Release of soluble transferrin receptor from the surface of human leukemic HL60 cells

Information regarding transferrin (Tf) receptor degradation is largely incomplete. HL60 cells were shown to release to their growth medium a Tf-binding protein which could be immunoprecipitated by anti-Tf receptor monoclonal antibodies (MoAbs) B3/25 and OKT9. Soluble Tf receptor was detected in the medium within one hour of replating of cells, and its release was inhibited at 4 degrees C. The affinity of Tf for the soluble receptor released by cells (kd = 2.3 x 10(-10) mol/L) was slightly lower than its affinity for the detergent-solubilized cellular receptor (kd = 1.2 x 10(-10) mol/L). 125I-Tf internalized and released by cells subsequently bound to Tf receptor released by the same cells, and soluble Tf receptor in the conditioned medium (CM) inhibited 125I-Tf binding to intact cells. The soluble Tf receptor isolated from the CM was smaller (78,000 daltons) than the cell surface receptor (94,000 daltons) when analyzed by gel electrophoresis under reducing conditions. Isolated cell membranes readily released soluble receptor; however, this release could be blocked by protease inhibitors. The soluble Tf receptor may represent the extracytoplasmic domain of the cellular Tf receptor released from the surface of HL60 cells through proteolytic cleavage by a membrane-based protease.     

Development of neoplastic phenotype following transfection of HNF cells with sarcoma DNA

DNA isolated from chondrosarcoma cells effectively transformedNIH-3T3 cells and human foreskin fibroblasts. The transfectedNIH-3T3 cells, directly implanted three or four passages later,formed progressively growing tumors ( 2.0 cm in diameter) subcutaneouslyin nude mice. No metastasis was evident upon pathological examinationof the tumor bearing mice. Transfected human foreskin fibroblaststhat exhibited anchorage independent growth formed only smalltumors in nude mice (<0.6 cm in diameter). The transfectedhuman cells which exhibited anchorage independent growth reactedwith the monoclonal antibody 345.134S, specific for an epitopeexpressed by human sarcoma cells. The transfected NIH-3T3 cellsdid not exhibit reactivity with the same monoclonal antibody.Southern blot analysis of the DNA prepared from the transfectedNIH-3T3 cells, that developed as a progressively growing tumorin a nude mouse, revealed the presence of human repetitive DNAsequences.     

Localization of a gene for otosclerosis to chromosome 15q25-q26

Among white adults otosclerosis is the single most common cause of hearing impairment. Although the genetics of this disease are controversial, the majority of studies indicate autosomal dominant inheritance with reduced penetrance. We studied a large multi- generational family in which otosclerosis has been inherited in an autosomal dominant pattern. Five of16 affected persons have surgically confirmed otosclerosis; the remaining nine have a conductive hearing loss but have not undergone corrective surgery. To locate the disease- causing gene we completed genetic linkage analysis using short tandem repeat polymorphisms (STRPs) distributed over the entire genome. Multipoint linkage analysis showed that only one genomic region, on chromosome 15q, generated a lod score >2.0. Additional STRPs were typed in this area, resulting in a lod score of 3.4. STRPs FES (centromeric) and D15S657 (telomeric) flank the 14. 5 cM region that contains an otosclerosis gene.      

Peritoneal fluid concentrations of interleukin-8 in women with endometriosis: relationship to stage of disease

There is increasing evidence that immunological mechanisms play a role in the pathogenesis and pathophysiology of endometriosis. It was therefore of interest to study interleukin-8 (IL-8), a chemokine, in the peritoneal fluid and peripheral blood of women undergoing laparoscopic procedures. The presence and concentrations of IL-8 in relation to endometriosis, infertility and abdominal pain were evaluated. Samples of peritoneal fluid (n = 49) and peripheral blood (n = 50) were obtained from 50 consecutive patients undergoing laparoscopic surgery for various gynaecological indications (abdominal pain, infertility, sterilization). IL-8 was present in the peritoneal fluid of most women (87%). The concentration of IL-8 in the peritoneal fluid was higher in women with endometriosis compared to women without (P = 0.02). This difference was more pronounced in early (stage 1) endometriosis (P = 0.001). IL-8 concentrations in the peritoneal fluid were also higher in women with early endometriosis compared to women with later stages of the disease (P = 0.003). Peripheral blood concentrations did not correlate with peritoneal fluid concentrations of IL-8 and/or the presence of endometriosis. We conclude that IL-8 is an important factor that may contribute to the pathogenesis of endometriosis possibly by promoting neovascularization. This information can be a guide in the development of new therapeutic approaches for the treatment of endometriosis.      

HLA-D restriction of antigen-specific proliferative T cell responses

The specificity of the inhibitory activity of anti HLA-DR antisera on the proliferative response of human T cells to soluble antigens has been recently challenged. Furthermore, there are conflicting reports about the effect of antisera to the stimulatory antigen. Therefore, we investigated the inhibitory activity of different antisera on the proliferative response of T lymphocytes from sensitized donors to the antigens HSV and PPD. Alloantisera to DR specificities shared between antigen presenting cells (APC) and T cells displayed a strong inhibitory activity: alloantisera to HLA-DR specificities expressed only by APC or the T cell donor displayed lower or no inhibitory activity. Monoclonal antibodies to monomorphic determinants of HLA-DR molecules were inhibitory, but only when used at a high concentration. Antisera to HLA-ABC molecules and to HSV displayed little if any inhibition. These findings provide further evidence that the HLA-DR molecules as such in the APC membrane, which were also present during initial sensitization, are restriction elements for T cells.     

Loss of LMP and TAP expression in human melanoma cells

The goal of this study was to determine the frequency of LMP2, LMP7, TAP1 and/or TAP2 loss in melanoma cell lines and in surgically removed melanoma lesions. Cell extracts from control and IFN-γ treated melanoma cell lines were tested in Western blotting with antisera elicited with LMP2, LMP7, TAP1 and TAP2-specific peptides. LMP2 and LMP7 were not detected in 7 out of 9 cell lines. Expression of both LMP subunits was induced in 5 of the 7 negative cell lines by treatment with IFN-γ. TAP1 and TAP2 were not detected in 3 out of 9 cell lines and were induced in one of them by IFN-γ. Surgically removed lesions were stained in the immunoperoxidase reaction with antibodies purified from the antisera by affinity chromatography on the immunizing peptides. LMP2 and LMP7 were not detected in 20% and 6%, respectively, of 32 primary lesions and in 40% and 12%, respectively, of 25 metastatic lesions. TAP1 and TAP2 were not detected in 15% and 18%, respectively, of 32 primary lesions and in 20% and 24%, respectively, of 25 metastatic lesions. Moreover, the frequency of TAP loss is increased in primary lesions from patients with recurrence of their disease, suggesting a potential role in the course of the disease and a negative impact on the outcome of T cell-based immunotherapy.