INTRODUCTION: Monoclonal serum Free Light Chains (FLCs) are present in the serum and urine of many patients with plasma cell proliferation diseases. The physical-chemical characteristics of FLCs vary among patients and to date the different tissue damage potential is not fully understood. FLCs are bioactive molecules that induce antigen specific hypersensitivity and interact with several cell lines by activating different intracellular trafficking patterns. GOAL: The mechanism of internalization and trafficking of FLCs is not well understood, therefore, we aimed to characterize this pathway. We have developed an experimental protocol to identify the FLCs membrane tropism and their pathogenicity by analysing FLCs interaction with different cell lines (endothelial, epithelial and myocardial). METHODS: In order to identify a correlation between the FLCs phenotype and pathogenicity, we have investigated the internalization rate of different lambda and kappa FLCs (from hospitalized patients) in different cell lines. The mechanisms of internalization in different cellular subtypes might be an important key in the study of different pathological conditions strictly correlated to FLCs (systemic and not only kidney related) metabolism. Moreover we performed a sucrose density gradient protocol to isolate, quantify, and characterize vesicles containing paraproteins. RESULT: Our results highlight that the monoclonal FLCs of every patient behave differently in different cellular lines and demonstrate that cells other than epithelial cells are able to process paraproteins. We could show that FLCs distributes in differently sized intracellular vesicles: the FLCs colocalized with Lamp1 (Lisosomal associated membrane protein ) and c-Src , a proto-oncogene that belongs to a family of non-receptor tyrosine kinases involved in the Nf-KB activation pathway.
Immunoglobulin Free Light Chains Trafficking Pathways
DI NOTO, Giuseppe;PAOLINI, Lucia;RICOTTA, Doris
2012-01-01
Abstract
INTRODUCTION: Monoclonal serum Free Light Chains (FLCs) are present in the serum and urine of many patients with plasma cell proliferation diseases. The physical-chemical characteristics of FLCs vary among patients and to date the different tissue damage potential is not fully understood. FLCs are bioactive molecules that induce antigen specific hypersensitivity and interact with several cell lines by activating different intracellular trafficking patterns. GOAL: The mechanism of internalization and trafficking of FLCs is not well understood, therefore, we aimed to characterize this pathway. We have developed an experimental protocol to identify the FLCs membrane tropism and their pathogenicity by analysing FLCs interaction with different cell lines (endothelial, epithelial and myocardial). METHODS: In order to identify a correlation between the FLCs phenotype and pathogenicity, we have investigated the internalization rate of different lambda and kappa FLCs (from hospitalized patients) in different cell lines. The mechanisms of internalization in different cellular subtypes might be an important key in the study of different pathological conditions strictly correlated to FLCs (systemic and not only kidney related) metabolism. Moreover we performed a sucrose density gradient protocol to isolate, quantify, and characterize vesicles containing paraproteins. RESULT: Our results highlight that the monoclonal FLCs of every patient behave differently in different cellular lines and demonstrate that cells other than epithelial cells are able to process paraproteins. We could show that FLCs distributes in differently sized intracellular vesicles: the FLCs colocalized with Lamp1 (Lisosomal associated membrane protein ) and c-Src , a proto-oncogene that belongs to a family of non-receptor tyrosine kinases involved in the Nf-KB activation pathway.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
