Surfactant protein A2 (SP-A2) variants expressed in CHO cells stimulate phagocytosis of Pseudomonas aeruginosa more than do SP-A1 variants.

Surfactant protein A (SP-A) enhances phagocytosis of Pseudomonas aeruginosa. Two functional genes, SP-A1 and SP-A2, encode human SP-A. As we showed before, baculovirus-mediated insect cell-expressed SP-A2 enhances the association of P. aeruginosa with rat alveolar macrophages (rAMs) more than does SP-A1. However, true phagocytosis (internalization) was not shown, and insect cell derived proteins lack or are defective in certain mammalian posttranslational modifications that may be important for SP-A1 and SP-A2 activity and specificity. Here we used SP-A1 (6A(2), 6A(4)) and SP-A2 (1A(0), 1A(1)) allele variants expressed by CHO (Chinese hamster ovary) mammalian cells to study their effect on association and/or internalization of P. aeruginosa by rAMs and/or human AMs (hAMs) and to study if phagocytosis can be modulated differentially and/or more effectively by CHO cell-expressed SP-A variants than by insect-cell expressed SP-A variants. For cell association and internalization assessments, light microscopy and fluorescence-activated cell sorter analyses were used, respectively. We found the following for the first time. (i) SP-A2 variants enhanced phagocytosis (cell association and/or internalization) of P. aeruginosa more than SP-A1 variants did, and the cell association correlated with internalization. (ii) Differences in the activities of SP-A variants were observed in the following order: 1A(1)>1A(0)>6A(2)>6A(4). (iii) rAMs, although more active than hAMs, are an appropriate model, as SP-A2 variants exhibited activity higher than that seen for SP-A1 variants with either rAMs or hAMs. (iv) CHO cell-expressed SP-A was considerably more active than insect cell-expressed variants. We conclude that SP-A2 variants stimulate phagocytosis of P. aeruginosa more effectively than SP-A1 variants and that posttranslational modifications positively influence the phagocytic activity of SP-A.

Recognition of apertures in overhead transparent barriers by leopard frogs.

Anurans have independent systems for detecting moving stimuli and stationary opaque objects. We have discovered that leopard frogs will also orient to, and spontaneously and accurately jump through, circular apertures in overhead transparent covers. When given a choice between one large aperture of 3.8 cm diameter, and three apertures of smaller but equal diameter (2.5 cm diameter, 1.9 cm diameter, or 1.3 cm diameter) they choose the larger diameter aperture at a frequency (64, 87 and 97%, respectively) that is statistically greater than chance. In only 1 of 255 attempts was there a jump to the overhead cover that was not directed at an aperture. Atectal frogs are still able to detect and jump accurately through transparent apertures. Frogs cannot distinguish between two apertures of equal diameter if one aperture is covered with clear plastic with high light transmissibility (92% of transmissibility of air). However, if the plastic covering of the aperture has a residue which reduces light through the cover from 92 to 87% of the transmissibility of air, frogs will jump to the uncovered aperture at a frequency that is statistically greater than chance. Our results show that leopard frogs have an extremely well developed ability to detect overhead apertures just as they can vertical obstacles. They are able to jump towards such openings with a small margin of error independent of the tectal visual system.