PMID- 16126965
OWN - NLM
STAT- MEDLINE
DCOM- 20060626
LR  - 20061115
IS  - 0077-8923 (Print)
IS  - 0077-8923 (Linking)
VI  - 1051
DP  - 2005 Jun
TI  - Generation and characterization of three monoclonal IgM antiphospholipid 
      antibodies recognizing different phospholipid antigens.
PG  - 240-54
AB  - Antiphospholipid antibodies (APLs) might be involved in the pathogenesis of the 
      antiphospholipid syndrome (APS). This study analyzes the structural 
      characteristics of monoclonal APLs derived from patients with this disease. 
      Patient-derived B cells were immortalized using Epstein-Barr virus transformation 
      and subsequent fusion to the myeloma cell line CB-F7. APL-producing hybridomas 
      were cloned to obtain cell lines producing monoclonal APL. DNA encoding the 
      variable region of heavy and light chains of the antibodies was sequenced and 
      analyzed regarding their usage within the V-gene family and the existence of 
      somatic hypermutation. Binding patterns of APL to various phospholipids and 
      beta-2-glycoprotein-I (beta2-GPI) were determined using ELISA, with special 
      regard to beta2-GPI dependency. As a result, three APL-producing hybridoma cell 
      lines from patients with APS were established: JGG9, HVA2, and HLC9. APLs were of 
      the IgM isotype and showed different binding patterns toward phospholipids and 
      beta2-GPI. One of them, JGG9, showed extensive somatic hypermutations in both the 
      CDR3 region and a framework region of the heavy chain. JGG9 bound to cardiolipin 
      in the presence of the protein cofactor beta2-GPI. In contrast, the antibodies 
      HVA2 and HLC9 (which also showed somatic hypermutations in the CDR3 region) 
      presented polyreactivity to several phospholipids-cardiolipin, 
      phosphatidyl-serine, -ethanolamine, -inositol, -choline, and sphingomyelin-but 
      not to beta2-GPI. In conclusion, JGG9 presents a high degree of mutation in the 
      CDR3 and framework region resulting from the deletions of nucleotides, and 
      affects amino acid composition. Polyreactivity and the absence of cofactor 
      dependency present HLC9- and HVA2-like natural antibodies that have no contact 
      with any antigen. Nonetheless, these natural antibodies show somatic 
      hypermutation of the heavy chain, indicating antigen-driven maturation. Regarding 
      the possible role of APL in infection, HVA2 in particular may represent a 
      pathogen-maturated antibody showing cross-reactivity between phospholipids and 
      infectious agents. Further experiments are needed to reveal the functional 
      activity of these antibodies.
FAU - Buschmann, Catharina
AU  - Buschmann C
AD  - Institute of Clinical Chemistry and Laboratory Medicine, Johannes Gutenberg 
      University of Mainz, Langenbeckstrasse 1, 55131 Mainz, Germany.
FAU - Fischer, Christian
AU  - Fischer C
FAU - Ochsenhirt, Viola
AU  - Ochsenhirt V
FAU - Neukirch, Carolin
AU  - Neukirch C
FAU - Lackner, Karl J
AU  - Lackner KJ
FAU - von Landenberg, Philipp
AU  - von Landenberg P
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Ann N Y Acad Sci
JT  - Annals of the New York Academy of Sciences
JID - 7506858
RN  - 0 (Antibodies, Antiphospholipid)
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Complementarity Determining Regions)
RN  - 0 (Glycoproteins)
RN  - 0 (Immunoglobulin M)
RN  - 0 (Immunoglobulin Variable Region)
RN  - 0 (Phospholipids)
RN  - 0 (beta 2-Glycoprotein I)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Antibodies, Antiphospholipid/genetics/*immunology
MH  - Antibodies, Monoclonal/biosynthesis/*immunology
MH  - Antiphospholipid Syndrome
MH  - Base Sequence
MH  - Cell Line
MH  - Complementarity Determining Regions
MH  - Enzyme-Linked Immunosorbent Assay
MH  - Glycoproteins/immunology
MH  - Humans
MH  - Immunoglobulin M/biosynthesis/*immunology
MH  - Immunoglobulin Variable Region/genetics
MH  - Male
MH  - Mice
MH  - Middle Aged
MH  - Molecular Sequence Data
MH  - Phospholipids/*immunology
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - beta 2-Glycoprotein I
EDAT- 2005/08/30 09:00
MHDA- 2006/06/27 09:00
CRDT- 2005/08/30 09:00
PHST- 2005/08/30 09:00 [pubmed]
PHST- 2006/06/27 09:00 [medline]
PHST- 2005/08/30 09:00 [entrez]
AID - 1051/1/240 [pii]
AID - 10.1196/annals.1361.065 [doi]
PST - ppublish
SO  - Ann N Y Acad Sci. 2005 Jun;1051:240-54. doi: 10.1196/annals.1361.065.