Limits...
A novel nanobody specific for respiratory surfactant protein A has potential for lung targeting.

Wang SM, He X, Li N, Yu F, Hu Y, Wang LS, Zhang P, Du YK, Du SS, Yin ZF, Wei YR, Mulet X, Coia G, Weng D, He JH, Wu M, Li HP - Int J Nanomedicine (2015)

Bottom Line: Here, we generated recombinant rat pulmonary SPA (rSPA) as an antigen and immunized an alpaca to produce two nanobodies (the smallest naturally occurring antibodies) specific for rSPA, designated Nb6 and Nb17.Using immunohistochemistry, we demonstrated that these anti-rSPA nanobodies selectively bound to rat lungs with high affinity.Collectively, we have generated and characterized lung-specific nanobodies, which may be applicable for lung drug delivery.

View Article: PubMed Central - PubMed

Affiliation: Department of Respiratory Medicine, Shanghai Pulmonary Hospital, School of Medicine, Tongji University, Shanghai, People's Republic of China.

ABSTRACT
Lung-targeting drugs are thought to be potential therapies of refractory lung diseases by maximizing local drug concentrations in the lung to avoid systemic circulation. However, a major limitation in developing lung-targeted drugs is the acquirement of lung-specific ligands. Pulmonary surfactant protein A (SPA) is predominantly synthesized by type II alveolar epithelial cells, and may serve as a potential lung-targeting ligand. Here, we generated recombinant rat pulmonary SPA (rSPA) as an antigen and immunized an alpaca to produce two nanobodies (the smallest naturally occurring antibodies) specific for rSPA, designated Nb6 and Nb17. To assess these nanobodies' potential for lung targeting, we evaluated their specificity to lung tissue and toxicity in mice. Using immunohistochemistry, we demonstrated that these anti-rSPA nanobodies selectively bound to rat lungs with high affinity. Furthermore, we intravenously injected fluorescein isothiocyanate-Nb17 in nude mice and observed its preferential accumulation in the lung to other tissues, suggesting high affinity of the nanobody for the lung. Studying acute and chronic toxicity of Nb17 revealed its safety in rats without causing apparent histological alterations. Collectively, we have generated and characterized lung-specific nanobodies, which may be applicable for lung drug delivery.

No MeSH data available.


Related in: MedlinePlus

Histological analysis of rat lungs, liver, spleen, kidneys by hematoxylin and eosin staining.Notes: Different reagents (Nb17 or control antibodies) were administered to rats for acute (1 week) or chronic (3 months) toxicity study. Then, the lung, liver, spleen, and kidney tissues were processed for hematoxylin and eosin staining.Abbreviation: Neg, negative.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4403696&req=5

f7-ijn-10-2857: Histological analysis of rat lungs, liver, spleen, kidneys by hematoxylin and eosin staining.Notes: Different reagents (Nb17 or control antibodies) were administered to rats for acute (1 week) or chronic (3 months) toxicity study. Then, the lung, liver, spleen, and kidney tissues were processed for hematoxylin and eosin staining.Abbreviation: Neg, negative.

Mentions: Serum alanine transaminase, aspartate transaminase, blood urea nitrogen, and serum creatinine were determined to assess the acute (1-week) and chronic (3-month) toxicity in rats as detailed earlier. No statistically significant differences were found between any of the four groups by multiple-comparison analysis (P>0.05, Table 3). Therefore, overall data indicated that Nb17 had not induced significant toxicity. We further analyzed the histological alterations caused by Nb17 using cryosections of the lung, liver, spleen, and kidney in acute toxicity, chronic toxicity, and control groups. Our results showed that there were no apparent histological alterations by morphological assessment in tissues of the lung, liver, spleen, or kidney of rats between the Nb17 groups and controls using hematoxylin and eosin staining (Figure 7).


A novel nanobody specific for respiratory surfactant protein A has potential for lung targeting.

Wang SM, He X, Li N, Yu F, Hu Y, Wang LS, Zhang P, Du YK, Du SS, Yin ZF, Wei YR, Mulet X, Coia G, Weng D, He JH, Wu M, Li HP - Int J Nanomedicine (2015)

Histological analysis of rat lungs, liver, spleen, kidneys by hematoxylin and eosin staining.Notes: Different reagents (Nb17 or control antibodies) were administered to rats for acute (1 week) or chronic (3 months) toxicity study. Then, the lung, liver, spleen, and kidney tissues were processed for hematoxylin and eosin staining.Abbreviation: Neg, negative.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4403696&req=5

f7-ijn-10-2857: Histological analysis of rat lungs, liver, spleen, kidneys by hematoxylin and eosin staining.Notes: Different reagents (Nb17 or control antibodies) were administered to rats for acute (1 week) or chronic (3 months) toxicity study. Then, the lung, liver, spleen, and kidney tissues were processed for hematoxylin and eosin staining.Abbreviation: Neg, negative.
Mentions: Serum alanine transaminase, aspartate transaminase, blood urea nitrogen, and serum creatinine were determined to assess the acute (1-week) and chronic (3-month) toxicity in rats as detailed earlier. No statistically significant differences were found between any of the four groups by multiple-comparison analysis (P>0.05, Table 3). Therefore, overall data indicated that Nb17 had not induced significant toxicity. We further analyzed the histological alterations caused by Nb17 using cryosections of the lung, liver, spleen, and kidney in acute toxicity, chronic toxicity, and control groups. Our results showed that there were no apparent histological alterations by morphological assessment in tissues of the lung, liver, spleen, or kidney of rats between the Nb17 groups and controls using hematoxylin and eosin staining (Figure 7).

Bottom Line: Here, we generated recombinant rat pulmonary SPA (rSPA) as an antigen and immunized an alpaca to produce two nanobodies (the smallest naturally occurring antibodies) specific for rSPA, designated Nb6 and Nb17.Using immunohistochemistry, we demonstrated that these anti-rSPA nanobodies selectively bound to rat lungs with high affinity.Collectively, we have generated and characterized lung-specific nanobodies, which may be applicable for lung drug delivery.

View Article: PubMed Central - PubMed

Affiliation: Department of Respiratory Medicine, Shanghai Pulmonary Hospital, School of Medicine, Tongji University, Shanghai, People's Republic of China.

ABSTRACT
Lung-targeting drugs are thought to be potential therapies of refractory lung diseases by maximizing local drug concentrations in the lung to avoid systemic circulation. However, a major limitation in developing lung-targeted drugs is the acquirement of lung-specific ligands. Pulmonary surfactant protein A (SPA) is predominantly synthesized by type II alveolar epithelial cells, and may serve as a potential lung-targeting ligand. Here, we generated recombinant rat pulmonary SPA (rSPA) as an antigen and immunized an alpaca to produce two nanobodies (the smallest naturally occurring antibodies) specific for rSPA, designated Nb6 and Nb17. To assess these nanobodies' potential for lung targeting, we evaluated their specificity to lung tissue and toxicity in mice. Using immunohistochemistry, we demonstrated that these anti-rSPA nanobodies selectively bound to rat lungs with high affinity. Furthermore, we intravenously injected fluorescein isothiocyanate-Nb17 in nude mice and observed its preferential accumulation in the lung to other tissues, suggesting high affinity of the nanobody for the lung. Studying acute and chronic toxicity of Nb17 revealed its safety in rats without causing apparent histological alterations. Collectively, we have generated and characterized lung-specific nanobodies, which may be applicable for lung drug delivery.

No MeSH data available.


Related in: MedlinePlus