bluuebunny/crossref_metadata_2025 huggingface dataset:
Synergistic effects of active specific immunotherapy and chemotherapy in guinea pigs with disseminated cancer.
Authors: Key M E , Brandhorst J S , Hanna M G
Published: December 2022

Sewall Wright strain 2 guinea pigs bearing pulmonary metastases of the syngeneic line 10 (L10) hepatocarcinoma were treated with a vaccine composed of 10(7) bacillus Calmette-Guérin admixed with 10(7) x-irradiated L10 tumor cells beginning 10 days after tumor inoculation. Although this treatment failed to cure most of the guinea pigs of their metastatic disease, histologic examination of the pulmonary tumors in the vaccinated guinea pigs provided evidence of a cell-mediated hypersensitivity response that disrupted the normally compact architecture seen in control tumors. When a monoclonal antibody against the L10 tumor was injected i.v. to evaluate the vascular permeability of the tumors, significantly more antibody localized in tumors of vaccinated guinea pigs than in tumors of untreated controls. These results suggested that blood-borne substances could be delivered more efficiently to L10 metastases after the tumor-bearing guinea pigs had been treated with vaccine. To determine whether such increased vascular permeability would enhance the antitumor effects of chemotherapeutic agents, combined immunotherapy and chemotherapy studies were performed. Although cyclophosphamide treatment by itself did not cure L10-bearing guinea pigs, cyclophosphamide used in conjunction with prior immunotherapy increased the survival rate of animals to more than twice that of animals treated with immunotherapy alone (74 vs 33%). These results suggest that one mechanism by which active specific immunotherapy enhances chemotherapy of disseminated tumors is by rendering tumor foci more permeable to subsequently administered cytotoxic drugs.
Inhibition of human natural killer activity by lysosomotropic agents.
Authors: Verhoef J , Sharma S D
Published: December 2022

We have examined the effect of three lysosomotropic amines on human NK cell activity. Dansylcadaverine (DCA), diphenylamine (DPA), and lidocaine (LID) inhibited NK activity of nylon wool-purified and large granular lymphocyte (LGL)-enriched cell preparations. Cadaverine (CAD), an analog of DCA that does not affect lysosomal function, had no effect on NK activity. Binding of the K562 target cells to effector cells, as assessed in a single cell assay, was not inhibited by DCA, DPA, or LID. Cytotoxicity was inhibited by DCA and DPA only when these drugs were added within 5 min after the initiation of NK assays. In contrast, LID inhibited NK activity even when added 60 min after the addition of effector cells to target cells. All three amines that inhibited NK activity also reduced the intracellular concentration of the lysosomal enzyme beta-glucuronidase without affecting the activity of the cytoplasmic enzyme lactate dehydrogenase. Kinetic analysis revealed that LID inhibited both the maximum velocity (Vmax) of the cytotoxicity reaction as well as the affinity constant (Km); whereas DCA and DPA only inhibited Vmax.
Macrophage-dependent activation of antigen-specific T cells requires antigen and a soluble monokine.
Authors: DeFreitas E C , Chesnut R W , Grey H M , Chiller J M
Published: December 2022

Antigen-induced T cell proliferation requires T cell interaction with antigen in the context of MHC I region-compatible accessory cells. The resulting activation and proliferation of T cells involves the production and utilization of several lymphokines or interleukins. This report describes experiments wherein these events could be separated into two phases, T cell activation and T cell proliferation. The first phase was achieved by stimulating antigen-specific T cell lines with antigen-pulsed ultraviolet light-irradiated accessory cells. T cell proliferation (second phase) could then be initiated by the addition of a soluble lymphokine with the characteristics of interleukin 1 (IL 1). These effects were only observed with homologous antigen and accessory cells syngeneic to the T cells at the I-A and E/C subregion of the MHC. This report has two applications in the study of lymphocyte-lymphokine interactions. First, T cell recognition of antigen and antigen-induced T cell proliferation can be examined as physically separate events. Secondly, this system may be used as a specific and sensitive means of measuring the effects of IL 1 on T cells.
Ontogeny of human hematopoietic cells: analysis utilizing monoclonal antibodies.
Authors: Rosenthal P , Rimm I J , Umiel T , Griffin J D , Osathanondh R , Schlossman S F , Nadler L M
Published: December 2022

Lymphoid and myeloid cells isolated from second trimester fetal lymphoid organs were characterized by utilizing a panel of monoclonal antibodies that define human lineage-restricted, differentiation, histocompatibility, and activation antigens. At distinct gestational stages, the appearance of morphologically identifiable lymphoid and myeloid cells paralleled the appearance of cells expressing definable lymphoid and myeloid antigens. The proportion of cells in fetal liver, bone marrow, and spleen that expressed histocompatibility, myeloid, and B cell antigens increased with fetal maturation. In contrast, even the earliest fetal thymuses studied were of a phenotype no different than that seen during later stages of ontogeny. Although the cellular lineage of most fetal hematopoietic cells could be identified by this panel of reagents, a considerable number of fetal liver and bone marrow cells did not express any of these antigens, suggesting the possibility that they might represent early hematopoietic progenitor cells. These studies support the notion that the adult cellular phenotype is the result of both an orderly acquisition of differentiation antigens and the migration of these primitive cellular populations to specific fetal organs. Identification of hematopoietic progenitors in fetal tissues may facilitate the identification and isolation of early lymphoid and myeloid progenitor cells in adults.
A rabbit B cell determinant for a conserved portion of myelin basic protein, rabbit encephalitogenic sequence 65-74.
Authors: Lazarus K J , Hashim G A , Varitek V A , Paterson P Y , Day E D
Published: December 2022

Synthetic peptide S24 (TTHYGSLPQKG) represents residues 65-74 of myelin basic protein (MBP) and contains the major determinant involved in the development of experimental allergic encephalomyelitis (EAE) in rabbits. This peptide is completely conserved in all nonprimate mammals for which sequence information is available. Although it is clear that peptides containing the S24 region are capable of inducing EAE, previous serologic studies have resulted in the conclusion that the determinant is "buried" or sequestered in intact MBP. Employing a liquid phase radioimmunoassay, we studied Ab responses to the S24 determinant in six rabbits injected with rat myelin. Two of the six animals developed small but measurable responses to the S24 determinant. In one of these rabbits, the response was boosted with a covalent conjugate of S82 and methylated BSA (MBSA). We also measured antibodies to the S24 determinant in rabbit antisera to human, monkey, dog, bovine, and the large and small forms of rat MBP. By nonequilibrium inhibition analysis, we determined that the antibody responses to these antigens were all directed to a determinant composed of residues 66-71 of MBP, and that intact MBP inhibits the binding of these antibodies to radiolabeled S24. The results demonstrate that the rabbit encephalitogenic region of myelin basic protein is exposed in the intact molecule both as an immunogen and as a reactant in liquid-phase assays; furthermore, they demonstrate that MBP antigenicity leading to B cell responses does not necessarily depend on sequence differences between the injected protein and its counterpart in the host species. The latter finding reinforces the contention of Atassi that autoantibody responses are not exclusive to "evolutionary hypervariable locations."
Biologic properties of homogeneous interleukin 3. I. Demonstration of WEHI-3 growth factor activity, mast cell growth factor activity, p cell-stimulating factor activity, colony-stimulating factor activity, and histamine-producing cell-stimulating factor activity.
Authors: Ihle J N , Keller J , Oroszlan S , Henderson L E , Copeland T D , Fitch F , Prystowsky M B , Goldwasser E , Schrader J W , Palaszynski E , Dy M , Lebel B
Published: December 2022

Interleukin 3 (IL 3) was initially defined as a factor in conditioned media from concanavalin A-stimulated lymphocytes (Con A CM) that induces the enzyme 20-alpha-hydroxysteroid dehydrogenase (20 alpha SDH) in cultures of nu/nu splenic lymphocytes. To determine the spectrum of additional "biologic" activities, IL 3 was purified to homogeneity and its properties were assessed. The protein preparation was judged to be homogeneous IL 3 by the following criteria: 1) elution of a peak of IL 3 with a constant specific activity in the last step of purification, 2) presence of a single protein by SDS-PAGE analysis, 3) receptor-binding activity against IL 3-dependent cell lines, 4) a specific activity of congruent to 0.2 ng/ml required for 50% of maximal biologic activity, and 5) the presence of a single amino terminal sequence. With the use of this preparation of IL 3, the dose-response curves for 20 alpha SDH induction were identical or similar to the dose-response curves for the activities of 1) WEHI-3 growth factor, 2) mast cell growth factor, 3) P cell-stimulating factor, and 4) histamine-producing cell-stimulating factor. In addition, homogeneous IL 3 had colony-stimulating factor activity, although only approximately 2% of the total CSF activity found in Con A CM was associated with IL 3. The major peak of CSF activity could be resolved from IL 3 by DEAE column chromatography and lacked many of the biologic activities associated with IL 3.
Biology and biochemistry of T cell-derived lymphokines. I. The coordinate synthesis of interleukin 2 and colony-stimulating factors in a murine T cell lymphoma.
Authors: Watson J D
Published: December 2022

Two cloned murine T cell lymphomas of identical lineage exhibit coordinate control of the synthesis of two lymphokines. One line, LBRM-33 5A4, growing in culture can be activated by phytohemagglutinin (PHA) to secrete Interleukin 2 (IL 2), a T cell growth factor, and colony-stimulating factor (CSF), a granulocyte/macrophage growth factor. In the absence of PHA, 5A4 cells do not produce IL 2 or CSF. The second line, LBRM-33 1A5, is incapable of IL 2 and CSF production after treatment with PHA. Stimulation of 1A5 cells with Interleukin 1 (IL 1) converts these cells to a state in which subsequent treatment with PHA leads to the production of IL 2 and CSF. When the 5A4 cell line is recloned, variant subclones can be isolated that do not produce IL 2 or CSF upon stimulation with PHA. The frequency of occurrence of nonproducing variants from 5A4 cells is approximately 1 in 20. Such clones appear to retain a stable nonproducer phenotype. Treatment of a number of these variant subclones with IL 1 prior to exposure to PHA results in the secretion of both IL 2 and CSF from cells. Thus, cell lines derived from cloned LBRM-33 tumor cells either produce both IL 2 and CSF upon stimulation with PHA or they do not produce either lymphokine. These results indicate that the synthesis of IL 2 and CSF in T cells may share common regulatory mechanisms. Further, the LBRM cells appear to be capable of oscillation between two phenotypes representing different states of lymphokine-producing cells.
Macrophage activation: synergism between hybridoma MAF and poly(I). Poly(C) delivered by liposomes.
Authors: Pidgeon C , Schreiber R D , Schultz R M
Published: December 2022

High concentrations of a murine T cell hybridoma culture supernatant containing macrophage-activating factor (MAF) rendered resident mouse peritoneal macrophages cytotoxic for P815 mastocytoma cells. The capacity of the hybridoma-derived MAF (MAFH) to induce tumoricidal activity increased 10(3) to 10(4)-fold when the lymphokine was encapsulated into liposomes. Combinations of MAFH and poly(I) X poly(C) acted synergistically to render macrophages potently cytotoxic. Subthreshold (nonactivating) concentrations of free or liposome-encapsulated MAFH increased the potency of free poly(I) X poly(C) and liposome encapsulated poly(I) X poly(C). Either as free agent or encapsulated in liposomes, single-stranded poly(I) or poly(C) did not activate macrophages in the presence or absence of MAFH. Double-stranded poly(I) X poly(C) was thus required for macrophage activation and synergism with MAFH.
Frequency of responsiveness to the H-Y antigen among the B10.W lines.
Authors: Vucak I , Raska K , Nagy Z A , Klein J
Published: December 2022

B10.W females were immunized against syngeneic male cells (via the footpad and also i.p. in some strains) and their spleen cells were then restimulated in vitro and tested in the cell-mediated lympholysis assay for H-Y-specific killing of target cells. Only seven of the 33 tested lines were anti-H-Y responders. The effector cells obtained from each of the responder lines were then tested against male and female cells of other B10.W lines, as well as a number of classic B10 congenic lines, and the MHC molecules providing the context for H-Y recognition were identified. They were: Kk, Kw3, Kw7, Kw17, Kw27, Dk, and Dp. None of the strains generated effector cells capable of recognizing the H-Y antigen simultaneously in the context of the K and D molecules. The WOA1 females generated effector cells by using the Kw7 molecule for context of recognition, whereas the WR7 females produced cells recognizing the H-Y antigen exclusively in the context of the Dk molecule despite the fact that both lines share the Kw7 gene. Some of the effector cells cross-reacted with both male and female cells of other strains and this cross-reactivity could be attributed to the recognition of allogeneic MHC molecules controlled by K or D region genes. Interestingly, STA39 females generated Dp- but not Kw3-restricted anti-H-Y responses, whereas SAA48 females generated Kw3- but not Dw3-restricted responses; the Kw3-restricted cells cross-reacted with the Dp molecule. This cross-reaction might explain why the STA39 females do not mount a Kw3-restricted anti-H-Y response. Because the Kw3 + H-Y combination resembles Dp, the anti-Kw3 + H-Y T cells are functionally eliminated when tolerance of Dp molecules is attained in the STA39 mice.
Variation in antigenic determinants of p53 transformation-related protein obtained from various species.
Authors: Rotter V , Friedman H , Katz A , Zerivitz K , Wolf D
Published: December 2022

p53 is a cellular-encoded transformation-related protein. It is synthesized at elevated levels in tumor cells but has also been detected at low concentrations in several types of nontransformed cells. The p53 of tumor cells is immunogenic and elicits specific antibody production. The antigenic determinants of the p53 protein were studied by specific binding to anti-p53 monoclonal antibodies obtained from the RA3-2C2, PAb122, and PAb421 established hybridoma cell lines, and their conservation was followed in various animal species. We found that whereas mouse p53 efficiently immunoprecipitated with all three anti-p53 monoclonal antibodies, human and rat p53 bound PAb122 and PAb421 but lacked a determinant binding RA3-2C2. The hamster p53 molecule represented a third category, which immunoprecipitated with polyclonal anti-p53 antibodies but failed to bind all three monoclonal antibodies analyzed here. Using these monoclonal antibodies, we detected no variations between p53 found in transformed and p53 found in nontransformed cells, within a given species. The results also showed that RA3-2C2, which recognizes a mouse-specific determinant, binds a site located at a proteolytic digestion fragment of the p53 molecule that differs from that containing PAb122 and PAb421 recognition site(s). p53 is a single protein that can be immunoprecipitated through different antigenic determinants that vary between species.
C1q binding and C1 activation by various isolated cellular membranes.
Authors: Storrs S B , Kolb W P , Olson M S
Published: December 2022

Cellular and subcellular membranes obtained from heart, liver, and brain tissue from human, baboon, bovine, rabbit, and rat bound highly purified, radioiodinated human C1q with a high affinity (Ka = 10(8) to 10(10) M-1). The majority of these membrane preparations were able to activate fully assembled C1 as evidenced by the conversion of 125I-C1s, incorporated into C1 complexes, to 125I-C1s. C1 activation by baboon heart mitochondrial membranes required an intact C1 complex and appeared to be mediated by the binding of the C1q subcomponent in that excess C1q completely blocked C1 activation. Several experiments suggested that the heart mitochondrial membrane binding site for C1q is an integral component of the mitochondrial membrane and that C1q interacted with the membrane binding site through its globular head regions. It is suggested that the binding of C1q and the activation of C1 by cellular and subcellular membranes may be involved in the initiation and/or enhancement of the inflammatory process after acute tissue damage.
Idiotypic heterogeneity of the cross-reactive idiotype associated with the anti-p-azophenylarsonate antibodies of BALB/c mice.
Authors: Brown A R
Published: December 2022

The cross-reactive idiotype (CRI) associated with the BALB/c antibody response against the p-azophenylarsonate (Ar) hapten was studied by analyzing hybridoma anti-Ar derived from BALB/c mice. The BALB/c CRI (CRIc) was previously thought to be a single idiotype as defined by rabbit anti-idiotype. CRIc has been resolved into at least two separate families of anti-Ar antibodies that are unrelated idiotypically to each other. Analysis of the 5AF6 family revealed considerable idiotypic heterogeneity, including a number of public idiotopes that appeared to be primarily associated with combinatorial idiotopes (requiring H + L chains) or L chain-specific idiotopes.
Iodination and identification of surface membrane antigens in procyclic Trypanosoma rhodesiense.
Authors: Gardiner P R , Finerty J F , Dwyer D M
Published: December 2022

Surface membrane proteins and glycoproteins of procyclic Trypanosoma rhodesiense were labeled with 125I by the use of the insoluble catalyst Iodo-Gen. Autoradiography of whole solubilized procyclic trypanosomes after sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showed a minimum of 25 surface components to have incorporated radioactivity. These components ranged in m.w. from approximately 10,000 to approximately 285,000. Immunoprecipitation with rabbit antisera of Triton X-100 extracts of radiolabeled trypanosomes revealed a subset of at least 14 surface antigens. Two of these antigens (m.w. of 63,000 and 96,000) showed heavy incorporation of label and may be major proteins of the procyclic membrane. Sera from trypanosome-infected mice recognized an overlapping but different subset of surface antigens, including a doublet of very high m.w. Lectin precipitation using antilectin conjugates or bead-bound lectins indicated that many of the labeled surface components are glycoproteins including the two major proteins precipitated by rabbit antisera. Radiolabeled glycoproteins identified by these methods bear alpha-methyl-mannopyranoside and/or galactose residues but not N-acetyl glucosamine or fucose residues in quantity. The use of these methods in identifying potentially pathogenic trypanosomal antigens is suggested.
B cell genotype determines interaction preference with T cells: no effect of maturation environment.
Authors: Vainio O , Toivanen A
Published: December 2022

B cell development in the bursa of Fabricius of the chicken was examined. We constructed neonatal bursa cell chimeras (F1 leads to parent, parent leads to F1) and studied the in vivo interaction of these chimeric B cells with host-derived T cells in adoptive cell transfer to determine whether there exists any environmental effect on B cells for MHC-restricted T-B cell interaction. The results indicate that F1 B cells that have developed in a parental host bursa still behave as normal F1 B cells and do not show any change in their MHC-restriction pattern. In addition, parent leads to F1 chimeric B cells were indistinguishable from normal parental B cells. B cells from all constructed chimeras, including fully allogeneic, responded well to the T-independent antigen Brucella. We conclude that the genotype of the B cell, and not the developmental environment, determines the MHC restriction phenotype of mature B cells.
Cell-mediated lysis of tumor targets directed by murine monoclonal antibodies of IgM and all IgG isotypes.
Authors: Ralph P , Nakoinz I
Published: December 2022

Seven murine monoclonal antibodies to antigens expressed on T lymphoma targets were tested for directing antibody-dependent cellular cytotoxicity (ADCC). Peptone-induced peritoneal exudate macrophages, the LPS-stimulated RAW264.10 cell line, and human blood nonadherent mononuclear leukocytes were used as effector cells. All six IgG monoclonals tested, representing the four murine IgG isotypes and directed against four antigens (Thy-1.2, H-2k, Ly-2.1, Ly-9.2), were all active in ADCC. In contrast, an IgM anti-Thy-1.2 showed no activity despite very high C-cytotoxic titers. Thus, there does not seem to be any restriction among IgG classes for directing ADCC to tumor targets mediated by murine macrophages or human K cells.
Ontogeny of interleukin 2 responsive cells in the fetal thymus.
Authors: Bocchieri M H , Batuman O A , Hauptman S P , Zitron I M , Smith J B
Published: December 2022

The ontogeny of IL 2-responsive cells in the thymus of CBA/J mice was examined in neonatal animals and in fetuses at 14, 16, and 18 to 20 days gestation. The thymocytes were tested for responsiveness to 2 micrograms/ml Con A, TCGF, IL 2, and co-stimulation by Con A plus TCGF or IL 2. These responses were compared with those of thymocytes of 6- to 8-wk-old CBA/J. Thymocytes (1 X 10(5)) were cultured, and the reaction was measured at maximum response (96 hr). Neonatal animals gave an unusually high response to TCGF or partially purified IL 2 alone, approximately five times greater than the adult. A low but significantly enhanced proliferation, stimulated by partially purified IL 2 alone, was observed with 14-day fetal thymocytes, even though cultures of these cells in medium alone had higher background proliferation than any other age tested. In the co-stimulator reaction, proliferation significantly above background was measured at 16 days of gestation with Con A plus TCGF. The magnitude of the co-stimulator reaction increased with age, especially between the 16th and 18th day of gestation and immediately after birth.
Specific suppression of human T lymphocyte function by leukotriene B4.
Authors: Payan D G , Goetzl E J
Published: December 2022

Leukotriene B4, but not leukotrienes C4, D4, E4, or 12(S)-leukotriene B4, inhibited in a concentration-dependent manner the capacity of PHA-stimulated human T lymphocytes to generate LIF activity and to proliferate, as assessed by the uptake of [3H]thymidine.
Maturation of cytotoxic T cells within sponge matrix allografts.
Authors: Ascher N L , Chen S , Hoffman R A , Simmons R L
Published: December 2022

Two parallel events may provide cytotoxic effector cells at the site of allograft challenge: 1) the migration of effector cells to the graft, and 2) the local maturation of effector cells. Various experimental conditions were manipulated to ascertain the relative role of local maturation of precytotoxic cells at the site of alloantigen challenge. A C57BL/6 allogeneic-coated sponge matrix allograft was placed in a BALB/c animal. The entire animal with the sponge was subjected to sublethal irradiation, or alternatively, just the animal was subjected to the same dose of irradiation, and the sponge was shielded. The data demonstrate that by day 5, precytotoxic cells directed against donor alloantigen appears within the sponge matrix allograft. These precytotoxic cells are in themselves x-ray sensitive, but in the absence of irradiation and in the absence of further contributions from the host can develop into mature cytotoxic cells with specificity limited to that of donor alloantigen. The relative role of local maturation vs continued influx of mature cytotoxic cells cannot be determined from these sets of experiments. These data, however, show that there is potential for local development of mature cytotoxic cells in the absence of other host influences.
Characterization of human T lymphocytes that express the C3b receptor.
Authors: Wilson J G , Tedder T F , Fearon D T
Published: December 2022

The presence of the C3b receptor (C3bR) on human peripheral blood T lymphocytes was recognized by the capacity of rabbit F(ab')2 anti-C3bR and tetramethylrhodamine isothiocyanate (TRITC)-conjugated goat F(ab')2 anti-rabbit F(ab')2 to stain 14.5 +/- 3.7% (mean +/- SEM; n = 5) of lymphocytes forming rosettes with sheep erythrocytes (E). The F(ab')2 anti-C3bR also blocked the capacity of peripheral blood lymphocytes stained with OKT11 to form rosettes with bovine E bearing C3b and immunoprecipitated a single membrane protein having a m.w. of approximately 250,000 from detergent lysates of 125I-labeled, purified T cells. Measurement by fluorescent flow cytometry of the quantitative expression of the C3bR indicated that T cells had slightly more antigenic sites/cell than did E and approximately 10-fold fewer sites than were present on B cells. The surface constituents of the peripheral blood T cells expressing the C3bR were assessed in an assay that employed simultaneously three markers: rosette formation with sheep E, TRITC staining with anti-C3bR and fluorescein isothiocyanate (FITC)-staining with a panel of monoclonal antibodies or with aggregated IgG. Among lymphocytes forming rosettes with sheep E and expressing the C3bR, 99.6 +/- 0.4%, 65.0 +/- 5.8%, 17.2 +/- 6.2%, and 15.3 +/- 5.0% of the cells expressed antigens detected by OKT3, OKT4, OKT8, and OKM1 monoclonal antibodies, respectively. Ninety-seven per cent of the C3bR-bearing T cells were also capable of specifically binding aggregated IgG, indicating the presence of Fc receptors for IgG (Fc gamma R) on these cells. The T cells expressing the C3bR had large nuclei, thin rims of basophilic cytoplasm and no azurophilic granules. Thus, the C3bR is present on some T cells, all of which have a typical lymphocyte morphology, the T3 antigen and the Fc gamma R.
The internal image of IgG in cross-reactive anti-idiotypic antibodies against human rheumatoid factors.
Authors: Fong S , Gilbertson T A , Carson D A
Published: December 2022

A network of idiotypes and anti-idiotypes has been hypothesized to modulate antibody production against exogenous antigens. Idiotypic antigens on autoantibodies have been studied because of their potential use for specific immunomodulation. The present studies describe the preparation and characterization of rabbit anti-idiotypic antibody against human IgM anti-IgG autoantibodies (rheumatoid factors, RF) that bear the "internal image" of the human IgG-Fc fragment, and hence react specifically with the majority of RF from patients with rheumatoid arthritis. The anti-idiotype was isolated from rabbit anti-RF antisera by either immunodepletion of anti-immunoglobulin antibodies, or more simply by a single affinity purification step on a rabbit anti-human IgG Fc column. As measured by an enzyme-linked immunoassay, the anti-idiotype prepared by both methods bound to plates coated with purified IgM RF, but not to plates coated with non-RF IgM proteins. The anti-idiotype dose dependently blocked the binding to IgG of IgM-RF in 83% of sera from multiple patients with rheumatoid arthritis, Sjogren's syndrome, and macroglobulinemia. The anti-idiotype did not inhibit the activity of human IgM antibodies against DNP, tetanus toxoid, or thyroglobulin. The antigen recognized by the cross-reactive anti-idiotype was not apparently associated with a particular light or heavy chain amino acid sequence, but rather was intrinsic to most immunoglobulins with RF activity. Broadly cross-reactive anti-idiotypes with the "internal image" of IgG are simple to generate, and react with most RF. They may facilitate studies on the specific regulation of the human anti-IgG autoantibody response.
Modulation of surface antigens of a human monocyte cell line, U937, during incubation with T lymphocyte-conditioned medium: detection of T4 antigen and its presence on normal blood monocytes.
Authors: Moscicki R A , Amento E P , Krane S M , Kurnick J T , Colvin R B
Published: December 2022

The human monocyte line, U937, derived from an individual with histiocytic lymphoma, undergoes morphological and functional changes when incubated with medium conditioned by lectin-stimulated cloned human T lymphocytes. Using monoclonal antibodies and flow cytometry, we therefore analyzed alterations in surface components that might accompany these morphological changes, in comparison with components present on normal blood monocytes. The U937 cells possess three surface antigens in common with blood monocytes, detected with OKM1, 4F2, and anti-monocyte.2 (the last monocyte specific). DR antigen was not detectable on U937 cells with three anti-DR framework antibodies but was detected on blood monocytes. Unexpectedly, OKT4, a monoclonal antibody to T4 antigen previously believed to be restricted to helper T lymphocytes, also reacted with U937 cells. Six monoclonal antibodies to other epitopes on T4 also reacted with U937 cells. None of these could be inhibited by blocking of Fc receptors. T4 with its various epitopes were also expressed on normal human blood monocytes. Other lymphocyte surface markers (T3, T8, T6) and fibronectin were not detectable on U937 cells or monocytes. An individual, whose lymphocytes lacked the epitope detected with OKT4 but had epitopes detected with OKT4 A, B, C, and D, had monocytes with identical reactivity, evidence that the T4 on monocytes and lymphocytes are products of the same structural gene. Stimulation of U937 cells for 24 hours with supernatants from Con A-stimulated T lymphocyte clones caused an increase in expression of OKM1 and Fc receptor activity and a decrease in expression of T4, consistent with a more mature phenotype of blood monocytes. Although the function of the T4 molecule is unknown, it is notable that it is displayed by two cells of distinct lineage which interact in the response to soluble antigens.
A novel cell surface antigen (T305) found in increased frequency on acute leukemia cells and in autoimmune disease states.
Authors: Fox R I , Hueniken M , Fong S , Behar S , Royston I , Singhal S K , Thompson L
Published: December 2022

Monoclonal antibody T305, prepared by immunizing mice with the T-ALL derived cell line RPMI-8402, immunoprecipitates a single chain glycoprotein with m.w. 160,000 daltons (under reducing conditions) or 180,000 daltons (under nonreducing conditions). In immunofluorescence assays, antibody T305 reacted with a subpopulation of T cells in normal blood (22 +/- 6%), thymus (28 +/- 11%), and lymph node (24 +/- 6%). Increased frequency of T cells reactive with antibody T305 was found in peripheral blood of patients with infectious mononucleosis (greater than 80%), graft-vs-host disease after bone marrow transplantation (65 +/- 11%), acquired immunodeficiency syndrome (53 +/- 12%). The T cells in synovial fluid of patients with rheumatoid arthritis had increased frequency of antibody T305 reactive cells (59 +/- 8%) as compared to their peripheral blood (18 +/- 7%). Two color immunofluorescent studies demonstrated that the T305+ T cells predominantly co-stained with antibody Leu 2a (suppressor/cytotoxic subset) in both normals and disease state blood. After cell sorting to obtain T305+ and T305- subpopulations, we demonstrated that a) natural killer and antibody-dependent cellular cytotoxicity activity in normal blood was in the T305+ but not T305- T cells; b) cytotoxic T cells induced by mixed lymphocyte reaction were predominantly T305+; c) T305- T cells could be induced in vitro to express T305 antigen by mitogens or allogeneic B cells; d) the DNA content of T305+ and T305- T cells in normal blood was similar (greater 95% of cells with G0/G1 level); e) after mitogen stimulation, T305 antigen induction on previously T305- cells occurs before S-phase; and f) significantly more [3H]-thymidine after mitogen stimulation was incorporated by originally T305- cells than by originally T305+ cells (p less than 0.001). The T305 antigen was not restricted to T cells because it was also found on myeloid precursors in bone marrow but was not present on polymorphonuclear leukocytes, red blood cells, platelets, muscle, liver, skin, kidney, lung, or brain. Antibody T305 was found on 24/25 cases of acute leukemia (6 T-ALL, 10/11 cALL, 7 AML, and 1 AMOL) but not on 18 cases of chronic leukemia (B-CLL, T-CLL, null CLL, CML). The importance of the T305 antigen is that it is present on a high number of T cells in certain autoimmune diseases and on virtually all acute leukemia cells. Its distribution on immature and in vitro activated cells suggests that it may represent a receptor for signals related to cellular replication or differentiation.
The effects of interleukin 1 on human B cell activation and proliferation.
Authors: Falkoff R J , Muraguchi A , Hong J X , Butler J L , Dinarello C A , Fauci A S
Published: December 2022

The precise role of B cell surface immunoglobulin (slg) in the activation of B cells is unclear at present. In particular, it is uncertain whether ligands interacting with the B cell slg suffice to induce proliferation, or simply induce a state of activation in which the B cell becomes responsive to growth factors made by accessory cells. We have examined the effects of two ligands, Staphylococcus aureus Cowan strain I (SAC) and antihuman mu chain (anti-mu), which interact with B cell slg on highly purified human peripheral blood and tonsillar B cells cultured at low cell concentrations. The effects on B cell proliferation of these ligands alone or in combination with highly purified interleukin 1 (IL 1) or a supernatant of a human T-T hybridoma containing a B cell growth factor (BCGF) were studied. SAC with its high cell wall content of protein A triggered maximal B cell proliferation which was not increased further by IL 1 or BCGF. High concentrations of soluble F(ab')2 fragments of goat anti-mu chain also induced significant B cell proliferation. Lower concentrations of anti-mu resulted in little or no B cell proliferation but activated the B cell to a state of responsiveness to both IL 1 and BCGF. IL 1 by itself had no effect on the proliferation of unstimulated B cells or on the proliferation of in vivo-activated B cells which responded to BCGF in vitro, but demonstrated clear synergy with low concentrations of anti-mu antibody. BCGF alone augmented the proliferation of unstimulated B cells, presumably by acting on B cells which had undergone some degree of activation in vivo. In addition, it showed marked synergy with anti-mu antibody, which resulted in proliferation similar in magnitude to that induced by SAC. This synergy was far greater than that seen between anti-mu antibody and IL 1, and the resulting proliferative response was only slightly increased by the presence of IL 1. We conclude that the importance of accessory cell factors for the initial rounds of B cell proliferation depends on the strength of the initial slg-mediated activation signal. When this is strong, the response is maximal and independent of accessory cells or accessory cell factors. When it is suboptimal, a moderate synergy is seen with IL 1 and a dramatic synergy with BCGF.
Degeneracy of antibody specificity.
Authors: Sperling R , Francus T , Siskind G W
Published: December 2022

Evidence was obtained, by direct binding assays (radioimmunoassays) and by inhibition of binding assays, that after immunization some of the antibody molecules produced are degenerate in that they bind not only the immunizing antigen, but also unrelated ligands. It can be concluded that the exquisite specificity of the immune response is not necessarily a property of any given antibody molecule but is, at least to some extent, due to the summation of specificities held in common by the population of antibody molecules produced during the response.
Ultrastructural, biochemical, and functional characteristics of histamine-containing cells cloned from mouse bone marrow: tentative identification as mucosal mast cells.
Authors: Sredni B , Friedman M M , Bland C E , Metcalfe D D
Published: December 2022

Two histamine-containing granulated cell lines were cloned from mouse bone marrow by using sequential soft agar and limiting dilution techniques. Concanavalin A-stimulated mouse spleen cell supernatants were required for cell proliferation. Cloned cell lines were Lyt-1.2, Lyt-2.2, Thy-1.2, surface Ig negative, I-A positive, and failed to ingest latex particles. Cells expressed 6.4 to 9.0 X 10(4) IgE high affinity receptors and contained approximately 1 to 2 pg of histamine per cell. Electron microscopy revealed granule and surface membrane features characteristic of mast cells. The cloned histamine-containing granulated cell lines synthesized a proteoglycan containing glycosaminoglycan side chains with an estimated average m.w. of 40,000. The glycosaminoglycan side chains consisted only of chondroitin sulfates identified by charge characteristics and through the use of selective polysaccharidases. These cells degranulated to immunologic stimuli and the calcium ionophore A23187 but not to compound 48/80. Taken together, the above findings suggest that these histamine-containing granulated cell lines that are cloned from bone marrow are mucosal (atypical) mast cells.
Modulation of rat serosal mast cell biochemistry by in vivo dexamethasone administration.
Authors: Marquardt D L , Wasserman S I
Published: December 2022

To examine steroid-induced biochemical alterations in the mast cell secretory process, rats were injected with intramuscular dexamethasone or saline for 4 days, and serosal mast cells and lung tissue were obtained from each group. Radioligand binding studies utilizing 1-[propyl-1,2-3H]dihydroalprenolol (3H-DHA) demonstrated a 23.1 +/- 0.8% increase in rat lung beta-adrenergic receptors in steroid-treated rats, but the mast cell beta-adrenergic receptors were unaffected. Neither resting mast cell cyclic adenosine 3':5'-monophosphate (cAMP) levels nor the degree of cAMP augmentation induced by isoproterenol were changed by steroid administration. Mast cells from rats treated with dexamethasone released only 48.6 +/- 8.9 and 58.8 +/- 6.0% of the beta-hexosaminidase released from saline-treated rat mast cells when sensitized with anti-dinitrophenyl (DNP) IgE and challenged with DNP-bovine serum albumin antigen or the calcium ionophore A23187, respectively. [3H]serotonin release in cells from steroid-treated rats was 41.8 +/- 7.9 and 87.6 +/- 2.6% of control release stimulated by antigen or A23187, respectively. [14C]arachidonic acid incorporation into mast cell phospholipids followed by antigen or A23187 challenge revealed that cells from dexamethasone-treated rats release 61.3 +/- 15.6% and 62.1 +/- 11.8% of labeled metabolites, respectively, compared to controls. The addition of exogenous arachidonic acid 5 min prior to antigen challenge caused a similar decrease in mediator release in cells from saline- and steroid-treated rats (36.7 +/- 6.1 and 38.4 +/- 0.9%, respectively). When arachidonic acid was added to sensitized cells after specific antigen, no significant changes in beta-hexosaminidase release were noted in either group. Chronic in vivo dexamethasone administration markedly decreases mast cell mediator release without changing resting cAMP levels. The release of arachidonic acid metabolites is reduced in steroid-treated cells, possibly through the inhibition of phospholipases. Exogenous arachidonic acid cannot overcome this inhibition, suggesting that an earlier step in phospholipid metabolism, perhaps involving phospholipase C, may be important.
Resistance to macrophage-mediated killing as a factor influencing the pathogenesis of chronic cutaneous leishmaniasis.
Authors: Scott P , Sacks D , Sher A
Published: December 2022

Cutaneous leishmaniasis can be either a spontaneously healing or chronic disease, depending upon the strain of parasite and the immunological status of the host. We have investigated parasite factors responsible for the variable pathogenesis observed in leishmanial infections by testing the sensitivity of several leishmanial strains to intracellular killing in lymphokine (LK) activated mouse macrophages. Significant microbicidal activity against Leishmania tropica, a strain which heals in C57BL/6 (B6) mice, was found. In contrast, a strain (Maria) which has previously been shown to induce chronic nonhealing cutaneous lesions in B6 mice was resistant to killing in activated macrophages. This resistance to killing was observed in macrophages activated by LK obtained from either Bacille Calmette-Guérin-, L. tropica, or the Maria strain infected mice. The inability of LK activated macrophages to kill the Maria strain was shown not to be due to parasite induced inhibition of killing mechanisms, since Maria strain infected, LK treated macrophages exhibited tumoricidial activity similar to uninfected macrophages. Furthermore, LK activated macrophages simultaneously infected with the Maria strain and another intracellular pathogen, Toxoplasma gondii, killed Toxoplasma, but not the Maria strain. Temperature was also found to significantly influence the multiplication and killing of Leishmania parasites. As would be expected from their cutaneous nature, L. tropica and Maria strain parasites multiplied better at 35 degrees C than at 37 degrees C. Also consistent with the failure of cutaneous strains to visceralize in immunocompetent mice was the observation that the killing of leishmanial parasites was enhanced at the higher temperature. Thus, the temperature dependent growth capacity and sensitivity to killing of a given leishmanial strain in macrophages may be important factors influencing the pathogenesis of cutaneous leishmaniasis.
Immunoregulation in experimental disseminated histoplasmosis: flow microfluorometry (FMF) studies of the Thy and Lyt phenotypes of T lymphocytes from infected mice.
Authors: Watson S R , Miller T B , Redington T J , Bullock W E
Published: December 2022

Previous studies have shown that mice infected i.v. with 6 X 10(5) yeast phase Histoplasma capsulatum (Hc) develop suppressed immune responses during weeks 1 to 4 of infection but that by weeks 8 to 12 of infection these responses return to normal. In this study total and differential cell counts showed that as early as the third day of infection there was a marked reduction in the number of lymphocytes recovered from the peripheral blood, bone marrow, and thymus of infected animals. Concomitantly, there was an increase in the number of splenic lymphocytes. By day 28 both the total and differential cell counts were similar in both infected and normal animals. Flow microfluorometric (FMF) studies comparing the Thy-1.2, Lyt-1, Lyt-2, and surface immunoglobulin (slg) phenotypes of lymphocytes from normal and infected mice were performed. Between days 5 and 7 the thymocytes from infected mice displayed a higher relative fluorescence intensity (RFI) of the Thy-1.2 marker than normal thymocytes, whereas at day 10, the RFI was less than that of normal thymic lymphocytes. Between days 7 and 10 of infection the RFI of the Lyt-2 marker was less on thymocytes from Hc-infected mice; however, there was no change in the Lyt-1 marker. Examination of these lymphocyte markers in blood, spleen, and mesenteric lymph nodes showed that there were decreases in the RFI of both the Thy-1.2 and Lyt-2 between days 5 and 10 of infection. No changes were observed in the Lyt-1 or slg markers. By day 28 there were no differences between the normal and infected mice with respect to any surface marker in any of the organs studied. In other experiments, the effect of adrenalectomy before infection on these surface markers was studied. Absolute numbers of Thy-1.2+, Lyt-1+, and Lyt-2+ cells were significantly increased in the spleen and significantly decreased in the thymus and peripheral blood of infected mice relative to normal controls. These studies suggest that there is a migration of cells from the thymus, blood, and bone marrow to the spleens of mice with disseminated Hc infection.
Modulation of F1 cytotoxic potentials by GvHR. Host- and donor-derived cytotoxic lymphocytes arise in the unirradiated F1 host spleens under the condition of GvHR-associated immunosuppression.
Authors: Kubota E , Ishikawa H , Saito K
Published: December 2022

As an approach to dissect complex cellular events that lead to GvHR-associated immune disorders, we followed cytotoxic activities, including NK cytotoxicity, in the spleens of unirradiated F1 hosts undergoing GvHR induced by parental spleen cells. Spleen cells of (B10 X DBA/2)F1 or (B10 X AKR/J)F1 hosts undergoing GvHR induced by parental B10 spleen cells displayed a prompt and marked increase in NK cell activity within 36 hr, and the heightened activity lasted until day 8. The activity then declined abruptly and disappeared on day 12 of GvHR. Inversely, donor B10-derived CTL specifically directed to the opposite parental alloantigens of the F1 hosts emerged in these F1 host spleens on day 8, and the CTL activity reached a peak on day 12 when the host NK cell activity disappeared. During the period that the donor-derived anti-host CTL were present, these F1 host spleen cells lost not only NK cell activity but also the ability to mount in vitro CTL responses. In contrast, the respective F1 strain mice undergoing GvHR induced by the parental DBA/2 or AKR/J spleen cells showed only transient but marked increases in NK cell activity during the initial 36 hr, and then the activity decreased gradually to return to the normal level on day 10. In such GvHR F1 host spleens, donor-derived CTL could never be detected, and the spleen cells showed normal in vitro CTL responsiveness during the entire observation period of 16 days. These results are discussed from the viewpoint of genetically defined cellular events that lead to the GvHR-associated immune disorders.
Definition of a "trigger" stage in the NK cytolytic reaction sequence by a monoclonal antibody to the glycoprotein T-200.
Authors: Targan S R , Newman W
Published: December 2022

The monoclonal antibody 13.1 recognizes an epitope on the T-200 glycoprotein and blocks natural killer (NK) lysis of the erythroleukemia target K562, but not of the acute lymphoblastic leukemia T cell target Molt-4. The inhibitory effect is at the killer cell level and not the target cell level, which suggests that 13.1 may react with a receptor on NK cells. This hypothesis was tested in assays to delineate precisely where in the NK cytolytic reaction sequence 13.1 interferes with lysis; 13.1 did not block initial NK-target cell interaction as measured in a target binding cell assay. With the use of a Ca++ pulse technique, 13.1 did not block any events occurring during Ca++-dependent programming. If the antibody was added after conjugate formation but before the addition of CaCl2 to initiate programming, however, full inhibition of NK lysis occurred. Therefore, 13.1 antibody defines a distinct stage in the NK reaction sequence that links target binding to the initiation of calcium-dependent programming events. NK cell binding alone is not sufficient to trigger lytic events, and the presence of a second structure or a distinct portion on a single structure is required to trigger lysis. We show that 13.1 blocks the ability of K562 to inhibit NK killing of Molt-4 in a cold target inhibition assay. Therefore, despite the fact that 13.1 does not disrupt conjugate formation, NK specificity may exist at a post-binding site rather than at the initial NK-target binding interaction. Our data suggest that the T-200 glycoprotein on NK cells triggers the initiation of the lytic events.