All members of the family
Chlamydiaceae
have lipopolysaccharides (LPS) that possess a shared carbohydrate trisaccharide antigen, 3-deoxy-
d
-manno-oct-2-ulosonic acid (Kdo) that is functionally uncharacterized. A single gene, genus-specific epitope (
gseA
), is responsible for attaching the tri-Kdo to lipid IVA. To investigate the function of Kdo in chlamydial host cell interactions, we made a
gseA
-null strain (L2Δ
gseA
) by using TargeTron mutagenesis.
The Chlamydiae are obligate intracellular pathogens that develop and multiply within a poorly characterized parasitophorous vacuole (the inclusion) during growth.
Chlamydia abortus
is a major pathogen of sheep and other ruminants, and its inclusion development is poorly characterized. We used immunofluorescence microscopy, quantitative culture, and qPCR to examine
C. abortus
inclusion development and to examine the interaction of
C. abortus
inclusions with those formed by other species. Antibodies used in these studies include sera from ewes from production facilities that were naturally infected with
C. abortus
. Multiple inclusions are often found in
C. abortus
-infected cells, even in populations infected at very low multiplicity of infection. Labeling of fixed cells with sera from infected sheep revealed fibrous structures that extend away from the inclusion into the cytoplasm of the host cell.
C. abortus
inclusions fused with
C. caviae
and
C. psittaci
inclusions in coinfected cells. Inclusions formed by
C. abortus
and
C. caviae
did not fuse with inclusions formed by
C. trachomatis
,
C. pneumoniae
, or
C. pecorum
. The ability of inclusions to fuse was correlated with the overall genomic relatedness between species, and with sequence similarity in the inclusion membrane protein IncA. Quantitative PCR data demonstrated that
C. abortus
grows at a decreased rate during coinfections with
C. caviae
, while
C. caviae
growth was unaffected. The collected data add depth to our understanding of inclusion development in this significant zoonotic veterinary pathogen.
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