Abstract

We have developed a method for short-term culture of Pneumocystis carinii using the CCL 64 mink lung cell line as a feeder cell layer. This culture method has enabled us to obtain a relatively pure Pneumocystic carinii trophozoite population in culture. This population has been surface labeled using a 125I Iodogen method. A 120 Kilodalton glycoprotein (p120) has been identified and purified. It is a major surface protein. We propose to obtain the primary structure of this protein. We will use this information to synthesize a DNA consensus probe. This probe will be used to identify and study the structure of the p120 gene and will also be used in a hybridization assay for P. carinii. We have produced antibodies in rabbits to the p120 protein and propose to develop a quantitative ELISA assay to measure the p120 protein in pulmonary lavage fluid. Pneumocystis pneumonia is the commonest fatal complication of the Acquired Immunodeficiency Syndrome AIDS. There are at present no quantitative assays for P. carinii.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI024194-02
Application #
3136983
Study Section
Tropical Medicine and Parasitology Study Section (TMP)
Project Start
1987-09-30
Project End
1990-08-31
Budget Start
1988-09-01
Budget End
1989-08-31
Support Year
2
Fiscal Year
1988
Total Cost
Indirect Cost

  Institution

Name
Yale University
Department
Type
Schools of Medicine
DUNS #
082359691
City
New Haven
State
CT
Country
United States
Zip Code
06520

  Publications

Koziel, H; Phelps, D S; Fishman, J A et al. (1998) Surfactant protein-A reduces binding and phagocytosis of pneumocystis carinii by human alveolar macrophages in vitro. Am J Respir Cell Mol Biol 18:834-43
Narasimhan, S; Armstrong, M; McClung, J K et al. (1997) Prohibitin, a putative negative control element present in Pneumocystis carinii. Infect Immun 65:5125-30
Narasimhan, S; Armstrong, M Y; Rhee, K et al. (1994) Gene for an extracellular matrix receptor protein from Pneumocystis carinii. Proc Natl Acad Sci U S A 91:7440-4
Williams, D J; Radding, J A; Dell, A et al. (1991) Glucan synthesis in Pneumocystis carinii. J Protozool 38:427-37
Armstrong, M Y; Smith, A L; Richards, F F (1991) Pneumocystis carinii pneumonia in the rat model. J Protozool 38:136S-138S
Armstrong, M Y; Koziel, H; Rose, R M et al. (1991) Indicators of Pneumocystis carinii viability in short-term cell culture. J Protozool 38:88S-90S
Radding, J A; Armstrong, M Y; Ullu, E et al. (1989) Identification and isolation of a major cell surface glycoprotein of Pneumocystis carinii. Infect Immun 57:2149-57