RESUMO
We have established a ligation based typing method to detect HLA-B*27 alleles at the DNA level. The method requires amplification of exon 2 of the HLA-B locus from genomic DNA by the polymerase catalyzed chain reaction (PCR) using group specific primers. An aliquot of the PCR amplification product, heat stable ligase and a pair of oligonucleotide probes, designed to hybridize adjacently to HLA-B*27 specific sequences of the amplified DNA are subsequently thermocycled. If the probes are perfectly complementary they become ligated otherwise they stay separated. The ligation of probes can be detected through their different labels by an enzyme linked immunosorbent assay (ELISA). Ligation based detection of beta-actin sequences which have been co-amplified serves as positive control for each PCR reaction. We observed complete concordance when typing 76 HLA-B*27 positive and 107 HLA-B*27 negative individuals either by serology or by the ligation based approach. We conclude that ligation based typing is a reliable tool for the DNA based detection of HLA-B*27 alleles. The procedure allows automation to a large extent and should be easily applicable to the typing of other HLA-class I alleles.
Assuntos
Antígeno HLA-B27/genética , Biologia Molecular/métodos , Sequência de Bases/genética , Genótipo , Humanos , Leucócitos Mononucleares , Reação em Cadeia da Polimerase/métodosRESUMO
The results of the serological typing and the patterns of restriction fragment length polymorphisms for the detection of HLA-DR gene products are compared. The data shown demonstrate that the use of DNA typing gives a clearer definition of the HLA-DR antigens and that the HLA-DR polymorphism is greater than detected by serology.
Assuntos
Antígenos HLA-DR/imunologia , Polimorfismo de Fragmento de Restrição , Sondas de DNA , Desoxirribonuclease EcoRI , Epitopos , Antígenos HLA-DR/genética , HumanosRESUMO
Analysis of class I, class II, and class III gene products of the human MHC in a Caucasoid family with four children gave evidence for a double crossing over event in monozygotic twins between the C2, Bf/C4A, C4B gene loci. In view of the small genetic distance between C2 and C4, a point mutation within the Bf locus must be considered alternatively.
Assuntos
Troca Genética , Recombinação Genética , Feminino , Antígenos HLA/classificação , Humanos , Complexo Principal de Histocompatibilidade , Masculino , MutaçãoRESUMO
In ten families with 65 children (with 64 informative meioses), a close linkage between the monocyte antigen locus (HMA) and HLA was found with a recombination frequency of 1.56%.
Assuntos
Mapeamento Cromossômico , Cromossomos Humanos Par 6 , Isoantígenos/genética , Monócitos/imunologia , Ligação Genética , Antígenos HLA/genética , Humanos , LinhagemRESUMO
138 sera from renal transplant recipients were screened for the presence of monocyte-specific antibodies. Most of the sera contained antibodies against monocytes, T- and/or B-lymphocytes. One serum was identified which defined a monocyte-specific antigen, MOLI. This serum was investigated in intensive population and family studies for the estimation of the formal genetic criteria of this 'new' monocyte antigen. A gene frequency of 0.0614 was obtained by population analysis. Family investigations conveyed the information that the gene coding for MOLI was transmitted in linkage with HLA genes. A positive linkage disequilibrium of MOLI and HLA-B17 was found.
Assuntos
Antígenos/imunologia , Monócitos/imunologia , Anticorpos/imunologia , Antígenos/genética , Linfócitos B/imunologia , Frequência do Gene , Ligação Genética , Humanos , Transplante de Rim , Linhagem , Linfócitos T/imunologiaRESUMO
A simple method of cryopreservation (TTK) of lymphocytes is presented. The functional properties of TTK-lymphocytes are examined by the lymphocyte toxicity micro test and in the mixed lymphocyte culture (MLC). 51Cr-release technique is performed as a measure for reversible and irreversible cell damages in the course of the Cryopreservation process.
