Recurrent miscarriage and variant alleles of mannose binding lectin, tumour necrosis factor and lymphotoxin alpha genes

Abstract

Variant alleles of the mannose binding lectin (MBL) gene are associated with increased susceptibility to infection and polymorphisms of tumour necrosis factor and lymphotoxin alpha genes (TNF, LTA) are associated with increased severity of infection. Studies have associated recurrent miscarriage with low serum mannose binding lectin concentrations and premature membrane rupture and preterm delivery with elevated maternal and fetal levels of TNF and the TNF (- 308) polymorphism. In this study the frequencies of variant MBL, TNF and LTA alleles in 76 Caucasian couples with idiopathic recurrent miscarriage were compared with those in 69 Caucasian control couples with no history of miscarriage and at least one previous live birth. A new assay based on hybridization to immobilized sequence-specific oligonucleotides (SSO) was used to rapidly detect nine MBL, two TNF and two LTA sequence variants. The assay genotyped all the structural and promoter MBL variants known to influence serum MBL concentrations. This assay was more reliable than restriction digestion or nested allele-specific PCR for the structural variants at codon 54 or 52, respectively. Reliability for codon 57 alleles was not assessed because of the low frequency in this population. The MBL haplotype frequencies in antenatal controls were similar to those reported in other control populations. The frequencies of structural variant MBL genes and of low, medium and high MBL level haplotypes were similar in the recurrent miscarriage and control couples. The TNF and LTA haplotype frequencies were similar in the recurrent miscarriage and control couples. In this carefully defined population no association has been found between recurrent miscarriage and variant alleles of the MBL, TNF or LTA genes.

Fig. 1

Map of 5′ flanking sequence and exon 1 of the MBL gene. The positions of the variable sites in the 5′ flanking sequences are shown as H/L, M/N, F/G, J/K, deletion -324 to -329, Y/X and P/Q. The positions of the structural variants in exon 1 at codons 52, 54 and 57 are shown. Consensus sequences for glucocorticoid-responsive elements (GRE) and a heat-shock promoter element (HSE) are present in the 5′ sequences.

Fig. 2

Immobilized sequence-specific oligonucleotide assay which rapidly genotypes nine MBL, two TNF and two LTA sequence variants. Typical results for six strips are shown. The strips are aligned to the Ref.line on a reference grid and the genotypes of the DNA sample are read off.