title: Virulence of feline Chlamydia psittaci in mice is not a function of the major outer membrane protein (MOMP).
abstract: The virulent strain of feline Chlamydia psittaci, the Cello strain, produces conjunctivitis and upper respiratory disease in cats. This same strain produces a lethal disease in mice when inoculated intraperitoneally (i.p.). In this study we have shown that the Baker strain of feline C. psittaci is attenuated in the mouse model system. Intraperitoneal inoculation of mice with the Baker strain produced no disease but did stimulate an immune response that protected the mice from subsequent produced i.p. inoculation with the virulent Cello strain. To determine if the difference between these two strains was in the major outer membrane protein (MOMP), the omp1 gene which codes for this protein was sequenced for both the Baker and Cello strains. The MOMP was chosen to study because in Chlamydia trachomatis this protein has been shown to contain neutralizing epitopes and has been shown to play a role in cell attachment. These functions make it a likely structural component capable of mutating and causing altered cell tropism and virulence. The DNA sequence of the omp1 was determined by amplifying the gene with PCR, cloning the PCR product into the pCR-II cloning vector and determining the DNA sequence of the inserted gene using primers to sites in the plasmid vector. From the DNA sequence, the deduced amino acid sequence of MOMP was determined for both the attenuated Baker and the virulent Cello strains of feline C. Psittaci. The results indicated that the omp1 gene of both strains contained 1179 base pairs which coded for a protein 392 amino acids. The DNA sequences of the omp1 gene of the two strains differed by only two base pairs which resulted in two amino acid changes in the MOMP. The Baker strain had a serine instead of a tryptophan at amino acid 7 and a tyrosine instead of an aspartic acid at amino acid 125 of the uncleaved protein. Neither amino acid change was in an area of the MOMP which could logically account for the difference in biological activity. Amino acid 7 was in the leader sequence which is cleaved from the authentic MOMP and is not present in the infectious elementary body. Amino acid 125 was in a conserved hydrophobic area of one of the constant regions of the protein. A change at this location was not likely surface exposed and thus could not affect cell adhesion, tissue tropism or neutralizing epitopes. Therefore, the differences in the primary structure of the MOMP from the Baker and Cello strains of feline C. psittaci could not account for the attenuation of the Baker strain for mice. The molecular basis of their difference is yet to be determined.
authors.affiliation: Department of Veterinary and Biomedical Sciences, University of Nebraska-Lincoln 68583, USA.
authors.forename: S W
authors.initials: SW
authors.lastname: May
authors.forename: C L
authors.initials: CL
authors.lastname: Kelling
authors.forename: M
authors.initials: M
authors.lastname: Sabara
authors.forename: J
authors.initials: J
authors.lastname: Sandbulte
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics; D001483:Base Sequence; D002371:Cat Diseases; D002415:Cats; D002460:Cell Line; D002691:Chlamydophila psittaci / Q000235:genetics / Q000472:pathogenicity* / Q000502:physiology; D017931:DNA Primers; D004285:Dogs; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D009956:Psittacosis / Q000382:microbiology / Q000662:veterinary*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity; D014774:Virulence
chemical_list: D001425:Bacterial Outer Membrane Proteins; D017931:DNA Primers; C069991:outer membrane protein, Chlamydia psittaci 6BC