Immunoassay of human Neisseria meningitidis serogroup A antibody.

An enzyme immunoassay is described which quantitates antibodies to Neisseria meningitidis serogroup A capsular polysaccharide in human sera. Modifications of a previously developed two-day assay by Carlone et al. were made to permit analysis in one day and to be compatible with automation. The allowable variations in assay conditions and the areas in which stringent control must be maintained for consistent assay performance are described. Antigen-coating parameters, the kinetics of primary and secondary antibody incubation steps, the buffer compositions, including detergents, serum requirements, and the need for blocking steps were examined. Our modified one-day assay showed excellent agreement with the standardized method of Carlone, with a correlation coefficient between the two methods of 0.989. This assay is adaptable within a permissible range of parameters thus facilitating the implementation of the standardized assay. This will maximize the consistency of results from serum analysis for conjugate vaccine trials related to serotype A Neisseria meningitidis.

Immunoassay of human Neisseria meningitidis serogroup A antibody.

An enzyme immunoassay is described which quantitates antibodies to Neisseria meningitidis serogroup A capsular polysaccharide in human sera. Modifications of a previously developed two-day assay by Carlone et al. were made to permit analysis in one day and to be compatible with automation. The allowable variations in assay conditions and the areas in which stringent control must be maintained for consistent assay performance are described. Antigen-coating parameters, the kinetics of primary and secondary antibody incubation steps, the buffer compositions, including detergents, serum requirements, and the need for blocking steps were examined. Our modified one-day assay showed excellent agreement with the standardized method of Carlone, with a correlation coefficient between the two methods of 0.989. This assay is adaptable within a permissible range of parameters thus facilitating the implementation of the standardized assay. This will maximize the consistency of results from serum analysis for conjugate vaccine trials related to serotype A Neisseria meningitidis.

Evaluation of purified UspA from Moraxella catarrhalis as a vaccine in a murine model after active immunization.

Moraxella catarrhalis causes otitis media, laryngitis, and respiratory infections in humans. A high-molecular-weight outer membrane protein from this bacterium named ubiquitous surface protein A (UspA) is present on all isolates. A monoclonal antibody (MAb) to UspA that recognizes a conserved epitope of this protein has been shown to promote pulmonary clearance of bacteria in passively immunized mice. In the present study, M. catarrhalis heterologous isolates were screened by dot blot with a panel of four additional MAbs specific for surface-exposed epitopes of UspA from M. catarrhalis isolate 035E. Three of the MAbs were specific for 035E, and the fourth reacted with 17 (74%) of the 23 isolates tested. Thus, UspA contains highly conserved, semiconserved, and variable surface-exposed epitopes. The UspA was purified from the 035E isolate by ion-exchange and size-exclusion chromatography, formulated with the adjuvant QS-21, and used to immunize BALB/c mice. Upon pulmonary challenge with either 035E or the heterologous isolate TTA24, significantly fewer bacteria were recovered from the lungs of immunized mice 6 h postchallenge than from control mice. The immune sera from mice or guinea pigs contained high titers of antibodies to the homologous isolate and heterologous isolates in a whole-bacterial-cell enzyme-linked immunosorbent assay. Sera against UspA, whether prepared in mice or guinea pigs, had complement-dependent bactericidal activity toward homologous and 11 heterologous M. catarrhalis isolates. These results indicate that the conserved epitopes of the UspA are highly immunogenic and elicit broadly reactive and biologically functional antibodies. UspA may offer protection against M. catarrhalis infections and is being further evaluated as a vaccine candidate.

Information about morality and behaviour of victims.

According to the Italian Law of Procedure (1988) documents which inform about the morality of the guilty, the witness and the victim are not admitted; those about the victim are admitted only when the criminal event needs to be evaluated in relation to his/her behaviour and moral qualities. In our opinion, in this case, generic moral qualities do not have any real significance; only information about the victim's objective behaviour should be admitted: for instance, in the case of the crime of "corruption of a minor", a type of objective behaviour can be habitual prostitution. Moreover, the norm at issue contrasts with the general principle in the Italian law that the rights concerning the personality of an individual, particularly of a minor, are to be protected, and with the fundamental norm in the law of Procedure according to which criminological examinations are not allowed during the trial. It is useful, however, to acquire elements in order to evaluate the credibility of the witness.

Deontological aspects in recombinant DNA researches.

Legal problems exist in regard to patent rights for new biotechnologies. Ethical problems arise in connection with the application of the technique to patients and with the possibility to cure embryos. The potentialities of preventive medicine have affected even hiring criteria, in that individuals who present enzymic deficiencies can be considered at risk when working in contact with certain substances. For example, firms could decide not to advance certain individuals to positions of responsibility on the grounds that they present the gene of familial hypercholesterolemia. It can be anticipated that the diagnostic potentialities of the recombinant DNA technique will be applied to the evaluation of risk for the stipulation of life insurance, and that further developments, permitting to determine the genetic predisposition for most diseases, would altogether nullify the basic principles of health- and life-insurance policies. An international committee will have to discuss these issues and to formulate deontological rules to regulate both the sphere of occupation and that of insurance.