The C. parapsilosis sensu lato group involves three closely related species, C. parapsilosis sensu stricto,
C
. orthopsilosis
and
C
. metapsilosis
. Although their overall clinical importance is dramatically increasing, there are few studies regarding the virulence properties of the species of the psilosis complex. In this study, we tested 63 C. parapsilosis sensu stricto, 12
C
. metapsilosis
and 18
C
. orthopsilosis
isolates for the ability to produce extracellular proteases, secrete lipases and form pseudohyphae. Significant differences were noted between species, with the
C
. metapsilosis
strains failing to secrete lipase or to produce pseudohyphae. Nine different clinical isolates each of C. parapsilosis sensu stricto,
C
. orthopsilosis
and
C
. metapsilosis
were co-cultured with immortalized murine or primary human macrophages. C. parapsilosis sensu stricto isolates showed a significantly higher resistance to killing by primary human macrophages compared to
C
. orthopsilosis
and
C
. metapsilosis
isolates. In contrast, the killing of isolates by J774.2 mouse macrophages did not differ significantly between species. However, C. parapsilosis sensu stricto isolates induced the most damage to murine and human macrophages, and
C
. metapsilosis
strains were the least toxic. Furthermore, strains that produced lipase or pseudohyphae were most resistant to macrophage-mediated killing and produced the most cellular damage. Finally, we used 9 isolates of each of the C. parapsilosis sensus lato species to examine their impact on the survival of
Galleria
mellonella
larvae. The mortality rate of
G
. mellonella
larvae infected with
C
. metapsilosis
isolates was significantly lower than those infected with C. parapsilosis sensu stricto or
C
. orthopsilosis
strains. Taken together, our findings demonstrate that
C
. metapsilosis
is indeed the least virulent member of the psilosis group, and also highlight the importance of pseudohyphae and secreted lipases during fungal-host interactions.