RTS,S malaria vaccine efficacy and immunogenicity during Plasmodium falciparum challenge is associated with HLA genotype.

Although RTS,S remains the most advanced malaria vaccine, the factors influencing differences in vaccine immunogenicity or efficacy between individuals or populations are still poorly characterised. The analyses of genetic determinants of immunogenicity have previously been restricted by relatively small sample sizes from individual trials. Here we combine data from six Phase II RTS,S trials and evaluate the relationship between HLA allele groups and RTS,S-mediated protection in controlled human malaria infections (CHMI), using multivariate logistic or linear regression. We observed significant associations between three allele groups (HLA-A∗01, HLA-B∗08, and HLA-DRB1∗15/∗16) and protection, while another three allele groups (HLA-A∗03, HLA-B∗53, and HLA-DRB1∗07) were significantly associated with lack of protection. It is noteworthy that these 'protective' allele groups are thought to be at a lower prevalence in sub-Saharan African populations than in the UK or USA where these Phase II trials occurred. Taken together, the analyses presented here give an indication that HLA genotype may influence RTS,S-mediated protective efficacy against malaria infection.

Genetic Diversity and Protective Efficacy of the RTS,S/AS01 Malaria Vaccine.

BACKGROUND: The RTS,S/AS01 vaccine targets the circumsporozoite protein of Plasmodium falciparum and has partial protective efficacy against clinical and severe malaria disease in infants and children. We investigated whether the vaccine efficacy was specific to certain parasite genotypes at the circumsporozoite protein locus. METHODS: We used polymerase chain reaction-based next-generation sequencing of DNA extracted from samples from 4985 participants to survey circumsporozoite protein polymorphisms. We evaluated the effect that polymorphic positions and haplotypic regions within the circumsporozoite protein had on vaccine efficacy against first episodes of clinical malaria within 1 year after vaccination. RESULTS: In the per-protocol group of 4577 RTS,S/AS01-vaccinated participants and 2335 control-vaccinated participants who were 5 to 17 months of age, the 1-year cumulative vaccine efficacy was 50.3% (95% confidence interval [CI], 34.6 to 62.3) against clinical malaria in which parasites matched the vaccine in the entire circumsporozoite protein C-terminal (139 infections), as compared with 33.4% (95% CI, 29.3 to 37.2) against mismatched malaria (1951 infections) (P=0.04 for differential vaccine efficacy). The vaccine efficacy based on the hazard ratio was 62.7% (95% CI, 51.6 to 71.3) against matched infections versus 54.2% (95% CI, 49.9 to 58.1) against mismatched infections (P=0.06). In the group of infants 6 to 12 weeks of age, there was no evidence of differential allele-specific vaccine efficacy. CONCLUSIONS: These results suggest that among children 5 to 17 months of age, the RTS,S vaccine has greater activity against malaria parasites with the matched circumsporozoite protein allele than against mismatched malaria. The overall vaccine efficacy in this age category will depend on the proportion of matched alleles in the local parasite population; in this trial, less than 10% of parasites had matched alleles. (Funded by the National Institutes of Health and others.).

Safety and immunogenicity profile of an experimental hepatitis B vaccine adjuvanted with AS04.

The reactogenicity and safety of an experimental hepatitis B (HB) vaccine containing adjuvant system (AS04) was compared with a licensed vaccine in a phase III, single-blind, randomised study in healthy volunteers >or=15 years of age. A total of 1303 subjects were enrolled to receive either two doses of HB-AS04 (0, 6 months) or three doses of the comparator vaccine (0, 1, 6 months). Two doses of HB-AS04 elicited seroprotection rates close to 100% and two-fold higher GMTs than the comparator vaccine. Results showed that both vaccines were well tolerated and the general safety profile of HB-AS04 was similar to that of the comparator vaccine.

Accidental ingestion of a zinc and copper sulfate preparation.

CASE REPORT: An 86-year-old woman accidentally ingested a preparation containing zinc and copper sulfate. At ninety minutes after ingestion, the peak plasma concentration was 1979 micrograms/dL for zinc and 209 micrograms/dL for copper, suggesting preferential absorption of zinc. The major complications were gastric and bronchial inflammation due to the corrosive properties of these compounds. Systemic manifestations also developed with cardiovascular failure and renal insufficiency, but the patient made a complete recovery. In addition to symptomatic treatment, chelation therapy with dimercaprol and D-penicillamine was given for 48 h. CONCLUSION: The available clinical and toxicokinetic data do not support the benefits of chelation in addition to supportive therapy.

Towards standardization of specific protein assays.

A new international reference preparation for proteins in human serum has been recently proposed in Europe from the Community Bureau of Reference (Brussels) under the name CRM 470 (Certified Reference Material) and in the United States from the College of American Pathologists under the name RPPHS (Reference Preparation for Proteins in Human Serum). This serum based material offers the opportunity to the laboratories to standardize their serum protein concentrations, and to improve dramatically the transferability of the results from studies to the local working conditions. The International Federation of Clinical Chemistry strongly recommends the use of this preparation by all laboratories. The Belgian Institute of Hygiene and Epidemiology expects an improvement in the agreement of the results of the three proteins (IgA, IgG, CRP) included in its external quality control of the clinical laboratories. These proteins were omitted from the controls of the second semester of 1994, but would reappear in early 1995, after the conversion of the laboratories to the new protein calibrators. Of course, this implies the assignment of new reference ranges for most of the serum proteins, some of them being significantly modified (alpha 1-antitrypsin, haptoglobin, transferrin, C3, IgM). The clinical chemists must inform the physicians of these changes.

Measurement of beta 2-microglobulin in serum by a particle-enhanced nephelometric immunoassay.

A particle-enhanced immunoassay of beta 2-microglobulin in serum is described. It is based on the agglutination of complexes formed between the serum beta 2-microglobulin and latex particles coated with F(ab')2 fragments of polyclonal anti-beta 2-microglobulin antibodies. The analytical range of the method is 0.50 to 16 mg/l; it can be extended by appropriate dilution to 0.12 to 80 mg/l with good precision (CV less than 5% over the whole range). The accuracy and the precision are confirmed by a good correlation with radioimmunoassay (n = 123, r = 0.993). No error due to antigen excess was observed, even up to 292 mg/l. The main advantages of the method are its simplicity, its low cost per test and its high sensitivity (final dilution of the sample at 1/1200) with no known interference. The calibration curve is stable for at least 2 weeks.

Clostridium difficile-associated diarrhea after norfloxacin.

Although theoretically the spectrum of antimicrobial quinolone antibiotics are at low risk of producing Clostridium difficile overgrowth and diarrhea, a patient developed this clinical problem while receiving norfloxacin. We review the antimicrobial activity of the quinolone antibiotics, with respect to their predisposition for producing C. difficile-induced diarrhea.

Virological examinations of patients with AIDS-related complex/Walter-Reed 5 enrolled in a double-blind placebo-controlled study with intravenous gammaglobulin administration. Prognostic value of anti-p24 determination. The ARC-IVIG Study Group.

Thirty patients with AIDS-related complex/Walter-Reed 5 enrolled in a placebo-controlled double-blind study with high-dose intravenous gammaglobulin administration were tested by quantitating HIV Western blot and other serological tests for viral antibodies. Furthermore, conventional virus isolation attempts were performed. Absence or loss of p24 antibodies during the study period was associated with progression to AIDS (p = 0.01) and thereby was an earlier prognostic parameter of a poor prognosis than T4 cell count. Neither changes in antibody patterns against other HIV polypeptides, HIV titers in the immunofluorescence test nor demonstration of HIV antigen were significantly associated with progression to AIDS. Cytomegalovirus (CMV) could be isolated from two duodenal biopsies of a patient who developed AIDS at the same time, but a concomitant serological diagnosis of CMV infection was not successful. Though signs in the serology of human herpesviruses (herpes simplex virus, CMV, Epstein-Barr virus), possibly indicating a reactivation of latent infections, could be observed in some instances, a correlation with clinical symptoms or the clinical outcome was not feasible, perhaps also because of a poor standardization of some of the test kits used. All patients were positive for IgG antibodies against the three herpesviruses when entering the study. High prevalence of hepatitis B virus (HBV) markers was found (83% anti-HBc positive), only 1 patient being chronically infected and highly infectious, as shown by HBV-DNA hybridization. No significant difference between treatment and placebo group was observed with the parameters tested in this study.

Medical and technical usefulness of measurement of kappa and lambda immunoglobulin light chains in serum with an M-component.

Kappa and lambda-immunoglobulin light chains were measured to evaluate their usefulness for identifying monoclonal components in serum. Reference concentrations of the Ig light chains were evaluated by the analysis of the serum of 50 blood donors. Two hundred and fifty patients were selected for the presence of an M-component in their serum, which was detected by serum electrophoresis on agarose. The heavy and light chains of the M-component were identified by immunofixation. The concentrations of the three major immunoglobulin classes and of the two light chain types were measured by immunonephelometry. On the basis of the Ig kappa/lambda ratio alone, the Ig light chain type was correctly identified in 76% of the M-components, and there were no misidentifications. By comparing the calculated concentrations of M-component and residual polyclonal Ig with the measured concentrations of the 3 major Ig classes, it was possible to identify the heavy chain type of 58% of the M-components. These observations suggest that M-components can often be identified by the measurement of Ig light chains, a procedure involving less work and time than immunofixation. Moreover, the follow up of gammapathies could benefit from quantitative parameters evaluated from the Ig light chain concentrations: the concentration of the M-component, the ratio of M-component vs total Ig of the same class, and the ratio of M-component vs the sum of the 3 major Ig classes.

Immunoassay of low concentrations of albumin in urine by latex particle counting.

We describe here a nonisotopic immunoassay, based on particle-counting technology, for the determination of urinary albumin. The assay takes only 35 min and has been fully automated on the IMPACT (Acade Diagnostic Systems, Brussels, Belgium) machine. The system measures albumin within a linear range between 6.25 and 50 mg/L and has a detection limit of 0.4 mg/L. Analytical recoveries at three concentrations ranged between 96% and 102%. Within-run precision ranged from 1.6% to 9.5%. The method was compared with a commercial nephelometric immunoassay system and a correlation coefficient of 0.996 was found for 216 urine samples. No antigen excess affects the shape of the curve in our system, whereas in nephelometry a 3 g/L solution of albumin starts to decrease the dose-response curve.