Individual VH genes detected with oligonucleotide probes from the complementarity-determining regions

• Published in: The Journal of immunology (1950) Vol. 145; no. 6; pp. 1934 - 1945
• Main Authors: Guillaume, T, Rubinstein, DB, Young, F, Tucker, L, Logtenberg, T, Schwartz, RS, Barrett, KJ
• Format: Journal Article
• Language: English
• Published: Bethesda, MD Am Assoc Immnol 15.09.1990; American Association of Immunologists
• Subjects: Amino Acid Sequence; Antibody Diversity; Base Sequence; Biological and medical sciences; Blotting, Northern; Blotting, Southern; Cell Line; Fundamental and applied biological sciences. Psychology; Genes, Immunoglobulin; Genes. Genome; Humans; Immunoglobulin gamma-Chains - genetics; Immunoglobulin Heavy Chains - genetics; Immunoglobulin mu-Chains - genetics; Immunoglobulin Variable Region - genetics; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; Multigene Family; Oligonucleotide Probes; RNA, Messenger - genetics; Human; Molecular hybridization; Immunogenetics; Immunoglobulins; Gene; Nucleotide sequence; heavy-Peptide chain; Germ line; Variable region; Oligonucleotide; Molecular probe
• ISSN: 0022-1767; 1550-6606
• DOI: 10.4049/jimmunol.145.6.1934

The germ-line and expressed Ig repertoire was examined with three oligonucleotide probes from the CDR regions of VH18/2, a VH gene from the largest human VH gene family, VHIII. Each oligonucleotide probe detected small numbers of germ-line bands (1-5) under conditions in which single base differences can be detected; more than half of these bands were polymorphic. The combined results from pairs of oligonucleotides from CDR1 and CDR2 identified a single band on Southern blots, as did a probe from the 5' end of CDR2. This band contains the 18/2 germ-line gene. The nucleotide sequence of expressed VH genes that hybridized to both CDR probes or to the 5' CDR2 probe were greater than or equal to 97% homologous to 18/2 in both the framework and CDR regions. This group of closely related VH genes, the 18/2 CDR family, appears to be overexpressed. The role of polymorphisms and differential expression of individual V genes in multigenic autoimmune diseases, as well as the organization and expression of individual V genes, can be examined with pairs of oligonucleotides from CDR1 and the 3' end of CDR2, or with probes from the 5' end of CDR2.