Important note: Information in this article was accurate in 1994. The state of the art may have changed since the publication date.
Production and characterization of monoclonal antibodies to human Pneumocystis carinii for the diagnosis of P. carinii pneumonia.
Eur J Med. 1992 Jun;1(3):132-8. Unique Identifier : AIDSLINE MED/94080314 Nato F; Contini C; Zamora-Zavala C; Viscardi P; Delia S; Mojon M; Mazie JC; Hybridolab, Departement des Biotechnologies, Institut Pasteur,; Paris, France.
Abstract:
OBJECTIVES: Monoclonal antibodies against human Pneumocystis carinii were produced by fusion of myeloma cells (X63-AG8.653) with splenocytes from Biozzi mice that had been immunized against P. carinii cysts isolated from infected human lung. The aim of this study was to evaluate the usefulness of these monoclonal antibodies for the diagnosis of P. carinii pneumonia by indirect immunofluorescence in comparison with a modified silver stain method and commercial kits. METHODS: One hundred fifty-seven specimens from 87 patients, infected or non-infected with human immunodeficiency virus, were examined for the presence of P. carinii. Specimens were either induced sputum samples or bronchoalveolar lavage fluids. Indirect immunofluorescence was performed with six stable clones obtained by limiting dilution. Four of the monoclonal antibodies were IgG1 isotypes, one was an IgG3 and one was an IgM. Their isoelectric points varied from 6.5 to 8.3. Tests were also performed with silver methenamine staining and with two commercially available monoclonal antibodies (Monofluo kit from Diagnostics Pasteur and MAb from Dako). RESULTS: The 6 antibodies all recognized P. carinii cysts in indirect immunofluorescence. No cross reactivity was observed with yeast or host cells. P. carinii antigens could not be identified with western immunoblotting suggesting that the monoclonal antibodies recognized native antigens. This result was confirmed by dot blot analysis. Spots were observed with native but not with denatured antigens. Inhibition studies showed that these 6 antibodies recognized the same or overlapping sites. The sensitivities of detection of P. carinii in sputum were 87% by silver stain and from 93.5 to 96.7% by immunofluorescence. The sensitivities of detection in bronchoalveolar lavage were 67.3% by silver stain and from 75.7% to 76.8% by immunofluorescence. CONCLUSION: Immunofluorescence was more sensitive than silver staining and the best results were obtained with E5-8 and A8-13 monoclonal antibodies and with Monofluo kit.
Keywords: Animal *Antibodies, Monoclonal/ISOLATION & PURIF Antigens, Fungal/IMMUNOLOGY Bronchoalveolar Lavage Fluid/IMMUNOLOGY Comparative Study Electrophoresis *Fluorescent Antibody Technique Human Immunoblotting Mice Pneumocystis carinii/*IMMUNOLOGY Pneumonia, Pneumocystis carinii/*DIAGNOSIS/IMMUNOLOGY Rabbits Reagent Kits, Diagnostic Sensitivity and Specificity Silver Staining Sputum/IMMUNOLOGY JOURNAL ARTICLE 940330
M9431022
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Important note: Information in this article was accurate in 1994. The state of the art may have changed since the publication date.
