Fusion of P3 X 63 Ag8 mouse myeloma cells with splenocytes obtained from mice immunized with cells derived from human colorectal carcinomas resulted in the production of antibody-secreting hybridomas. Two hybridomas (1083-17 and 1116-56) and their clones secreted antibodies binding specifically to human colorectal carcinoma cells either grown in culture or obtained from patients, but did not bind to normal colonic mucosa or other normal and malignant human cells. The binding specificity was consistent in three assays: radioimmunoassay, mixed hemadsorption, and immunofluorescence. Adsorption of these antibodies to colorectal carcinoma cell lines totally eliminated their specific binding.

https://www.pnas.org/content/pnas/76/3/1438.full.pdf

Colorectal carcinoma-specific antigen: detection by means of monoclonal antibodies

Literature regarding the biochemistry of aluminum and eight similar ions is reviewed. Close and hitherto unknown similarities were found. A hypothetical model is presented for the metabolism, based on documented direct observations of Al3+ and analogies from other ions. Main characteristics are low intestinal absorption, rapid urinary excretion, and slow tissue uptake, mostly in skeleton and reticuloendothelial cells. Intracellular Al3+ is probably first confined in the lysosomes but then slowly accumulates in the cell nucleus and chromatin. Large, long-lived cells, e.g., neurons, may be the most liable to this accumulation. In heterochromatin, Al3+ levels can be found comparable to those used in leather tannage. It is proposed that an accumulation may take place at a subcellular level without any significant increase in the corresponding tissue concentration. The possible effects of this accumulation are discussed. As Al3+ is neurotoxic, the brain metabolism is most interesting. The normal and the lethally toxic brain levels of Al3+ are well documented and differ only by a factor of 3-10. The normal brain uptake of Al3+ is estimated from data on intestinal uptake of Al3+ and brain uptake of radionuclides of similar ions administered intravenously. The uptake is very slow, 1 mg in 36 years, and is consistent with an assumption that Al3+ taken up by the brain cannot be eliminated and is therefore accumulated. The possibility that Al3+ may cause or contribute to some specific diseases, most of them related to aging, is discussed with the proposed metabolic picture in mind.

Metabolism and possible health effects of aluminum.

We describe a new monoclonal murine antibody that reacts with a 50,000-mol wt polypeptide that appears to be present on all E-rosetting cells. We conclude that this antigen is either identical to or closely associated with the E receptor because of (a) the high degree of concordance between E-rosette formation and 9.6 antigen expression, (b) the inhibition of rosette formation by preincubation of cells with 9.6 antibody, and (c) the observed failure of cells lysostripped of 9.6 antigen to form E-rosettes. This last finding suggests cocapping of 9.6 antigen and the E receptor.

/pdf/identification-of-a-human-t-lymphocyte-surface-protein-3lfyc4wiwx.pdf

Identification of a human T lymphocyte surface protein associated with the E-rosette receptor.

We describe here two new monoclonal antibodies that react with surface antigens of human lymphocytes. Antibody 7.2 identified a determinant on the framework region of the human Ia antigen. It was cytotoxic for all cultured B-cell lines, normal B cells, and monocytes. The antibody was not cytotoxic for normal T cells or for established T leukemic cell lines. In immune precipitation assays, the 7.2 antibody reacted with a bimolecular complex of two chains that resolved in polyacrylamide gels as polypeptides with molecular weights of 29000 and 34000 daltons. These precipitation results were analogous to those achieved with a rabbit antiserum prepared against human Ia antigens. Antibody 9.3 identified a determinant on the framework region of a T-cell antigen. It was cytotoxic for 50–80% of peripheral T cells and for 20–50% of thymocytes. It was not cytotoxic for cultured B-cell lines, normal B cells, or monocytes. In immune precipitation assays, the 9.3 antibody reacted with a single polypeptide with a molecular weight of 44000 daltons. Due to the expression of this antigen on a limited subpopulation of human T cells, we have designated the antigen HuLyt-1.

Monoclonal antibodies identifying a novel T-Cell antigen and Ia antigens of human lymphocytes

Conjugates of tumor-reactive antibody and toxins (immunotoxins) have been used to eradicate tumor cells in vitro and in vivo. Such immunotoxins are highly effective in killing murine leukemic cells...

Immunotoxins: A new approach to cancer therapy

Multiple lymphoid cell lines were derived from 35HLA-D homozygous donors by EB-viral transformation of B lymphocytes. The expression of Ia-like alloantigens (HLA-DR) was studied by microcytotoxicity and by absorption with alloantisera exchanged through the Seventh Histocompatibility Workshop. B-lymphoid lines expressed the same specificities as normal B lymphocytes. Workshop antisera representing DRw1, DRw2, DRw3, and DRw7 gave well-defined typing patterns with cell lines derived from donors of corresponding D-locus specificities. A more complex reaction pattern was seen for antisera representing DRw4, DRw5, and DRw6. The available reagents could not discriminate between lines from donors homozygous for Dw4, Dw10, or D-KH. All lines studied, except for those from one donor homozygous for a unique D-locus determinant (D-SPO), could be assigned one of the provisional DRw specificities. The advantage of obtaining multiple cell lines from a single donor was evident. One line could not be typed by microcytotoxicity because it was lysed in all human sera tested, and some other lines gave weak cytotoxic reactions. Absorption studies, however, did indicate similar expression of DRw antigens on these lines. The availability of multiple lines from the same donor circumvented these difficulties.

B-Lymphoid cell lines derived fromHLA-D homozygous donors

Chromosome analyses have revealed abnormalities in many cases of lymphoma, non-lymphocytic leukaemia and acute lymphoblastic leukaemia. Most studies on chronic lymphocytic leukaemia (CCL), however, have shown normal karyotypes1–4 although chromosomal abnormalities have been reported5,6 . In most of these studies specific surface marker analyses on leukaemic lymphocytes have not been carried out. It has been difficult to establish whether the metaphases seen in the majority of the CLL studies were from normal or from leukaemic cells. Recently, in one of our laboratories special interest has centred on cases of CLL with monoclonal serum protein bands; in two such cases with immunoglobulin M (IgM) bands, it was demonstrated with idiotypic antisera against the serum IgM band that the leukaemic lymphocytes were the precursors of plasma cells responsible for the monoclonal serum IgM7,8. In addition, the leukaemic lymphocytes of these patients could be stimulated in vitro to divide and to differentiate to plasma cells8. Epstein–Barr virus (EBV)-transformed lymphoblastoid cell lines were obtained from the two patients. With the idiotypic determinants as leukaemia specific markers, certain of these lines were shown to be derived from leukaemic cells9. Thus, rapidly proliferating populations of cells, which could be identified with certainty as being of CLL origin, were available for chromosome analysis. In the present study, conventional G- and Q-banding techniques for karyotypic analysis were carried out on the leukaemic lymphocytes induced to divide in vitro and on the cells of the leukaemic lymphoblastoid lines. Aneuploid karyotypes were found in the leukaemic B cells of both patients.

Chromosome abnormalities of leukaemic B lymphocytes in chronic lymphocytic leukaemia

Multiple lymphoblastoid cell lines have been derived from two patients with chronic lymphocytic leukemia with an associated monoclonal immunoglobulin (Ig) band. Idiotypic antisera raised against the monoclonal serum Ig bands were shown to be specific for the membrane Ig of the patients' leukemic cells. The idiotypic determinants in these patients thereby constitute tumor-specific antigens. Surface and intracellular immunofluorescence studies utilizing these idiotypic antisera were used to identify the cell lines of leukemic origin. These studies showed that certain cell lines from each patient were derived from the leukemic cells while other cell lines were derived from residual normal B lymphocytes. The leukemic cell lines were variable and contained different percentages of lymphoid cells with the idiotype-specific membrane Ig and, in addition, different percentages of plasma cells with intracellular Ig of the same specificity. Specific Ig synthesis was also demonstrated by hemagglutination-inhibition analysis of cell line supernatants. Aside from Ig specificity, no differences have been found between the leukemic cell lines and those derived from normal cells. One of the leukemic cell lines was cloned in soft agarose. All the clones were shown to be of leukemic origin.

http://www.pnas.org/content/75/11/5706.full.pdf

Lymphoblastoid cell lines from patients with chronic lymphocytic leukemia: identification of tumor origin by idiotypic analysis.

A group of very similar cell lines was established from peripheral blood or bone marrow of 12 patients with a variety of disorders. The cells in these cell lines were uniform and round in shape. They grew as single-cell suspensions or as aggregates of small numbers of cells in stationary culture. The most striking characteristic of these lines was the lack of cells with surface immunoglobulin or with demonstrable immunoglobulin synthesis. This lack of immunoglobulin synthesis and their special growth characteristics distinguished them from the lymphoblastoid B cell lines previously described. The cells of these unusual cell lines had strong Fc receptors and C3 receptors and expressed Ia antigens. They did not form rosettes with sheep erythrocytes and did not have detectable levels of terminal deoxynucleotidyltransferase. They did not secrete lysozyme and failed to stain for peroxidase. The presence of the Epstein-Barr virus nuclear antigen in the cells indicated the presence of Epstein-Barr viral genome. The possibility that these cells represent some type of precursor cell in the B cell lineage is discussed, but the exact cellular origin remains to be ascertained.

Human cell lines containing Epstein-Barr virus but distinct from the common B cell lymphoblastoid lines

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B-Lymphoid cell lines derived fromHLA-D homozygous donors

Chromosome abnormalities of leukaemic B lymphocytes in chronic lymphocytic leukaemia

Lymphoblastoid cell lines from patients with chronic lymphocytic leukemia: identification of tumor origin by idiotypic analysis.

Human cell lines containing Epstein-Barr virus but distinct from the common B cell lymphoblastoid lines