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Biomedical subjects

M Cohn

Publications and source records attributed to M Cohn.

At least 91 records · Page 5Linked to original sources

T lymphoma variants with specifically altered growth in semi-solid media.

The relationship between the tumorigenic potential and the cloning efficiency of T lymphoma BW5147 in semi-solid media has been studied. Two stable variants exhibiting a 30-fold decrease of their cloning efficiencies in agarose an methylcellulose media were independently isolated by negative selection with FuDr. These variants show no alteration of their growth properties in liquid medium and are still able to proliferate in liquid suspension over a bottom layer of agarose. This new phenotype is not correlated with any decreased tumorigenicity in syngeneic AKR/J mice.

Animals

Induction of lambda 1-immunoglobulin is determined by a regulatory gene (r lambda 1) linked (or identical) to the structural (c lambda 1) gene.

The cis-acting gene regulating specifically the inducibility of lambda 1-bearing B cells has been mapped within 2.9 cM of the structural gene. If the lambda 1lo-phenotype is due to the gly leads to val interchange in C lambda 1, then an argument can be made that (a) the lambda 1lo-phenotype is due to inefficient induction of lambda 1lo-bearing B cells and (b) B cell triggering is dependent upon a conformational change in the Ig receptor upon interaction with antigen. If the lambda 1lo-phenotype is due to a regulatory sequence linked to the structural C lambda 1-gene, then it must control the expression of the lambda 1-locus during development into adulthood, e.g., by an effect on methylation.

Animals

Evidence that cytotoxic T cells and natural cytotoxic cells use different lytic mechanisms to lyse the same targets.

There is evidence that natural cytotoxic (NC) cells are in the T cell lineage. To lyse targets, cytotoxic T cells (TK) must recognize any of the myriad antigens plus syngeneic major histocompatibility complex (MHC) determinants. In spite of the evidence which indicates TK and NC are in the same lineage, NC cells can recognize few determinants (perhaps only one) and do not require recognition of MHC determinants. In addition to differences in the requirement for target recognition, in this report we show that TK cells and NC cells also use different lytic mechanisms to lyse the same targets. NC effectors initiate a lytic mechanism in NC-sensitive and NC-resistant targets. This lytic mechanism requires approximately 4 h before target lysis is apparent in NC-sensitive targets; it is inactivated by a protein synthesis-dependent counterlytic mechanism in NC-resistant targets. In contrast the TK lytic mechanism causes a rapid release of 51Cr from both NC-sensitive and NC-resistant targets and is not inhibited by the NC counterlytic mechanism present in NC-resistant cells. These findings lead to the conclusion that the mechanism used by NC cells to lyse targets is fundamentally different from that used by TK cells.

Animals

The T-cell receptor mediating restrictive recognition of antigen.

Four facts characterize restrictive recognition of antigen. First, in large measure, allele-specific determinants on R are recognized when R is functioning either as a restricting element (RL) or as an allo-target (or even xeno-target) (RF). Second, there is a high frequency of virgin antigen-responsive t cells with alloreactivity, i.e. anti-RF. Third, there is a strict relationship between the class of effector function and the class of RL recognized (restrictive recognition of antigen, XF) but a relaxed relationship between class of effector function and class of RF recognized (alloreactivity). Fourth, the effector T cell functions anti-RL-dependently when XF is the target (restrictive recognition of antigen) and anti-RL-independently when RF is the target (alloreactivity). From these facts are derived the following conclusions. The T cell uses a dual recognitive, single receptor (Model I, Figure 1). A single germ-line VT locus specifying anti-allele-specific recognition of species R encodes both the anti-R and the anti-X combining sites. A "learning" process (occurring in the thymus) is required to establish the restriction specificity (anti-RL) as well as the effector function/class of RL relationship. The repertoire is derived by somatic mutation of all germ-line VT genes specifying anti-RF (Model IA, Table 3 and Figure 9). Given Model IA (Table 3 and Figure 9), we can account further for the existence of an extensive polymorphism of R and minimal polygeneism, for the high frequency of crossreactivity between anti-XF and RF, and for the physiology and genetics of cell-cell communication in immune responsiveness.

Animals

Monoclonal antibodies to murine immunoglobulin isotypes.

We have obtained cell lines producing monoclonal antibodies against mu, alpha, lambda, and lambda 1 by fusing the mouse myeloma P3X63Ag8.653 with spleen cells from Lewis rats immunized with several lambda 1 bearing myeloma proteins. The anti-lambda 1 antibody is particularly useful. It was shown to be suitable for radioimmunoassays, for ELISAs when coupled to alkaline phosphatase, and for indirect fluorescence staining of the membrane as well as the cytoplasm.

Animals

The WI-L2-729-HF2 human hybridoma system. Stable hybrids at high frequency.

A new system for the rapid production at high frequency (greater than 10(-5] of stable human hybridomas is described. This system is based on a high efficiency fusion variant human lymphoblastoid cell line designated WI-L2-729-HF2 and is comparable in many ways to commonly used murine hybridoma systems. WI-L2-729-HF2 cells fuse at high frequency with mitogen-stimulated human b cells resulting in the rapid appearance (within 2 weeks) of stable hybridomas in hypoxanthine/aminopterin/thymidine (HAT) medium. Whereas the WI-L2-729-HF2 cell line secretes only trace levels (50 to 100 ng/ml) of an IgG,kappa and has surface IgM,kappa, HAT-resistant hybridomas secrete high levels (10 to 20 micrograms/ml) of new human immunoglobulins, including immunoglobulins containing alpha or lambda chains not found in the tumor parental cell line. These hybridomas have remained stable for over four months in continuous culture, secreting high levels of new immunoglobulins in both conventional and serum-free medium, a feature which makes possible large-scale production and purification of human antibodies. This system has the potential to provide tools and reagents for investigations involving, for example, the diagnosis and treatment of human autoimmune disease and cancer.

B-Lymphocytes

An analysis of the sensitivity of somatic cell hybrids to natural killer cell- and natural cytotoxic cell-mediated lysis.

The analysis of the NK and NC sensitivity of somatic cell hybrids formed between parental cell lines that differ in their NK and NC sensitivity has shown the following. 1) The dominant expression of both NK and NC recognition determinants on target cells; 2) the dominant expression of two post-recognitive NC resistance mechanisms, one requiring protein synthesis and one being protein synthesis independent; and 3) the dominant expression of a post-recognitive NK resistance mechanism, which is protein synthesis independent. The post-recognitive protein synthesis-independent NC resistance mechanism confers no NK resistance and the post-recognitive NK resistance mechanism confers no NC resistance. Whether the post-recognitive protein synthesis-dependent NC resistance mechanism confers NK resistance remains open to question. The analysis of the hybrids indicates that transformed cells become sensitive to either NK- or NC-mediated lysis by losing their resistance to the lytic activity of these effector cells, and it appears that differentiation plays a role in determining whether NK or NC resistance will be lost upon transformation. A model is proposed in which the differentiation into a fibroblast associates the loss of NC resistance with transformation, whereas the differentiation into a lymphocyte associates the loss of NK resistance with transformation. Because the loss of NK resistance is not associated with the transformation of fibroblasts, they remain NK resistant, and because the transformation of lymphocytes is not associated with the loss of NC resistance, they remain NC resistant. This provides the basis for the target specificity exhibited by NK and NC effectors.

Animals

In vivo surveillance of tumorigenic cells transformed in vitro.

Any theory of surveillance against cancer requires that cells susceptible to host protective mechanisms exist as intermediates on the pathway from normal to cancer. The failure to demonstrate a significant frequency of such intermediates as a result of chemical carcinogenesis has cast serious doubt on the validity of the surveillance hypothesis. Here we report the conditions in which such intermediates can be identified as the major class of transformed cells resulting from in vitro chemical carcinogenesis of a cloned fibroblastic cell line.

Benzopyrenes

Immunological identification of cerebellar cell lines.

Spleen cells from BALB/c mice, previously immunized with rat cerebellar tissue, were fused to the mouse myeloma cell line SP2/0-Ag14 and the cerebellar cell type specificity of the resultant hybridomas determined. In this report we describe the specificity of one hybridoma, C4/12. Monoclonal antibodies secreted by this hybridoma recognize granule cell neurons in adult cerebellar frozen sections, and in primary cultures started from 3 to 5-day-old newborn rats. In addition, C4/12 recognizes a subclass of astrocytes when screened on primary cultures but not adult cerebellar tissue. Two temperature sensitive Rous sarcoma virus transformed cerebellar cell lines, previously shown to be either neuronal or glial, were screened for the presence of the antigen. Both cell lines are positive at the temperature permissive for transformation, whereas the glial line but not the neuronal line exhibits the antigen at the nonpermissive temperature. These results are discussed in light of the cell lines being representative of precursor cells.

Animals