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Updated: Aug 8, 2026
Identification of Alternative Splicing and Polyadenylation in RNA-seq Data
Published on: June 24, 2021
Identification and characterization of an alternatively spliced isoform of mouse Langerin/CD207
Elisabeth Riedl1, Yayoi Tada, Mark C Udey
1Dermatology Branch, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, Maryland 20892, USA. riedle@mail.nih.gov
Insights
Researchers identified mouse Langerin (CD207) and a variant, DeltaE3Langerin, involved in antigen uptake. Their expression and function, including Birbeck granule formation, were characterized in vitro and in vivo.
Area of Science:
- Immunology
- Cell Biology
- Molecular Biology
Background:
- Langerin (CD207) is a C-type lectin crucial for Langerhans cell function in antigen recognition.
- Identifying the mouse homologue of human Langerin is essential for understanding immune responses in mice.
Purpose of the Study:
- To isolate and characterize the mouse homologue of human Langerin (CD207).
- To investigate the expression and function of full-length and alternatively spliced variants of mouse Langerin.
Main Methods:
- Subtractive cloning and RACE to isolate mouse Langerin cDNA.
- RT-PCR to detect Langerin mRNA variants in dendritic cells and skin.
- Transfection of fibroblasts with Langerin cDNAs to study protein localization and function.
- Analysis of Birbeck granule formation in transfected cells.
Main Results:
- Mouse Langerin cDNA was isolated, and an alternatively spliced variant (DeltaE3Langerin) lacking the neck domain was identified.
- Both Langerin isoforms were expressed in vitro by dendritic cells, with broader tissue expression observed.
- Full-length mouse Langerin bound mannan and induced Birbeck granule formation, while DeltaE3Langerin did not.
- Recombinant DeltaE3Langerin localized to transferrin compartments in fibroblasts.
Conclusions:
- Mouse Langerin exists in full-length and a neck-domain-deleted variant, DeltaE3Langerin.
- Langerin isoform expression is developmentally regulated, potentially controlling its role in antigen uptake and processing.
- The neck domain is critical for mannan binding and Birbeck granule formation by mouse Langerin.
Abstract:
The mouse homologue of human Langerin (CD207), a novel Langerhans cells (LC)-restricted C-type lectin that likely participates in antigen recognition and uptake, has been recently identified. In this study, we isolated the mouse Langerin cDNA from murine fetal skin-derived dendritic cells (FSDDC) by subtractive cloning and rapid amplification of cDNA ends (RACE). An alternatively spliced variant of mouse Langerin that lacked the extracellular neck domain (DeltaE3Langerin) was detected in RNA derived from FSDDC and epidermal LC by RT-PCR. In vitro-generated FSDDC and epidermal LC expressed both full-length and DeltaE3Langerin mRNA, but tissue expression was not restricted to skin. Mouse Langerin protein isoforms were readily detected in fibroblasts transfected with cDNAs encoding epitope-tagged Langerin and DeltaE3Langerin. Recombinant DeltaE3Langerin protein localized with transferrin-containing compartments in transfected fibroblasts. Full-length mouse Langerin-bound mannan, whereas DeltaE3Langerin and soluble bacterial recombinant Langerin protein lacking the neck domain did not. Fibroblasts transfected with mouse Langerin cDNA contained typical Birbeck granules (BG) and cored tubules, whereas DeltaE3Langerin cDNA did not induce BG or cored tubule formation in transfected fibroblasts. Developmentally regulated expression of Langerin isoforms provides a mechanism by which Langerin involvement in antigen uptake and processing could be regulated.
