Development of a rapid diagnostic test for pertussis: direct detection of pertussis toxin in respiratory secretions.

Monoclonal antibodies (MAb) were produced against the specific Bordetella pertussis antigen pertussis toxin (PT). In preliminary studies, one MAb (IB12) was selected and used in an enzyme-linked dot blot immunoassay to evaluate the ability of the method to detect known amounts of PT in control experiments and to test its potential for direct detection of PT in nasopharyngeal secretion (NP) specimens from patients with confirmed cases of whooping cough. The dot blot assay was able to detect PT at levels as low as 10 ng per dot in either buffer or control NP specimens. The assay demonstrated specificity, reacting only with dot blots of whole B. pertussis and not Bordetella bronchiseptica, Bordetella parapertussis, or other bacterial strains. In preliminary studies, NP aspirate, swab, and wash specimens were compared. The specimen of choice was found to be the NP aspirate, for which 100% positive results were found in the assay. These initial studies suggest that the dot blot immunoassay in which a MAb is used for direct detection of PT in NP specimens may be useful as a rapid diagnostic test for pertussis.

Regional differences in quantities of histochemically detectable mucosubstances in nasal, paranasal, and nasopharyngeal epithelium of the bonnet monkey.

Inhaled irritants induce secretory cell hyperplasia in nasal epithelium of animals. To characterize this response histochemically it is first important to know the histochemical character and distribution of epithelial mucosubstance in the normal nasal cavity. An automated image analyzing method was used to detect and quantitate acidic, neutral, and sulfated mucosubstances in the epithelium lining the nasal and paranasal airways of eight bonnet monkeys. Tissue sections 2 micron thick from defined regions of these airways were stained with either alcian blue/periodic acid-Schiff to demonstrate acid and neutral mucosubstances or high iron diamine to demonstrate sulfated mucins. Respiratory epithelium covering maxilloturbinates had the largest volume of stainable mucosubstance per unit surface area of basal lamina, whereas the maxillary sinus epithelium had the least. There was a general anteroposterior increase in the quantity of total epithelial mucosubstance along the septal and lateral walls of the nasal cavity, and there was more acidic than neutral mucosubstance in the posterior nasal airway than in the anterior. Epithelial mucosubstance in the maxillary sinus was predominantly neutral. Therefore, we conclude that there are substantial regional quantitative differences in stainable mucosubstances in the primate nasal epithelium which must be considered when examining nasal mucosa for irritant-induced changes in epithelial mucins.

Different patterns of cytokeratin expression in the normal epithelia of the upper respiratory tract.

The distribution and type of cytokeratins present in the normal human epithelia of the nasopharynx, oropharynx, tongue, palatine tonsil, epiglottis, vocal cord, and laryngeal ventricle were studied using immunohistochemical techniques and by gel electrophoresis of cytoskeletal proteins microdissected from frozen tissues. Noncornifying stratified epithelia covering the oropharynx, tongue, surface of the palatine tonsil, pharyngeal surface of the epiglottis, and vocal cord were all found to contain cytokeratins nos. 4, 5, 6, 13, 14, and 15, together with minor amounts of cytokeratin no. 19, i.e., a pattern similar to that previously reported for esophageal epithelium. The immunohistochemical reaction with KA4, an antibody specific for cytokeratins nos. 14, 15, 16, and 19, revealed reactivity confined to the basal epithelial cells of the tongue, oropharynx, pharyngeal epiglottis, and two out of five samples of vocal cords. This same antibody reacted with the entire thickness of three out of the five true vocal cords which were shown by gel electrophoresis to also contain cytokeratins nos. 16 and 17. Gel electrophoresis revealed that the pseudostratified columnar epithelium covering the laryngeal ventricle was more complex, in that it contained cytokeratins nos. 5, 13, 14, 15, and 17, which are typical of stratified epithelia, as well as cytokeratins nos. 7, 8, 18, and 19, which are characteristic of simple epithelia. This pattern is similar to that found in bronchial epithelium. The laryngeal surface of the epiglottis exhibited cytokeratins nos. 4, 5, 7, 8, 13, 14, 15, 17, 18, and 19, i.e., a pattern combining features of both esophageal- and bronchial-type epithelia. The reaction of these epithelia containing columnar cells with antibody RGE-53, which is specific for cytokeratin no. 18, revealed a staining reaction confined to the superficial columnar cells, whereas KA1 stained only the basal cells of these epithelia. The results of our study make it possible to distinguish two types of noncornifying stratified squamous epithelium, namely the 'esophageal type' which covers the tongue, oropharynx, and pharyngeal surface of the epiglottis, and another type which overlies the vocal cords and the transitional zone between the pharyngeal and laryngeal surfaces of the epiglottis. Furthermore, there appear to be variants of pseudostratified columnar epithelium, i.e., the usual bronchial type lining the laryngeal ventricle, and a type with a thicker subcolumnar cell compartment that is found on the laryngeal surface of the epiglottis. The patterns of expression of cytokeratins in the respiratory tract are compared with those of other epithelia.

Neutralizing influenza antibodies, IgA and total protein in the nasopharyngeal secretions of subjects vaccinated by nasal route with the inactivated influenza vaccine prepared in the "Stefan S. Nicolau" Institute of Virology.

Intranasal administration of two doses of the inactivated influenza vaccine prepared in the "Stefan S. Nicolau" Institute of Virology was followed by rises in the level of neutralizing secretory influenza antibodies in 82% of the cases. The concomitant study of secretory antibody, IgA and total protein levels, as well as of the serum HAI influenza antibodies demonstrated that their evolution was parallel only in 23% of the vaccinees. The percentage of secretory antibody conversion was similar to the rate of protection conferred by the vaccine.

Fetal lung maturatio. III. Amniotic fluid phosphatidic acid phosphohydrolase (PAPase) and its relation to the lecithin/sphingomyelin ratio.

The purpose of this study was to establish 1) the sequential changes in specific activity of phosphatidic acid phosphohydrolase (PAPase) in amniotic fluid and its relation to the lecithin/sphingomyelin (L/S) ratio; and 2) the origin of amniotic fluid PAPase. the increase in the amniotic fluid L/S ratio is preceded by an increase in PAPase activity, rising from 15 nmoles of phosphate released per milligram of protein per hour at 30 weeks to 100 nmoles at 37 weeks. The mean PAPase activity in the nasopharyngeal fluid of the infant is 456 nmoles of phosphate released per ml per hour, the amniotic fluid mean PAPase activity at delivery being 129 nmoles (P less than 0.01). These findings are consistent with the view that amniotic fluid PAPase originates, in part, from the fetal lung and likely participates in the regulation of the synthesis of lecithin.

New Japanese rubella vaccine: comparative trials.

A total of 142 seronegative volunteers were given one of the following rubella vaccines: Cendehill, HPV77. DE-5, RA27/3, or a new Japanese vaccine, To-336. To-336 vaccine produced a slightly higher geometric mean antibody titre (G.M.T.) (65.7) than did the HPV77. DE-5 (63.1) or RA27/3 vaccine (61.9) but the G.M.T. induced by Cendehill vaccine was much lower (39.3).Reactions, particularly joint symptoms, occurred least commonly after vaccination with To-336 vaccine. Joint symptoms occurred within seven days of menstruation in 30 out of 37 (81%) vaccines (P <0.01); their incidence was not related to oral contraception.Though there is evidence to suggest that Japanese virus strains may be non-teratogenic further data on the incidence of congenitally acquired infection in Japan must be collected before this conclusion can be supported on epidemiological grounds.