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B E Elliott

Publications and source records attributed to B E Elliott.

At least 37 records · Page 2Linked to original sources
PubMed1988

Expression of epithelial-like markers and class I major histocompatibility antigens by a murine carcinoma growing in the mammary gland and in metastases: orthotopic site effects.

A spontaneous murine mammary carcinoma, designated SP1, grew more aggressively in the mammary gland than in the subcutis exhibiting a 10-fold lower 50% lethal tumor dose and the ability to metastasize spontaneously from the orthotopic mammary gland site. The appearance of metastasis could be abrogated by resection of the primary tumor up to 21 days postinjection, arguing against the possibility that metastasis occurred due to trauma of the injection and/or healing processes. In addition, tumor cells recovered from lung metastases exhibited an increased ability to metastasize when reinjected into either the s.c. or mammary sites. Tumor cells from lung metastases showed low levels of Class I major histocompatibility (MHC) antigens, like the parental SP1 cells, but were found to express differentiation markers typical of normal basal and luminal mammary epithelium. SP1 tumors expressed increased Class I MHC antigens, as well as high levels of basal and luminal breast epithelial markers, within 7 days of implantation into the mammary gland. On the other hand, SP1 tumors growing in the subcutis never expressed increased Class I MHC levels and expressed the epithelial marker antigens at lower levels and not until at least 21 days of growth. Removal of host epithelium by cauterization of the mammary bud at 3 weeks had no effect on the increased growth, metastasis and acquired heterogeneity of MHC and epithelial associated antigens, suggesting that the mammary gland stroma was responsible for the observed phenomenon. These findings suggest that the mammary gland either selects distinct tumor subpopulations, or induces a phenotypic change leading to tumor progression and the generation of metastatic subpopulations.

Animals↗
PubMed1988

Heritable high frequency modulation of antigen expression in neoplastic cells exposed to 5-aza-2'-deoxycytidine or hydroxyurea: analysis and implications.

A study was undertaken to determine the ability of the DNA hypomethylating drug 5-aza-2'-deoxycytidine (5-azadCyd) to induce antigen expression in a high proportion of treated tumor cells and to evaluate if this effect could be mimicked by the drug hydroxyurea (HU) which is a genomic DNA hypermethylating agent. Induction of heritable changes in gene expression by 5-azacytidine or the 2'-deoxy analogue (5-azadCyd), at least in some cases, may not be necessarily due to their hypomethylating properties, but to some other induced high frequency genetic change which occurs when DNA synthesis or repair is perturbed. A comparison of 5-azadCyd and HU effects on human and murine tumors was chosen for this study. The phenotypic properties examined with the above treatments were (a) induction of a Mr 110,000 antigen, detected with M111 antibody, in a variant subpopulation (SMeLus7) of human melanoma cells which fail to maximally express this antigen (M111). The parent cell line, MeWo, and other MeWo-derived variant cell cell lines do not demonstrate a similarly inducible phenotype; and (b) induction of class I major histocompatibility complex antigens in a population of mouse mammary adenocarcinoma cells which normally fail to express these antigens. The results showed that 5-azadCyd was effective in inducing antigen expression in both systems whereas HU was effective (and equally so) only in the human melanoma cell line system. In these treatments 5-azadCyd was demonstrated to transiently hypomethylate the same human melanoma cell line whereas HU hypermethylated genomic cytosines. The results suggest that some of the reported effects of 5-azacytidine or 5-azadCyd in inducing very high frequency heritable phenotypic alterations may not necessarily be related to the drugs' ability to cause DNA hypomethylation. The implications of these results are discussed in terms of the use of 5-azacytidine or 5-azadCyd and the effects of chemotherapy on tumor progression and metastasis.

Animals↗
PubMed1987

In situ augmentation of class I major histocompatibility antigen expression on immunogenic variants of a spontaneous murine mammary carcinoma.

The relationship between expression of cell surface glycoproteins encoded by the major histocompatibility complex (MHC) and immunogenicity of a recently obtained spontaneous murine mammary adenocarcinoma (designated CBA.SP1) was examined. Immunogenic and nonimmunogenic variant clones were isolated from a subclone of the parent tumor after treatment with the mutagen N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) or the DNA hypomethylating agent and "gene activator," 5-aza-2'-deoxycytidine (5-aza-dCyd). All clones from the untreated tumor population were tumorigenic in normal syngeneic recipients. In contrast, immunogenic variant clones, isolated at high frequencies after drug treatment [ranging from 5% (5-aza-dCyd treated) to greater than 90% (MNNG treated)], were rejected in normal syngeneic mice but grew progressively in T-cell deficient nude mice. Consistent with our previous report (J. Natl. Cancer Inst., 75: 291, 1985), all 5-aza-dCyd induced immunogenic clones expressed elevated levels of class I (particularly Dk) MHC antigens. However some (three out of nine) nonimmunogenic clones also showed enhanced class I MHC expression, implying that not all high MHC expressors were immunogenic. In contrast to 5-aza-dCyd induced variants, only 50% of MNNG induced immunogenic variants showed elevated levels of Dk or Dk and Kk antigens in vitro. Strong augmentation of class I MHC antigens in situ was observed on all immunogenic, but not nonimmunogenic, clones following transplant into syngeneic mice; no increase in MHC expression on variants during progressive growth in athymic nude mice occurred. Although no class II (Ak or Ek) antigens were detected on the parent line or any of the immunogenic variants, a strong infiltration of host I-A bearing cells occurred during immune rejection of SP1 variants. These results are consistent with the hypothesis that induction of class I MHC antigen expression on certain low MHC expressing tumors, although not the sole requirement for immunogenicity, can facilitate immune rejection of the SP1 tumor and, conversely, that the reduced level of MHC observed in certain clinical cancers may significantly affect the immunological aspects of the tumor-host relationship.

Adenocarcinoma↗
PubMed1985

Enhanced expression of class I major histocompatibility complex gene (Dk) products on immunogenic variants of a spontaneous murine carcinoma.

Both immunogenic and nonimmunogenic variant clones were isolated from a recently obtained spontaneous murine adenocarcinoma after treatment (xenogenization) with either the mutagen ethyl methanesulfonate or the DNA hypomethylating agent, and "gene activator," 5-azacytidine. Clonal analysis of the untreated tumor population confirmed that immunogenic variants arose as a consequence of the xenogenization protocol. At a dose of 10(6) cells per mouse, nonimmunogenic variants, like the parental tumor line, grew progressively in normal syngeneic recipients. In contrast, immunogenic variants were rejected in normal syngeneic mice and grew progressively only in T-cell-deficient nude mice. Serologic analysis of the respective clonal variants revealed that immunogenic variants expressed substantially elevated (fourfold to tenfold) levels of class I H-2Dk antigen relative to parental or nonimmunogenic cell lines. Two variants exhibiting marginal immunogenicity expressed high and low levels of major histocompatibility complex (MHC) antigen, respectively suggesting that elevated MHC expression, although possibly a contributing factor, did not account for the immunogenic phenotype in all cases. Finally, the immunogenic phenotype of two variants decayed with time in culture. Clones in the process of reversion lost their elevated Dk gene expression and became progressively more tumorigenic in normal syngeneic mice. Together, these data are consistent with a hypothesis that elevated MHC expression can contribute to the immunogenic phenotype of originally low MHC-expressing tumors and that the reduced level of MHC observed in certain clinical cancers may have significant implications with regard to immunologic aspects of the tumor-host relationship.

Animals↗
PubMed1984

Adoptive immune therapy in mice bearing poorly immunogenic metastases, using T lymphocytes stimulated in vitro against highly immunogenic mutant sublines.

MDW3, a highly immunogenic and non-tumorigenic (tum-) mutant of the poorly immunogenic metastatic murine tumor called MDAY-D2, has been employed in an immune therapy scheme for the treatment of widespread established visceral MDAY-D2 metastases in syngeneic mice. MDW3 was selected from a mutagenized population of MDAY-D2 cells for the ability to grow in the presence of toxic concentrations of wheat-germ agglutinin (WGA) in vitro. The mutant expresses a common tumor-associated antigen (TAA) present on MDAY-D2 as well as a new antigen whose presence enhances the anti-TAA cell-mediated immune response in vivo and in mixed lymphocyte tumor cultures (MLTC) in vitro. For immune therapy, spleen cells from DBA/2 mice which had rejected an inoculum of MDW3 cells were restimulated in MLTC and injected i.v. into MDAY-D2 tumor-bearing mice. Two protocols were used. In the first, mice were given an i.v. injection of 10(3) MDAY-D2 cells ("artificial metastasis") and subsequently treated with 400 R whole-body irradiation and MDW3-stimulated T cells. Such mice had a 75% long-term survival rate, whereas 400 R alone, or no treatment, resulted in 25% and 0% long-term survivors, respectively. In the second protocol, treatment of mice bearing a 12-day-old subcutaneous MDAY-D2 tumor by surgical removal of the solid tumor, 400 R whole-body irradiation, and systemic administration of MDW3-stimulated spleen cells, resulted in a 75-100% survival rate, whereas omitting any part of the treatment resulted in 0-50% survival rates. The treatment increased splenic anti-TAA CTL activity, and the mice acquired immunity against the new antigen on MDW3, suggesting that the injected lymphocytes were proliferating in the host. The optimal combination of resection, whole-body irradiation and passive infusion of MDW3-stimulated spleen cells was ineffective when used on mice bearing a tumor-antigen-loss variant of MDAY-D2, suggesting that success of our immune therapy protocol required specific recognition of the tumor-associated antigen of MDAY-D2.

Animals↗
PubMed1984

Flow cytometric analysis of specific binding of soluble Ia by I-region restricted alloactivated T cells.

An antigen binding assay has been developed for quantitation by flow cytometry of vesicular and soluble Ia binding by alloactivated T cells. Binding of stimulator membrane vesicles was detected by anti-Ly-6.2 or anti-Ia monoclonal antibodies coupled to fluorescent latex beads. Vesicle binding by an I-Ak specific A.TH anti-A.TL T cell line occurred via I-Ak molecules, in that (a) vesicles expressing I-Ak molecules bound much more effectively than vesicles of H-2b,q strains, and (b) inhibition of H-2k vesicle binding occurred with anti-I-Ak, but not anti-Kk, anti-Ek, or anti-Dk antibodies. T cell receptor/Ia interactions were directly studied by inhibition of H-2k vesicle binding by T cells with partially purified Ia glycoproteins. Inhibition of binding occurred via Ia molecules since (a) affinity column partially purified allogeneic I-Ak molecules inhibited binding much more effectively than syngeneic I-As molecules and (b) depletion of I-Ak but not Ek molecules in Iak containing glycoprotein fractions abrogated the inhibitory effect. The ability of this method to detect specific binding of soluble Ia with antigen activated T cells makes it a useful tool for studying interaction of membrane free major histocompatibility complex (MHC) products with native T cell receptor.

Animals↗
PubMed1984

Possible epigenetic mechanisms of tumor progression: induction of high-frequency heritable but phenotypically unstable changes in the tumorigenic and metastatic properties of tumor cell populations by 5-azacytidine treatment.

Treatment of a variety of highly tumorigenic mouse lines in vitro with chemical mutagens, such as ethyl methane sulfonate (EMS) or N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), can result in extraordinarily high frequencies (sometimes in excess of 90%) of strongly immunogenic clones unable to grow progressively in normal syngeneic hosts. These clones will, however, grow in immunosuppressed hosts and gradually regain tumorigenic ability in normal mice if maintained in long-term (several months-1 year) culture, i.e., they are often phenotypically unstable. These features--phenotypic drift and high frequency--make it unlikely that point mutations are the underlying mechanism involved in the generation of the variants. Results presented here demonstrate that these observations can be reproduced on the same tumor lines using 5-azacytidine--an analogue of cytidine which can be incorporated into DNA causing subsequent extensive hypomethylation of cytosine residues in the absence of any significant mutagenic effects. Furthermore, 5-azacytidine treatment of a nonmetastatic mouse mammary tumor led to the emergence of a small number of heritable but unstable tumor clones capable of spontaneous metastatic spread. Because it is known that DNA hypomethylation can lead to transcriptional activation of normally silent genes, that altered methylation patterns can be somatically replicated with a high but not perfect fidelity, and that mutagens can cause DNA hypomethylation, we propose that DNA hypomethylation followed by de novo methylation represents a plausible mechanism to account not only for the induction of the nontumorigenic variants but for a number of aspects of tumor progression and tumor heterogeneity, as well. In particular, we refer to heritable phenotypic alterations in tumor cell populations which occur at very high frequency but which are not necessarily stable over very long periods of time.

Animals↗
PubMed1984

Receptor specificity of Ia-restricted T lymphoblasts activated against trinitrobenzene sulfonate-coupled spleen cells: recognition of distinct trinitrophenyl and Ia moieties.

The receptor specificity of H-2-restricted T lymphoblasts activated against trinitrobenzene sulfonate (TNBS)-coupled spleen cells was examined using an antigen binding assay. A population of Lyt-1+,2-T lymphoblasts acquired syngeneic Ia determinants during 4 days of primary culture with hapten-coupled stimulator cells. Syngeneic Ia was not reexpressed after trypsin treatment of the T cells, but was found after incubation with soluble Ia shed from lipopolysaccharide-activated blasts. Self-Ia binding was specific in that Lyt-1+,2- but not Lyt-1-,2+ cells acquired the antigen, and in that self-Ia bound more effectively than allogeneic Ia material. To determine the relationship of self-Ia binding to the recognition of foreign antigen, the binding of trinitrophenyl (TNP)-coupled plasma membrane vesicles by TNP-specific T cells was studied. TNP-vesicle binding occurred via TNP and H-2(Ia) molecules on the vesicles in that binding was inhibited with antibodies against TNP or H-2(Ia) molecules but not non-major histocompatibility complex (e.g., Ly-6.2) molecules on the vesicles. Complete inhibition of TNP-vesicle binding by an Iak-restricted TNP-specific T-cell line occurred with soluble TNP-lysine, but not an unrelated hapten, N-iodoacetyl-N-(5-sulfonic-1-naphthyl)ethylenediamine (I-AED)-cysteine. Conversely, I-AED-cysteine, but not TNP-lysine, inhibited binding of I-AED-coupled B6 vesicles by B6 anti-I-AED T cells. Significant, but weak inhibition of TNP-vesicle binding by the anti-TNP line was observed with glycoprotein preparations containing partially purified self-Ia molecules. However, inhibition was specific for I-Ak molecules, in that inhibition was lost after removal of I-Ak molecules from the glycoprotein preparation, and very little inhibition occurred with soluble glycoproteins prepared from thymocytes which contained very little Ia material or from LPS blasts of an unrelated H-2 haplotype. These results suggest a recognition model in which TNP and Ia determinants are recognized by neighboring receptor combining sites.

Animals↗
PubMed1983

Detection of Fc receptors.

There are an almost bewildering array of assays one can use to detect Fc-receptors, but we have focused, for the most part, on relatively rapid and convenient methods--those that have become standard, e.g., EA rosetting procedures or uptake of soluble antigen-antibody complexes. For most types of Fc-receptor studies these methods are quite adequate. We have also tried to stress the newer methodology of detecting Fc receptors, i.e., the use of anti Fc-receptor antibodies, including monoclonal antibodies. The advent of monoclonal antibody technology, described in detail by Galfrè and Milstein, is changing the approach of many immunological procedures, and detection of Fc receptors is no exception. There are, it should be noted, many newer Fc-receptor methods, e.g., a solid-phase radioimmunoassay and a quantitative fluorometric assay, and methods such as these may be the most appropriate ones to choose in a particular experimental situation. In addition, we have not stressed "functional" Fc-receptor detecting assays, for example, antibody-dependent cell-mediated cytotoxicity (ADCC), which can be used to monitor the activity of Fc receptors in a whole cell population, rather than in single cells.

Animals↗
PubMed1983

Idiotypes and allotypes on Ia-binding alloactivated Lyt-1+,2-,3- T cells are coded for by genes linked to the igh-1 allotype locus.

Xenogeneic antisera, designated 5936 and 6036, have been used in this laboratory to define 5936-idiotypes (Id) and 6036-T cell receptor (Tcr) allotype determinants, respectively, on populations of B6 anti-CBA MLC T blasts. The present experiments were carried out to determine a) whether T lymphocytes that bear 5936-Id also express antiserum 6036-defined Tcr allotypes; b) if so, what is the Lyt phenotype and c) specificity of this T lymphocyte population, and d) whether the genes coding for 5936-Id and 6036-allotypes are linked to immunoglobulin (Ig) allotype genes. The results showed that 5936-Id-bearing T lymphocytes were included in a subset of cells that expressed antiserum 6036-defined Tcr allotypic determinants and the Lyt-1+,2-,3- phenotype. By genetic segregation analysis it was shown that both 5936-Id and antiserum 6036-defined allotypic determinants on T cells were linked to Igh-1b allotypes. 5936-Id determinants were expressed on T lymphocytes of Igh-1b allotype strains, specific for allogeneic I-Ak but not on those specific for I-Ak/Ek molecules. In contrast, 6036-Tcr allotype determinants were present on T lymphocytes from Igh-1b allotype strains, specific for either I-Ak or I-Ak/Ek molecules. The results support the hypothesis that 5936-Id determinants are present only on a subset of anti-I-Ak T cell receptors, whereas 6036-Tcr allotype determinants are shared among T cell receptors of several unrelated specificities from Igh-1b allotype-bearing strains.

Animals↗
PubMed1982

An enzyme-linked immunosorbent assay (ELISA) for detergent solubilized Ia glycoproteins using nitrocellulose membrane discs.

An enzyme-linked immunosorbent assay has been developed for the quantitative detection of detergent solubilized murine Ia. Nitrocellulose membrane discs were used to bind membrane glycoproteins applied in solutions containing detergent. The bound antigen was detected by monoclonal antibodies and horseradish-peroxidase-coupled anti-IgG. The assay produced a linear response with respect to antigen concentration, and could readily detect partially purified Ia derived from 10(3) to 10(4) mitogen stimulated spleen cells. Nitrocellulose discs efficiently bound protein in the presence of deoxycholate, taurocholate, and octylglucoside. Less binding occurred in the presence of Triton X-100 or Tween 80, but 90% binding efficiency was obtained in 0.01% solutions of these detergents. The association of protein with the discs was stable under normal conditions for antigen detection, but could be further stabilized by briefly fixing with glutaraldehyde for more rigorous procedures. The ability of this method to detect antigen in detergent solutions makes it useful in monitoring fractions during the purification of cell membrane proteins.

Animals↗
PubMed1982

T cell idiotypes recognizing self-major histocompatibility complex molecules: H-2 specificity, allotype linkage, and expression on functional T cell populations.

An anti-idiotypic serum (antiserum 5936, B. Rubin et al., J. Exp. Med. 1979. 150: 307) was used to demonstrate receptor sites for self-major histocompatibility complex (MHC) antigens on T lymphocytes. The antiserum was raised by injecting rabbits tolerant to mouse Ig with a B6 anti-CBA (anti-H2k) alloantibody. It recognized a large proportion of T cells from H-2k strains carrying the b, c, d or e allele at the Igh-1 locus, but only a few T cells from H-2k strains with Igh-1 alleles a, f and j. Allotype linkage of the 5936 idiotype was also demonstrated by segregation analysis. The antiserum did not recognize either H-2k B cells or T cells from other H-2 haplotypes despite the presence of a permissive Igh-1 allele. The 5936 idiotype was found to be associated with several different antigen specificities, indicating that it is not located on the binding site for foreign antigen. Furthermore, the 5936 antiserum inhibited the binding of soluble Ik antigens by H-2k, Igh-1b, T cells, and, in the presence of complement, eliminated T cells responding to different antigens in an I-Ak-restricted fashion. Collectively, the data indicate that the structure bearing the 5936 idiotype is a receptor for I-Ak antigens, expressed by strains carrying the I-Ak allele and a permissive allele at the Igh-1 locus. The relevance of this finding to the MHC-restricted recognition of antigens by T cells is discussed.

Animals↗
PubMed1982

Comparative analysis of cell surface markers on murine NK cells and CTL target-effector conjugates.

The rosetting of sheep erythrocytes (SRBC) coated with non-haemagglutinating monoclonal antibodies rather than conventional haemagglutinating antisera revealed readily detectable FcR on most splenic natural killer (NK) cells since 76% of splenic lymphocytes forming conjugates with YAC also rosetted with SRBC coated with high concentrations of monoclonal anti-SRBC antibody of the IgG2b subclass and since Ficoll depletion or enrichment of splenic lymphocytes rosetting with IgG2b-coated SRBC resulted in a corresponding 4-fold decrease or increase in conjugate-forming cells and a 10-fold decrease or increase in NK cytolytic activity. NK cells bound much less readily to monoclonal IgG2a and not at all to monoclonal IgG1 or IgM, but the degree of binding was directly proportional to the amount of antibody on the erythrocytes and was not isotope-restricted. In addition, immunofluorescent studies revealed that YAC-1-conjugated lymphocytes were Lyt-1-, Lyt-2-, partially Thy-1+ (60%), asialo(GM1+ (80%). Qa-4+ (77%), Qa-5+ (79%), and Ly-5+ (94%). In comparison, a proportion (39%) of alloimmune peritoneal exudate cells which conjugated with P815-2 also stained by immunofluorescence with anti-asialo GM1 antisera. Most (greater than 90%) P815-conjugated cells were Thy-1+, Lyt-2%, and a subpopulation of Lyt-1+2+ conjugates was observed (25%). Qa-5 and Ly-5 were also expressed on most (two-thirds) cytolytic T lymphocytes (CTL) conjugates, whereas Qa-4 and FcR for IgG2b were not detected. The best phenotype distinctions between NK cells and CTL were therefore based on the presence or absence of Lyt-2, Qa-4, and FcR for IgG2b on most effector cells. Anti-asialo-GM1 or monoclonal anti-Qa-4 and complement treatment greatly diminished both the frequency of NK conjugates and the percentage of conjugates with detectable IgG2b FcR or asialo-GM1. These results confirm that NK cells co-express asialo-GM1 and Fc receptors, at the single-cell level, and provide a simple method for greatly enriching NK populations at least 10-fold.

Animals↗
PubMed1981

Receptor specificity, functional characteristics, and cell-surface phenotype of a highly selected anti-I-Ab-specific, long-term T-cell line.

A highly selected alloreactive T-cell line was developed by repeated restimulation of B10.D2/n lymph-node cells with irradiated C57BL/10Sn (B10) spleen cells in long-term MLC for up to 21/2 years. Continuous growth of the line requires restimulation every 2 to 4 weeks with fresh H-2b stimulator cells. The line proliferates strongly against H-2b but not against H-2d, H-2f, H-2q, H-2r, or H-2s stimulators. Analysis of recombinant mouse strains showed that the proliferative response is directed against I-Ab, but not Kb or Db determinants. During the growth period of the line, strong cross-reactivity with H-2p (B10.P) and weak cross-reactivity with H-2k strains (e.g., CBA/J and B10.BR) was observed. A clone with exquisite specificity for I-Ab, but with no cross-reactivity with H-2p or H-2k was isolated from the line; thus clonal heterogeneity of the line still exists despite the highly selective growth conditions.--The majority of T cells from the line or clone were shown to bind I-Ab but not Kb or Db determinants either spontaneously during restimulation with fresh B10 stimulator cells or via membrane vesicles expressing I-Ab determinants. No killing activity by the line in either specific or nonspecific cytolytic T-cell assays was observed nor was the T 145 glycoprotein, characteristic of killer T cells, detected.

Animals↗
PubMed1980

Surface receptors on lymphoreticular cells: sensory devices for host recognition of foreign antigens and neoplasia.

Distinct types of cells in the lymphoreticular system regulate an individual's immunologic homeostasis and response to disease. By means of multiple receptors the cell membrane transfers signals between the environment and the cell. This article reviews the important surface antigens and receptors on normal and activated macrophages (e.g., receptors for the crystallizable fragment [Fc] of immunoglobulin), on lymphocytes derived from the bone marrow (e.g., immunoglobulins and immune-associated [Ia] antigens), on thymus-derived lymphocytes (e.g., Thy-1 antigens) and on "null' cells. Although many of these markers were originally defined in rodents and birds, analogous markers in humans have proved extremely useful in characterizing lymphoreticular cell populations in healthy and sick individuals. Established and postulated functions of the markers in host defence mechanisms are discussed.

Antigens, Neoplasm↗