Intraclonal Variability of VH Genes in Follicular Lymphoma Patients Who Have Received Anti-Idiotypic Immunotherapy

Abstract

Subclonal heterogeneity can affect idiotypic determinants present in the clonotypic immunoglobulin of B-cell follicular lymphomas (FLs) and may limit the effect of antilymphoma treatments performed by immunization of patients with their own tumor-associated idiotypic immunoglobulin. Idiotype-secreting hybridomas were obtained by fusion of tumor cells from 5 patients with FL, and the K6H6/B5 human heteromyeloma and rearranged VH genes from tumor samples and hybridomas were amplified, cloned, and sequenced. Sequences were aligned with germline genes and somatic mutations, intraclonal heterogeneity and genealogic relations of the B-cell clones in the different biopsy specimens were determined. The VH sequence of the progenitor clone was determined in samples of the tumoral population. Further diversification resulted in the presence of 2 to 6 subclones in 4 of the 5 samples studied. Only in 1 patient did the hypermutation mechanism introduce differences among most of the potential idiotopes present in individual subclones. The VH sequence of the hybridoma that provided the idiotypic-vaccine was identified in one of the tumor subclones in all cases. No relapse has been demonstrated in 3 of the 4 vaccinated patients (follow-up: 29–103 months). We conclude that despite potential differences in the idiotypic region expressed by individual tumor cells, at least some potential idiotopes may be preserved among all the tumor subclones in most cases studied. All vaccinated patients developed immune responses against the autologous tumor idiotypic immunoglobulin. Polyclonal anti-idiotypic immune responses induced with a vaccine obtained from 1 hybridoma may be effective against all the idiotypic variants present in the tumor population.