Biologic and genetic characterization of the novel amyloidogenic lambda light chain-secreting human cell lines, ALMC-1 and ALMC-2.
Primary systemic amyloidosis (AL) is a rare monoclonal plasma cell (PC) disorder characterized by the deposition of misfolded immunoglobulin (Ig) light chains (LC) in vital organs throughout the body. To our knowledge, no cell lines have ever been established from AL patients. Here we describe the establishment of the ALMC-1 and ALMC-2 cell lines from an AL patient. Both cell lines exhibit a PC phenotype and display cytokine-dependent growth. Using a comprehensive genetic approach, we established the genetic relationship between the cell lines and the primary patient cells, and we were also able to identify new genetic changes accompanying tumor progression that may explain the natural history of this patient's disease. Importantly, we demonstrate that free lambda LC secreted by both cell lines contained a beta structure and formed amyloid fibrils. Despite absolute Ig LC variable gene sequence identity, the proteins show differences in amyloid formation kinetics that are abolished by the presence of Na(2)SO(4). The formation of amyloid fibrils from these naturally secreting human LC cell lines is unprecedented. Moreover, these cell lines will provide an invaluable tool to better understand AL, from the combined perspectives of amyloidogenic protein structure and amyloid formation, genetics, and cell biology.
Arendt
Bonnie K
BK
Department of Immunology, Mayo Clinic, Rochester, MN 55905, USA.
Ramirez-Alvarado
Marina
M
Sikkink
Laura A
LA
Keats
Jonathan J
JJ
Ahmann
Gregory J
GJ
Dispenzieri
Angela
A
Fonseca
Rafael
R
Ketterling
Rhett P
RP
Knudson
Ryan A
RA
Mulvihill
Erin M
EM
Tschumper
Renee C
RC
Wu
Xiaosheng
X
Zeldenrust
Steven R
SR
Jelinek
Diane F
DF
D000595:Amino Acid Sequence; D000682:Amyloid / Q000096:biosynthesis* / Q000737:chemistry / Q000648:ultrastructure; D000686:Amyloidosis / Q000235:genetics* / Q000276:immunology* / Q000473:pathology; D001483:Base Sequence; D002460:Cell Line; D002869:Chromosome Aberrations; D016207:Cytokines / Q000494:pharmacology; D017931:DNA Primers / Q000235:genetics; D005260:Female; D005803:Genes, Immunoglobulin; D006801:Humans; D007146:Immunoglobulin lambda-Chains / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D017404:In Situ Hybridization, Fluorescence; D046529:Microscopy, Electron, Transmission; D008875:Middle Aged; D008958:Models, Molecular; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D010950:Plasma Cells / Q000187:drug effects / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid
D000682:Amyloid; D016207:Cytokines; D017931:DNA Primers; D007146:Immunoglobulin lambda-Chains