A chemically inert drug can stimulate T cells in vitro by their T cell receptor in non-sensitised individuals

Drugs can interact with T cell receptors (TCR) after binding to peptide-MHC structures. This binding may involve the formation of a stable, covalent bond between a chemically reactive drug and MHC or the peptide embedded within. Alternatively, if the drug is chemically inert, the binding may be non-covalent and readily reversible. Both types of drug presentation account for a substantial number of adverse side effects to drugs. Presently no tests are available to predict the ability of chemically inert drugs to stimulate an immune response. Here we present data on the successful induction of a primary T cell immune response in vitro against a chemically inert drug using blood from healthy individuals, previously not exposed to the drug. Blood lymphocytes were stimulated by the chemically inert drug sulfamethoxazole and the protein-reactive drug-metabolite sulfamethoxazole-nitroso in the presence of IL-2. 9/10 individuals reacted in response to sulfamethoxazole-nitroso, but only three reacted to the chemically inert compound sulfamethoxazole. Drug reactive T cells could be detected after 14-35 days of cell culture by drug-specific proliferation or cytotoxicity, which was MHC-restricted. These cells were CD4, CD8 positive or CD4/CD8 double positive and T cell clones generated secreted Th0 type cytokines. Drug interaction lead to down-regulation of specific TCR. These data confirm the ability of chemically inert drugs to stimulate certain T cells by their TCR and may provide the opportunity to screen new drugs for their ability to interact with TCRs.     

Preclinical to Clinical Translation of CNS Transporter Occupancy of TD-9855, a Novel Norepinephrine and Serotonin Reuptake Inhibitor

Background:

Monoamine reuptake inhibitors exhibit unique clinical profiles that reflect distinct engagement of the central nervous system (CNS) transporters.

Methods:

We used a translational strategy, including rodent pharmacokinetic/pharmacodynamic modeling and positron emission tomography (PET) imaging in humans, to establish the transporter profile of TD-9855, a novel norepinephrine and serotonin reuptake inhibitor.

Results:

TD-9855 was a potent inhibitor of norepinephrine (NE) and serotonin 5-HT uptake in vitro with an inhibitory selectivity of 4- to 10-fold for NE at human and rat transporters. TD-9855 engaged norepinephrine transporters (NET) and serotonin transporters (SERT) in rat spinal cord, with a plasma EC₅₀ of 11.7ng/mL and 50.8ng/mL, respectively, consistent with modest selectivity for NET in vivo.Accounting for species differences in protein binding, the projected human NET and SERT plasma EC₅₀ values were 5.5ng/mL and 23.9ng/mL, respectively. A single-dose, open-label PET study (4–20mg TD-9855, oral) was conducted in eight healthy males using the radiotracers [¹¹C]-3-amino-4- [2-[(di(methyl)amino)methyl]phenyl]sulfanylbenzonitrile for SERT and [¹¹C]-(S,S)-methylreboxetine for NET. The long pharmacokinetic half-life (30–40h) of TD-9855 allowed for sequential assessment of SERT and NET occupancy in the same subject. The plasma EC₅₀ for NET was estimated to be 1.21ng/mL, and at doses of greater than 4mg the projected steady-state NET occupancy is high (>75%). After a single oral dose of 20mg, SERT occupancy was 25 (±8)% at a plasma level of 6.35ng/mL.

Conclusions:

These data establish the CNS penetration and transporter profile of TD-9855 and inform the selection of potential doses for future clinical evaluation.     

T-cell reaction to local anaesthetics: relationship to angioedema and urticaria after subcutaneous application — patch testing and LTT in patients with adverse reaction to local anaesthetics

BACKGROUND: Local anaesthetics are known to elicit T-cell reactions after epicutaneous application, namely contact dermatitis. In addition, adverse reactions like urticaria and angioedema are rather common after submucosal or subcutaneous injection. The pathogenesis of these side-effects, which appear frequently hours after application, is unknown, but thought to be not immunoglobulin E-mediated, since immediate skin tests are mostly negative. OBJECTIVES: We investigated whether patients who developed urticaria and angioedema after subcutaneous application have a T-cell sensitization to local anaesthetics, which might be responsible for the symptoms. METHODS: Twenty patients with generalized and/or local cutaneous reactions after LA were examined with intradermal testing using a standard panel of six LAs and patch testing using between seven and nine LAs in vaseline and four LAs in PBS. In 10 patients, a lymphocyte transformation test (LTT) was performed. RESULTS: Only 2/20 patients had an immediate skin reaction (positive intradermal test), whereas 6/20 patients had a positive delayed skin reaction (positive patch test). In 6/10 subjects the LTT was positive. CONCLUSIONS: Delayed appearance of urticaria and angioedema after subcutaneous application of local anaesthetics may be related to a T cell- mediated sensitization, which might be detected by patch testing or LTT.     

Measurement of T1, T2, and magnetization transfer properties during embryonic development at 7 Tesla using the chicken model

Purpose

To evaluate whether techniques of high field magnetic resonance imaging may be used to characterize embryonic tissue during proliferation and differentiation.

Materials and Methods

Thirteen chicken embryos with incubation times between 5 days and 16 days have been measured in a small animal magnetic resonance imager (ClinScan, Bruker) at 7 Tesla using the built‐in resonator. T1, T2‐, and magnetization transfer imaging was performed using fast spin‐echo with inversion recovery, half acquisition single shot turbo spin‐echo, and spoiled gradient‐echo sequences with and without off‐resonance pulse, respectively. T1, T2, and magnetization transfer ratio (MTR) maps were calculated on a pixel‐by‐pixel basis.

Results

T1‐, T2‐, and MTR maps showed good image quality allowing for delineation of embryonic organs. During embryonic development, a decrease of T1 and T2 relaxation times was found, whereas, embryonic tissue typically showed an increase of magnetization transfer, for example, liver properties at day 5: T1 = 2431 ± 163 ms, T2 = 122 ± 12 ms, MTR = 9.2 ± 4.2%; liver properties at day 16: T1 = 1763 ± 89 ms, T2 = 71 ± 4 ms, MTR = 16.9 ± 2.2%.

Conclusion

Embryonic tissues show changing relaxation and magnetization transfer properties during development, therefore, high field MRI seems suitable for characterization of tissue replacement derived from embryonic stem cells. J. Magn. Reson. Imaging 2008;28:1510–1514. © 2008 Wiley‐Liss, Inc.     

Hypophosphatemia during parenteral nutrition of patients with renal insufficiency (author's transl)]

Hypophosphatemia is a much commoner condition than generally recognized from investigations on this subject so far. Hypophosphatemia may cause ill-defined disturbances in the course of illness in patients with renal insufficiency. On the basis of our results we recommend the addition of 5--10 mmol phosphate (155--310 mg phosphorus) per 1000 kcal. of the nutrient solution right from the start of parenteral nutrition in patients with chronic renal insufficiency. The phosphate dosage must be further increased, at least temporarily, in hypophosphatemic patients with acute renal insufficiency. Serum phosphate determination should be obligatory in patients with renal insufficiency at the time when the patient is first seen. It should also be performed at least every second day during the first week of parenteral nutrition. Thereafter, the determination of serum phosphate twice weekly should suffice to control the dosage of phosphate required by the patient.