David Read Johnson

INTERFERON-GAMMA RAPIDLY INCREASES PEPTIDE TRANSPORTER (TAP) SUBUNIT EXPRESSION AND PEPTIDE TRANSPORT CAPACITY IN ENDOTHELIAL CELLS

Human cytotoxic T lymphocytes (CTL) recognize specific complexes of HLA class I molecules and peptides, which assemble when nascent class I molecules bind peptides transported from the cytoplasm into the endoplasmic reticulum by the heterodimeric transporter associated with antigen processing (TAP). Increased class I molecule expression on the cell surface increases the efficiency of CTL lysis. The kinetics of interferon (IFN)-γ induction of TAP, peptide transport capacity, and HLA class I molecule expression was determined in endothelial cells, which are targets of CTL following transplantation or viral infection. TAP mRNAs are induced rapidly, increasing 20-fold (TAP1) or 10-fold (TAP2) by 12 h, whereas HLA class I mRNA is induced more slowly, increasing 10-fold in 24 h. TAP1 and TAP2 proteins are also induced rapidly, increasing 10-fold in 24 h, whereas HLA class I heavy chain proteins and surface expression increase more slowly. Peptide transport capacity in endothelial and HeLa cells increases within 6 h of IFN-γ treatment, suggesting that the IFN-γ-induced TAP heterodimers are functional. Therefore, the IFN-γ-induced increase in TAP proteins is accompanied by an increased peptide transport capacity, which may be important in supporting the subsequent rise in HLA class I protein expression.

Neurobiology of Post-Traumatic Stress Disorder

In the general population, post-traumatic stress disorder (PTSD) has a 1% lifetime prevalence (Heizer et al., 1987). Estimates among war veterans are much higher, with 15% of Vietnam veterans currently meeting criteria for PTSD and 30% meeting lifetime criteria. Since their tour in Vietnam, another 20% of veterans have suffered from partial PTSD, having at least some symptoms disruptive of normal social functioning (Kulka et al., 1990).

The Impact of the Homecoming Reception on the Development of Posttraumatic Stress Disorder: The West Haven Homecoming Stress Scale (WHHSS)

This study reports on the development of a self-report measure of the homecoming experience among Vietnam veterans with posttraumatic stress disorder (PTSD). The West Haven Homecoming Stress Scale (WHHSS), measuring Frequency of Events, Intensity of Feelings, and Level of Support during the first 6 months after return from overseas, and within the past 6 months, was collected from 247 veterans who were receiving inpatient treatment for PTSD. Homecoming Stress was the most significant predictor of current PTSD symptomatology superseding combat exposure, childhood and civilian traumas, and stressful life events. A factor analysis resulted in four orthogonal factors: Shame, Negative Interpersonal Interaction, Social Withdrawal, and Resentment. Homecoming Stress was unchanged over the course of a 4 month inpatient program.

Locus-Specific Constitutive and Cytokine-Induced HLA Class I Gene Expression

Cytokine induction of the MHC class I genes increases the nascent molecules available for binding potentially antigenic peptides. The human H chain loci, HLA-A, -B, and -C, encode highly homologous and polymorphic mRNAs. Here, these transcripts were resolved and measured by competitive PCR of cDNA using locus-specific primers. Endothelial cells expressed many HLA-A and -B, but fewer HLA-C, transcripts. In contrast, HeLa cells expressed many HLA-A and -C, but fewer HLA-B, transcripts. The inflammatory cytokines TNF-α, IFN-β, and IFN-γ induced HLA-B strongly, but HLA-A and -C weakly in both cell types. Combined treatment with IFNs and TNF further increased HLA-A and -B, but not HLA-C transcripts. The constitutive and inducible activities of transfected promoters correlated well with mRNA levels. The weak IFN response of the HLA-A2 promoter was not due to variations in the IFN consensus sequence, the site α, or a 3-bp insertion between them. The HLA-Cw6 promoter was less TNF responsive due to a variant κB enhancer, which also reduced the IFN responses. The NF-κB subunit RelA strongly activated the HLA-A2 and -B7 promoters but only weakly activated the HLA-Cw6 promoter due to the variant κB. Cotransfecting NF-κB1 with RelA further increased activity of the HLA-A2 and -B7, but not HLA-Cw6, promoters. All three promoters were activated by MHC class II trans-activator, but not CREB-binding protein, whereas IFN regulatory factor-1 and -2 weakly activated the HLA-B7 and -Cw6, but not HLA-A2, promoters. These studies illustrate common and locus-specific mechanisms that may be targeted to modulate immune reactions.

Long-term course of treatment-seeking Vietnam veterans with posttraumatic stress disorder: mortality, clinical condition, and life satisfaction.

This study is a 6-year longitudinal study of 51 treatment-seeking male veterans with combat-related posttraumatic stress disorder. Measures of PTSD and psychiatric symptomatology, social functioning, and program impact were assessed at admission to an inpatient treatment program, at 18 months, and 6 years later. Previous studies had shown that the treatment program’s impact on course of illness had been negligible. The sample showed an extremely high mortality rate of 17% over 6 years. The remaining veterans showed improvement in violence and alcohol and drug use, but an increase in hyperarousal symptoms and social isolation. Nearly three-fourths had had an inpatient hospitalization. Veterans’ self-ratings, in contrast, indicated significant improvement in all areas of functioning except employment, as well as an overall positive view of the impact of the program on their lives. Results indicate that the majority of the veteran sample had experienced some improvement in their ability to cope with their chronic illness, decreasing their use of violence and substance abuse but still were experiencing high levels of symptomatology. The extremely high mortality rate, however, provides a somber reminder of the seriousness of this disorder.

Effect of mycophenolic acid on TNFα-induced expression of cell adhesion molecules in human venous endothelial cells in vitro

1 Mycophenolic acid (MPA) is an inhibitor of inosine‐5′‐monophosphate dehydrogenase and therefore interferes with cellular GTP biosynthesis. Recently, MPA has been used as an antiproliferative and immunosuppressive agent. In the present study, the effect of MPA on the expression of the endothelial cell adhesion molecules (CAMs), intercellular (I) CAM‐1, vascular (V) CAM‐1 and endothelial (E)‐selectin, was investigated in tumour necrosis factor‐α (TNFα)‐activated cultured human venous endothelial cells (EC). 2 Surface expression of CAMs was measured by flow cytometry and mRNA expression by Northern blot analysis. Transcriptional activation of CAMs by the nuclear factor NF‐κB was determined by an electromobility shift assay. The function of CAMs was studied by a static adhesion assay with human monocyte‐like undifferentiated U937 cells. 3 Pretreatment of TNFα‐ (5  ng ml−1, 12 h) activated EC with MPA (10 μM, 24 h) increased the binding of U937 cells, which had not been treated with MPA, by ∼amp;2 fold. MPA‐pretreatment of EC did not affect TNFα‐induced surface expression of ICAM‐1. However, VCAM‐1 and E‐selectin were increased 2–3 fold and remained elevated up to 24 h, by which time TNFα‐activated control EC had returned to baseline levels of expression. The effect of MPA on the surface expression of CAMs was half‐maximal at ∼amp;1 μM and required 12 h of pretreatment. Guanosine (0.3 mM), a precursor of GTP, did not prevent the effect of MPA on the expression of CAMs in TNFα‐activated EC. 4 Kinetics of mRNA expression of CAMs mirrored protein expression: mRNA for ICAM‐1 was unaffected, whereas TNFα‐induced mRNA expression for E‐selectin and VCAM‐1 was prolonged and increased by MPA. This effect was not due to increased transcription mediated by the nuclear transcription factor NF‐κB. However, half‐life for E‐selectin mRNA was increased 10 fold by MPA, whereas ICAM‐1 mRNA half‐life was unchanged. 5 The data demonstrate that apart from its antiproliferative effects on lymphocytes, MPA enhances TNFα‐induced VCAM‐1 and E‐selectin surface expression on EC by selectively increasing the mRNA‐stability of these cell adhesion molecules. This effect of MPA on EC appears to be independent from inhibition of inosine‐5′‐monophosphate dehydrogenase.

Interferon Induction of TAP1: The Phosphatase SHP-1 Regulates Crossover Between the IFN-α/β and the IFN-γ Signal-Transduction Pathways

Interferon (IFN)-gamma and IFN-alpha/beta induction of the transporter associated with antigen processing-1 (TAP1) promoter was compared in HeLa cells and endothelial cells (ECs). In HeLa cells, IFN-gamma acts through Stat1alpha/Stat1alpha homodimers binding to the gamma activating sequence (GAS) and IFN-alpha/beta acts through Stat1/Stat2/p48 binding to the IFN-stimulated response element (ISRE). In ECs, however, IFN-gamma and IFN-alpha/beta act through both the GAS and ISRE. The basis of the IFN signaling crossover in ECs was investigated. HeLa and ECs contain similar ratios of Stat1alpha to Stat2 proteins, and IFN-alpha/beta also activates the same Janus kinases (JAKs) (Jak1 and tyrosine kinase (Tyk) 2 but not Jak2). However, IFN-alpha/beta activates more Stat1alpha than does IFN-gamma in ECs, whereas the reverse occurs in HeLa, and expression of the IFN-alpha/beta receptor-associated phosphatase SHP-1 is much lower in ECs than HeLa cells. Overexpression of SHP-1 in ECs blocks IFN-alpha/beta signaling through GAS, and expression of a dominan negative SHP-1 in HeLa cells permits IFN-alpha/ss signaling through GAS, demonstrating a role for SHP-1 in regulating crossovers between the IFN-alpha/beta and IFN-gamma signaling pathways.

The theory and technique of transformations in drama therapy

This paper presents the theoretical and technical applications of Transformations, an advanced form of the Developmental Method (Johnson, 1982, 1984, 1986, 1991), in the treatment of motivated, neurotic clients in individual psychotherapy. First, I will present the basic theoretical concepts of this approach and then articulate the goals of therapy that are derived from this theoretical perspective. After a brief review of the developmental method, transformations will be described in terms of the basic session format, the role of the therapist, types of interventions, and stages of therapy. A detailed case example will then be presented and discussed.

Developmental approaches in drama therapy

Drama therapy, like other creative arts therapies, has attracted a number of different conceptual frameworks which have been used to organize complex clinical material and to provide theoretical coherence to relatively uncharted psychological territory. These frameworks, nearly all imported from outside the creative arts themselves, include psychoanalytic theory, gestalt therapy, developmental psychology, existential thought, and Jungian analysis. Each of these perspectives has contributed greatly toward a fuller understanding of the therapeutic functions of drama and action. This paper will elucidate the outlines of a developmental approach to drama therapy, and attempt to show how both drama and development are intimately linked to the concept of transformation: the flexible alteration of self in response to the ever-changing world about one. The writer will first describe the developmental paradigm and then examine five basic developmental processes which bring some measure of coherence to the theory and technique of drama therapy. These five processes will be illustrated by specific clinical examples. Finally, the writer will discuss how a developmental approach aids in the evaluation of specific techniques for specific populations.

Differential expression of human major histocompatibility class I loci: HLA-A, -B, and -C.

Expression of the human class I MHC loci, HLA-A, -B, and -C, was examined by reverse transcription and competitive PCR with locus-specific primers. This approach allows unambiguous identification of target sequences by analysis of the amplified products. JY and Pala lymphoblastoid B cells express more HLA-A than HLA-B mRNAs and little HLA-C mRNA. Raji Burkitt lymphoma and HeLa carcinoma cells express approximately equal amounts of HLA-A and HLA-C mRNAs but less HLA-B mRNA. Jar trophoblast cells express no HLA class I mRNAs. Surprisingly, K562 leukemia cells express significant amounts of HLA-C mRNA. However, K562 cells contain no detectable HLA-A or -B mRNAs, suggesting that these loci are regulated independently. Furthermore, cultured endothelial cells and smooth muscle cells express low, approximately equal amounts of HLA-A, -B, and -C mRNAs, whereas donor-matched, EBV transformed B cells express much more HLA-B mRNA, suggesting that cell type dependent regulation underlies differential locus expression. Finally, expression of HLA class I molecules on the cell surface correlates with total HLA mRNAs but not with mRNAs encoded by any one locus. Differential expression of these HLA class I loci may contribute to cell-type dependent immune reactions by preferentially presenting distinct peptides to T cells.