Jane E. Holliday

Psychosocial modifiers of immunocompetence in medical students.

&NA; This study addressed the effects of a naturally occurring stressor on components of the immune response. Blood was drawn twice from 75 first‐year medical students, with a baseline sample taken one month before their final examinations and a stress sample drawn on the first day of final examinations. Median splits on scores from the Holmes—Rahe Social Readjustment Rating Scale and the UCLA Loneliness Scale produced a 2 X 2 X 2 repeated measures ANOVA when combined with the trials variable. Natural killer (NK) cell activity declined significantly from the first to the second sample. High scorers on stressful life events and loneliness had significantly lower levels of NK activity. Total plasma IgA increased significantly from the first to second sample, while plasma IgG and IgM, C‐reactive protein, and salivary IgA did not change significantly.

Stress, loneliness, and changes in herpesvirus latency

This study used a prospective design to examine the influence of examination stress and loneliness on herpesvirus latency as measured by changes in antibody levels to three herpesviruses, Epstein-Barr virus (EBV), Herpes simplex type I (HSV-1), and cytomegalovirus (CMV). Three blood samples were obtained from 49 first-year medical students, with the first sample drawn 1 month before final examinations, the second on the first day of final examinations, and the third during the first week after their return from summer vacation. A median split on the UCLA Loneliness Scale divided subjects into high- and low-scoring loneliness groups. There were significant changes in the antibody titers to all three herpesviruses across the sample points, with the lowest levels found in the third (low stress) sample. High-loneliness subjects had significantly higher EBV antibody titers than low-loneliness subjects. These data suggest that stress-related immunosuppression can significantly modulate herpesvirus latency.

Modulation of Cellular Immunity in Medical Students

This study assessed the psychosocial modulation of cellular immunity in 34 medical-student volunteers. The first blood sample was obtained 1 month before examinations, and the second on the day of examinations. There were significant declines in the percentage of helper/inducer T- lymphocytes, in the helper/inducer-suppressor/cytotoxic-cell ratio, and in natural killer-cell activity in the blood samples obtained on the day of examinations. Half of the subjects were randomly assigned to a relaxation group which met between sample points; the frequency of relaxation practice was a significant predictor of the percentages of helper/inducer cells in the examination sample. Three biochemical nutritional assays (albumin, transferrin, and total iron-binding protein) were within normal limits on both samples. Data from the Brief Symptom Inventory showed significantly increased global self-rated distress associated with examinations in the no-intervention group, compared to nonsignificant change in the relaxation group. Clinical and theoretical implications are discussed.

Stress-related impairments in cellular immunity

The percentages of total T-lymphocytes (OKT-3+), helper T-cells (OKT-4+), and suppressor T-cells (OKT-8+) were significantly lower in blood samples obtained from 40 medical students during examinations, compared to baseline values obtained 6 weeks earlier. In addition, the response of T-lymphocytes to stimulation by phytohemagglutinin and concanavilin A was also significantly lower during examinations, compared to baseline. Self-report data documented significantly greater distress associated with examinations. The data have implications for immunosuppressive disorders and stress-related illnesses.

Stress and the Transformation of Lymphocytes by Epstein-Barr Virus

Although various stressors appear to influence herpesvirus infections, the underlying mechanisms have not been studied. A prospective design was used to examine the effects of examination stress and loneliness on the transformation of B lymphocytes in mixed cultures of T and B lymphocytes by Epstein-Barr virus (EBV). Three blood samples were drawn from 42 EBV-seropositive medical students, with the baseline sample taken 1 month before their final examinations, the stress sample drawn on the first day of final examinations, and the third sample taken the first week after their return from summer vacation. A median split on the UCLA Loneliness Scale divided the subjects into high- and low-scoring loneliness groups. There were significant effects for change over trials, with the lowest transformation levels (i.e., more virus required to transform cells) found in the stress sample. There was also a significant main effect for loneliness, in which high loneliness was associated with lower transformation levels. Possible immunological pathways for the observed changes are discussed.

Induction of a deoxyuridine triphosphate nucleotidohydrolase activity in Epstein-Barr virus-infected cells

Superinfection of Raji cells with Epstein-Barr virus (EBV) or chemical induction of HR-1 cells with 12-O-tetradecanoyl-phorbol-13-acetate (TPA) results in the induction of a deoxyuridine triphosphate nucleotidohydrolase (dUTPase) which is not observed in mock-treated cells or TPA-treated EBV genome-negative BJAB cells. The EBV-induced dUTPase could be distinguished from the host dUTPase based upon differences in their migration in polyacrylamide gels and sensitivity to the 5-mercurithioguanosine derivitive of dUTP. The expression of the EBV-specified dUTPase is prevented by phosphonoacetic acid indicating that its expression is dependent upon EBV-DNA replication.

Functional mapping of the Epstein-Barr virus genome: Identification of sites coding for the restricted early antigen, the diffuse early antigen, and the nuclear antigen

Attempts were made to functionally map antigenic expression of the Epstein-Barr virus (EBV) to specific regions on the EBV genome, using the B95-8 strain. Experiments were performed to map the expression of early antigen (EA), both restricted and diffuse (R and D, respectively), and the EBV nuclear antigen (EBNA), using intact B95-8 DNA, cloned BamHI fragments or Charon 4A fragments. DNA preparations were microinjected into two EBV genome-negative epithelial tumor cell lines. Expression of EBV antigens was monitored using precharacterized human sera, as well as monoclonal antibodies to EA-R and EA-D. The data suggest that EA-R maps to the BamHI H fragment, and EA-D maps to the Charon 4A fragment 7. A previous report that BamHI K is associated with the expression of a nuclear neoantigen tentatively identified as EBNA (W.P. Summers, E.A. Grogan, D. Shedd, M. Robert, C.R. Liu, and G. Miller, Proc. Nat. Acad. Sci. USA 79, 5688-5692, 1982) was also confirmed.

Identification of a region of the Epstein-Barr virus (B95-8) genome required for transformation

In a previous study the identification of a region(s) of the Epstein-Barr virus (EBV) genome, which is associated with transformation, was attempted by marker rescue. A transforming EBV was rescued from D98/HR-1 hybrid cells, which contain the non-transforming HR-1 EBV genome, after transfection with specific BamHI and Charon 4A fragments (J. Stoerker and R. Glaser, Proc. Nat. Acad. Sci. USA 80, 1726-1729, 1983). In this study, characterization of the EBV DNA in four human lymphoblastoid cell lines (LCL) transformed with rescued virus was performed. It was found that recombination between the transfected fragments, BamHI H,F,X and the Charon 4A fragment (EB-26-36) which is equivalent to the BamHI H,F,X region, and the endogenous HR-1 EBV genome in the D98/HR-1 cells took place. This recombination resulted in the formation of transforming EBV. The EBV DNA in the four LCLs are similar to each other and to HR-1 EBV DNA. However, the EBV DNA in all four LCLs also contain the U2 region plus additional sequences of B95-8 DNA. The U2 region is deleted in HR-1 EBV DNA which is associated with HR-1 cells and the D98/HR-1 hybrid cells. Thus, transforming activity of the HR-1-like viruses rescued from D98/HR-1 cells was concomitant with the recombination of the 0.26-0.36 region of the EBV genome, suggesting that this region is necessary for at least the initiation of transformation.

Identification of EBV-Specific Antigens Following Microinjection of Subgenomic DNA Fragments

The regions of the Epstein-Barr Virus (EBV) genome which code for proteins within the early antigen (EA) and viral capsid antigen (VCA) complex were identified by indirect immunofluorescence (IF) 2–4 days after microinjection of subgenomic cloned fragments of EBV DNA. Two new regions have been identified as part of the early antigen (EA) complex, namely, the Charon 4A cloned fragments which cover map units 38–47 and 83–93 respectively. One DNA fragment from map units 45–54, produces a protein in human cells after microinjection which reacts with EA-VCA+ human sera. Attempts to transform human B-lymphocytes from cord blood with a variety of EBV DNA fragments is described.