title: Type II mixed cryoglobulinaemia as an oligo rather than a mono B-cell disorder: evidence from GeneScan and MALDI-TOF analyses.
abstract: OBJECTIVE
  To identify and characterize rheumatoid factor (RF)-producing B-cells and cryoprecipitate immunoglobulin (Ig) M in hepatitis C virus (HCV)-positive patients.
  
  
  METHODS
  We purified and characterized, by peptide mass fingerprinting integrated with an NCBI IgBlast data bank search, the IgM component of cryoprecipitate and analysed the VDJ pattern of bone marrow B-cells by gene scan analysis of 17 HCV-positive patients with type II mixed-cryoglobulinaemia.
  
  
  RESULTS
  IgM purified from all of the patients presented an RF specificity. In three of these patients a high and predominant B-cell clone (>or=30%) was found in the bone marrow. B-cell-receptor sequences were determined and immunophenotyping of these clones was performed. Peptide masses originating after tryptic digestion of the B-cell-receptor combinatory regions and those originating by tryptic digestion of the cryoprecipitated IgM from the same patient were comparable. In the remaining patients an oligoclonal/polyclonality was found. However, in some of these patients we were able to find peptides that matched with the B-cell-receptor sequences of overexpanded B cells, indicating that, even in the absence of a clear monoclonal expansion, a fraction of total cryoprecipated IgM may derive from overexpanded B-cell clones found in patients' bone marrow.
  
  
  CONCLUSIONS
  In the majority of mixed cryoglobulinaemia-HCV-positive patients, both in the serum and in B cells from the bone marrow, an oligoclonal pattern is the main molecular picture. When a monoclonal B-cell clone is found, its B-cell-receptor shows an antigen-binding fragment identical to that of cryoprecipitable RF-IgM. Phenotypically, B cells are CD20-positive but CD5-negative, suggesting that the B-1 B-cell subset is not likely to produce high-affinity IgM-RF molecules.
authors.affiliation: Division of Experimental Oncology I, Centro di Riferimento Oncologico Via Pedemontana Occidentale 12, 33081 Aviano (PN), Italy. vdere@cro.it
authors.forename: V
authors.initials: V
authors.lastname: De Re
authors.forename: S
authors.initials: S
authors.lastname: De Vita
authors.forename: D
authors.initials: D
authors.lastname: Sansonno
authors.forename: D
authors.initials: D
authors.lastname: Gasparotto
authors.forename: M P
authors.initials: MP
authors.lastname: Simula
authors.forename: F A
authors.initials: FA
authors.lastname: Tucci
authors.forename: A
authors.initials: A
authors.lastname: Marzotto
authors.forename: M
authors.initials: M
authors.lastname: Fabris
authors.forename: A
authors.initials: A
authors.lastname: Gloghini
authors.forename: A
authors.initials: A
authors.lastname: Carbone
authors.forename: F
authors.initials: F
authors.lastname: Dammacco
authors.forename: M
authors.initials: M
authors.lastname: Boiocchi
mesh_terms: D000328:Adult; D000368:Aged; D000595:Amino Acid Sequence; D001402:B-Lymphocytes / Q000276:immunology* / Q000473:pathology; D001854:Bone Marrow Cells / Q000276:immunology; D002999:Clone Cells / Q000276:immunology / Q000473:pathology; D003449:Cryoglobulinemia / Q000235:genetics / Q000276:immunology* / Q000821:virology; D005260:Female; D005803:Genes, Immunoglobulin; D006526:Hepatitis C / Q000150:complications / Q000276:immunology; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007147:Immunoglobulin Light Chains / Q000235:genetics; D007075:Immunoglobulin M / Q000235:genetics / Q000276:immunology; D016130:Immunophenotyping; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D009154:Mutation; D012217:Rheumatoid Factor / Q000276:immunology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007075:Immunoglobulin M; D012217:Rheumatoid Factor