Biomedical subjects

A L Epstein

Publications and source records attributed to A L Epstein.

At least 109 records · Page 6Linked to original sources

PubMed1985

Use of novel chemical supplements in the establishment of three human malignant lymphoma cell lines (NU-DHL-1, NU-DUL-1, and NU-AMB-1) with chromosome 14 translocations.

Three new cell lines have been established from patients with malignant lymphoma utilizing a human diploid feeder layer, pooled human serum, and the chemical supplements L-cysteine, iron-saturated transferrin, and bathocuproine disulfonate, a copper chelator. After a short period of growth, the 3 cell lines were successfully weaned from the feeder layers but continued to require human serum and the chemical supplements for up to 9 months of culture. The cell lines are currently grown in RPM1-1640 medium and fetal calf serum without further supplementation. The NU-DHL-1 cell line was established from the involved lymph node of a 73-year-old White male with diffuse large-cell lymphoma. The cell line expresses cytoplasmic IgM/lambda heavy and light chains, is Epstein-Barr virus (EBV)-negative, and is positive for several B-cell markers, indicating that it is derived from a mature-B-cell neoplasm. The NU-DUL-1 cell line was established from the cerebrospinal fluid of a 42-year-old White male with undifferentiated lymphoma, non-Burkitt's type, who initially presented with a mediastinal mass and had subsequent involvement of the central nervous system. The cell line is EBV-negative, but surprisingly it is positive for early B-cell markers. The NU-AmB-1 cell line was established from the abdominal mass of a 12-year-old Hispanic male with undifferentiated lymphoma, Burkitt's type. The cell line is EBV-positive and expresses early B-cell markers. All 3 cell lines are aneuploid or pseudodiploid and contain chromosome 14q+ abnormalities including a newly described complex translocation t(?;1;8;14) in the NU-AmB-1 cell line. The establishment of these cell lines was made possible by refinements in the cell culture of the human malignant lymphomas. The availability of well-characterized lymphoma cell lines with specific chromosomal translocations will aid molecular and cellular studies designed to identify the biological significance of genomic rearrangements.

Cell Line↗

PubMed1985

Histiocytic lymphoma cell lines: immunologic and cytogenetic studies.

Cell lines were established from 15 patients with diffuse histiocytic lymphoma (DHL) of the intermediate grade, diffuse large cell (class G), and high-grade, large cell immunoblastic (class H) types. Immunologic studies indicated that 11 of the 15 DHL cell lines were B cell in origin, 2 were histiocytic, and 2 were null cell. Cytogenetic studies revealed 1 hypodiploid, 11 hyperdiploid, and 3 near-tetraploid cell lines. Chromosome #7 was trisomic in 3 lines, chromosomes #12 in 4 lines, and chromosome #13 in 3 lines. Chromosome #2 was monosomic in 3 lines, chromosome #8 was monosomic in 5 lines, chromosome #14 in 4 lines, and chromosome #22 in 6 cell lines. This is of special interest, as chromosomes #2, #8, #14, and #22 are clearly concerned with rearrangements in Burkitt's lymphoma and immunoglobulin expression. The most common rearrangement in the DHL cell lines involved chromosome #14 at band 14q32. However, in contrast to Burkitt's lymphoma, the pattern of translocation in DHL is between chromosome #14 and usually chromosome #11 or chromosome #18. The 14;18 translocation is not restricted to patients with low-grade follicular, small cleaved cell lymphomas, as has been reported. The 14q+ chromosome is characteristic of lymphoid malignancies in general. It is due, invariably, to a translocation with the breakpoint in band 14q32, which is the locus of the immunoglobulin heavy chain genes. We propose that in each translocation, for example, chromosomes #11 or #18, an oncogene may be transposed onto chromosome #14, and that each 14q+ translocation in DHL represents an event that transposes an oncogene from another chromosome to chromosome #14.

Cell Line↗

PubMed1985

A method for simultaneous nuclear immunofluorescence and DNA content quantitation using monoclonal antibodies and flow cytometry.

A preparative technique for the two-parameter flow cytometric study of nuclear antigen expression is reported. This method employs a brief sequential treatment of cells at 4 degrees C first with 0.5% paraformaldehyde and second with 0.1% Triton X-100 in phosphate-buffered saline followed by cellular staining with indirect immunofluorescence and propidium iodide. Using this technique, cellular morphology is preserved, cell clumping is minimized, and high-quality indirect immunofluorescence and DNA staining are obtained with a minimum of nonspecific labeling. Utilizing nuclear antigen-specific monoclonal antibodies in conjunction with this technique, the cell-cycle phase-dependent expression of such antigens is examined. From these data, the utility of two-parameter flow cytometry in the identification and quantification of cell-cycle-dependent modulation of nuclear antigens is discussed.

Antibodies, Monoclonal↗

PubMed1985

Cloning the chromosomal breakpoint of t(14;18) human lymphomas: clustering around JH on chromosome 14 and near a transcriptional unit on 18.

Specific chromosomal translocations found in distinct neoplasms suggest that genes that flank such breakpoints play a critical role in transformation. We have characterized the t(14;18)(q32;q21) chromosomal translocation present in over 60% of human follicular lymphomas. We exploited an unexpected rearrangement of an Ig heavy-chain gene to clone the chromosomal breakpoint. An element isolated from 18q21 mediated translocations in all four t(14;18) bearing cell lines and in six of 11 follicular lymphomas, but did not normally rearrange in other B or non-B cells. The breakpoints clustered within a small 4.3 kb region on chromosome 18. The breakpoints on chromosome 14 were focused within or immediately 5' to JH. These breakpoints retained the Ig enhancer region close to a new transcriptional unit identified on chromosome segment 18q21. Since none of the cellular oncogenes are known to map to 18q21, cloning this element provides an opportunity to characterize a potentially new transforming gene.

Cell Line↗

PubMed1985

Monoclonal antibodies. Their promise for tumor diagnosis, staging, and therapy.

Monoclonal antibodies will have utility as tools for diagnosis, staging, and therapy of malignant disease. Investigators have produced monoclonal antibodies that are directed against tumor-associated antigens and have varying degrees of cross-reactivity against normal tissues. These reagents have homogeneous molecular structure, recognize specific antigenic sites, can be produced in mass quantities, and are easily purified. Clinical trials with monoclonal antibodies are in progress and suggest modest toxicity and potential therapeutic efficacy.

Antibodies, Monoclonal↗

PubMed1985

Susceptibility to herpes simplex virus type 1 infection of non-permissive rat XC(HPRT-) x permissive mouse L(TK-) hybrid cells.

Somatic cell hybrids between rat XC(HPRT-) cells, non-permissive for herpes simplex virus type 1 (HSV-1) infection, and permissive mouse L(TK-) cells were constructed and karyotyped. Infection of these hybrid cells by HSV-1 strains F and MP revealed that they were susceptible to the virus. The amounts of virus produced by the hybrid cells, as well as the cytopathic effect observed, was very similar to that of the parental L(TK-) cells. Our results suggest that failure of HSV-1 to replicate in XC cells is more likely to be due to the absence of cellular elements required for efficient virus multiplication rather than to the presence of blocking or inhibiting factors.

Animals↗

PubMed1985

Expression of maturation-specific nuclear antigens in differentiating human myeloid leukemia cells.

The expression of three myeloid-specific nuclear antigens was studied by indirect immunofluorescence with murine monoclonal antibodies in human myeloid (HL-60, ML-2, KG-1, and B-II) leukemia cells treated with chemical inducers of cell differentiation. Treatment of the promyelocytic HL-60 cells with dimethyl sulfoxide or 1,25-dihydroxyvitamin D3 induced the cells to acquire a phenotype that resembled that of granulocytes and monocytes-macrophages, respectively. These phenotypes were characterized by changes in cell growth, cell morphology, expression of specific cell surface antigens, and activities of lysozyme and nonspecific esterase enzymes. Induction of these differentiation markers in the HL-60 cells was associated with induction of the myeloid-specific nuclear antigens. The ML-2 cells, which are arrested at the myeloblast-promyelocyte stage, were also susceptible to the induction of cell differentiation and to changes in the expression of the nuclear antigens, but the degree of susceptibility was less than in the HL-60 cells. The less-differentiated KG-1 and B-II myeloid cells were either not responsive or responded only in a limited degree to the induction of cell differentiation or to changes in the expression of the nuclear antigens. We suggest that the reactivity of cells with monoclonal antibodies to specific nuclear antigens can be used as a maturational marker in cell differentiation studies. Furthermore, nuclear antigens expressed early in cellular differentiation may provide information about changes in regulatory elements in normal and malignant cells.

Antibodies, Monoclonal↗

PubMed1985

Immunohistochemical analysis of human lymphomas with monoclonal antibodies to B cell and Ia antigens reactive in paraffin sections.

Monoclonal antibodies (MoAbs) to B cell- and T cell-specific antigens uniformly have been restricted in their use to cell suspension or frozen section techniques because the antigens that they identify are either masked or lost in the fixation or paraffin-embedding processes. Antiimmunoglobulin antisera, although readily identifying cytoplasmic immunoglobulin in paraffin sections, has not been as useful as originally hoped since the majority of B cell lymphomas express surface immunoglobulins, which requires cell suspensions or frozen sections for detection. Because cell suspension procedures disrupt tissue architecture and frozen section techniques grossly distort morphology, neither method allows the combination of optimal morphologic and immunologic classification of lymphomas. We recently reported two MoAbs, LN-1 and LN-2, that react with B cells in paraffin sections. LN-1 reacts with the surface membrane and cytoplasm of germinal center B cells. LN-2 reacts uniquely with the nuclear membrane and cytoplasm of mantle zone and germinal center B cells and interdigitating histiocytes. We have also identified a new MoAb, LN-3, that reacts with the HLA-DR antigen in paraffin sections. We now report the use of LN-1, LN-2, and LN-3 in the analysis of paraffin sections from 58 non-Hodgkin's lymphomas and 15 cases of Hodgkin's disease. The types of cells reactive with these MoAbs in neoplastic lymphoid proliferations largely recapitulate their benign morphologic and immunologic counterparts. As a panel, LN-1, LN-2, and LN-3 were reactive with 98% of B cell lymphomas, and LN-1 and LN-2 were negative on all T cell lymphomas. In addition to identifying the cell of origin of these malignant proliferations, these MoAbs were also useful for identifying architectural features in neoplastic lymph nodes. Thus, these reagents provide the ability to assess the immunologic phenotype of neoplastic lymphocytes in conjunction with the critical morphologic criteria requiring paraffin embedding.

Antibodies, Monoclonal↗

PubMed1984

Infection of a restrictive cell line (XC cells) by intratypic recombinants of HSV-1: relationship between penetration of the virus and relative amounts of glycoprotein C.

The ability to induce the synthesis of virus polypeptides in nonpermissive XC cells by several strains, variants, and intratypic recombinants of HSV-1 has been investigated. Our results show that (i) whereas HSV-1 strains mP and MP, and recombinants F(MP)A through D (Ruyechan et al., J. Virol. 29, 677-697, 1979) and HFEMtsB5/MP3 (Manservigi et al., Proc. Nat. Acad. Sci. USA 74, 3913-3917, 1977) induced in XC cells the synthesis of several virus polypeptides (the "positive" viruses), in the case of HSV-1 strain F, variants HFEMtsB5 and mPtsHA1 (both tested at 33 degrees), and recombinants F(MP)E and F(MP)F, no virus specific polypeptides could be detected in these cells (the "negative" viruses). (ii) Failure of the "negative" viruses to synthesize polypeptides in XC cells could be explained by failure of the virions to penetrate the cells since polyethylene glycol, a fusion promotor agent, enabled these viruses to overcome the blockage of their expression. (iii) The ability to modulate penetration into XC cells is genetically determined. A locus affecting this function maps between coordinates 0.70 to 0.82 of HSV-1 DNA and is closely linked to Cr, a locus controlling the synthesis or accumulation of virus glycoprotein C (gC) (Ruyechan et al., 1979). (iv) A decrease in the amount of gC, relative to gB, is associated with an enhanced ability to enter XC cells, suggesting that gC may control penetration into these restrictive cells by negatively modulating the gB promoted fusion between host cell and virion membranes.

Animals↗

PubMed1984

Identification of a nuclear protein component of interchromatin granules using a monoclonal antibody and immunogold electron microscopy.

A monoclonal antibody, designated 780-3, has been generated which preferentially recognizes an antigenic component of interchromatin granules in human cells. By indirect immunofluorescence procedures, monoclonal antibody 780-3 produces a cell cycle-specific speckled nuclear staining pattern in adult human fibroblasts which is dramatically altered during metaphase. In contrast, transformed cells appear to express this antigen throughout the cell cycle in increased quantities. Immunogold electron microscopy revealed that the nuclear antigen is intimately associated with interchromatin granules in human cells. Analysis by immunoblot procedures showed that monoclonal antibody 780-3 recognizes two polypeptides of 105 and 41 kD. From these data, a possible nucleolar derivation of interchromatin granules is discussed. These studies demonstrate for the first time that monoclonal antibodies may be used in combination with immunogold electron microscopy to identify the ultrastructural location of nuclear antigens.

Antibodies, Monoclonal↗

PubMed1984

Expression of immediate-early genes in herpes simplex virus type 1-infected XC cells: lack of ICP22 (68K) polypeptide.

The expression of immediate-early (IE) genes of herpes simplex virus type 1 (HSV-1, MP strain) in non-permissive rat XC cells was analysed and compared with the expression of IE genes in permissive HEp-2 cells by the following three methods: analysis of virus polypeptide synthesis in infected cells, Northern blot hybridization between poly(A) nuclear or cytoplasmic RNA and in vitro labelled virus DNA or plasmid-cloned fragments corresponding to IE genes, and ability of poly(A) cytoplasmic RNAs to direct synthesis of virus polypeptides in vitro. ICP4 (175K), ICP0 (110K) and ICP27 (62K) were synthesized in XC cells although in smaller amounts than in HEp-2 cells; ICP4 is functional since early and late polypeptides could be observed. Their corresponding mRNAs were present at low levels in nuclei and in cytoplasm and are functional since the polypeptides were synthesized in a rabbit reticulocyte system. ICP22 (68K) was not detectable in infected XC cells; its mRNA was present in nuclei and in cytoplasm, but it is not functional since the corresponding polypeptide was not synthesized in a rabbit reticulocyte system. This suggests some structural differences in the ICP22 mRNA molecules in infected XC and HEp-2 cells and implicates cellular determinants in the control of the expression of HSV-1 IE genes.

Cell Nucleus↗

PubMed1984

Immunobiochemical characterization with monoclonal antibodies of Epstein-Barr virus-associated early antigens in chemically induced cells.

Five monoclonal antibodies which are reactive to early antigens of Epstein-Barr virus have been produced by using somatic cell hybridization techniques. The specificity of the monoclonal antibodies to early antigens was demonstrated by indirect immunofluorescence, which showed that the antigens were localized to the nucleus of early antigen-induced Raji cells. Additional indirect immunofluorescence studies showed that like patient antisera to diffuse-staining early antigen, the monoclonal antibodies gave positive staining reactions after methanol fixation. One of the antibodies, 1150-4, was positive by the anti-complement immunofluorescence technique but differed with Epstein-Barr virus-associated nuclear antigen-positive patient sera in that it only stained induced cells. Different fixation methods were found to alter dramatically the appearance of the nuclear staining reactions produced by the monoclonal antibodies. Immunoprecipitation and immunoblot experiments revealed that monoclonal antibodies 1108-1 and 1129-1 recognized two polypeptides of 55,000 and 50,000 daltons (p55;50), 1173-6 and 1180-2 recognized just p50, and 1150-4 identified a 65,000-dalton nuclear protein. Immunobiochemical characterization of these viral antigens showed that p55 is a phosphoprotein, and p55;50 has strong DNA-binding activity preferentially to single-stranded DNA. Elucidation of the role of these nuclear proteins in Epstein-Barr virus infection and the events associated with Epstein-Barr virus-directed lymphocyte transformation may provide significant information on the pathogenicity of this important human virus.

Animals↗

PubMed1984

Use of immunogold electron microscopy and monoclonal antibodies in the identification of nuclear substructures.

A cytochemical technique for the ultrastructural localization of unique nuclear antigens is reported. Using a post-embedding indirect immunogold labeling procedure, nuclear antigens in electron-dense regions of the nucleus are localized with a minimum of nonspecific staining. Using this technique and indirect immunofluorescence, a panel of antinuclear monoclonal antibodies is shown to recognize preferentially cell cycle-dependent nuclear substructures. The antigenic domains recognized include specific regions in condensed chromatin, interchromatin granules, euchromatin, and chromosomes. The specificity of antigen recognition is demonstrated with qualitative and quantitative immunogold electron microscopy and immunoblot analysis. These results reveal the existence of previously undefined supramolecular organization within the nucleus and demonstrate the utility of the immunogold procedure when monoclonal antibodies are used.

Animals↗

PubMed1984

Use of a newly established human cell line (SU-CCS-1) to demonstrate the relationship of clear cell sarcoma to malignant melanoma.

A new tumor cell line, designated SU-CCS-1, was established from the malignant pleural effusion of a 16-year-old Caucasian girl with clear cell sarcoma. Morphological studies at the light- and electron-microscopic levels revealed similar features between the SU-CCS-1 cells and the primary tumor. Ultrastructural and cytochemical techniques showed that both the SU-CCS-1 cell line and the original tumor were amelanotic in nature. The malignant derivation of the SU-CCS-1 cell line was demonstrated by intracranial and s.c. heterotransplantation in the nude, athymic mouse and by cytogenetic analysis which showed that the cell line had a hypodiploid chromosome number and several karyotypic abnormalities. Live-cell radioimmunoassay procedures using a large panel of monoclonal antibodies directed against tumor-associated antigens revealed that, phenotypically, SU-CCS-1 closely resembled melanoma tumor cell lines. Immunological assays for the detection of neuroendocrine-associated peptides, hormones, and enzymes revealed that, like melanoma, the SU-CCS-1 cell line was actively producing alpha-melanotropin, S-100 antigen, and nerve growth factor. A notable difference between these tumor types was the capacity of SU-CCS-1 to produce bombesin, an active neuropeptide whose synthesis has been found in cell lines from patients with small cell carcinoma of the lung. From these studies, we concluded that the SU-CCS-1 cell line is phenotypically similar to melanoma, yet displays unique characteristics which distinguishes it from other sarcomas. The availability of an established clear cell sarcoma cell line will greatly facilitate further studies aimed at uncovering the histogenesis of this rare cancer.

Adolescent↗

PubMed1984

Two new monoclonal antibodies (LN-1, LN-2) reactive in B5 formalin-fixed, paraffin-embedded tissues with follicular center and mantle zone human B lymphocytes and derived tumors.

Two monoclonal antibodies (LN-1, LN-2) reactive with B lymphocytes in B5 formalin-fixed, paraffin-embedded tissue sections have been produced by utilizing cell extracts from pokeweed mitogen-stimulated peripheral blood lymphocytes and diffuse histiocytic lymphoma SU-DHL-4 cells, respectively. Both monoclonal antibodies were initially identified by indirect immunofluorescence screening techniques on paraformaldehyde-acetone-fixed cell preparations. Specificity screens with 36 well-characterized human lymphoma and leukemia cell lines showed that both LN-1 and LN-2 stained cell lines of B cell lineage but were unreactive with those of T cell or, with one exception, myeloid derivation. Null cell acute lymphoblastic leukemia cell lines were found to be LN-2+ but LN-1-. The B cell specificity of these reagents was confirmed on 15 lymphoma and 17 leukemia biopsy specimens by using indirect immunofluorescence techniques. Immunoperoxidase staining of sections from B5-fixed, paraffin-embedded human lymphoid tissues showed that LN-1 bound to the cell membrane and cytoplasm of germinal center cells whereas LN-2 stained the nuclear membrane and cytoplasm of germinal center and mantle zone B lymphocytes as well as interfollicular histiocytes and thymic medullary dendritic cells. Both monoclonal antibodies failed to stain cortical thymocytes, lymph node T cells, and peripheral blood T and myeloid cells. Immunoperoxidase staining of 20 nonlymphoid human organs and tissues revealed that LN-1 reacted positively with red blood cell precursors of the bone marrow, ciliated epithelial cells of the bronchus, distal tubular cells of the kidney, and ductal cells from several organs including the breast and prostate. In contrast, LN-2 was unreactive with all human nonlymphoid organs and tissues including the bone marrow. Indirect immunofluorescence staining of a panel of 26 solid tumor cells lines showed that LN-1 was reactive with the majority of epithelium-derived cell lines, glioblastomas, and astrocytomas but was unreactive with neuroblastomas, small cell carcinoma of the lung, and sarcomas. LN-2 was unreactive with 25 of 26 of the solid tumor cell lines by these techniques. Immunobiochemical studies have shown that LN-1 recognizes a cell surface sialoantigen whereas LN-2 is directed against a 35,000 dalton nuclear membrane protein. Because of their high specificity for B cell tumors and their ability to stain B5-fixed, paraffin-embedded tissues, LN-1 and LN-2 are useful reagents for the diagnosis and classification of the human lymphomas and leukemias.

Antibodies, Monoclonal↗

PubMed1984

Phenotypic analysis of established diffuse histiocytic lymphoma cell lines utilizing monoclonal antibodies and cytochemical techniques.

Ten diffuse histiocytic lymphoma (DHL) cell lines were extensively characterized with monoclonal antibodies and histochemical techniques. The original biopsy specimens, representing nine of ten cases from which the cell lines were derived, were reviewed utilizing the International Working Formulation. Eight of ten cell lines reacted with anti-immunoglobulin reagents and/or a subset of B-lymphocyte surface markers, supporting a B-cell derivation. Only U-937, a monocytoid DHL cell line reactive with OKT4 and 6, displayed any T-cell markers. Cytochemical analysis alone proved to be of little value in the subclassification of the DHLs. The pathologic review revealed that, despite disparate immunologic phenotypes, five of the diffuse large cell lymphomas were subclassified as large, noncleaved lymphomas. Our analysis confirms the phenotypic diversity of this subgroup of malignant lymphomas and underscores the value of monoclonal reagents for the immunologic evaluation of the hematologic malignancies. These well characterized cell lines constitute a valuable resource for the laboratory investigation of the lymphomas.

Adult↗

PubMed1983

Virus polypeptide synthesis induced by herpes simplex virus in non-permissive rat XC cells.

The synthesis of virus polypeptides in rat XC cells infected with herpes simplex virus type 1 (HSV-1; 13VB4tsC75) was studied. At the permissive temperature the virus induced the synthesis, in a cascade fashion, of significant amounts of several early polypeptides (ICP 6, 8 and 39) and those late polypeptides that are relatively resistant to inhibition by phosphonoacetic acid in HEp2 cells (ICP 5, 11, 25, 29, 43 and 44). The infectious cycle appeared to become arrested in XC cells at about 7 to 9 h postinfection, because the relative concentrations of early and latest polypeptides labelled thereafter remained constant and the levels of several of the late virus polypeptides were severely reduced (ICP 2, 10, 24 and 26) or not synthesized at all (ICP 32, 34 and 37). When XC cells were infected at a very high m.o.i., only a small amount of virus DNA synthesis could be detected; the synthesis of cellular DNA was not impaired and the infected XC cells continued to replicate for several weeks at least. When XC cells were infected at the non-permissive temperature, only the immediate-early (IE) ICP 4 could be detected while IE ICP 0 and 22 were not observed. Infection of XC cells with HSV-1 (MP) also resulted in the production of early and late viral polypeptides. On the other hand, in XC cells infected with HSV-1 (F) and HSV-1 (HFEM), the synthesis of virus polypeptides could not be detected.

Animals↗

PubMed1983

Estrogen receptor analysis in chronic lymphocytic leukemia.

Estrogen receptor (ER) determinations were performed on cytosol preparations of Ficoll-Hypaque density separated mononuclear cells from 11 patients with chronic lymphocytic leukemia (CLL). The presence of ER was noted in 8 of 11 specimens (73%). ER ranged from 431 fmole/mg to 4.3 fmole/mg cytosol protein. Two types of receptor subunits were observed at the 8S and 4S region of the sucrose gradient. In addition, 1 of 3 Epstein-Barr virus (EBV) transformed B-lymphoblastoid cell lines from healthy donors had a measurable amount of ER. Patient R.L., who was refractory to standard chemotherapy and radiation and was ER positive, experienced a minor response to Tamoxifen therapy, with subsequent loss of ER. The demonstration of ER in CLL suggests that this malignancy may have a hormone-dependent subpopulation of cells.

Aged↗