Receptor Phenotype Research Articles

Reply to Verheyden and Demanet

1 Giebel S, Nowak I, Wojnar J, Krawczyk-Kulis M, Holowiecki J, Kyrcz-Krzemien S et al. Association of KIR2DS4 and its variant KIR1D with leukemia. Leukemia 2008; 22: 2129–2130. 2 Verheyden S, Bernier M, Demanet C. Identification of natural killer cell receptor phenotypes associated with leukemia. Leukemia 2004; 18: 2002–2007. 3 Verheyden S, Demanet C. Susceptibility to myeloid and lymphoid leukemia is mediated by distinct inhibitory KIR-HLA ligand interactions. Leukemia 2006; 20: 1437–1438. 4 Verheyden S, Demanet C. NK cell receptors and their ligands in leukemia. Leukemia 2008; 22: 249–257. 5 Ross OA, Maxwell LD, Rea IM, Curran MD. Hypothetical soluble KIR2DS4 natural killer cell receptor molecule does not associate with successful ageing in the Irish. Exp Gerontol 2004; 39: 801–805. 6 Middleton D, Gonzalez A, Gilmore PM. Studies on the expression of the deleted KIR2DS4*003 gene product and distribution of KIR2DS4 deleted and nondeleted versions in different populations. Hum Immunol 2007; 68: 128–134. 7 Whang DH, Park H, Yoon JA, Park MH. Haplotype analysis of killer cell immunoglobulin-like receptor genes in 77 Korean families. Hum Immunol 2005; 66: 146–154.

Long-Term Synaptic Plasticity in Cerebellar Stellate Cells

Inhibitory transmission controls the action potential firing rate and pattern of Purkinje cell activity in the cerebellum. A long-term change in inhibitory transmission is likely to have a profound effect on the activity of cerebellar neuronal circuits. However, little is known about how neuronal activity regulates synaptic transmission in GABAergic inhibitory interneurons (stellate/basket cells) in the cerebellar cortex. We have examined how glutamate released from parallel fibers (the axons of granule cells) influences postsynaptic alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptors in stellate cells and modulates gamma-aminobutyric acid (GABA) release from these neurons. First, we found that burst stimulation of presynaptic parallel fibers changes the subunit composition of post-synaptic AMPA receptors from GluR2-lacking to GluR2-containing receptors. This switch reduces the Ca(2+) permeability of AMPA receptors and the excitatory postsynaptic potential amplitude and prolongs the duration of the synaptic current, producing a qualitative change in synaptic transmission. This switch in AMPA receptor phenotype can be induced by activation of extrasynaptic N-methyl-D: -aspartate (NMDA) receptors and involves PICK1 and the activation of protein kinase C. Second, activation of presynaptic NMDA receptors triggers a lasting increase in GABA release from stellate cells. These changes may provide a cellular mechanism underlying associative learning involving the cerebellum.

4.5 Defective phosphorylation of the interleukin 18 receptor beta-chain causes impaired NK cell function in systemic onset Juvenile Idiopathic Arthritis

Systemic onset Juvenile Idiopathic Arthritis (SoJIA) is an autoimmune disease characterised by arthritis and systemic features. Cytokines like IL1, IL6 and IL18 are thought to be responsible for clinical symptoms. IL18 can drive NK cell function. Recently we showed high IL18 levels in plasma correlate with disease activity in SoJIA [1]. Since SoJIA patients have diminished NK cell function we investigated the functionality of these cells in relation to IL18. All cell culture experiments were performed with IL18 in the presence or absence of IL12. SoJIA patients (n = 15) show significant lower NK cells present in peripheral blood and a differential NK cell receptor phenotype is present compared with pJIA patients (n = 15) and healthy controls (n = 10). No difference in IL18 receptor expression was observed. Furthermore IL18 in plasma of SoJIA patients is able to drive NK cell function of healthy controls and is able to bind the IL18 receptor. However, SoJIA patients fail to upregulate cell mediated killing molecules, such as perforin and IFNγ after IL18 stimulation, nor does IL18 induce phosphorylation of several MAP kinases (ERK1/2 and JNK). Both pJIA patients and healthy controls show phosphorylation of MAP kinases which results in upregulation of cell mediated killing molecules such as IFNg and perforin after stimulation with IL18. We were able to pinpoint the mechanism of the impaired NK cell – IL18 axis to an IL18 receptor defect. This important observation has major implications for understanding the immunepathogenesis of SoJIA.

Receptor Activity-Modifying Proteins Differentially Modulate the G Protein-Coupling Efficiency of Amylin Receptors

Receptor activity-modifying proteins (RAMPs) 1, 2, and 3 are prototypic G protein-coupled receptor accessory proteins that can alter not only receptor trafficking but also receptor phenotype. Specific RAMP interaction with the calcitonin receptor (CTR) generates novel and distinct receptors for the peptide amylin; however, the role of RAMPs in receptor signaling is not understood. The current study demonstrates that RAMP interaction with the CTRa in COS-7 or HEK-293 cells leads to selective modulation of signaling pathways activated by the receptor complex. There was a 20- to 30-fold induction in amylin potency at CTR/RAMP1 (AMY1) and CTR/RAMP3 (AMY3) receptors, compared with CTR alone, for formation of the second-messenger cAMP that parallels an increase in amylin binding affinity. In contrast, only 2- to 5-fold induction of amylin potency was seen for mobilization of intracellular Ca++ or activation of ERK1/2. In addition, in COS-7 cells, the increase in amylin potency for Ca++ mobilization was 2-fold greater for AMY3 receptors, compared with AMY1 receptors and this paralleled the relative capacity of overexpression of Galphaq proteins to augment induction of high affinity 125I-amylin binding. These data demonstrate that RAMP-complexed receptors have a different signaling profile to CTRs expressed in the absence of RAMPs, and this is likely due to direct effects of the RAMP on G protein-coupling efficiency.

International Union of Pharmacology. LXIX. Status of the Calcitonin Gene-Related Peptide Subtype 2 Receptor

Historically, calcitonin gene-related peptide (CGRP) receptors have been divided into two classes, CGRP(1) and CGRP(2). After the cloning of calcitonin receptor-like receptor (CLR) and receptor activity-modifying proteins (RAMPs), it became clear that the CGRP(1) receptor was a complex between CLR and RAMP1. It is now apparent that the CGRP(2) receptor phenotype is the result of CGRP acting at receptors for amylin and adrenomedullin. Accordingly, the term "CGRP(2)" receptor should no longer be used, and the "CGRP(1)" receptor should be known as the "CGRP" receptor.

A Critical Evaluation of Adenosine A2A Receptors as Potentially “Druggable” Targets in Huntingtons Disease

Huntington's disease (HD) is a dominantly inherited neurodegenerative disorder caused by the expansion of a polymorphic CAG trinucleotide repeat encoding a poly-glutamine tract within the Huntingtin protein. GABAergic enkephalin neurons of the basal ganglia, which show the highest levels of expression of adenosine A(2A) receptors, are the most vulnerable in HD. Such a selective neuronal vulnerability, which occurs despite ubiquitous expression of mutant and normal Huntingtin, has suggested that adenosine A(2A) receptors might play a pathogenetic role in HD. In agreement, changes in A(2A) receptor expression and signaling have been reported in various experimental models of HD. The interpretation of the functional significance of the aberrant A(2A) receptor phenotype in HD mice is however complicated by the conflicting data so far reported on the potential neuroprotective and neurodegenerative effects of these receptors in the brain, with some data suggesting a potential pathogenetic role and some other data suggesting activation of trophic or protective pathways in neurons. The same complex profile has emerged in experimental models of HD, in which both A(2A) receptor agonists and antagonists have shown beneficial effects. The main aim of this review is to critically evaluate whether adenosine A(2A) receptors may represent a suitable target to develop drugs against HD.

Estrogen Receptor–Negative Breast Cancer Is Less Likely to Arise among Lipophilic Statin Users

Preclinical studies have shown the anticancer potential of HMG-CoA reductase enzyme inhibitors (statins), whereas epidemiologic studies remain controversial. Because lipophilic statins show preclinical anticancer activity against hormone receptor [estrogen receptor (ER)/progesterone receptor (PR)]-negative breast cancer models, we explored the hormone receptor phenotype of breast cancers that arise in statin users. We did a retrospective cohort analysis via electronic pharmacy records from the Kaiser Permanente Northern California Cancer Registry on 2,141 female patients listed in 2003 as incident cases of breast malignancy. Measures included tumor grade, stage, and receptor phenotype in statin users versus nonusers and controlled for hormone replacement therapy and race. 387 of the 2,141 breast cancer patients used lipophilic statins [lovastatin (85%), simvastatin, and atorvastatin]. Fifty-one women developed ER/PR-negative tumors. The age-adjusted odds ratio (OR) of developing an ER/PR negative tumor was 0.63 (95% confidence interval, 0.43-0.92; P = 0.02) for statin use >or=1 year before breast cancer diagnosis compared with statin use <1 year (including nonuse). Breast cancers in patients with >or=1 year of statin use were more likely to be low grade (OR, 1.44) and less invasive stage (OR, 1.42). Breast cancer patients with exposure to statins have proportionately fewer ER/PR-negative tumors that are of lower grade and stage. Although our data set cannot address whether statins affect the incidence of breast cancer, we show that statin use may influence the phenotype of tumors. This suggests a new potential strategy for breast cancer prevention, that of combining statins with agents that prevent ER-positive cancer (tamoxifen, aromatase inhibitors).

Standardized method to minimize variability in a functional P2X7 flow cytometric assay for a multi‐center clinical trial

Flow cytometric analysis of human P2X(7) pore activity segregates variant from common P2RX7 genotypes and may serve as a biomarker for cancer, pain, inflammation, and immune responses to infection. Standardization is needed to accommodate variable sample age and instrumentation differences in a multicenter clinical trial. CD14-PE-stained whole blood samples were treated with YO-PRO-1 combined with a P2X(7) agonist (BzATP) or control, followed by the addition of PI after closure of the P2X(7) pore. Recalled instrument settings from previous publications were used to adapt a standardized fluorescent particle-adjusted set-up method. Experiments were performed to compare the two methods while evaluating components of systematic variability and facilitating reliable processing of samples with varied ages. The median YO-PRO-1 fluorescence of BzATP-treated samples had less variability when collected by the bead-adjusted method and was less influenced by the compensation strategy used. The average day-to-day coefficient of variance for assessments of P2X(7) pore activity by this method was 0.11 +/- 0.04, and the exclusion of nonviable cells was found to accommodate samples aged up to 4 days after phlebotomy. The bead-adjusted set-up method produced measurements differing by only 2.0% +/- 1.5% on two analog cytometers, and within similar decades when comparing analog to digital instruments. These results provide a standardized method for quantitative flow cytometric analysis of P2X(7) receptor phenotypes in blood monocytes with minimal intralaboratory variation and potential for interlaboratory comparisons that can greatly facilitate multicenter functional genomic clinical studies.

Association of Androgen Receptor CAG Repeat Polymorphism and Polycystic Ovary Syndrome

Genetically determined heightened androgen sensitivity may influence the phenotype of polycystic ovary syndrome (PCOS). To date, studies of the androgen receptor exon 1 polymorphic CAG repeat have produced conflicting results in PCOS. We tested the hypothesis that a lower number of CAG repeats is associated with increased odds of PCOS. We also compared X-chromosome inactivation between cases and controls. Women with and without PCOS were genotyped for the CAG repeat and assessed for X-chromosome methylation. Association analyses were performed. Subjects were recruited from the reproductive endocrinology clinic at the University of Alabama at Birmingham; controls were recruited from the surrounding community. Genotyping took place at Cedars-Sinai Medical Center in Los Angeles. Participants included 330 women with PCOS and 289 controls (77% white, 23% black). Androgen receptor genotype, X-chromosome methylation, and phenotyping for PCOS were measured. A smaller biallelic mean of CAG repeats was associated with increased odds of PCOS. X-chromosome inactivation was not different comparing cases with controls; however, in the subset with nonrandom inactivation, the chromosome bearing the shorter CAG allele was preferentially active in PCOS women. Association of shorter CAG repeats with PCOS is consistent with in vitro functional studies demonstrating higher activity of androgen receptors expressed from alleles with fewer CAG repeats, suggesting inherited alteration in androgen sensitivity may contribute to PCOS. In some women, such heightened sensitivity may also result from preferential expression of androgen receptors with shorter alleles. Thus, genetic and epigenetic changes may be involved in the pathogenesis of PCOS.

Cutaneous γ/δ T-Cell Lymphoma Treated with Radiation and Denileukin Diftitox

Cutaneous γ/δT-cell lymphoma (CGD-TCL) is a rare and usually fatal lymphoma that is generally resistant to chemotherapy and radiation. It has recently been included as a provisional entity in the combined 2005 World Health Organization—European Organization for Research and Treatment of Cancer consensus classification. Subcutaneous panniculitic T-cell lymphomas with a γ/δ T-cell receptor phenotype are now distinguished from the less aggressive α/β panniculitic T-cell lymphoma. We describe a patient with deep ulcers initially diagnosed as pyoderma gangrenosum, which was reclassified as CGD-TCL. Local radiation followed by weekly infusions of denileukin diftitox put the patient into remission.

Cerebral vasoconstriction after subarachnoid hemorrhage - Role of changes in vascular receptor phenotype

The pathological constriction of cerebral arteries known as cerebral vasospasm (CVS) is with a delay of 4 to 10 days linked to subarachnoid hemorrhage. Several agents have been suggested as being responsible; amongst these perhaps 5-hydroxytryptamine (5-HT) and endothelin-1 (ET-1) are the most prominent given their ability to elicit powerful constriction of cerebral arteries. Investigating both 5-HT and ET receptors we have observed that there are distinct changes in receptor phenotype after experimental SAH, namely upregulation of the ETB and 5-HT1B receptors, and that this upregulation is linked to a higher sensitivity to the endogenous agonists. It has also been shown that reduction in regional cerebral blood flow (CBF) is associated with receptor upregulation and interventional animal experiments have shown a benefit from inhibiting the PKC and MAP kinase pathways on receptor upregulation, CBF and neurological outcome.

Human Th17 Cells Share Major Trafficking Receptors with Both Polarized Effector T Cells and FOXP3+ Regulatory T Cells

It is a question of interest whether Th17 cells express trafficking receptors unique to this Th cell lineage and migrate specifically to certain tissue sites. We found several Th17 cell subsets at different developing stages in a human secondary lymphoid organ (tonsils) and adult, but not in neonatal, blood. These Th17 cell subsets include a novel in vivo-stimulated tonsil IL17+ T cell subset detected without any artificial stimulation in vitro. We investigated in depth the trafficking receptor phenotype of the Th17 cell subsets in tonsils and adult blood. The developing Th17 cells in tonsils highly expressed both Th1- (CCR2, CXCR3, CCR5, and CXCR6) and Th2-associated (CCR4) trafficking receptors. Moreover, Th17 cells share major non-lymphoid tissue trafficking receptors, such as CCR4, CCR5, CCR6, CXCR3, and CXCR6, with FOXP3+ T regulatory cells. In addition, many Th17 cells express homeostatic chemokine receptors (CD62L, CCR6, CCR7, CXCR4, and CXCR5) implicated in T cell migration to and within lymphoid tissues. Expression of CCR6 and CCR4 by some Th17 cells is not a feature unique to Th17 cells but shared with FOXP3+ T cells. Interestingly, the IL17+IFN-gamma+ Th17 cells have the features of both IL17-IFN-gamma+ Th1 and IL17+IFN-gamma- Th17 cells in expression of trafficking receptors. Taken together, our results revealed that Th17 cells are highly heterogeneous, in terms of trafficking receptors, and programmed to share major trafficking receptors with other T cell lineages. These findings have important implications in their distribution in the human body in relation to other regulatory T cell subsets.

114. Androgen receptor protein expression influences freeze sensitivity of prostate cancer cells

Introduction A major challenge in prostate cancer (CaP) treatment is evident due to molecular-based transformations resulting in the loss of response to and/or expression of the androgen receptor phenotype. It is generally recognized that many CaPs transform from androgen-dependent/sensitive (AS) to androgen-independent/insensitive (AI) phenotypes with age. Further, is now appreciated that androgen independent CaP is relatively refractory to in situ therapies. Recently, the presumption that freezing serves as a ubiquitous lethal stress in prostate cancer (CaP) has been challenged by anecdotal clinical experience and experimental evidence demonstrating time–temperature related cell death dependencies. As such, in this study we investigate the difference in freeze sensitivity between AI and AS CaP cell lines to determine the impact of CaP AR responsiveness on cryoablation efficacy. We hypothesized that molecular changes resulting in a more aggressive phenotype following androgen ablation treatments is responsible, in part, for the observed increase in cell tolerance to cryotherapy. Methods A series of studies were designed to identify the cellular mechanisms contributing to variations in freeze response. Survival estimates of AS and AI CaP cell lines following freezing exposure were determined using cell viability assays and fluorescent microscopy. Cell death and stress pathway involvement was detailed through molecular analysis and inhibition studies of select key proteins. Results LNCaP (passaged multiple times) cells demonstrated a phenotypic shift to freeze resistance that matched the freeze response of AI PC-3 cells along with a transformation to androgen independence, while PC-3 cells stably expressing a functional androgen receptor matched the freezing sensitivity of the AS LNCaP cell line. Importantly, AI cell lines survived and recovered freezing exposure to temperatures to −40 °C, whereas AS cell lines exposed to temperatures colder than −20 °C did not survive. Cell death assays using an apoptotic inhibitor revealed that apoptosis was significantly increased in both AS cell lines following freezing compared to AI counterparts. These results were further confirmed through caspase activity assays and apoptotic fluorescent probes. Protein analysis indicated the androgen receptor is manipulated post-freezing exposure. Finally, AKT inhibition significantly increased the freezing sensitivity of AI CaP cells. Conclusions These observations are the first to describe a phenotypic variation in CaP cellular response to freezing. Our study demonstrates that AS cancers are far more sensitive to a freezing insult, which might support an interpretation that a younger patient pool may prove to be ideal candidates for cryoablation as a primary therapy. Specifically, CaP cryoablation may prove most effective in CaPs that have not yet progressed to a more resistant AI phenotype. In addition, a further understanding of the role of the AR in freezing responses may increase options for neo-adjunctive treatments targeting the AR signaling pathway.

Prevention of acute graft-versus-host disease by blocking T-cell entry to secondary lymphoid organs

In acute graft-versus-host disease (aGVHD), donor T cells attack the recipient's gastrointestinal tract, liver, and skin. We hypothesized that blocking access to distinct lymphoid priming sites may alter the specific organ tropism and prevent aGVHD development. In support of this initial hypothesis, we found that different secondary lymphoid organs (SLOs) imprint distinct homing receptor phenotypes on evolving alloreactive effector T cells in vivo. Yet preventing T-cell entry to specific SLOs through blocking monoclonal antibodies, or SLO ablation, did not alter aGVHD pathophysiology. Moreover, transfer of alloreactive effector T cells into conditioned secondary recipients targeted the intestines and liver, irrespective of their initial priming site. Thus, we demonstrate redundancy of SLOs at different anatomical sites in aGVHD initiation. Only prevention of T-cell entry to all SLOs could completely abrogate the onset of aGVHD.

Arg389Gly-β1-adrenergic receptors determine improvement in left ventricular systolic function in nonischemic cardiomyopathy patients with heart failure after chronic treatment with carvedilol

Administration of the beta-adrenergic receptor blocker carvedilol to patients with chronic heart failure leads to clinically significant benefits, including improvement in left ventricular systolic function in some, but not all, patients. We sought to determine the basis of the variable effect obtained with carvedilol in patients with heart failure. Carvedilol blocks both beta1-adrenergic and beta2-adrenergic receptors, and both receptors exist as polymorphisms. We aimed to determine whether these polymorphisms contribute to variability in response to carvedilol in patients with chronic heart failure. We retrospectively and prospectively investigated 135 patients with nonischemic cardiomyopathy and chronic stable heart failure (New York Heart Association class II, III) treated with carvedilol. Baseline echocardiography was obtained before introduction of carvedilol and repeated after stabilization of a maximally tolerated dose of carvedilol (50-100 mg/day) for at least 1 year. Polymerase chain reaction and restriction fragment length polymorphism analysis were used to genotype beta1-adrenergic and beta2-adrenergic receptor polymorphisms. When grouped according to receptor polymorphisms patients were well matched for severity of heart failure, comorbidity and treatment. No significant difference was observed in baseline left ventricular ejection fraction (LVEF) between groups (P>0.05). After 1.5 years of treatment with carvedilol patients with Arg389Arg-beta1-adrenergic receptors had a significantly greater improvement in LVEF compared with Gly389 carriers (Arg389Arg 18.8%; Arg389Gly 9.4%; Gly389Gly 6.0%; P<0.001) whereas there were no differences attributable to other beta1-adrenergic and beta2-adrenergic receptor polymorphisms (P>0.05). In patients with nonischemic dilated cardiomyopathy, carvedilol leads to a significantly greater improvement in LVEF in patients with the Arg389Arg-beta1 adrenergic receptor phenotype.

The effects of C-terminal truncation of receptor activity modifying proteins on the induction of amylin receptor phenotype from human CTb receptors

Receptor activity modifying proteins (RAMPs) interact with calcitonin receptors to produce novel amylin receptor phenotypes. We have recently demonstrated that the short intracellular C-terminus of RAMPs plays a key role in the function of amylin receptors derived from the CTa calcitonin receptor through the use of chimeric RAMPs and RAMPs that are truncated at the C-terminus [15, Udawela M, Christopoulos G, Morfis M, Christopoulos A, Ye S, Tilakaratne N, Sexton PM. A critical role for the short intracellular C terminus in receptor activity modifying protein function. Mol Pharmacol 2006;70:1750–60., 18, Udawela M, Christopoulos G, Tilakaratne N, Christopoulos A, Albiston A, Sexton PM. Distinct receptor activity-modifying protein domains differentially modulate interaction with calcitonin receptors. Mol Pharmacol 2006;69:1984–89.]. The calcitonin receptor in humans is expressed as two major alternatively spliced isoforms termed CTa and CTb. Relatively little is known about how alternate splicing of the receptor affects the interaction between calcitonin receptors and RAMPs. We have examined the effect of RAMP truncation, through use of mutant constructs that delete the last 8 amino acids of each of the 3 known human RAMPs, and characterised these for interaction with CTb receptors through co-expression in COS-7 cells. As seen with the CTa receptor isoform, RAMP truncation caused a marked loss in induction of AMYb receptor phenotypes as characterised by 125I-rat amylin radioligand binding assays and cAMP accumulation assays; the latter as a marker of receptor signalling. The effect was most pronounced for RAMP1 and RAMP2 deletion mutants, but attenuated responses were also observed with co-expressed RAMP3 deletion mutants. These data support a direct role for the RAMP C-terminus in the interaction of RAMP/calcitonin receptor complexes with intracellular accessory proteins involved in signalling and/or receptor trafficking.

The Transcription Factor Runx3 Represses the Neurotrophin Receptor TrkB during Lineage Commitment of Dorsal Root Ganglion Neurons

Runx3, a Runt domain transcription factor, determines neurotrophin receptor phenotype in dorsal root ganglion (DRG) neurons. Molecular mechanisms by which Runx3 controls distinct neurotrophin receptors are largely unknown. Here, we show that RUNX3 abolished mRNA induction of TRKB expression, and concomitantly altered the neurotrophin response in a differentiating neuroblastoma cell line. In contrast, RUNX3 did not play a significant role in TRKC regulation even under the relevant BMP signaling pathway. We identified putative regulatory elements of Ntrk2/NTRK2 (a gene that codes for TrkB) using an unbiased computational approach. One of these elements was a highly conserved intronic sequence that contains a cluster of Runx binding sites. In a primary culture of DRG neurons, endogenous Runx3 bound to the consensus cluster, which had repressor activity against the Ntrk2 promoter under the control of NT-3 signaling. Consistent with these findings, Runx3-deficient embryos showed an increased number of trkB+ DRG neurons and failed to maintain trkC expression. Taken together, Runx3 determines TrkC positive sensory neuron identities through the transcriptional repression of TrkB when Trk-BTrkC double positive neurons differentiate into TrkC single positive neurons.

Close Resemblance between Chemokine Receptor Expression Profiles of Lymphoproliferative Disease of Granular Lymphocytes and Their Normal Counterparts in Association with Elevated Serum Concentrations of IP-10 and MIG

T-cell large granular lymphocyte (T-LGL) leukemia and chronic natural killer (NK) cell lymphocytosis (CNKL) are major subtypes of lymphoproliferative disease of granular lymphocytes (LDGL). To clarify the mechanism of LGL proliferation and the relationship with the chemokine system in LDGL, we enrolled 22 T-LGL leukemia patients and 8 CNKL cases, analyzed the expression profiles of chemokine receptors, and measured the serum concentrations of the corresponding chemokines. There were no significant differences in chemokine receptor expression profiles between T-LGL leukemia patients and healthy donors. An association of CCR5 and CXCR3 expression levels on LGLs was recognized in T-LGL leukemia patients (r = 0.84; P < .001). Among the chemokines, serum IP-10 and MIG levels were significantly higher in LDGL patients than in healthy donors (P < .05, and P < .001, respectively), and MIG expression was associated with the number of circulating LGLs (r = 0.73; P < .01). The chemokine receptor phenotypes of LDGL cells are essentially similar to those of normal T-cells and NK cells. The roles of IP-10 and MIG in the pathophysiology of LDGL need further examination.

Hindbrain glutamatergic control of meal size: Evidence for participation of specific N-methyl- D-aspartate receptor (NMDAR) phenotypes on myelinated vagal afferent neurons.

Previously published results from our laboratory indicate that injection of NMDA receptor antagonists into the dorsal hindbrain increase food intake and suggest that capsaicin-resistant (myelinated) vagal afferents terminals that express NMDA-type glutamate receptors participate in control of meal size. NMDA receptors are heteromeric ion channels incorporating distinct subunits, including NR1, NR2A, NR2B, NR2C and NR2D. The NMDA channel's electrical and signaling properties are determined by its NR2 subunit phenotype. In the present study we tested the hypothesis that myelinated vagal afferent neurons express different NMDAR NR2 subunits than non-myelinated neurons. Six-week-old male Sprague-Dawley rats received intraperitoneal injections of capsaicin or vehicle. Nodose ganglia were processed for with primary antibodies against the capsaicin receptor (VR1), and NR1, NR2B, NR2C, or NR2D NMDAR subunits. In vehicle-treated rats, at least 60% of all nodose vagal afferents were immunopositive for VR-1 and capsaicin treatment reduced VR1 immunoreactivity to 10%. Furthermore, capsaicin reduced the number of NMDA NR1 and NR2B subunit expressing neurons by at least 50%, while resulting in no detectable loss of NMDA NR2C or NR2D immunopositive neurons. Our results indicate that NMDA NR2C and NR2D subunit immunoreactivity in the nodose ganglia is limited to capsaicin-resistant myelinated vagal afferents. Taken collectively with previous behavioral findings, we hypothesize that NMDA NR2C and/or NR2D expressing vagal afferents may mediate increased food intake evoked by injection of NMDA antagonists into the hindbrain and may contribute to the control of meal size. Supported by NIH Grants DK52849 and NS20561 and Poncin Scholarship 2006.

The alternation of estrogen receptor (ER) and progesterone receptor (PgR) expression in primary breast cancer patients treated with neoadjuvant chemotherapy (NAC)

21165 Background: Currently, neoadjuvant chemotherapy (NAC) for the operable primary breast cancer has become standard treatment strategy. Primary treatment could affect the status of Estrogen receptor (ER) and Progesterone receptor (PgR) expressions in the tumor, and might have impact on the choice of adjuvant hormone therapy. Methods: We evaluated consecutive 165 primary breast cancer cases those treated with 4 cycles of FEC (500–100–500 mg/m2, q3wks) followed by 4 cycles docetaxel (75 mg/m2, q3wks) as their neoadjuvant treatment in our institution from January 2000 to February 2006. Except for the bilateral breast cancer cases and patients achieved pCR, 107 pairs of core needle biopsy before treatment and tumor block after surgery were evaluated by immunohistochemistry (IHC) for receptor status and analyzed by Allred scoring. Score 4 or more was designated as positive. Results: Median age of patients was 51 (23–71) years old. Hormone receptor (HR) phenotypes before treatment were following; ER+/PgR+:62(57.9%), +/ -:23(21.5%), -/ +:2(1.9%), -/ -:20(18.7%). As the HR status, 94.4 % of patients did not show any alternation for their HR evaluation. Only 4.7% of patients experienced changes from HR+ to HR-, and HR- to HR+ were extremely rare (0.9%). Although ER positivity was not changed in most of cases (95.1%), conversion from PgR + to - were observed in 35.5% of ER+/PgR+ cases. Particularly, the rate of negative conversion of PgR were found more frequently in patients with age under 50 (51.7% in &lt;50y.o. vs. 22.6% in =50 y.o., p=0.03). Whereas, in older patients, negative conversion of PgR correlated to favorable pathological response. Conclusions: In most of patients, neoadjuvant chemotherapy did not affect the evaluation of HR status. Alternation of PgR expression seems to reflect the chemotherapy induced amenorrhea in younger patients. In the older patients, negative conversion of PgR may have some impact on pathological response. No significant financial relationships to disclose.