Coexpression of cell-surface immunoglobulin (sIg), terminal deoxynucleotidyl transferase (TdT) and recombination activating gene 1 (RAG-1): two cases and derived cell lines.

While it is generally agreed that in the lymphoid differentiation of B lineage cells there is no stage in which cell-surface immunoglobulin (sIg) and terminal deoxynucleotidyl transferase (TdT) are expressed simultaneously, a few B cell acute lymphoblastic leukemia (B-ALL) cases with this phenotype have been reported. Two such cases and the derived cell lines are reported here, in which the expression of recombination activating gene-1 (RAG-1) was also detected. One case was a CD19+ CD22+ HLA-DR+ sIg+ (gamma, kappa) B-ALL. The cell line (Bay9I) also expressed CD10. Karyotypic analysis revealed t(14;18)(q32;q21) and additional aberrations. In the other case, the fresh leukemia cells expressed CD19, CD24 and HLA-DR antigen. The derived cell line (Tree92) also expressed CD22 and sIg (mu, lambda). The karyotype of the Tree92 cells was t(8;14)(q24;q32) with additional aberrations. Tree92 is the first established cell line having both t(8;14)(q24;q32) and TdT. TdT was detected by Northern blotting as well as indirect immunofluorescence analysis. In addition, both Bay9I and Tree92 expressed RAG-1, as detected by Northern blot analysis. Cross-linking of sIg on Tree92 cells with anti-mu antibody led to significant down-regulation of RAG-1 expression. It seems that there is a sIg+ TdT+ RAG-1+ B lineage differentiation stage, and that signaling through sIg can modulate RAG-1 expression.

[Haemophilus aphrophilus isolated from the blood of a patient with infective endocarditis].

On July 1994, a 62-year-old female, having a history of mitral regurgitation, was admitted because of high fever, hematuria and conjunctival petechiae. She was diagnosed as having infective endocarditis with mitral valve vegetation proved by ultrasonic cardiography. The gram negative rods were isolated from blood cultures performed five times, performed prior to the administration of antibiotics. The isolates were identified as strains of H. aphrophilus. After two days of treatment with PCG (12 million units/day), the organism became undetectable from the blood. Since the minimal inhibitory concentrations (MICs) of PCG and ABPC were ranged between 0.06-2.0 micrograms/ml and 0.06-0.5 microgram/ml, respectively, ABPC was selected as a first choice antibiotic instead of PCG. ABPC was given 12 g/day for the first 3 days, then 6 g/day for 28 days, followed by 3 g/day for 7 days. The patient recovered and was discharged after the 55 hospital days. H. aphrophilus grew on BTB lactose agar, chocolate agar and sheep blood agar, but failed to grow on MacConkey agar. H. aphrophilus produced smooth transparent nonhaemolytic micro colonies after 48 hours on sheep blood agar and chocolate agar plates. Atmosphere with 5% CO2 failed to enhance their growth. All the five strains of H. aphrophilus isolated, required neither factors V nor X. Positive synthesis of porphyrin from delta-aminolevlinic acid confirmed their ability to grow without X factor. For the correct identification of H. aphrophilus strains, fermentation test of glucose, lactose, maltose and sucrose in either phenol red broth or CTA medium are necessary.(ABSTRACT TRUNCATED AT 250 WORDS)

Leukemic cells from a chronic T-lymphocytic leukemia patient proliferated in response to both interleukin-2 and interleukin-4 without prior stimulation and produced interleukin-2 mRNA with stimulation.

Recently, interleukin-4 (IL-4) has been clarified as having T-cell growth factor activity; therefore, it becomes of interest whether IL-4, as well as interleukin-2 (IL-2), affects the proliferation of leukemic cells derived from mature T cells. In the present study, we describe a case of chronic T-lymphocytic leukemia (T-CLL) with monoclonal proliferation of human T-lymphotropic retrovirus (HTLV)-I or HTLV-II negative CD3(+)4(+)8(-) T cell expressing IL-2 receptors without stimulation. Radiolabeled IL-2 binding assay revealed 750 high-affinity and 6,750 low-affinity binding sites per cell. In accordance with the expression of high-affinity IL-2 receptors, the leukemic cells proliferated in response to exogenous IL-2 without prior stimulation. In addition, exogenous IL-4 also induced their proliferation. Moreover, IL-2 and IL-4 exerted a synergistic effect on the leukemic cell proliferation. Although the expression of IL-2 or IL-4 mRNA was not detected in fresh leukemic cells, the expression of IL-2 mRNA, but not IL-4 mRNA, was induced by phytohemagglutinin stimulation, and the leukemic cells proliferated. These findings suggest that not only IL-2, but also IL-4 are involved in the proliferation of leukemic cells of T-CLL.

UVB/PUVA-induced suppression of human natural killer activity is reduced by superoxide dismutase and/or interleukin 2 in vitro.

We have evaluated the effects of ultraviolet irradiation or PUVA treatment [8-methoxypsoralen (8-MOP) plus long-wave ultraviolet (UVA) irradiation] on natural killer (NK) activity of human peripheral blood mononuclear cells (PBMC). In vitro exposure of PBMC to UVB (280-320 nm, 1-30 mJ/cm2) or PUVA [8-MOP, 0.1 microgram/ml; UVA (320-400 nm), 0.5-5 J/cm2] resulted in a dose-dependent suppression of NK activity, whereas UVA (0.5-5 J/cm2) or 8-MOP (0.1 microgram/ml) treatment alone did not have the inhibitory effects. This suppressive effect of UVB/PUVA irradiation was successfully reduced in the presence of superoxide dismutase (SOD) (100 or 1000 U/ml) during the irradiation. The addition of interleukin 2 (IL-2) (100 U/ml) markedly enhanced the NK activity of both irradiated and nonirradiated PBMC. Combination treatment with both SOD and IL-2 to UVB/PUVA-irradiated PBMC resulted in a more remarkable improvement of NK suppression than with either SOD or IL-2 treatment alone.

[The prognosis of Graves' disease after antithyroid drug treatment: clinical significance of the T3 suppression test and abnormal thyroid stimulators].

The prognostic significance of both the triiodothyronine (T3) suppression test and the detectability of thyroid stimulating immunoglobulins in patients with Graves' disease who had been treated with antithyroid drugs was evaluated. Eighty-three patients underwent a T3 suppression test after having been euthyroid for at least 6 months. In 33 patients, the human thyroid stimulator (HTS) was assayed by measuring cyclic AMP increase in cultured thyroid adenoma cells, and TSH-binding inhibitor immunoglobulins (TBII) were measured by using the radioreceptor assay of TSH. Among 43 patients who had discontinued the drug treatment, 37 patients were under observation for 6-42 months. When a fall in 30-minute thyroid 99mTcO4- uptake of 50% or more after T3 administration was defined as positive suppression, the relapse rate was 30% in non-suppressive cases and 26% in suppressive cases. The relapse rate was lower in cases whose pre-suppression uptake was less than 3.0% (3 out of 17 patients) or in cases whose uptake after T3 administration was less than 0.8% (1 out of 9 patients). Of 15 patients with negative suppression, 5 (33.3%) were positive in HTS and 4 (26.7%) were positive in TBII. On the other hand, two (11.1%) each of 18 patients with positive suppression were positive in HTS and TBII respectively. Neither HTS nor TBII had been detected at the cessation of therapy in any of the 12 patients who remained euthyroid during the follow-up period. On the other hand, four (44.4%) out of 9 patients who relapsed had been positive in either HTS of TBII. Thus the Graves' disease specific immunoglobulins were found to be significantly associated with the relapse of the disease (p less than 0.05). The above data indicates that regardless of the suppressibility of 99mTcO4- uptake after T3 administration, the rate of recurrence is high when the uptake after T3 is more than 0.9% and/or when either HTS or TBII are positive.

Effect of corynebacterium liquefaciens on a C3Hf mouse squamous cell carcinoma.

The antitumor effect of anaerobic Corynebacterium liquefaciens was compared with that of specific immunization. Experimental tumors were fourth or fifth generation isotransplants of a NR-Sl squamous cell carcinoma that arose spontaneously in a C3Hf/He female mouse. Specific immunization failed to exhibit an antitumor effect, whereas a single administration of the bacterium markedly inhibited the growth of the tumor. This growth inhibition was most effective when C. liquefaciens was administered 2 to 4 days before transplantation of tumor cells, but marked inhibition was also observed when this agent was administered after transplantation. The inhibitory effect was independent of dose within a range of 0.1 to 2.0 mg/mouse; a single dose of less than 0.05 mg/mouse did not exhibit antitumor effect. Multiple administrations of large doses, if given with short treatment intervals, were no more effective than one small dose. Multiple doses given at 14-day intervals resulted in marked growth retardation. The dose of cells that produced 50% tumor takes in C. liquefaciens-treated animals was not significantly different from that in nontreated animals, indicating that this bacterium exhibited no lethal effect on the tumor cells studied.

[Effect of continuous bleomycin treatment and of oil-suspended bleomycin on experimental tumor growth (author's transl)].

Effects of continuous administration of bleomycin solution and of intralesional injection of sesame oil-suspended bleomycin on tumor growth were studied. Experimental animal tumors were 3 rd generation isotransplants of a spontaneous C3H mouse mammary carcinoma. Bleomycin treatments were started when transplanted tumors reached 8 mm in diameter and the measurement of tumor volume was followed. Dose administered was fixed as 100 mg/kg in all the groups. Bleomycin solution was given intralesionally in a single or 4 daily doses, or intraperitoneally by continuous infusion. The latter method inhibited tumor growth most effectively, while the single injection was the last effective. Intralesional injection of oil-suspended exhibited similar effectiveness as the continuous infusion, and it was independent of the number of fractions. These results were interpreted by the several features in the response of mammalian cells to the antibiotic.

Combined use of bleomycin in radiotherapy of a mouse mammary carcinoma.

Experiments were carried out to determine TCD50 (50% tumor control dose) of 3rd generation isotransplants of a C3H mouse mammary carcinoma treated or not treated with Bleomycin. If the antibiotic was injected 30 min before a single X-ray dose, TCD50 was reduced. This reduction in TCD50 was independent of Bleomycin dose of more than 15 mg/kg, because of the upward-concave nature of Bleomycin dose-cell survival curve. The combined effect, when tested by TCD50 assays, appeared less than additive. This effect was further examined by a series of TD50 assays which revealed that these tumor cells were capable of repairing the potentially lethal damage induced by X-rays and that induced by Bleomycin. It was also found that the potentially lethal damage after combined X-ray and Bleomycin treatments was repaired. These findings indicated that the combined X-ray and Bleomycin treatment resulted in additive effect if the repair of potentially lethal damage in tumor cells were taken into account.

Repair of potentially lethal radiation damage in acute and chronically hypoxic tumor cells in vivo.

The ability of animal tumor cells to repair potentially lethat damage was studied in vivo. Fifth-generation isotransplants of a spontaneous mouse squamous-cell carcinoma were irradiated under tourniquet-induced hypoxia or in air. Tumors were removed either immediately or 6 hours after irradiation and dose-response curves were determined by TD50 assays. Repair was attributed to cells in the hypoxic cell component for animals irradiated in air. Extensive repair was also noted for those irradiated under typoxic conditions. Implications of these results are discussed.