PubMed Health⌕ Search

Biomedical subjects

E Lakow

Publications and source records attributed to E Lakow.

7 recordsLinked to original sources
PubMed1984

Isolation of highly malignant Thy-1-positive revertants from cultured cloned Thy-1-negative lymphoma cells of low tumorigenicity.

We have used a positive immunoselection method to isolate a Thy-1-positive revertant of a Thy-1-negative line. The technique makes use of the ability of surface bound antigen-antibody complexes containing the enzyme catalase to protect cells from the toxic effects of hydrogen peroxide. The revertant cells produce a Thy-1 molecule indistinguishable from that produced by the original parent line, and they resemble that line in being far more tumorigenic than the Thy-1-negative variant.

Animals↗
PubMed1983

Cell separation using positive immunoselective techniques.

Positive immunoselection is the direct selection and recovery of cells which express a given specificity from among a heterogeneous group of contaminating cells. A variety of methods are available to effect such separations. The principles of affinity chromatography, using solid-phase matrices or cellular immunoadsorbents, are extensively used. Liquid-phase positive immunoselection can also be performed using either a fluorescence-activated cell sorter or by using 'cellular engineering' to protect a cell from an otherwise noxious environment. The enzyme catalase coupled to specific antibody has been used for this purpose and renders cells resistant to hydrogen peroxide. The various positive immunoselection techniques available are reviewed and evaluated in the following report.

Animals↗
PubMed1983

Human T cell hybridomas specific for Epstein Barr virus-infected B lymphocytes.

A mutant of the Jurkat human T lymphoblastoid cell line deficient in hypoxanthine phosphoribosyltransferase, and resistant to ouabain, was fused with peripheral blood T lymphocytes primed in vitro with Epstein Barr virus- (EBV) transformed autologous B lymphocytes. After selection of somatic cell hybrids and cloning, hybridoma cell lines were obtained that reacted with autologous EBV-infected B lymphocytes, as detected by the release of interleukin 2 into the culture medium. The hybridomas did not react with i) EBV-uninfected autologous or allogeneic B lymphocytes, ii) three out of four allogeneic EBV-transformed cell lines, or iii) two established EBV-negative B cell lines. These functional hybridomas may ultimately prove useful in dissecting the means by which human T lymphocytes recognize and regulate EBV infection in vivo.

B-Lymphocytes↗
PubMed1982

Possible metabolic basis for the different immunodeficient states associated with genetic deficiencies of adenosine deaminase and purine nucleoside phosphorylase.

An inherited deficiency of adenosine deaminase (Ado deaminase; adenosine aminohydrolase, EC 3.5.4.4) causes severe combined immunodeficiency disease in humans. A similar deficiency in purine nucleoside phosphorylase (Puo phosphorylase; purine-nucleoside:orthophosphate ribosyltransferase, EC 2.4.2.1) engenders a selective cellular immune deficit. To elucidate the possible metabolic basis for the contrasting immunologic phenotypes, we compared the toxicity toward mature resting human lymphocytes of the Ado deaminase substrates deoxyadenosine and adenosine and the Puo phosphorylase substrate deoxyguanosine. When Ado deaminase was inhibited, micromolar concentrations of deoxyadenosine progressively killed nondividing helper and suppressor-cytotoxic T cells, but not B cells. The toxicity required phosphorylation, with subsequent dATP formation. The deoxyadenosine analogs 2-chlorodeoxyadenosine, 2-fluorodeoxyadenosine, and adenine arabinonucleoside also killed resting T cells. Cell death was unrelated to inhibition of adenosylhomocysteinase (EC 3.3.1.1) but was preceded by a gradual decline in ATP levels. As much as 1 mM deoxyguanosine did not impair resting lymphocyte viability, despite the synthesis of dGTP. The combination of 200 microM adenosine plus 500 microM homocysteine thiolactone killed dividing lymphocytes but had no discernible toxic effect toward resting T cells, which accumulated adenosylhomocysteine over a 4-hr period but thereafter excreted the nucleoside into the culture medium. The different clinical syndromes associated with genetic deficiencies of Ado deaminase and Puo phosphorylase may be explained by the ability of dATP to kill mature resting T lymphocytes by depleting ATP levels.

Adenosine↗
PubMed1981

Positive immunoselection using antibody-enzyme complexes.

We have developed an immunoselection technique using catalase-anti-catalase complexes coupled to specific antibodies to protect antigen positive target cells from the lethal effects of H2O2. The antibody-enzyme complexes are bound to the target cells through an antibody bridge with specificity for both the complexes and an anti-target cell antibody. In a model system, Thy-1 positive (RL male 1-3) cells were protected by incubation with rabbit anti-brain-associated theta antigen (BAT), sheep anti-rabbit IgG [F(ab)'2] and catalase-anti-catalase complexes. The amount and composition of the complexes adhering to the cells were measured by dual radiolabeling of the catalase and anti-catalase immunoglobulin. This technique provides a means of identifying and isolating large numbers of cells bearing any antigen for which specific antisera are available.

Animals↗