Abstract: Objective To obtain in situ pairing of the variable region genes of the immunoglobulin heavy and light chains of B cells from vitiligo patients.Methods Fifty vitiligo patients were screened by a live melanocyte enzyme-linked immunoabsorbent assay.The sera from seven patients were proved to be strongly positive.Then CD19⁺ B cells were isolated from the peripheral blood lymphocytes of the 7 patients,then fixed in 10% buffered formaldehyde and permeabilised by Proteinase K.The mRNA was amplified within the cells by reverse transcription-polymerase chain reaction (RT-PCR) with specific primers.The immunoglobulin VH and VL DNA assembled within the same cells using the Cre/loxP system.Nested primers were designed to amplify the known major human VH and VL gene familes.Result A unique 800bp band was obtained corresponding in size to single chain Fv fragments.Conclusion The in situ pairing of the variable region genes of the immunoglobulin heavy and light chains of B cells is obtained from vitiligo patients by in-cell PCR.

Key words: Vitiligo, Polymerase chain reaction, Auto antibodies