Astrocyte cytolysis by MHC class II-specific mouse T cell clones.

The brain is "immunologically privileged," in part because class I and II MHC antigens are not normally present on glia or neurons. However, under certain conditions such as transplantation, glial cells express MHC proteins at levels sufficient for glia to become targets of immune responses. Cultured astrocytes expressing very low levels of MHC class I protein are killed efficiently by MHC class I antigen-specific CTL. Mouse brain allografts, however, are rejected by CD4+ T cells that are likely to be class II MHC-specific. The level of expression of MHC class II antigen needed to trigger specific killing of astrocytes by CD4+ T cells, independent of exogenous antigen, has not been studied. We examined the role of glial class II MHC in the lysis of cultured neonatal mouse astrocytes by an alloreactive murine CD4+ CTL alone. IFN-gamma induced functionally relevant increases in MHC class II antigen on target cells. Astrocytes were lysed by the CD4+ clone only when class II MHC antigens reached levels readily detectable by flow cytometry. MHC class II expression and lysis increased when astrocytes were coincubated with IFN-gamma and TNF-alpha. Conversely, lysis decreased when class II expression was downregulated by IFN-alpha/beta or dbcAMP. Cytolysis by CD4+ clones was blocked by antibodies to CD4 and LFA-1 on T cells, and to ICAM-1 and class II molecules on astrocytes. The role of LFA-1 in CD4+ cell-mediated lysis was greater than that of LFA-1/ICAM-1 in CD8+ T cell-mediated lysis. CD4+ cells may lyse activated astrocytes when the immune privilege of the brain is compromised as in transplantation, tumors, and inflammatory diseases.

Trends in spinal pain management injections in academic radiology departments.

BACKGROUND AND PURPOSE: There is a paucity of information present in the current literature with regard to the role of SPMI performance in academic radiology centers. Our aim was to evaluate the current practice patterns for the performance of SPMIs in academic radiology departments. MATERIALS AND METHODS: A survey of 186 academic radiology departments in the United States was conducted between March 2009 and May 2009. The survey included questions on departmental demographics, recent trends in departmental SPMI performance, type of physicians who refer to radiology for SPMI performance, types of SPMIs offered, the fraction of total institutional SPMI volume performed by radiologists, and the current state of resident and fellow SPMI training proficiency. RESULTS: Forty-five of the 186 (21.4%) surveys were completed and returned. Twenty-eight of the 45 responding departments stated that they performed SPMIs; the other 17 stated that they did not. Among the 28 responding departments that perform SPMIs, 6 (21.4%), 5 (17.9%), and 8 (28.6%) stated that the number of departmental SPMIs had, respectively, increased, decreased, or remained stable during the past 5 years. SPMI referrals to radiology were made by orthopedic surgeons, neurologic surgeons, neurologists, psychiatrists, anesthesiologists, and internal medicine physicians. CESIs, SNRBs, facet injections, and synovial cyst aspirations are the most frequently performed injections. Fellows and residents become proficient in 88.5% and 51.9%, respectively, of SPMI-performing departments. Most departments perform <50% of the SPMI volume of their respective institutions. CONCLUSIONS: Most responding academic radiology departments perform SPMIs. Most fellows and just more than half of residents at SPMI-performing departments achieve SPMI proficiency. For the most part, the number of SPMIs performed in responding departments has been stable during the past 5 years.

CT-guided cervical transforaminal steroid injections: where should the needle tip be located?

BACKGROUND AND PURPOSE: The aim of CT-guided CTSI is to inject medication into the foraminal region where the nerve root is inflamed. The optimal location for needle placement and therapeutic delivery, however, remain uncertain. The purpose of this study was to investigate how needle positioning and angle of approach impact the transforaminal distribution of injectate. MATERIALS AND METHODS: We retrospectively reviewed fluoroscopic images from 90 CT-guided CTSI procedures for needle-tip location, needle angle, and contrast distribution. Needle-tip position was categorized as either foraminal zone, junctional, or extraforaminal. Distribution of contrast injected immediately before steroid administration was categorized as central epidural, intraforaminal, or extraforaminal in location. Needle-tip location and angle were correlated with contrast distribution. RESULTS: The needle tip was most commonly placed in the junctional position (36 cases, 40%), followed by foraminal (30 cases, 33%) and extraforaminal (24 cases, 27%) locations. Intraforaminal contrast distribution was highest when the needle location was foraminal (30/30, 100%) or junctional (35/36, 97%), compared with extraforaminal (7/24, 29%) (P value <.0001). There was no relationship between needle angle and contrast distribution. CONCLUSIONS: Needle-tip location at the outer edge of the neural foramen (junctional location) correlated well with intraforaminal distribution of contrast for CT-guided CTSI and compared favorably with injectate distribution following foraminal zone needle positioning. Junctional needle positioning may be preferred over the foraminal zone by some proceduralists. Extraforaminal needle positioning resulted in less favorable contrast distribution, which may significantly diminish the therapeutic efficacy of CTSI.

Whole-cell chloride currents in rat astrocytes accompany changes in cell morphology.

Astrocytes can change shape dramatically in response to increased physiological and pathological demands, yet the functional consequences of morphological change are unknown. We report the expression of Cl- currents after manipulations that alter astrocyte morphology. Whole-cell Cl- currents were elicited after (1) rounding up cells by brief exposure to trypsin; (2) converting cells from a flat polygonal to a process-bearing (stellate) morphology by exposure to serum-free Ringer's solution; and (3) swelling cells by exposure to hypo-osmotic solution. Zero-current potentials approximated the Nernst for Cl-, and rectification usually followed that predicted by the constant-field equation. We observed heterogeneity in the activation and inactivation kinetics, as well as in the relative degree of outward versus inward rectification. Cl- conductances were inhibited by 4, 4-diisothiocyanostilbene-2,2'-disulfonic acid (200 microM) and by Zn2+ (1 mM). Whole-cell Cl- currents were not expressed in cells without structural change. We investigated whether changes in cytoskeletal actin accompanying changes in astrocytic morphology play a role in the induction of shape-dependent Cl- currents. Cytochalasins, which disrupt actin polymers by enhancing actin-ATP hydrolysis, elicited whole-cell Cl- conductances in flat, polygonal astrocytes. In stellate cells, elevated intracellular Ca2+ (2 microM), which can depolymerize actin, enhanced Cl- currents, and high intracellular ATP (5 mM), required for repolymerization, reduced Cl- currents. Modulation of Cl- current by Ca2+ and ATP was blocked by concurrent whole-cell dialysis with phalloidin and DNase, respectively. Phalloidin stabilizes actin polymers and DNase inhibits actin polymerization. Dialysis with phalloidin also prevented hypo-osmotically activated Cl- currents. These results demonstrate how the expression of astrocyte Cl- currents can be dependent on cell morphology, the structure of actin, Ca2+ homeostasis, and metabolism.

Cytoskeletal actin gates a Cl- channel in neocortical astrocytes.

Increases in astroglial Cl- conductance accompany changes in cell morphology and disassembly of cytoskeletal actin, but Cl- channels underlying these conductance increases have not been described. We characterize an outwardly rectifying Cl- channel in rodent neocortical cultured astrocytes and describe how cell shape and cytoskeletal actin modulate channel gating. In inside-out patch-clamp recordings from cultured astrocytes, outwardly rectifying Cl- channels either were spontaneously active or inducible in quiescent patches by depolarizing voltage steps. Average single-channel conductance was 36 pS between -60 and -80 mV and was 75 pS between 60 and 80 mV in symmetrical (150 mM NaCl) solutions. The permeability ratio (PNa/PCl) was 0.14 at lower ionic strength but increased at higher salt concentrations. Both ATP and 4, 4-diisothiocyanostilbene-2,2'-disulfonic acid produced a flicker block, whereas Zn2+ produced complete inhibition of channel activity. The frequency of observing both spontaneous and inducible Cl- channel activity was markedly higher in stellate than in flat, polygonally shaped astrocytes. In addition, cytoskeletal actin modulated channel open-state probability (PO) and conductance at negative membrane potentials, controlling the degree of outward rectification. Direct application of phalloidin, which stabilizes actin, preserved low PO and promoted lower conductance levels at negative potentials. Lower PO also was induced by direct application of polymerized actin. The actions of phalloidin and actin were reversed by coapplication of gelsolin and cytochalasin D, respectively. These results provide the first report of an outwardly rectifying Cl- channel in neocortical astrocytes and demonstrate how changes in cell shape and cytoskeletal actin may control Cl- conductance in these cells.