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J Immunol ; 142(11): 4027-33, 1989 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-2497186

RESUMO

RNA sequences for the V regions of human hybridoma-produced autoantibodies were determined by primer extension with reverse transcriptase. The sequencing of IgM autoantibodies from a leprosy patient revealed examples of recurrent use of V region gene segments in different autoantibodies from this patient and a previously studied patient with SLE. Moreover, several gene segments used in these autoantibodies show little alteration from germ-line sequences. mAb TH3, from a patient with leprosy, binds denatured DNA and poly(dT). The center of its H chain CDR35 has a sequence identical to that found previously in two anti-DNA antibodies from a lupus patient; these identities and their overlapping with two other published sequences define a human D-gene segment of approximately 25 nucleotides. Autoantibody TH9, from a leprosy patient, does not bind DNA. Its VH sequence has 87% identity with a VHI anti-DNA antibody, but differs from it markedly in the CDR1 region. TH9 also has a different H chain CDR3. The closely related JH4 or JH5 gene segments are expressed in five lupus or leprosy autoantibodies. In four of the antibodies, examples of V kappa 1, V kappa 3, or V kappa 4 and J kappa 2, or J kappa 5 segments were found. Two distinct leprosy-derived anti-DNA antibodies, 8E10 and TH3, share a completely identical V kappa sequence. This sequence differs in only two positions from that of a germ-line RF L chain gene. Several gene segments that are close to the germ line in sequence encode Ig V regions with autoantibody reactivity. These results provide a base line for determining whether these genes are precursors of more highly diversified antibodies that may be pathogenic in patients with SLE.


Assuntos
Anticorpos Antinucleares/genética , Imunoglobulina M/genética , Região Variável de Imunoglobulina/genética , Sequência de Aminoácidos , Anticorpos Antinucleares/isolamento & purificação , Doenças Autoimunes/genética , Doenças Autoimunes/imunologia , Sequência de Bases , Sítios de Ligação de Anticorpos , DNA/imunologia , Humanos , Hibridomas/análise , Cadeias Pesadas de Imunoglobulinas/genética , Cadeias Pesadas de Imunoglobulinas/isolamento & purificação , Imunoglobulina M/isolamento & purificação , Região Variável de Imunoglobulina/isolamento & purificação , Cadeias kappa de Imunoglobulina/genética , Cadeias kappa de Imunoglobulina/isolamento & purificação , Hanseníase/genética , Hanseníase/imunologia , Dados de Sequência Molecular
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