ayw, was analyzed for the presence of the major epitopes of HBsAg. Both a cyclic form, produced by introduction of an intrachain disulfide bond, and a linear form of the peptide were characterized. A panel of monoclonal antibodies with defined specificity for the cross-reactive group a antigenic determinant(s) and for the y and w subtype specificities was used for this analysis. The cyclic, but not the linear, form of SP1 reacted with five of 14 anti-a monoclonal antibodies, demonstrating that the cyclic peptide contains a conformation-dependent a epitope. Only one anti-a antibody was found to react with both cyclic and linear forms of SP1. Because SP1 failed to react with the remaining 8 anti-a monoclonal antibodies, it was concluded that the a antigenic reactivity associated with HBsAg contains an additional epitope(s) unrelated to that expressed on SP1. Both cyclic and linear SP1 reacted with three of three anti-y monoclonal antibodies, indicating that a sequential y epitope is also present on SP1; no w reactivity was detected. Analysis of the idiotypes associated with the monoclonal antibodies showed those that combined with cyclic SP1 also inhibited the binding of a common human anti-HBs (CHBs) idiotype with its rabbit anti-idiotype serum, whereas a monoclonal antibody that did not react with the cyclic SP1 epitope failed to inhibit the CHBs idiotype-anti-idiotype reaction. Thus, the conformational a epitope present on cyclic SP1 appears to contain the predominant epitope recognized by humans in response to a natural HBV infection.
Interleukin 2 dependence of human natural killer (NK) cell activity.
Authors: Domzig W , Stadler B M , Herberman R B
Published: December 2022

When purified populations of human natural killer (NK) cells were tested for cytotoxic activity in the presence of partially purified preparations of human interleukin 2 (IL 2), a definite, dose-dependent linear increase in reactivity was observed. To determine whether such augmentation by IL 2 might reflect an important aspect of the physiologic regulation of NK activity, we examined the effects of monoclonal antibodies against human IL 2 on spontaneous NK activity. The presence of such antibodies during the 4-hr cytotoxicity assay resulted in significant inhibition of NK activity, and when the NK cells were pretreated for 16 to 20 hr with anti-IL 2, little or no activity remained. These data suggest that the spontaneous cytotoxic activity of NK cells is dependent on their continued exposure to IL 2. The reduction in NK activity resulting from treatment with anti-IL 2 could be at least partially restored by exposure to only low amounts of partially purified IL 2. These data have provided the basis for formulating a novel model of NK cell activation.
Suppression of humoral immunity in chickens prevents transient paralysis caused by a herpesvirus.
Authors: Parker M A , Schierman L W
Published: December 2022

Marek's disease virus (MDV)3 is a highly oncogenic herpesvirus that usually causes visceral lymphomas and lymphoid infiltration of the peripheral nerves in chickens. A relatively rare encephalitic condition, first found in farm flocks and referred to as transient paralysis (TP), is also caused by MDV(1). TP symptoms occur 9 to 11 days after MDV inoculation and range from mild ataxia to profound coma. Most birds recover by 24 to 72 hr after onset of symptoms, although severely affected birds may die within the same time period. Previous studies in this laboratory (2) showed that susceptibility to TP is a recessive trait controlled by major histocompatibility complex (MHC) genes (i.e., B complex genes of chickens). Inbred line G-B1 chickens (B13/B13) are resistant to TP, whereas chickens from related inbred lines G-B2 (B6/B6) and G-B3 (B15/B15) are highly susceptible. In this study chickens were immunosuppressed by neonatal cyclophosphamide (CY) treatment or surgical bursectomy (BX) to determine the possible role of antibodies in the pathogenesis of TP.
Immunoregulatory T cell abnormalities in mucocutaneous lymph node syndrome.
Authors: Leung D Y , Chu E T , Wood N , Grady S , Meade R , Geha R S
Published: December 2022

We recently demonstrated that during the acute phase of mucocutaneous lymph node syndrome (MCLS)3 there was a significant reduction in circulating T8+ suppressor/cytotoxic T cells and an increased number of Ia/Dr-bearing T4+ T cells, which suggests the presence of circulating activated helper T cells (1). Furthermore, the vast majority of patients with acute MCLS had a significantly elevated number of circulating B cells spontaneously secreting IgG and IgM. In the present study, the possible role of the immunoregulatory T cell abnormalities in the polyclonal B cell activation was investigated by assaying the ability of T cells and T cell factors from patients with acute MCLS to induce immunoglobulin production by normal B lymphocytes. We also examined the capacity of normal T cells to suppress immunoglobulin production by activated B cells from patients with acute MCLS.
T cell-replacing activity of C8-derivatized guanine ribonucleosides.
Authors: Goodman M G , Weigle W O
Published: December 2022

The capacity of the C8-substituted guanine ribonucleosides to provide T cell-like signals to cultures of splenic B cells was evaluated. We showed previously that these low m.w. nucleoside derivatives traverse the cell membrane and induce their effects from an intracellular location. The current studies clearly demonstrate that 8 mercaptoguanosine (8MGuo), when added to cultures of B cells and macrophages in the presence of antigen, is capable of supplying a "second signal" for B cells, enabling them to generate high numbers of specific plaque-forming cells against the immunizing antigen. This effect is duplicated in cultures of spleen cells from congenitally athymic mice. Inhibition of interleukin 2 (IL 2) generation by cyclosporin A, such that the antibody response of normal spleen cells is entirely abrogated, has minimal effects on the T cell-replacing activity of 8MGuo. Additivity studies with MLC supernatants as well as kinetic analyses with IL 2-associated lymphokines substantiate that these factors act by a mechanism distinct from that of 8MGuo and 8BrGuo. These observations establish these nucleoside activators as exciting new probes for T helper cell activity and an effective non-T cell source of T cell-like signals.
Analysis of true anti-hapten cytotoxic clones in limit dilution microcultures after correction for "anti-self" activity: precursor frequencies, Ly-2 and Thy-1 phenotype, specificity, and statistical methods.
Authors: Good M F , Boyd A W , Nossal G J
Published: December 2022

By the use of split culture techniques we have been able to demonstrate conclusively that the "anti-self" activity and the spontaneous anti-hapten activity within an apparent anti-hapten cytotoxic T cell response is a clonal phenomenon and is not caused by cross-reactivity between anti-self and true anti-hapten clones. With the knowledge of this clonality we have been able formally to prove that the true hapten-generated, hapten-specific response can be obtained by subtracting the response generated by "self" but directed against modified targets from the response generated by modified self and directed against modified targets. Unbiased statistical estimators, which do not make assumptions about the origin (0 responder cells, 100% negative cultures), have been developed to plot accurately limit dilution data (maximal likelihood estimator and minimal chi-square estimator), and the analysis demonstrates that all the components of an apparent anti-hapten response obey zero order (single hit) kinetics. By specifically identifying true anti-hapten and true anti-self clones, we have been able to study the phenotype of their precursors as well as the effectors themselves at the clonal level. Precursors of true anti-hapten and anti-self clones are Thy-1+, Ly-2+. However, anti-hapten and anti-self effector cells show marked clonal variation with respect to Ly-2, as some clones are almost completely insensitive to anti-Ly-2 and complement whereas others show minimal to complete sensitivity. All anti-hapten clones are completely sensitive to anti-Thy-1 and complement, whereas about one-third of anti-self clones show only partial sensitivity, with the most lytic clones showing the most sensitivity. Hapten-specificity and anti-self-specificity have been examined clonally. Ten percent of anti-TNP clones recognize NIP, 10% of anti-NIP clones recognize TNP, and 20% of anti-self clones recognize an allo-target. These figures are in accordance with the overall specificity of effectors generated in bulk culture in the presence of CAS (concanavalin A-stimulated spleen cell conditioned medium). However, hapten specificity and H-2 restriction of bulk generated effectors are improved if CAS is omitted from cultures.
Ir gene control of immune responses to insulins. II. Phenotypic differences in T cell activity among nonresponder strains of mice.
Authors: Bucy R P , Kapp J A
Published: December 2022

Immune responses by mice to heterologous insulins are controlled by H-2 linked Ir genes. In studies to determine the mechanisms responsible for nonresponsiveness, we found that although pork insulin failed to stimulate antibody or proliferative responses in H-2b mice, it did prime T cells that can express helper activity in adoptive recipient mice. This helper activity was insulin-specific in both elicitation and expression. In studies presented in this paper, we have extended this analysis to the response patterns of helper T cells stimulated by sheep, horse, and rat insulins in mice bearing different H-2 haplotypes. The results demonstrate that nonresponder forms of insulin, including rat insulin, prime T cells in H-2b and H-2d, but not H-2k, mice. These results suggest that regulation of nonresponsiveness to insulin appears to be through different pathways in mice bearing different H-2 haplotypes.
Heterogeneity of an anti-H-2 I-A response as determined by cloned T cell reactivity.
Authors: Peck A B , Smith R T , Jadus M R
Published: December 2022

T lymphocytes activated against the I-Aq gene product in primary mixed leukocyte culture (MLC) exhibit an unusually strong cross-reactivity against all third-party stimulating strains tested in secondary MLC. Through the development of anti-I-Aq clones, this cross-recognition has been shown to result from the activation of a heterogeneous T lymphocyte population bearing receptors that recognize the I-Aq gene product differently, yet specifically. The determination of functional activities of the anti-I-Aq cloned cells, e.g., lymphokine production and ability to induce lethal graft-vs-host disease, suggested even further heterogeneity in T cells grouped according to their similar cross-reactive pattern.
The role of suppressor cells in the induction of murine photoallergic contact dermatitis and in its suppression by prior UVB irradiation.
Authors: Granstein R D , Morison W L , Kripke M L
Published: December 2022

Induction in mice of marked photoallergic contact dermatitis (PCD) to 3,3',4',5-tetrachlorosalicylanilide (TCSA) with UVA (320 to 400 nm) radiation requires pretreatment with cyclophosphamide (CY). Attempts to induce photoallergic contact dermatitis without CY result in only a small degree of sensitivity, accompanied by significant net splenic suppressor cell activity. These suppressor cells are antigen specific, inhibit the induction but not the elicitation of photoallergic contact dermatitis to TCSA, and are T lymphocytes. Exposure of mice to UVB (280 to 320 nm) radiation at a site distant from that of sensitization, before CY administration and sensitization, inhibits the development of photoallergic contact dermatitis. This is analogous to the suppression of allergic contact dermatitis (ACD) observed in mice after exposure to UVB radiation; such suppression is accompanied by the formation of antigen-specific splenic suppressor cells. However, in contrast to the findings with allergic contact dermatitis, splenic suppressor cells are not detected in mice that are treated with UVB radiation before CY administration and sensitization to TCSA. This is presumably because CY prevents their formation. This provides evidence that UVB-irradiated mice have a second form of anergy that is not mediated by suppressor cells.
Spontaneous in vitro occurrence and long-term culture of murine B lymphoblast cell lines.
Authors: Braun J
Published: December 2022

Under simple culture conditions, in vitro culture of T cell-depleted splenic mononuclear cells reliably promotes the outgrowth of B lymphoblasts. Such cells, termed BLY1, can be propagated as long-term B cell lines but are nontumorigenic in syngeneic recipients. The phenotype of BLy1 cells is unusual. They express certain B cell markers, including surface mu-lambda, I-Ak, and C3 and Fc receptors; however, delta heavy chain and kappa light chain production is absent. Although the cells do not express Thy-1 or Lyt-2, Lyt-1 is readily demonstrable. These distinctive features of BLy1 cells thus resemble a recently described B cell subset prominent early in ontogeny, and in the B cell defect of NZB mice.
Behaviorally conditioned suppression of the immune response by antilymphocyte serum.
Authors: Kusnecov A W , Sivyer M , King M G , Husband A J , Cripps A W , Clancy R L
Published: December 2022

Previous studies have shown that cyclophosphamide, a drug with a broad spectrum of cytotoxic activity and one that produces noxious gastrointestinal side effects, can elicit taste aversion conditioning when paired with a non-immunosuppressive oral stimulus (saccharin) resulting in immunosuppression after subsequent exposure to the paired stimulus (1). The study reported here indicates that rabbit anti-rat lymphocyte serum (ALS) which is selectively cytotoxic for lymphocytes and does not produce sensory side effects can similarly induce taste aversion conditioning of the immune response. Rats were exposed to oral saccharin paired with ALS injection. Upon subsequent reexposure to saccharin alone the immunosuppressive effects of ALS were reenlisted and the primary mixed lymphocyte culture response of conditioned rats to allogeneic lymphocytes was suppressed by 35% compared to controls. The demonstrated influence of psychologic factors on the immune response has far reaching implications, especially to human medicine, and their role in the course of disease and recovery in man demands further investigation.
Human natural killer cells analyzed by B73.1, a monoclonal antibody blocking Fc receptor functions. I. Characterization of the lymphocyte subset reactive with B73.1.
Authors: Perussia B , Starr S , Abraham S , Fanning V , Trinchieri G
Published: December 2022

We describe the production of the monoclonal antibody B73.1, reacting with a subset of human lymphocytes and, in about one-half of the donors, with neutrophilic polymorphonuclear leukocytes. In the peripheral blood from normal adult donors, 14.6 +/- 8.5% of the lymphocytes react with B73.1 antibody. The B73.1(+) lymphocyte subset does not bear markers of typical T or B cells and corresponds to the lymphocyte subset containing antibody-dependent killer (K) and natural killer (NK) cells. We demonstrate that: a) virtually all lymphocytes with K/NK cytotoxic activity are found in the lymphocyte subpopulation bearing the B73.1-defined antigen; b) the B73.1(+) lymphocyte subset bears the combination of antigens known to be present on K/NK cells; and c) there is a positive correlation between the level of cytotoxicity and the actual number of B73.1(+) lymphocytes in individual donors. We also report the distribution of B73.1(+) lymphocytes according to donor age and tissue types. The use of the B73.1 antibody in quantitating the actual number of K/NK cells and in performing functional studies on spontaneous cytotoxicity is discussed.
The fate of interleukin-2 after in vivo administration.
Authors: Donohue J H , Rosenberg S A
Published: December 2022

Interleukin 2 (IL 2) activity was measured in the serum of mice, using a bioassay, following various methods of IL 2 administration. IL 2 has a serum half-life of 3.7 min +/- 0.8 min (mean +/- SD) in mice after i.v. injection, and a serum IL 2 titer of 2 units/ml could be sustained for less than 30 min after injection of highly concentrated IL 2 solutions. Intraperitoneal (i.p.) and subcutaneous (s.c.) injection of similar amounts of IL 2 prolonged the duration of serum IL 2 activity at greater than 2 units/ml for 2 and 6 hr, respectively. Administration of IL 2 in a gelatin base was capable of sustaining serum IL 2 levels at greater than 2 units/ml for up to 16 hr. The reason for the short in vivo half-life of i.v. injected IL 2 was studied. No inhibitors of IL 2 could be detected in our cell growth assay of IL 2 activity. Exposure of IL 2 to mouse blood or serum had no impact on IL 2 titers. To evaluate the possibility that IL 2 was binding to lymphoid cells in vivo, the fate of IL 2 was studied in nude mice, in mice 4 days after 1000 rads total body irradiation, and in splenectomized mice. The serum half-life in these modified mice was identical to that in normal mice. The kidney appeared to be the main site of IL 2 clearance. The rates of IL 2 disappearance in mice rose from a control T 1/2 of 2.5 to 3.5 min in sham-operated animals to up to 84 min in mice with ligated renal pedicles. IL 2 was not excreted in an active form in the urine and ureteral ligation had only a small effect on the serum half-life of IL 2, implying probable renal tubular catabolism of filtered IL 2.
Positively selected Lyt-2+ and Lyt-2- mouse T lymphocytes are comparable, after Con A stimulation, in release of IL 2 and of lymphokines acting on B cells, macrophages, and mast cells, but differ in interferon production.
Authors: Guerne P A , Piguet P F , Vassalli P
Published: December 2022

Purified mouse T lymphocytes were separated into Lyt-2+ and Lyt-2- populations by the procedure of panning, in which a monoclonal rat anti-Lyt-2 antibody and dishes coated with affinity-purified mouse anti-rat Ig antibodies were used. The populations obtained were 95 to 99% pure as determined by immunofluorescence. Graded doses of these T cells were cultured with optimal mitogenic doses of concanavalin A and the 0 to 24 and 24 to 48-hr culture supernatants were collected. The dose-curve assays of the supernatants of Lyt-2+ and Lyt-2- cells showed comparable activity in interleukin 2 (IL 2) and T cell-replacing factor (TRF), assayed on antigen-stimulated culture of T-depleted spleen cells. Limiting dilution assays of IL 2-secreting precursor cells stimulated by Con A showed a high frequency of precursors in both populations, slightly higher among Lyt-2- cells. The supernatants also contained comparable levels of IPA (inducer of plasminogen activator production by the macrophages), MAF (macrophage-activating factor, assayed by induction of their cytolytic function), and MCGF (mast cells growth factor, assayed on a mast cell line). IPA and MAF were not produced with the same kinetics and in the same T cell concentration conditions as IL 2 and TRF. In contrast, interferon was principally produced by the Lyt-2+ cells.
Effect of interferon on thoracic duct lymphocyte output: induction with either poly I:poly C or vaccinia virus.
Authors: Korngold R , Blank K J , Murasko D M
Published: December 2022

Systemic inoculation of the interferon (IFN) inducer polyinosinic-polycytidylic polyribonucleotide (poly I:poly C) into CBA/J mice produces a significant decrease in the number of thoracic duct lymphocytes (TDL) collected 6 to 22 hr after injection. The number of TDL is restored to normal levels by 48 to 64 hr. Residual TDL collected during the inhibition period of poly I:poly C treatment showed a phenotypic profile similar to phosphate-buffered saline- (PBS) treated control groups when examined for Lyt-1.1, Lyt-2, Thy-1.2 and Iak surface markers. Interferon is implicated as the mediator of this phenomenon since: 1) Inoculation of exogenous IFN can induce a similar suppression in the number of recoverable TDL. 2) Pretreatment of mice with sheep anti-murine IFN serum can block this effect of poly I:poly C. Similar experiments with vaccinia virus demonstrate that although live virus can partially suppress the TDL output, almost complete suppression is achieved with ultraviolet (UV)-inactivated virus. Vaccinia virus-induced suppression of the number of TDL also appears to be caused by IFN since: 1) UV-inactivated virus induces significantly higher serum levels of IFN in comparison to live virus. 2) The suppression of TDL output by either live or UV-inactivated vaccinia virus can be blocked by pretreatment with anti-murine IFN. These findings suggest that the immunosuppression often associated with viral infections may be at least partially due to a decrease in lymphocyte recirculation mediated by the IFN initially released in response to the virus.
Assignment of antigenic determinants to separated I-A kappa chains.
Authors: Kupinski J M , Plunkett M L , Freed J H
Published: December 2022

The alpha- and beta-chains of the I-A kappa antigen from the AKTB-1b B cell lymphoma were separated by ion-exchange chromatography on CM-Sephadex in the presence of propionic acid and urea. Removal of the denaturants by dialysis produced isolated chains that regained a significant amount of their native configuration. These materials were used with a battery of monoclonal antibodies in a direct binding assay to localize specific alloantigenic determinants to the A alpha kappa or A beta kappa chains. This method allowed the assignment of the nominal specificity Ia. 17 and at least one epitope of the specificity Ia.2 to the A beta kappa chain. Finally, the I-A kappa antigen from the B cell lymphoma AKTB-1b was shown to be identical, by the criterion of tryptic peptide analysis, to that derived from normal B10.BR splenocytes. This constitutes the first demonstration that the polypeptide portion of a tumor-derived class II MHC antigen is identical to that derived from a normal tissue.
Monoclonal antibodies for carcinoembryonic antigen and related antigens as a model system: determination of affinities and specificities of monoclonal antibodies by using biotin-labeled antibodies and avidin as precipitating agent in a solution phase immunoassay.
Authors: Wagener C , Clark B R , Rickard K J , Shively J E
Published: December 2022

A general method is described for the determination of affinity constants and antigen cross-reactivities of monoclonal antibodies. The method employs biotin-labeled antibody, radiolabeled antigen, and avidin as a precipitating agent in a homogeneous phase, competitive radioimmunoassay. This method eliminates incomplete or variable precipitation of antigen-antibody complexes often encountered in immunoassays in which monoclonal antibodies are employed. Using this assay system, we were able to rapidly determine the affinity constants for a number of monoclonal antibodies elicited to carcinoembryonic antigen (CEA). In the preceding paper it was shown that five of the monoclonal antibodies recognized distinct epitopes on CEA. In antigen-binding experiments with these five monoclonal antibodies, the percent of radiolabeled CEA bound in antibody excess ranged from 30 to 92%. The CEA cross-reacting antigens, normal cross-reacting antigen (NCA), and tumor-extracted, CEA-related antigen (TEX) were significantly bound by one, and to a lesser degree, by two of the five antibodies. Two antibodies did not bind significant amounts of NCA or TEX. In inhibition studies, the amount of unlabeled CEA leading to 50% inhibition of 125I-labeled CEA-binding was in the range of 3.7 to 760 ng per tube. The amount of TEX showing the same degree of inhibition was 23-fold greater than the amount of CEA for two antibodies and 351-fold greater than the amount of CEA for a third antibody. The affinity constants for CEA were in the range of 1.0 x 10(8) to 5.1 x 10(10) M-1. The affinity constants for NCA and TEX, determined for one of the antibodies, were three orders of magnitude lower in comparison to CEA. The heterogeneity of radiolabeled CEA as indicated by the low fraction bound by one of the monoclonal antibodies is shown to be most probably an artifact resulting from radioiodination damage. The application of the approach described in this report should eliminate the problems most commonly encountered in the determination of affinity constants for monoclonal antibodies or the use of monoclonal antibodies in competitive, homogeneous-phase immunoassays.
Monoclonal antibodies for carcinoembryonic antigen and related antigens as a model system: a systematic approach for the determination of epitope specificities of monoclonal antibodies.
Authors: Wagener C , Yang Y H , Crawford F G , Shively J E
Published: December 2022

A systematic approach for the determination of epitope specificities of monoclonal antibodies to a complex antigen system is described. After initial screening to identify antigen-binding monoclonal antibodies, one or more of the clones are isolated by limiting dilution cloning, grown in ascites, and the resulting antibodies secreted into the ascitic fluid are affinity purified on Sepharose-bound protein A, radiolabeled, and cross-compared with antibodies from other clones by a solid-phase competitive immunoassay. In this work, BALB/c mice were immunized with either purified carcinoembryonic antigen (CEA) or the CEA-producing cell line HC 84S. Spleen cells were fused with the mouse myeloma cell line Sp2/0-Ag14. The supernatants from 25 hybrids showed a significant binding of 125I-CEA (greater than or equal to 15%). Nine hybrids were cloned, resulting in 33 different clones. The antibodies produced by the different cloned hybrids and the remaining uncloned hybrids recognized a total of five different epitopes on CEA. All of the epitopes reside on the protein moiety of the molecule as determined by antibody binding to deglycosylated CEA. The monoclonal antibodies with five different epitope specificities were reacted with tissue sections of normal and cancerous tissues and with peripheral blood smears. Each of the five monoclonal antibodies reacted with tissue sections from colonic, gastric, lung, and mammary carcinomas, as well as from a benign colonic polyp and a resection margin from a colonic carcinoma. Four monoclonals reacted with normal liver tissue. Granulocytes in peripheral blood smears bound three antibodies strongly and one antibody weakly, and one antibody was not bound. One monoclonal antibody that reacted with normal liver tissue was not bound by granulocytes. The ability of these five monoclonal antibodies to differentially detect three different CEA-related antigens in normal and malignant tissues may have clinical utility.
Use of monoclonal anti-gp70 antibodies to mimic the effects of the Rfv-3 gene in mice with Friend virus-induced leukemia.
Authors: Britt W J , Chesebro B
Published: December 2022

During the course of progressive Friend virus-induced leukemia in Rfv-3r/s mice, antiviral antibody caused a marked reduction in the frequency of leukemic spleen cells releasing infectious virus. We investigated the mechanism of this antibody-induced alteration of leukemia cell phenotype in a series of passive transfer experiments using monoclonal antiviral antibodies. Our results indicated that two IgG2a anti-gp70 cytotoxic antibodies could reduce the frequency of virus-producing cells within the leukemic spleen as well as maintain the virus-nonproducing phenotype once it was established. IgG2a and IgG2b monoclonal anti-p 15 antibodies, IgM, and IgA anti-gp70 antibodies, and an IgG3 anti-p 15(E) antibody were not effective. The mechanism of this phenotypic alteration appeared to involve an antibody-mediated cytostasis of virus-producing leukemia cells with the subsequent over-growth of virus-nonproducing cells. The maintenance of the virus-nonproducing phenotype was dependent on the presence of anti-gp70 antibodies capable of neutralizing and clearing infectious F-MuLV in vivo. The presence of these neutralizing antibodies appeared to prevent reinfection of virus-nonproducing cells and therefore interfered with reversion to the virus-producing phenotype.
Ten percent of normal B cells and plasma cells share A VH determinant(s) (J606-GAC) with a distinct subset of murine VHIII plasmacytomas.
Authors: Basta P , Kubagawa H , Kearney J F , Briles D E
Published: December 2022

Our findings indicate that a subset of VHIII antibodies, which we refer to as J606-GAC, contains a determinant(s) that is present on 5 to 15% of normal splenic B cells and plasma cells as detected by immunofluorescence. This subpopulation is detected by purified antibody, 0-1, which was prepared against a murine anti-group A carbohydrate (anti-GAC) hybridoma antibody. The J606-GAC subset includes the beta 2, 1 fructosan myelomas J606, EPC109, W3082, ABPC4, and UPC61, as well as 13 anti-GAC hybridomas. The 0-1 antiserum failed to react with hybridoma and myeloma immunoglobulins from murine VH groups I and II or other VHIII antibodies. By Western blot analysis, it was observed to react with isolated heavy, but not light, chains of J606-GAC-bearing antibodies. 0-1 failed to react with myelomas XRPC44 and J539, which have the same J region as J606 but a very different VH region. These observations indicate that 0-1 is detecting a VH region determinant. The J606-GAC marker recognized by 0-1 was expressed as early as 4 days after birth and was expressed at similar frequencies in germfree and conventional mice. Immunoprecipitation of both surface and biosynthetically labeled proteins from spleen cells or J606-GAC-positive hybridoma cell lines, respectively, confirmed that 0-1 was recognizing an immunoglobulin determinant.
Sezary syndrome. I. Specific skin-directed migration of peripheral blood lymphocytes.
Authors: Laroche L , Papiernik M , Bach J F
Published: December 2022

The in vitro migration of peripheral blood lymphocytes (PBL) from Sezary syndrome (SS) patients and from controls was studied using a migration assay under agar. Only PBL from SS patients migrate to normal allogeneic skin. This migration is organ specific. It requires the integrity of lymphocyte membrane receptors. Serum factors are not involved. Studies of the nature of these migrating lymphocytes using monoclonal antibodies suggest that they belong to a subset of helper T cells. These data indicate a functional impairment of SS lymphocytes, which enables them to respond to a skin-derived signal.
Proliferating and helper T lymphocytes display distinct fine specificities in response to human fibrinopeptide B.
Authors: Peterson L B , Wilner G D , Thomas D W
Published: December 2022

The fine specificity of T cell responses involved in the generation of help for antibody production and proliferation was examined by using the 14 amino acid peptide human fibrinopeptide B (hFPB, B beta 1-14) and its synthetic peptide homologues B beta 1-14(Lys14), B beta 1-13, and B beta 3-14. Peritoneal exudate or lymph node T cells from C57BL/10 and B10.BR mice immunized with hFPB or its synthetic homologues were used to measure in vitro proliferative responses. T cells from hFPB-immunized B10.BR mice showed specific proliferation to hFPB, but were unresponsive to B beta 1-14(Lys14), B beta 1-13, and B beta 3-14. B10.BR mice immunized with B beta 1-14(Lys14), B beta 1-13, or B beta 3-14 were unresponsive to all peptides tested. T cells from C57BL/10 mice showed no specific proliferation after immunization and challenge with any of the peptide antigens. In contrast to the patterns of T cell proliferation, immunization of both B10.BR and C57BL/10 mice with hFPB, B beta 1-14(Lys14), B beta 1-13, or B beta 3-14 primed for significant helper T cell activity, as assessed by the augmentation of a primary in vitro B cell IgM anti-FITC plaque-forming cell response after culture with B beta 1-1(Lys14)-FITC. Significant peptide-specific helper activity was observed when the FITC moiety was conjugated to the carboxyl terminal lysine (B beta 1-14(Lys14)-FITC) as well as FITC substitution at the amino terminus (FITC-B beta 3-13 or FITC-B beta 3-14). These results suggest that the fine specificity of T cell responses to peptide antigens are different for helper and proliferating T cells and that responsiveness by one T cell subpopulation does not predict the response pattern of other functional subpopulations.
Effect of rat alveolar lining material on macrophage receptors.
Authors: Coonrod J D , Yoneda K
Published: December 2022

Resident alveolar macrophages of many rat strains lack detectable C receptors and cannot carry out C-mediated bacterial attachment and phagocytosis. It has not been established whether this lack of functional receptors is innate or is acquired by contact with the alveolar microenvironment. In the present studies, peritoneal macrophages treated with lavage under conditions designed to maintain viability and adherence to glass showed loss of detectability of C and Fc receptors. Depending on the concentration of lavage and the time allowed for recovery, this effect appeared to be reversible. Continued incubation of peritoneal macrophages with lavage fluid led to loss of adherence to glass and cell death. Lavage-treated 51Cr-labeled macrophages rapidly released radiolabel. Although transmission electron micrographs of lavage-treated peritoneal macrophages appeared normal, micrographs prepared with freeze-fracture techniques revealed clumping and asymmetric distribution of the intramembrane particles. Similar membrane abnormalities were present in untreated resident alveolar macrophages. The anti-receptor effect of rat lavage was not species-specific and lung lavage fluid from dogs, rabbits, and mice affected receptor detectability on rat macrophages. The anti-receptor factor in rat lavage was localized to the surfactant-containing fraction and was stable on heating at 60 degrees C. It was resistant to trypsin and partitioned into chloroform in the Bligh-Dyer extraction procedure, suggesting that it was a lipid. Purified diacyl phospholipids had no effect on macrophage receptors, but purified lysophospholipids mimicked many of the effects of surfactant. We conclude that lipids in the alveolar lining material affect alveolar macrophage membranes and receptor function.
Experimental allergic orchitis: the isolation and partial characterization of an aspermatogenic polypeptide (AP3) with an apparent sequential disease-inducing determinant(s).
Authors: Teuscher C , Wild G C , Tung K S
Published: December 2022

An aspermatogenic polypeptide (AP3) capable of inducing experimental allergic orchitis (EAO) in the guinea pig (GP) was purified from GP testes by sequential delipidation, acid extraction, pH precipitation, ammonium sulfate fractionation, trichloroacetic acid precipitation, gel filtration on Sephadex G-75, preparative isoelectric focusing from pH 3-10 followed by isoelectric focusing from pH 7-10, gel filtration on Sephadex G-75 Superfine under reducing conditions, and reduced acid urea gel electrophoresis. Approximately 250 micrograms (BSA equivalents) of AP3 were obtained from 500 g wet weight of GP testes. On 15% reduced acid urea polyacrylamide gels, AP3 appeared as a single band with an Rf of 0.19. SDS-PAGE showed a single band with a mobility corresponding to a m.w. of 12,500 +/- 1500. The isoelectric point, determined during purification, was 9.90 +/- 0.50. Amino acid analysis of AP3 indicates it is a protein. Gas liquid chromatographic analysis failed to reveal the presence of either hexose or hexosamine, indicating that AP3 is probably not a glycopeptide. Two to 5.0 micrograms (BSA equivalents) of AP3 are capable of inducing severe EAO in 100% of GP tested; 1 to 2.0 micrograms (BSA equivalents) induced EAO in 60% of GP tested. Because AP3 appears to be nonglycosylated and the aspermatogenic activity of AP3 is highly resistant to various denaturing conditions including reduction and alkylation, the primary sequence of the polypeptide rather than higher ordered structure may be more important in defining the determinant(s) responsible for its aspermatogenic activity.
Macrophage procoagulant activity as a measure of cell-mediated immunity in the mouse.
Authors: Geczy C L , Farram E , Moon D K , Meyer P A , McKenzie I F
Published: December 2022

Thioglycollate-induced peritoneal exudate cells (TG-PEC) developed increased procoagulant activity after incubation with lymphokine and lipopolysaccharide (LPS). Dilutions of up to 1/1000 for insoluble Con A and 1/200 for periodate-induced lymphokine supernatants were active in enhancing macrophage procoagulant activity (MPCA), which was detected after a 2-hr incubation period and steadily increased over 20 hr. MPCA could also be induced by antigen; peritoneal cells from sensitized B6AF1 mice with strong footpad reactions to ovalbumin (OVA) responded to as little as 0.1 microgram/ml OVA in the MPCA test in an antigen-specific manner. By contrast, PEC from sensitized CBA/J mice that gave poor in vivo responses to OVA only reacted with high concentrations of the antigen in vitro. Production of the lymphokine responsible for induction of MPCA required an Ly-1+2- T cell, a nylon wool-adherent cell, and an la-17-bearing adherent cell. The MPCA induced by lymphokine or LPS did not appear to be a serine esterase and was not inhibited by phospholipase C. Coagulation of human factor-deficient plasma with activated TG-PEC indicated a requirement for Factor X.
Suppression of ACTH-induced steroidogenesis by supernatants from LPS-treated peritoneal exudate macrophages.
Authors: Mathison J C , Schreiber R D , La Forest A C , Ulevitch R J
Published: December 2022

The results from a number of clinical and experimental studies have suggested that during endotoxemia, suppression of adrenocortical steroidogenesis may occur. We have examined the possibility that macrophages are the source of a factor that suppresses adrenocortical steroidogenesis. Resident and peptone-elicited peritoneal exudate macrophages (PEM) from C3HeB/FeJ mice were incubated for 4 hr at 37 degrees C in the presence or absence of T cell hybridoma-derived lymphokine (LK) that contained high concentrations of MAF activity (assessed by induction of nonspecific tumoricidal activity in PEM). The LK was removed by rinsing, and fresh medium was added, followed by Salmonella minnesota R595 LPS (final concentration 10 micrograms/ml). After 18 hr at 37 degrees C the PEM supernatants and control medium from flasks without cells were harvested and stored at -20 degrees C. Explanted rabbit adrenocortical cells in 96-well plates were exposed to 30 microliters of PEM supernatant or control medium and ACTH (10 or 100 mU/ml) in a final volume of 120 microliters for 3 consecutive days. The adrenocortical cell supernatants were harvested each day, followed by replenishment of medium, PEM supernatant, and ACTH. Fluorogenic steroid production in wells that received control medium or supernatants from PEM not treated with LPS was normal (0.22 microgram +/- 0.010 (SD) per 5 X 10(4) cells). However, as much as 75 to 95% suppression of steroidogenesis was observed in wells that received supernatants from PEM treated with LK and LPS, compared to 40% suppression in wells that received supernatant from PEM treated with LPS alone. Continued exposure (over 3 days) of adrenocortical cells to supernatants from LPS-treated PEM resulted in progressively decreasing response to ACTH. Comparable suppressive activity was observed in supernatants from LPS-treated bone marrow-derived macrophages. In further experiments, suppression was observed in wells that were pretreated (22 hr) with the appropriate PEM supernatant, and evidence was obtained that the suppressive activity was not due to carry-over LPS. Finally, results from control experiments demonstrated that suppressive PEM supernatants neither inactivate ACTH nor interfere with the assay of fluorogenic steroids. Thus, these results suggest that during endotoxemia, products from LPS-stimulated macrophages may suppress adrenocortical function.
Surface-dependent modulation by H of C5 cleavage by the cell-bound alternative pathway C5 convertase of human complement.
Authors: Fischer E , Kazatchkine M D
Published: December 2022

Regulation by H of formation of the C3 and C5 alternative pathway convertases of complement on cells is dependent on such chemical characteristics of the cell surfaces as their membrane content in sialic acid. Properdin-stabilized C5 convertase sites were assembled on the non-activating cells of the alternative pathway, sheep erythrocytes (Es), and on the activating cells, desialated Es and rabbit erythrocytes (Er). C5 hemolytic sites were revealed by incubation of the convertase-bearing cells with limiting C5 and excess C6-C9. H inhibited generation of C5 hemolytic sites in a dose-related fashion on Es, Er, and desialated Es at molar ratios of H/C5 of 0.03 to 0.5. H similarly inhibited C5 utilization by the cell-bound C5 convertase on Es and desialated Es regardless of the cell membrane sialic acid content; however, H was three to five times less effective on Er. Kinetic experiments also suggested that C5 hemolytic sites are generated more rapidly on Er than on Es and desialated Es. The inhibition effect of H was independent of the number of C5 convertase sites per cell on all cell types; two to three times more residual hemolytic sites were found on convertase-bearing Es that had been incubated with C5 and H as compared with cells that had been decayed by H before incubation with C5. Furthermore, H also inhibited C5 interaction with a preformed classical pathway C5 convertase. These results suggest that H interacts with C5 so as to alter C5 binding and/or cleavage by the cell-bound C5 alternative pathway convertase. Sialic acid-independent modulation by H of C5 cleavage by the C5 convertase represents an additional regulatory step in the activation of the human alternative complement pathway.
Immune responses during human schistosomiasis mansoni. X. Production and standardization of an antigen-induced mitogenic activity by peripheral blood mononuclear cells from treated, but not active cases of schistosomiasis.
Authors: Gazzinelli G , Katz N , Rocha R S , Colley D G
Published: December 2022

Peripheral blood mononuclear cells (PBMN) from patients with active schistosomiasis mansoni are generally induced to proliferate upon exposure to a soluble worm antigenic preparation (SWAP). In contrast, only 25% of such patients responded to SWAP exposure by the production of detectable levels of a mitogenic factor (MF) activity capable of stimulating proliferation of resting cultures of allogeneic PBMN. When former schistosomal patients (chemotherapeutically cured 10 to 40 yr previously) were tested for these two responses to SWAP, all manifested blastogenesis; 80% also responded by the production of MF activity. Only 9% of control subjects with no history of schistosomiasis produced MF activity when exposed to SWAP. The production and assay of MF activity by former schistosomiasis patients was standardized. Reliable, optimal production was achieved by using 6 X 10(6) PBMN in 2-ml cultures upon exposure to 15 micrograms protein SWAP/ml. The cells were incubated for 20 hr in the presence or absence of SWAP, at which time they were washed and recultured in fresh medium for at least 28 more hours. Subject to subject variability occurred in the optimal time of detection of MF activity in culture supernatant fluids, but detection was assured between 24 and 48 hr after the wash. The assay system for MF detection required a 5 to 7-day culture. The consistent production of MF activity by former schistosomal patients emphasized the longevity of immunologic memory to SWAP, and may indicate a post-treatment decline of immunoregulatory mechanisms that are operative in patients with active schistosome infections.
Human monoclonal antibodies to a genus-specific chlamydial antigen, produced by EBV-transformed B cells.
Authors: Rosén A , Persson K , Klein G
Published: December 2022

Stable B cell lines producing human monoclonal antibodies to Chlamydia were established from salpingitis patients in the early convalescence phase. The antibody-producing cells were immortalized by Epstein Barr virus (EBV) transformation. Specific antibody-secreting clones were enriched by a stepwise microtiter plate cloning procedure. The selected B cell clones showed stable antibody production for more than 1 yr in continuous culture. Serologic specificity was demonstrated by micro-immunofluorescence (micro-IF) tests against a panel of Chlamydia reference strains. The antibodies were of the IgG1 subclass, and complement fixation could be demonstrated for one clone. There was no cross-reactivity against a large number of other bacteria. The monoclonal antibodies are directed against a common genus-specific surface antigen of the Chlamydia organism. Infected McCoy cells showed a brilliant, punctuated fluorescence surrounded by an inclusion membrane. Compared with conventional antisera, the monoclonal antibodies showed a clearer fluorescence pattern with very low background.
Characterization of an antigen expressed by human natural killer cells.
Authors: Griffin J D , Hercend T , Beveridge R , Schlossman S F
Published: December 2022

A monoclonal antibody, anti-N901, was produced by fusing NS-1 myeloma cells with spleen cells of a mouse immunized with human CML cells. This antibody was reactive with a subpopulation of peripheral blood LGL, including the natural killer cells. Monocytes, granulocytes, B cells, T cells (T3+ cells), erythrocytes, and platelets were nonreactive. The N901-positive cells in the peripheral blood were heterogeneous with respect to expression of other cell surface antigens. The majority of N901+ cells co-expressed T11, Mo1, and HNK-1, whereas a smaller percentage expressed T8. Ia, T3, T4, Mo2, or B1 antigens were very uncommon on N901+ cells. The heterogeneity of the N901+ LGL was further investigated by examining the expression of N901 antigen on a series of cloned normal human NK cell lines. N901 antigen was expressed by each of the NK cell lines tested, and by a minority of cloned T cell lines without NK activity. Anti-N901 does not block NK activity and can be used to rapidly purify functional NK cells for further study.