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Biomedical subjects

J Lustgarten

Publications and source records attributed to J Lustgarten.

11 recordsLinked to original sources
PubMed1995

Specific elimination of IgE production using T cell lines expressing chimeric T cell receptor genes.

B cells that are destined to secrete IgE express a membrane-bound form of IgE (mIgE) on their cell surface. Thus, elimination of such mIgE-positive cells should result in the suppression of IgE production, thereby alleviating the symptoms of IgE-mediated allergy. In this study, we examined, in a model system, whether IgE-specific effector T cells can be used specifically to eradicate IgE-producing B cells. To this end, we endowed T cells with anti-IgE specificity using chimeric T cell receptors (cTCR) containing the variable region domain (Fv) of the 84.1c non-anaphylactic anti-mouse IgE monoclonal antibody (mAb). Two configurations of chimeric receptor were used: in the first, we combined the heavy and light variable region chains of 84.1c with the constant (C) regions of the TCR alpha and beta chains. The second construct consisted of a chimeric single-chain receptor (scFvR), composed of a single-chain Fv region of the 84.1c antibody and the C beta domain of the TCR. Following transfection of the cTCR or the scFvR genes into the murine MD.45 cytotoxic T cell hybridoma or the Jurkat human T cell line, functional expression of IgE-specific chimeric receptors was detected on the cell surface. The transfected cells secreted interleukin-2 upon stimulation with immobilized IgE or fixed IgE-producing hybridoma cells. Moreover, cytotoxic T cell hybridomas expressing the chimeric receptor genes specifically eliminated IgE-secreting B cells in vitro, resulting in isotype-specific suppression of IgE production.

Animals↗
PubMed1994

The use and clinical importance of a substrate-specific electrode for rapid determination of blood lactate concentrations.

OBJECTIVE: To determine the validity and clinical importance of a newly developed amperometric, enzymatic, substrate-specific electrode for the rapid measurement of circulating lactate concentrations. DESIGN: A prospective multiexperiment study. SETTING: The critical care medicine research laboratory, intensive care unit (ICU), emergency department (ED), and general wards of a university-affiliated hospital. PATIENTS: A total of 1218 patients and control subjects were studied on one or more occasions. INTERVENTIONS: Blood lactate concentrations, descriptive data, physiological parameters, and outcome results were determined in various patient populations. MAIN OUTCOME MEASURES AND RESULTS: Experiment 1: Lactate determinations performed with the new substrate-specific electrode were compared with two laboratory reference methods. Blood samples from 80 ICU patients and 165 ED patients formed the basis of this first experiment. There was excellent agreement between the test instrument and the two reference methods as reflected by bias (with reference method 1, 0.19 mmol/L; reference method 2, 0.09 mmol/L), precision (with reference method 1, +/- 0.47 mmol/L; reference method 2, +/- 0.34 mmol/L), and correlation data (with reference method 1, r = .92; reference method 2, r = .98). Experiment 2: The new test microchemistry instrument was used to analyze blood samples from 927 patients. The mean (SE) blood lactate concentrations in the various patient populations were 1.26 (0.04) mmol/L for control subjects (n = 85), 1.52 (0.03) mmol/L for general ward patients (n = 489; P < .001 vs normal subjects), 2.34 (0.15) mmol/L for ICU patients (n = 180; P < .001 vs normal subjects and general ward patients), and 2.44 (0.15) mmol/L for ED patients (n = 173; P < .001 vs normal subjects and general ward patients). None of the normal subjects and only one (0.2%) of 489 nonhypotensive general ward patients had a blood lactate value greater than 4 mmol/L. Circulating lactate concentrations greater than 4 mmol/L were 98.2% specific in predicting the need for hospital admission in patients presenting to the ED. Furthermore, lactate concentrations greater than 4 mmol/L were 96% specific in predicting mortality in hospitalized nonhypotensive patients. Experiment 3: Blood samples from 46 hypotensive ICU and ED patients and from 353 nonhypotensive ICU and ED patients (the latter samples were derived from experiment 2) were analyzed. A statistically significant difference was noted between the mean (SE) lactate concentration in hypotensive patients in the ICU and ED (4.75 [0.75] mmol/L) when compared with nonhypotensive ICU and ED patients (2.28 [0.10] mmol/L; P < .001). Furthermore, blood lactate values greater than 4 mmol/L were 87.5% specific in predicting mortality in hypotensive patients. CONCLUSIONS: Lactate determinations performed using the new test instrument are precise and accurate. Blood lactate concentrations greater than 4 mmol/L are unusual in normal and noncritically ill hospitalized patients and warrant concern. In hospitalized (non-ICU) nonhypotensive subjects, as well as in critically ill patients, a blood lactate concentration greater than 4 mmol/L may portend a poor prognosis.

Blood Chemical Analysis↗
PubMed1994

Control of microwave heating of peritoneal dialysis solutions.

OBJECTIVE: To determine if microwave heating of dialysis solutions to 37 degrees C produced focal overheating (hot spots) and caramelization of dextrose. DESIGN: In vitro determination of conditions for controlling time, temperature, and procedures. Bags had been stored at ambient room temperature. MAIN OUTCOME MEASURES: Solution and external bag surface temperature determinations. Dextrose degradation products determined spectrophotometrically. Microscopy for potential caramel precipitates. RESULTS: A microwave oven with no rotation tray produced uneven heating of bags of two commercially available concentrations of dialysis solutions. The greatest hot spots were evident in spike ports. External bag surface temperatures were within 0.20 degrees C of reservoir temperatures. Initial solution temperatures correlated with temperatures of the solutions after microwave heating (r = 0.895). No statistically significant differences were found between dextrose degradation product concentrations of unheated and heated solutions, including hot spots. No precipitates were observed microscopically. CONCLUSIONS: Despite the presence of solution hot spots in bag infusion ports, 37 degrees C temperatures were achievable in the bag reservoirs with no evidence of increased glucose degradation. This outcome is assured if the initial temperature and the microwave conditions (procedure, time, mixing of solution) are held constant, and the external bag temperatures are measured after heating.

Dialysis Solutions↗
PubMed1992

Six week safety study of 2% MK-927 administered twice daily to ocular hypertensive volunteers.

Ocular hypertensive patients were enrolled in a 6-week double-masked safety study of 2% MK-927 (27 patients), a topically active carbonic anhydrase inhibitor, administered bilaterally b.i.d.; 9 additional patients received 0.5% timolol as the control agent. Intraocular pressure (IOP) was measured weekly prior to a.m. drug administration; twelve hour diurnal curves were performed prestudy and at 3 and 6 weeks. The mean reduction of IOP prior to a.m. drug administration ranged from 1.2 +/- 4.4 mm Hg (SD) to 3.0 +/- 4.2 mm Hg with MK-927 and from 4.7 +/- 3.9 mm Hg to 8.8 +/- 0.6 mm Hg with timolol. Mean outflow facility measured tonographically prestudy and on days 33 to 42 four hours after a.m. drug administration was unchanged in both groups. Corneal sensitivity (Cochet-Bonnet), corneal thickness (ultrasound pachymetry), Schirmer tear testing, and extensive ophthalmologic and medical examinations, and hematologic studies were not substantially altered throughout the study. In this longest chronic administration study to date, MK-927 did not cause adverse ocular or systemic side effects.

Adult↗
PubMed1992

Functional assembly of chimeric T-cell receptor chains.

We have generated cytotoxic T-cell hybridomas expressing chimeric T-cell receptors (cTCR) with an antibody-type specificity for the TNP hapten. Transfectants expressing the cTCR genes could mediate specific lysis of haptenated tumor cell lines of various types and secrete IL-2 upon stimulation with TNP modified cells. In a previous report, we showed that double-gene transfectants expressing either VHC alpha and VLC beta or VHC beta and VLC alpha could be activated by TNP-modified stimulator cells or TNP proteins immobilized on plastic. Single-chain transfectants (expressing VHC alpha or VHC beta alone) could be mainly activated by TNP-cells. We now report that transfection of chimeric VHC alpha gene into an alpha-chain-defective mutant restores the surface expression of the TCR/CD3 complex. In parallel, such transfectants regained the ability to respond to mitogen and anti-CD3 antibodies and responded weakly to TNP cells. Double gene transfectants, bearing 2 complementary chimeric chains, expressed high amounts of cTCR on their surface, sufficient to acquire sound anti-TNP reactivity. Cells expressing the VHC beta gene only were not functional and had no detectable surface TCR chains. Taken together, our results suggest that chimeric VHC alpha chains can pair with endogenous V beta C beta chains, but that there is preferential association between complementary chimeric chains, resulting in higher functional expression of the chimeric TCR.

Animals↗
PubMed1991

CD4 and CD8 accessory molecules function through interactions with major histocompatibility complex molecules which are not directly associated with the T cell receptor-antigen complex.

Both the subset-specific, CD4 and CD8 T cell accessory molecules and the antigen-specific T cell receptor (TcR) interact with major histocompatibility complex (MHC) class I and class II molecules on the surface of antigen-presenting cells. We analyzed whether the CD4/CD8 molecules exert their accessory function through binding with the same MHC molecules which participate in the TcR-antigen-MHC complex. We utilized a CD4-, CD8-, class I-allospecific T cell hybridoma which functionally manifests both cytotoxic T lymphocyte (CTL) and T helper1 (Th1) phenotypes, and rendered it bispecific by transfecting it with genes encoding either a class II-restricted, 2,4,6-trinitrophenyl (TNP)-I-Ad-specific TcR or a non-MHC-restricted chimeric TcR, composed of a variable part of an anti-TNP antibody. Expression of either CD4 or CD8 transgenes in these hybridomas enhanced and augmented their reactivity towards the appropriate target cells regardless of the type of TcR-MHC interaction. Thus, class I-specific responses could be enhanced through CD4-class II interactions, and class II-restricted responses could be augmented through CD8-class I interactions. Furthermore, these accessory molecules also potentiated TNP-specific responses by the chimeric TcR which is MHC unrestricted. The accessory molecules facilitated both interleukin 2 (IL2) production and cytolytic activity by shortening the activation time and rendering the cells responsive to lower antigenic stimuli. The degree of activity of the T cell hybridomas correlated with the level of accessory molecule expression and was not related to the effector function mediated by the cells. Anti-CD4 or -CD8 antibodies completely inhibited the activity of transfectants expressing the corresponding accessory molecule, regardless of the MHC type of the TcR interaction. Such antibodies blocked direct TcR stimulation provided by either anti-T3/Ti antibodies or lectins, but could not inhibit the activation through agents that bypass the TcR such as phorbol 12-myristate 13-acetate plus ionophore. Taken together, these studies demonstrate that the CD8/CD4 molecules can exert their accessory function through interactions with MHC molecules which are not directly associated with the TcR-Ag-MHC complex, and that this accessory effect is associated with TcR-mediated triggering at an early stage of the signaling process and is not related to the effector mechanism assigned to the CD4 and CD8 T cell subsets.

Animals↗
PubMed1987

Southern blot analysis of BII cell line--a putative variant of HL-60.

Southern blot analysis of various genes was used to compare the human promyelocytic leukemia cell line HL-60 and the BII cell line, which reportedly arose as a spontaneous differentiation inducer-resistant variant from an HL-60 culture. Granulocyte-macrophage colony stimulating factor gene restriction fragment polymorphism, due to a partial deletion of one of the alleles of this gene in HL-60, was not observed in the BII cells. Furthermore, the p53 oncogene, most of which is deleted in the HL-60 cell line, was found to be intact in the BII cell line. Human leukocyte antigen typing revealed that the two cell lines shared the A locus but differed at the B locus. Several unique restriction fragments hybridizing to human leukocyte antigen class I and DR beta gene probes were observed in the DNA digests of each cell line. Altogether these data provide definitive evidence that BII represents a human cell line of different origin than HL-60. Further lineage determination of this cell line could add a useful member to the group of leukemic cell lines.

Antigens, Neoplasm↗
PubMed1984

Laser trabeculoplasty. A prospective study of treatment variables.

Forty-five phakic eyes with open-angle glaucoma and uncontrolled intraocular pressure underwent laser trabeculoplasty. Each eye was assigned randomly to one of three treatment groups: group 1, 100 spots over 360 degrees; group 2, 50 spots over 180 degrees; or group 3, 50 spots over 360 degrees. A 50-micron spot was aimed at the anterior meshwork; power and time were varied to achieve a blanch. Forty-four eyes were followed up for at least four weeks without further intervention. The mean IOP before therapy and the initial IOP elevation were similar in all groups. After four weeks, the mean IOP reductions in 15 eyes in group 1, 15 eyes in group 2, and 14 eyes in group 3 were not significantly different. However, significantly more eyes in group 1 demonstrated a greater than 12 mm Hg reduction in IOP than eyes in the other groups. Group 2 tended to have the fewest eyes with reduced medications.

Aphakia↗
PubMed1984

Continent, nonrefluxing urinary reservoir constructed from ileum: report of two cases.

Most procedures for diverting urine sacrifice continence, but Kock's modification of the ileal conduit promises the ideal of appliance-free urine storage with voluntary control of emptying. The authors report on two men who underwent this procedure. One was a paraplegic who had total urinary incontinence after a sphincterotomy. The other had a carcinoma of the rectosigmoid obstructing the right ureter. He had congenital bladder exstrophy and had undergone ureteric implantation into the sigmoid colon at 3 years of age. In both patients, the reservoir was created from an isolated 60-cm segment of the middle portion of small bowel. Two nipple valves were created - one to prevent reflux and one to provide continence of urine. Follow-up was 27 and 17 months respectively. One patient had his outlet valve revised because of sliding and incontinence. He is now completely continent of urine and he intubates his reservoir three to four times daily. Postoperatively, the second patient had a leak at the ureteroileal anastomosis, but this healed spontaneously. The continent urinary diversion is a major intestinal operation and should not be performed in conjunction with exenteration. It should be confined to a few centres where greater experience can be accumulated.

Adult↗
PubMed1981

Serum hormones and lipoproteins in benign breast disease.

Seventeen young women with clinically confirmed mammary dysplasia and six age-matched controls were treated with alpha-tocopherol. Serum samples collected during the luteal phase of each woman at monthly intervals for the 4-month duration of the study were analyzed for serum luteinizing hormone, follicle-stimulating hormone, and prolactin concentrations by radioimmunoassay and for lipoprotein levels by a combination of precipitation, ultracentrifugation, and enzymatic techniques. Fifteen patients showed objective and subjective remission from disease. While prolactin levels did not change significantly, elevated levels of luteinizing and follicle-stimulating hormones were decreased to normal levels. Ratios of serum cholesterol to high-density lipoprotein cholesterol decreased; high-density lipoprotein and free cholesterol associated with low-density lipoproteins increased as a result of therapy. The results suggest that alpha-tocopherol may serve as an effective agent not only to treat patients with benign breast disease but also to normalize abnormal hormone and lipid levels in subjects at high risk for breast cancer.

Breast Diseases↗