Topically applied manganese-porphyrins BMX-001 and BMX-010 display a significant anti-inflammatory response in a mouse model of allergic dermatitis.

In this study, we topically administered two antioxidant compounds, the manganese-porphyrin-derivatives BMX-001 and BMX-010, in a mouse model of allergic dermatitis and compared the efficacy for reduction of itch and inflammation. In vitro effects of BMX-001 and BMX-010 on keratinocytes, bone marrow derived dendritic cells (BMDCs) and T-cells were initially analysed. For assessment of scratching behaviour, BMX-001 and BMX-010 (0.01 and 0.1 %) were topically applied 16 h and/or 1 h before compound 48/80 or toluene-2,4,-diisocyanate (TDI) challenge in a TDI induced mouse dermatitis model. Additionally, assessment of allergic skin inflammation was performed in a similar manner in the TDI model. Post-treatment ear thickness was measured 24 h after TDI challenge and compared to basal values. The mice were sacrificed and the ear auricle was removed for further analysis. In vitro, both BMX substances significantly inhibited cytokine production of keratinocytes as well as of BMDC and T-cell proliferation. Topical treatment with BMX cream resulted in a significant decrease in scratching behaviour in the compound 48/80 model, but not in the TDI model. Mice treated with BMX-001 and BMX-010 showed a moderate dose dependent decrease in ear thickness, and interestingly, the concentration of the cytokines IL-1ß and IL-4 in inflamed skin was reduced by 80-90 % by all treatment options. These first results suggest the potential benefit of a BMX-001 and BMX-010 cream for the treatment of allergic-inflammatory skin diseases.

Regulation of surfactant protein D in the rodent prostate.

BACKGROUND: Surfactant protein D (SP-D) is an innate immune protein that is present in mucosal lined surfaces throughout the human body, including the male reproductive tract. In the present study, we characterized the regulation of SP-D expression in the mouse and rat prostate. METHODS: Real time reverse transcriptase polymerase chain reaction (RT-PCR) and immunostaining were used to characterize SP-D mRNA and protein in the mouse male reproductive tract. In order to evaluate the effects of testosterone on SP-D gene expression, we measured SP-D mRNA levels via real time RT-PCR in prostates from sham-castrated mice and castrated mice. In addition, we used a rat prostatitis model in which Escherichia coli was injected into the prostate in vivo to determine if infection influences SP-D protein levels in the prostate. RESULTS: We found that SP-D mRNA and protein are present throughout the mouse male reproductive tract, including in the prostate. We determined that castration increases prostate SP-D mRNA levels (~7 fold) when compared to levels in sham-castrated animals. Finally, we demonstrated that infection in the prostate results in a significant increase in SP-D content 24 and 48 hours post-infection. CONCLUSION: Our results suggest that infection and androgens regulate SP-D in the prostate.

Regulation of surfactant protein D in the mouse female reproductive tract in vivo.

Surfactant protein D (SP-D) plays a role in innate immunity in the lung and is expressed at many other mucosal surfaces throughout the human body. In this study, we show that SP-D mRNA and protein are present in the murine female reproductive tract; i.e. in the vagina, cervix, uterus and oviduct. SP-D protein is primarily localized to epithelial cells lining the genital tract and is also present in secretory material within the lumen of the uterus and cervix. The levels of SP-D mRNA in the uterus vary by a factor of 10 during the estrous cycle with peak levels present at estrus and the lowest levels at diestrus. In contrast, SP-D mRNA levels in the lung do not change during the estrous cycle. Since SP-D is an innate host defense protein present in the mouse reproductive tract, we studied the influence of infection on SP-D levels in vivo. We found that Chlamydia muridarum infection caused an increase in the SP-D protein content of reproductive tract epithelial cells. These data are suggestive that SP-D may play a role in innate immunity in the female reproductive tract in vivo.

A role for surfactant protein D in innate immunity of the human prostate.

OBJECTIVES: Surfactant protein D (SP-D) is a member of the collectin family of proteins, which are involved in host defense mechanisms in the lung. In the present study, we found that SP-D is produced in the human prostate where it may play a role in innate immunity. METHODS AND RESULTS: Using reverse-transcriptase PCR and Western blot analysis, we demonstrate that SP-D mRNA and protein are present in human prostate tissue. In situ hybridization and immunohistochemistry revealed that SP-D mRNA and protein are localized in epithelial cells of prostate glands. Prostate glands that are surrounded by inflammatory cells produce increased amounts of SP-D protein. We also show that SP-D inhibits the infection of LNCaP and P69SV40T prostate epithelial cells by Chlamydia trachomatis in an in vitro infection assay. Furthermore, using truncated human SP-D mutants, we demonstrate that SP-D binds to Chlamydia trachomatis via its carboxy-terminal lectin domains. CONCLUSIONS: Our in vitro studies suggest that SP-D protects the prostate from infection by pathogens. SP-D protein levels are increased at sites of inflammation in the prostate, suggesting SP-D may also contribute more generally to inflammatory regulation in the prostate.

Surfactant protein D is present in the human female reproductive tract and inhibits Chlamydia trachomatis infection.

Surfactant protein D (SP-D) is a lung collectin involved in innate host defence mechanisms in the lung. SP-D is also expressed at other mucosal sites throughout the human body. In the present study, we show that SP-D mRNA and protein are expressed in the human female reproductive tract. SP-D protein was localized in the apical portion of the reproductive epithelial cells. We also demonstrate that endometrial and endocervical cell lines and primary endocervical cells in culture produce SP-D mRNA and protein. Chlamydia trachomatis is an intracellular pathogen that infects the female reproductive tract, primarily the cervix, and is responsible for the most prevalent infectious disease in the USA. Untreated chlamydial infections of the female reproductive tract often result in sterility of the infected woman. Since SP-D protein is produced in cervical glands, we examined the effect of SP-D on chlamydial infection of cervical epithelial cells in vitro. We found that SP-D protein inhibits the infection of HeLa cells (an endocervical epithelial cell line) by C. trachomatis in a dose-dependent manner. We further demonstrate that the SP-D lectin-binding domain is involved in inhibiting infection of HeLa cells by Chlamydia. In conclusion, we detected SP-D in the female reproductive tract and determined that one of the functions of the SP-D protein may be to protect cervical epithelial cells from infection by C. trachomatis.

Surfactant proteins A and D enhance the phagocytosis of Chlamydia into THP-1 cells.

Chlamydiae are intracellular bacterial pathogens that infect mucosal surfaces, i.e., the epithelium of the lung, genital tract, and conjunctiva of the eye, as well as alveolar macrophages. In the present study, we show that pulmonary surfactant protein A (SP-A) and surfactant protein D (SP-D), lung collectins involved in innate host defense, enhance the phagocytosis of Chlamydia pneumoniae and Chlamydia trachomatis by THP-1 cells, a human monocyte/macrophage cell line. We also show that SP-A is able to aggregate both C. trachomatis and C. pneumoniae but that SP-D only aggregates C. pneumoniae. In addition, we found that after phagocytosis in the presence of SP-A, the number of viable C. trachomatis pathogens in the THP-1 cells 48 h later was increased approximately 3.5-fold. These findings suggest that SP-A and SP-D interact with chlamydial pathogens and enhance their phagocytosis into macrophages. In addition, the chlamydial pathogens internalized in the presence of collectins are able to grow and replicate in the THP-1 cells after phagocytosis.

Recombinant human SP-A1 and SP-A2 proteins have different carbohydrate-binding characteristics.

Surfactant protein (SP)-A is a member of the collectin family of proteins and plays a role in innate host defense of the lung. SP-A binds to the carbohydrates of lung pathogens via its calcium-dependant carbohydrate-binding domain. Native human alveolar SP-A consists of two distinct gene products: SP-A1 and SP-A2; however, only SP-A2 is expressed in the submucosal glands of the conducting airways. The function of the isolated SP-A2 protein is unknown. We hypothesized that SP-A1 and SP-A2 might have different carbohydrate-binding properties. In this study, we characterized the carbohydrate-binding specificities of native human alveolar SP-A and recombinant human SP-A1 and SP-A2 in the presence of either 1 or 5 mM Ca(2+). We found that all of the SP-A proteins bind carbohydrates but with different affinities. All of the SP-A proteins bind to fucose with the greatest affinity. SP-A2 binds with a higher affinity to a wider variety of sugars than SP-A1 at either 1 or 5 mM Ca(2+). These findings are suggestive that SP-A2 may interact with a greater variety of pathogens than native SP-A.