Measles virus replication in cells of myelomonocytic lineage is dependent on cellular differentiation stage.

Measles virus (MV)-infected monocytes may have a central role in virus-induced immunosuppression. Our understanding of MV replication in monocytic cells is, however, incomplete. In this work we have investigated MV replication in cells of human myelomonocytic lineage with different maturation stages in order to study the effect of cellular maturation on virus infection. MV was able to infect human bone marrow myeloid granulocyte-macrophage colony-forming cells (CFC-GM) as well as monocytes and macrophages, but the replication cycle seemed to be regulated by the maturation stage of the cells. Virus infection in CFC-GM was productive, unlike in monocytes and macrophages, where an extensive viral RNA synthesis occurred and high amounts of proteins were synthesised without a remarkable release of infectious virus. Efficiency of viral macromolecular synthesis in macrophages was comparable to that of promonocytic cell line U-937 and human epithelial cell line A549, but in contrast to macrophages the cell lines highly supported productive infection. On the other hand, chemically induced maturation of the human promyelocytic and promonocytic cell lines HL-60, THP-1, and U-937 to more mature macrophage-like forms did not markedly alter the replication cycle of MV in these cell lines. Our results showed that MV replication in myelomonocytic cells varied depending on the maturation stage of the cells. The immature myelomonocytic cells supported productive virus infection, but the maturation process lead to cellular changes that caused a restriction of MV replication cycle partly at posttranscriptional and partly at posttranslational level. The metabolic milieu of monocytes and macrophages as such was sufficient to support extensive viral macromolecular synthesis.

Daily measurements of blood CD34+ cells after stem cell mobilization predict stem cell yield and posttransplant hematopoietic recovery.

The value of daily monitoring of the blood CD34+ cell concentration as a guide to the optimal timing of stem cell harvests was studied in 60 patients who underwent 66 stem cell mobilizations and 189 leukaphereses. There was a highly significant correlation between the blood CD34+ count and the CD34+ cell content in the apheresis product of the same day (r = 0.904, p < 0.01). Thus, the target yield of 4 x 10(6) CD34+ cells/kg can be harvested in one or two leukaphereses when the blood CD34+ cell count exceeds 50 x 10(6)/L. However, an insufficient harvest is to be expected when the blood CD34+ cell count is below 20 x 10(6)/L. The data from 35 autologous blood cell transplantations with a minimum CD34+ cell yield of 1.5 x 10(6)/kg showed that the recovery of blood neutrophil counts to 1.0 x 10(9)/L occurred in all patients within 9-14 days, but the time to recovery of the platelet counts to 20 x 10(9)/L may exceed 14 days, especially if the CD34+ cell content is below 4 x 10(6)/kg. Daily monitoring of blood CD34+ cell counts is a rapid and reliable means to guide the timing of stem cell collections. The count predicts well the CD34+ cell content of the harvests, the number of leukaphereses needed, and the speed of hematopoietic recovery.

Susceptibility of human bone marrow cells and hematopoietic cell lines to coxsackievirus B3 infection.

Viremia is commonly observed in association with enterovirus infections, and during this phase viruses can be transmitted to secondary target organs in the body. It is not known, however, whether blood cells play a role in the pathogenesis of enterovirus infection supporting virus replication. Our earlier work (T. Vuorinen, R. Vainionpää, H. Kettinen, and T. Hyypiä, Blood 84:823-829, 1994) demonstrated that coxsackievirus B3 is able to replicate in representatives of B- and T-cell lines but not in a monocytic cell line or peripheral blood mononuclear cells, indicating that virus replication may depend on the differentiation and maturation stages of the cells. Therefore, we have broaden our studies and analyzed the susceptibility of granulocyte-macrophage CFU and hematopoietic cell lines with various differentiation and maturation stages to coxsackievirus B3 infection. Virus replication was detected in B- and T-cell lines with no direct correlation to the maturation stage. Granulocyte-macrophage CFU were also able to support virus multiplication.

Androgen, estrogen and progestin binding in cytosols of benign gynecologic tumors and tumor-like lesions.

Specific androgen (Kd 0.27 +/- 0.06 nM), estrogen (Kd 0.19 +/- 0.04 nM) and progestin (Kd 0.22 +/- 0.07 nM) binding were investigated in benign gynecologic tumors and tumor-like lesions. The simultaneous presence of androgen and progestin receptors was a common finding. In one endometrioma, specific androgen, estrogen and progestin binding was observed simultaneously.

Pitfalls in the Dextran-coated charcoal assay of estrogen receptors in breast cancer tissue.

This study investigated the influence of the degree of concentration of breast tumor cytosols on the apparent estrogen receptor content as measured by the Dextran-charcoal assay. It was found that the dilution of cytosols to 1-2 mg protein/ml frequently but not always causes highly underestimated receptor concentrations. This could not be explained by the protein loss through adsorption to the charcoal. The effect was also studied in the presence of gelatin, sodium molybdate or with limited trypsinization of the incubation mixture. Addition of 1 mg/ml gelatin in the Dextran-charcoal suspension was very useful in most cases in preventing dilution induced losses in receptor sites. Both trypsinization and addition of sodium molybdate produced increases in receptor concentrations that were not as susceptible to inactivation through dilution of the cytosol. These data suggest that the observed high variability in the dilution induced receptor losses can be explained by receptor heterogeneity: some receptor form(s) are either readily absorbed to or "stripped" by the charcoal particles. As a conclusion we recommend that in order to optimize the estrogen receptor assay as regards both binding sites and affinities the cytosol concentrations should be maintained as high as possible and a protein expander be included in the Dextran-charcoal suspension. Though sodium molybdate frequently gives considerable increases in estrogen binding sites it occasionally has an opposite effect. For this reason we hesitate to recommend its use in routine assays of estrogen receptors.

Steroid receptors and response of ovarian cancer to hormones in vitro.

Oestrogen receptor (ER) and progesterone receptor (PR) content and the response in vitro to tamoxifen (T), medroxyprogesterone acetate (MPA) and to a combination of the two hormones were determined in 21 epithelial ovarian carcinomas. The response was assessed by the level of adenosinetriphosphate in the cells. ER and PR were detected in 62% and 57%, respectively, with significant variations between the different histopathological cancer types. ER and PR predicted the response in vitro in 62% of the tumours exposed to the combined hormones, and in 38% and 33% of those exposed to T and MPA, respectively. The value of steroid-receptor determinations in selecting the proper hormonal treatment in ovarian cancer is significantly reduced because of the high proportion of incorrect predictions.

Steroid receptors and response of ovarian cancer to cytostatic drugs in vitro.

Samples of 21 ovarian cancers were assayed for oestrogen receptor (ER) and progesterone receptor (PR) content, and the response in vitro to treatment with a combination of doxorubicin, diacetyldian hydrogalactitol and cisplatin was determined. The number of living cells after drug exposure was estimated by a new ATP-bioluminescence method and the tumours were considered responsive if cell survival was less than or equal to 50% of the value in a corresponding control culture. Of the 16 tumours that responded to drug exposure, nine were ER-positive, seven ER-negative and eight were PR-positive, eight PR-negative. The mean percentages of surviving cells ranged from 22.2% in PR-negative tumours to 30.9% in PR-positive tumours. There were no differences in the response rates or in the degree of response to the cytostatics in terms of either receptor status or tumour histology. The results were also compared with those obtained in the same tumour samples exposed to hormones, tamoxifen and medroxyprogesterone acetate. The average response of all tumours was better to cytostatics than to hormones (P less than 0.05); this was particularly marked in the ER-negative tumours. Cytostatics may be preferable to hormones as the primary drug treatment for ovarian cancers but steroid-receptor determinations appear not to help in formulating the optimum drug treatment.

Androgen, estrogen and progestin cytosol receptor concentrations in the normal human endometrium. Effects of intrauterine device.

The specific androgen, estrogen and progestin binding was studied in normal human endometrium during various phases of the menstrual cycle. The simultaneous presence of these three hormone receptors was a common finding. Androgen receptor concentrations did not show cyclic variations. A copper intrauterine device had no effect on the endometrial androgen, estrogen and progestin receptor concentrations.

Steroid hormone receptors in human squamous carcinoma cell lines.

Specific estrogen, progesterone and androgen receptors were determined in 19 well-characterized squamous-cell carcinoma (SCC) cell lines. Ten of the lines were derived from patients with SCC of the larynx and nine were from patients with SCC originating in other areas of the head and neck. Estrogen receptors (ER) were found in seven of 10 cell lines derived from squamous cancers of the larynx (70%) but in only one of the nine SCC cell lines from sites other than larynx. Progesterone receptors (PGR) were more evenly distributed. Eight of the 10 laryngeal carcinoma cell lines (80%) and five of the nine non-laryngeal SCC lines (55%) had progesterone receptors. Only one cell line, UM-SCC-10B (derived from a recurrent carcinoma of the larynx) was found to express androgen receptors (AR). Expression of specific estrogen receptors was not dependent on the sex of the patients since lines from both males and females contained receptors. These results establish that squamous carcinoma cell lines may express specific steroid hormone receptors and that cell lines from cancers of the larynx (an organ known to be androgen-responsive) are more likely to express estrogen receptors than androgen receptors. From this initial survey it appears that there is a striking difference in estrogen and progesterone receptor content between SCC cell lines originating from larynx cancers and cell lines established from squamous carcinomas of other head and neck regions. The presence of estrogen receptors in a high proportion of laryngeal carcinoma cell lines suggests that hormonal therapy may be a useful adjunctive therapy in selected patients with cancer of the larynx.

Estrogen receptors in mammary cancer: correlation with age, menopausal status, and response to therapy.

The concentration of estrogen receptor protein (ER) in 113 primary or metastatic breast cancers was studied. ER was found to be correlated with the age of the patient. The ER values were generally lower in premenopausal patients (5.6 fmol/mg cytosol protein) than in postmenopausal patients (32.8 fmol/mg cytosol protein). The ER values of perimenopausal patients (0-5 years since the last menstrual period) were heterogeneous but generally closer to those of the premenopausal patients. Use of the ER determination for allocation of the patients either to hormonal (tamoxifen or nandrolonedecanoate) or to cytostatic (adriamycin-cyclophosphamide or Cooper's regimen) therapy was shown to result in highly satisfactory clinical response rates (including complete and partial remissions and stabilized disease) of 68% and 67%, respectively. The practical limit of ER concentration for selection is between 3 and 10 fmol/mg cytosol protein in breast cancer.

Increase in the estrogen binding capacity of breast cancer cytosols following limited proteolysis with trypsin.

When small amounts of trypsin were added to prelabelled estrogen receptors in 24 human breast cancer cytosols there was a substantial increase in the binding capacity [79 +/- 11 (SE)%]. At the same time the affinity of the hormone receptor interaction was maintained at a very high level or even increased. This finding is discussed in relation to previous results where a diisopropylfluorophosphate (DFP) inhibitable protease activity was shown to cause a similar augmentation of estrogen binding sites in human myometrial cytosols. Addition of sodiummolybdate at or immediately after homogenization led to a similar increase in estrogen binding sites. Because these two effects were not additive we propose that the limited trypsin treatment reactivates the binding sites previously inactivated through a mechanism which can be inhibited by sodiummolybdate.