Cloning of Ig Fab genes of patients with chronic B lymphocytic leukemia and sequences analysis of their variable regions.
- Author:
Huifen ZHU
1
;
Feng WANG
;
Yue ZHANG
;
Guanxin SHEN
Author Information
- Publication Type:Journal Article
- MeSH: Base Sequence; Cloning, Molecular; DNA, Complementary; chemistry; Gene Rearrangement; Genes, Immunoglobulin; Humans; Immunoglobulin Fab Fragments; genetics; Immunoglobulin Heavy Chains; genetics; Immunoglobulin Light Chains; genetics; Immunoglobulin Variable Region; genetics; Leukemia, Lymphocytic, Chronic, B-Cell; genetics; immunology; Molecular Sequence Data; Polymerase Chain Reaction
- From: Chinese Journal of Hematology 2002;23(9):474-479
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVESequences analysis of Ig variable regions from the peripheral blood mononuclear cells (PBMC) of patients with chronic B lymphocytic leukemia.
METHODSTotal RNA was isolated from PBMC of patients with chronic B lymphocytic leukemia, oligo-dT-primed cDNA was synthesized from RNA. The cDNA was amplified by Taq DNA polymerase with a set of specific 5' primers corresponding to Ig FR1 and 3' primers corresponding to CH1 (C micro /C) or CL (Ckappa/Clambda), the PCR products of variable regions of Ig heavy (IgH) and light (IgL) chains were sequenced by ABI PRISM Dye terminator cycle sequencing ready reaction kit and ABI PRISM 310 Genetic Analyzer. The gene homology of variable regions of IgH and IgL chains was compared by using DNA tools 5.1 system and "the international immunogenetics database".
RESULTSFour light chains and 3 heavy chains were amplified from 4 and 3 patients respectively. Homology analysis of the sequences of 4 light chains and 3 heavy chains were performed by DNA tools system. The sequences of light chains are high homologous. And the sequences of heavy chains are quite different. The homologous analysis of the sequences of variable region by using "the international immunogenetics database" showed that the sequences were higher homologous to idiotype gene of some B lymphocytic leukemia. Four VL genes belong to human Ig Vkappa subgroup I, 2 of 3 VH genes belong to VH3 family and 1 belongs to VH5 family.
CONCLUSIONIg genes have idiotype and same disease may have same idiotype.