Charlene K. Edelman

RUBELLA VIRAEMIA AND ANTIBODY RESPONSES AFTER RUBELLA VACCINATION AND REIMMUNISATION

Eleven 4-13 year old schoolgirls, who were seronegative by haemagglutination inhibition (HI) and radioimmunoassay (RIA) tests despite having been given HPV77-DE5 vaccine 3-9 years previously, were revaccinated with RA27/3. They showed evidence of residual immunity since they had accelerated immune responses, little or no rubella-specific IgM, no viraemia, and no vaccine-induced reactions. In contrast, all but one of the five adult women who were primary vaccinees showed a more delayed immune response. Three of four women tested had viraemia and two had vaccine-induced reactions. Enhanced HI and enhanced RIA showed that many of the schoolgirls had antibody before challenge, as did a fifth adult, who also showed an accelerated immune response, yet became viraemic.

Human cytomegalovirus-stimulated peripheral blood mononuclear cells induce HIV-1 replication via a tumor necrosis factor-alpha-mediated mechanism.

Human cytomegalovirus (HCMV) is a potential cofactor in HIV-1 infection. To investigate the mechanism whereby HCMV promotes HIV-1 replication, a PBMC coculture assay which measures HIV-1 p24 antigen release was used as an index of viral replication. HCMV-stimulated PBMC were capable of inducing HIV-1 replication in cocultures with acutely infected PBMC; however, this occurred only when the PBMC were from HCMV-seropositive donors (598 +/- 207 versus 27 +/- 10 pg/ml p24 antigen with PBMC from HCMV-seronegative donors on day 6 of coculture). Upon stimulation with HCMV, PBMC obtained exclusively from HCMV-seropositive donors released tumor necrosis factor (TNF)-alpha (270 +/- 79 pg/ml at 18 h of culture). Monoclonal antibodies to TNF-alpha blocked the activity of HCMV-stimulated PBMC in cocultures both with acutely HIV-1-infected PBMC and with the chronically infected promonocytic line U1. Also, treatment of HCMV-stimulated PBMC with pentoxifylline, an inhibitor of TNF-alpha mRNA, markedly reduced HIV-1 replication in cocultures both with acutely and chronically infected cells. These results indicate that TNF-alpha is a key mediator of HIV-1 replication induced by HCMV-stimulated PBMC and support the concept that this cytokine plays an important role in the pathogenesis of HIV-1 infection.

Controlled trial of acyclovir for chickenpox evaluating time of initiation and duration of therapy and viral resistance

Background. Chickenpox is prevalent in the US despite the availability of an effective vaccine. Acyclovir treatment is limited by concerns about efficacy if given after the first day of rash and by concerns about induction of viral resistance. Objective. Evaluate initiation and duration of acyclovir treatment of chickenpox and its effect on viral resistance. Study design. Randomized, placebo-controlled, double blind trial in immunocompetent patients who were stratified by age at enrollment (children, 2 to 11 years; adolescents, ≥12 to 18 years; adults, ≥19 years) and duration of rash (≤24 h vs. >24 to 48 h). Lesions were staged, counted and cultured; temperatures and symptoms were recorded daily. Intervention. Subjects presenting within 24 h of rash onset (Group A) were randomly assigned to 5 or 7 days of oral acyclovir treatment, 80 mg/kg/day up to a maximum of 3200 mg/day in four divided doses. Subjects whose rash was >24 to 48 h old were randomized to receive 5 days of acyclovir treatment beginning on the first (Group B1) or second study day (Group B2). Matching placebos were used to ensure that subjects uniformly received 28 doses of study compound. Results. Of the 177 subjects recruited Group A patients who were treated on the first day of rash had the greatest number of significantly shortened event times with 5 days of therapy being equivalent to 7 days. There also were some shorter times to events for Group B1 patients who began therapy on the second day of rash vs. Group B2 patients who started acyclovir on the third. These included: time to maximum lesion formation (adolescents, P = 0.007; children, P = 0.03); 50% healing in adolescents (P = 0.005); and residual facial lesions in adults (P = 0.047). The probability of viral shedding was significantly reduced for Group A subjects vs. Group B1 subjects (P = 0.006). Viruses shed during therapy remained susceptible to acyclovir and retained normal thymidine kinase function. Conclusions. Immunocompetent children, adolescents and adults with chickenpox displayed a gradation in their clinical responses to acyclovir that correlated with the time from onset of rash to initiation of therapy. Five days of therapy is sufficient because a 7-day course provided no additional benefit. The susceptibility to acyclovir of viruses shed during treatment did not change; however, the effect of therapy on resistance of latent virus was not assessed.

Pharmacologic basis for high-dose oral acyclovir prophylaxis of cytomegalovirus disease in renal allograft recipients.

The incidence of cytomegalovirus disease, the most important infectious complication of renal transplantation, was reduced in renal allograft recipients by a regimen of prophylactic high-dose oral acyclovir. To analyze the pharmacologic aspects of our prophylactic approach, we evaluated safety, pharmacodynamics, and in vitro susceptibility data. One hundred four recipients of cadaveric renal allografts received either oral acyclovir (n = 53) in doses of up to 3,200 mg/day or a placebo (n = 51) for 12 weeks posttransplant. Leukocyte count and serum creatinine were selected as markers of laboratory safety and were evaluated pretransplant, at study midpoint (creatinine only), and at study completion. Concentrations of acyclovir in plasma were determined to verify the ability of the dosing strategy to achieve predicted values. Viral resistance was assessed by calculation of in vitro 50% inhibitory concentrations (IC50s) of acyclovir for the cytomegalovirus strains collected from the subjects. Our results showed no difference in leukocyte count or serum creatinine between the acyclovir and placebo recipients. Plasma acyclovir concentrations were maintained within the expected limits and did not differ between patients who developed cytomegalovirus disease and those who did not. The mean acyclovir IC50s for cytomegalovirus isolates were 42.6 mumol/liter in the acyclovir recipients and 48 mumol/liter in the placebo recipients. We conclude that the clinical benefit of high-dose oral acyclovir therapy occurred despite plasma drug concentrations below the mean IC50 for the patient viral isolates. Furthermore, the use of the regimen did not produce leukopenia, adversely affect renal function, or alter the susceptibility of cytomegalovirus strains to acyclovir. This approach and dose adjustment scheme may be appropriate for other immunocompromised patients at risk for cytomegalovirus infection and disease.

Laboratory studies of acute varicella and varicella immune status

We evaluated varicella-zoster virus (VZV) culture and serum antibody methods utilizing specimens from 620 children enrolled in protocols for prevention or treatment of varicella and samples routinely submitted to the Clinical Virology Laboratory. In a foreskin fibroblast tube culture system, we initially isolated VZV from only 29 (51%) of 57 children cultured on the first day of varicella. After modifying the method, the proportion of culture-positive children increased significantly to 36 (80%) of 45 (p less than 0.005 by corrected X2), and the median days-to-positivity were significantly shortened from 5.6 to 3.8 days (p less than 0.001, Wilcoxon rank-sum test). The Viran fluorescent antibody to membrane antigen (FAMA) assay was difficult to read and not reproducible. The standard FAMA was more sensitive than the Merck ELISA antibody test for detecting vaccine-induced antibody. The Whittaker ELISA did not detect vaccine-induced antibody but was comparable to FAMA for immune status testing (sensitivity, 95%; specificity, 92%) and for diagnosis of acute varicella.

Evaluation of centrifugation cultures of bronchoalveolar lavage fluid for the diagnosis of cytomegalovirus pneumonitis

Cytomegalovirus (CMV) pneumonitis is one of the most severe manifestations of CMV disease among immunocompromised patients. The diagnosis of CMV pneumonitis traditionally has required the use of invasive procedures such as lung biopsy. In this retrospective study, we evaluated a centrifugation culture method in samples of bronchoalveolar fluid for the noninvasive diagnosis of CMV pneumonitis. During a 9-mo period, 75 bronchoalveolar lavage samples were collected from 58 patients with pneumonitis. We analyzed the data from 21 patients in whom lung tissue samples were obtained within 14 days of the bronchoalveolar lavage. Centrifugation cultures of bronchoalveolar fluid were positive for CMV in 12 cases. CMV pneumonitis was confirmed in samples of lung tissue from five (42%) of the 12 patients, whereas no evidence of CMV pneumonitis was found in the remaining seven (58%) cases. Of nine patients with negative centrifugation cultures, CMV pneumonitis was confirmed in two (22%). When compared with conventional cultures, we found bronchoalveolar lavage fluid centrifugation cultures to be highly sensitive (100%) and specific (92%) for the detection of CMV infection. However, detection of CMV by centrifugation culture proved to be only moderately sensitive (71%) and nonspecific (50%) for the diagnosis of CMV pneumonitis.