Biomedical subjects

C A Lamb

Publications and source records attributed to C A Lamb.

6 recordsLinked to original sources

PubMed2001

Five novel hormone-responsive cell lines derived from murine mammary ductal carcinomas: in vivo and in vitro effects of estrogens and progestins.

We have developed an experimental model of mammary carcinogenesis in which the administration of medroxyprogesterone acetate (MPA) to female BALB/c mice induces progestin-dependent ductal metastatic mammary tumors with high levels of estrogen receptor (ER) and progesterone receptor (PR). Through selective transplants in untreated mice, we have obtained progestin-independent variants, still expressing high levels of ER and PR. Primary cultures of the MPA-induced carcinomas C4-HD and C7-HI were set up, and after 3-4 months, several different cell lines were obtained. Four of these, MC4-L1, MC4-L2, MC4-L3, and MC4-L5 were established from C4-HD and a fifth, MC7-L1, from C7-HI. All cells were of epithelial origin, as demonstrated by electron microscopy and by immunocytochemical identification of cytokeratin and cadherin. In vitro MC4-L1, MC4-L3, and MC4-L5 showed a typical epithelial morphology; when transplanted in vivo, they originated metastatic carcinomas with different degrees of differentiation. MC4-L2 and MC7-L1 deviated from the standard epithelial picture; they disclosed a spindle-shaped morphology in vitro and in vivo gave rise to a biphasic spindle cell/tubular carcinoma and an anaplastic carcinoma, respectively; both lines gave rise to metastases. This differential morphology correlated with a higher degree of aggressiveness, as compared with MC4-L1, MC4-L3, and MC4-L5. ERs and PRs were detected by binding, immunocytochemistry, and Western blot. In vitro, MC4-L2 and MC7-L1 were stimulated by MPA (nM to microM) and 17beta-estradiol (nM and 10 nM); no significant stimulation was observed in MC4-L1, MC4-L3, and MC4-L5 under the same experimental conditions. In vivo, MPA significantly stimulated tumor growth in all epithelioid lines but not in MC4-L2 and MC7-L1. A progestin-dependent growth pattern was confirmed for MC4-L1, MC4-L3, and MC4-L5 in successive transplants, whereas MC4-L2 and MC7-L1 behaved as progestin independent. This is the first description of mouse mammary carcinoma cell lines expressing ER and PR. The different in vitro hormone responses as compared with in vivo and the differential effects of 17beta-estradiol in the parental tumors and in cell lines render these lines useful tools for the in vitro and in vivo study of hormone regulation of tumor growth and metastases.

Animals↗

PubMed2001

What causes the isolation effect?

Events that are incongruent with their prevailing context are usually very well remembered. This fact often is described as the distinctiveness effect in memory, an effect that has served as explanation not only of memory phenomena but also of various other phenomena, including social judgment. The core laboratory paradigm for studying distinctiveness in memory research has long been the isolation paradigm. This paradigm, sometimes attributed to H. von Restorff, yields better memory for an item categorically isolated from surrounding items than for the surrounding items and a proper control item. The authors offer an interpretation of the isolation effect based on the analysis of the processing of similarities and differences among the items. Two experiments provide evidence for this interpretation. The results are discussed in the context of current theories of distinctiveness effects in memory. An appeal is made for a different conceptualization of distinctiveness effects, one that treats distinctiveness as a discriminative process in memory that requires processing of both similarities and differences among items.

Adult↗

PubMed1994

Transport properties of free and MHC class II-associated oligomers containing different isoforms of human invariant chain.

Major histocompatibility complex (MHC) class II molecules are heterodimers of alpha and beta subunits that associate intracellularly with the invariant chain. Human invariant chain exists in four forms, p33, p35, p41, and p43, generated by a combination of alternative initiation of translation and alternative splicing. The biological significance of the existence of the different forms of invariant chain is still unclear and to date no study has compared all four using one system. We have compared them for their transport characteristics and for their ability to transport associated MHC class II heterodimers into the endocytic pathway. Here we report that hetero oligomers containing p33 and p35 or p41 and p43 remain in the endoplasmic reticulum (ER) in the absence of class II alpha and beta chains. This is consistent with earlier reports suggesting that the N-terminal extension shared by p35 and p43 contains an ER retention signal. Homo oligomers containing only the p33 or p41 forms of invariant chain exit the ER and are sorted to endosomes following passage through the Golgi apparatus. Their accumulation leads to enlargement of the endosomes. Quantitation of the turnover rates of the p35/p33 forms with the alternatively spliced p43/p41 forms indicates that the latter are more stable, both in the ER and following transport through the Golgi apparatus. When class II molecules are co-expressed with p33 and p35, or p41 and p43, the assembled complex is efficiently transported to the endocytic pathway.

Amino Acid Sequence↗

PubMed1992

Assembly and transport properties of invariant chain trimers and HLA-DR-invariant chain complexes.

The MHC class II-associated invariant chain behaves as a resident endoplasmic reticulum protein in the absence of class II molecules. In humans, two predominant forms exist; one, p35, differs from the other, p33, by an N-terminal cytoplasmic extension of 16 amino acids that contains a strong endoplasmic reticulum-retention signal. Here we show that one mechanism for retention of p33 is its association with p35 in mixed invariant chain trimers. However, even for p33 homotrimers transport from the endoplasmic reticulum is inefficient. In an MHC class II-positive B cell line, the formation of invariant chain trimers is rapid and is the first intermediate in the assembly of a nine-chain alpha beta-invariant chain complex. With time, three higher molecular weight complexes are progressively formed. These correspond to an invariant chain trimer with one alpha beta dimer, two alpha beta dimers, and three alpha beta dimers, respectively. No free alpha beta dimers are detectable early in biosynthesis. However, beginning at 2 h of chase, alpha beta dimers begin to appear concomitant with the disappearance of the completely assembled alpha beta-invariant chain complex. This conversion is virtually complete by 4 h, and presumably reflects the proteolytic degradation of the invariant chain component of the alpha beta-invariant chain complex and the generation of endosomal alpha beta dimers capable of binding antigenic peptides.

Antigens, Differentiation, B-Lymphocyte↗

PubMed1991

Invariant chain targets HLA class II molecules to acidic endosomes containing internalized influenza virus.

The role of the HLA class II-associated invariant chain in the intracellular trafficking of HLA-DR molecules was examined in a transient expression system using HeLa cells. In the absence of alpha and beta polypeptides, invariant chain was retained in the endoplasmic reticulum (ER). In the absence of invariant chain, intracellular alpha beta heterodimers could be detected only in the ER and the Golgi apparatus. However, when alpha and beta subunits were coexpressed with invariant chain, HLA-DR molecules were detectable in peripheral cytoplasmic vesicles, which also contained invariant chain. In addition, an antibody directed to an acid-induced conformational determinant on the influenza hemagglutinin molecule detected internalized influenza virus in the HLA-DR-containing vesicles. These findings provide direct evidence that the invariant chain targets class II molecules to an acidic endosomal compartment and that this compartment, long suspected to be the site of antigen processing, is the site where class II molecules interact with natural antigen.

Antibodies↗

PubMed1990

Intracellular transport and peptide binding properties of HLA class II glycoproteins.

Protein antigens internalized by an antigen presenting cell are degraded into peptides, a subset of which binds to the class II glycoproteins encoded by the major histocompatibility complex to form epitopes recognized by specific T cells. Current evidence suggests that the immunogenic peptides are generated in an endosomal, acidic compartment containing internalized antigen, proteinases, and exocytic class II molecules. These exocytic class II glycoproteins are associated during transport from the endoplasmic reticulum to the endosomal compartment with an additional glycoprotein, the invariant chain. Proteolytic degradation of the invariant chain in the endosomal compartment dissociates it from the class II glycoproteins, which only then acquire the capacity to bind peptides. After peptide binding occurs, the class II-peptide complexes are transported to the antigen-presenting cell surface for recognition by T cells.

Biological Transport, Active↗