Hepatitis B Virus (HBV)

Tag: pregnancy

Null Allele Encodes Functional HLA-G Isoforms: DISCUSSION

After our study of the tolerogeneic role of HLA-G in maternal-fetal tolerance, we designed experiments to identify the structural and functional characteristics of proteins generated by the HLA-G*0105N allele. First, we used site-directed mutagenesis to construct the HLA-G*0105Nfrom the nucleotide sequence of HLA-G*010102 and transfected it into the M8 human cell line. Then we characterized […]

Null Allele Encodes Functional HLA-G Isoforms: RESULTS

Construction of HLA-G*0105N Null Allele Genomic DNA To study proteins encoded by the HLA-G null allele, we generated a DNA sequence identical to HLA-G*0105N in the pcDNA 3.1 expression vector. In vitro site-directed mutagenesis of HLA-G*010102 genomic DNA allowed us to delete the cytosine at position 815 from ATG (exon 1), yielding the genomic dNa […]

Null Allele Encodes Functional HLA-G Isoforms: MATERIALS AND METHODS

in Vitro Site-Directed Mutagenesis In vitro site-directed mutagenesis was carried out using the Stratagene QuickChange XL site-directed mutagenesis kit, which allows site-specific mutation in double-stranded plasmids. The plasmid DNA template (50 ng) HLA-G*010102-pcDNA3.1, isolated from the dam+ JM109 Escherichia coli strain, was replicated by PfuTurbo DNA polymerase (2.5 U) using two synthetic primers, G.Null S […]

Null Allele Encodes Functional HLA-G Isoforms: INTRODUCTION

Protection of the semiallogenic fetus against maternal immune recognition and attack has been attributed principally to the high and almost unique expression of human leukocyte antigen-G (HLA-G) on trophoblast cells at the fetal-maternal interface. The nonclassical HLA-G gene is characterized by alternative splicing that yields seven proteins, four membrane bound (HLA-G1 through -G4) and three […]

Pregnancy and Interferon: DISCUSSION(6)

The authors recognize that proteins other than UCRP could potentially immunoreact with the anti-UCRP antibody. However, a recent search of the NCBI-BLAST database revealed no known proteins with significant amino acid sequence identity to the UCRP peptide immunogen other than ubiquitin and huISG15 (see Fig. 1). Although unlikely, the existence of proteins that share antigenic […]

Pregnancy and Interferon: DISCUSSION(5)

Release of UCRP into the medium in response to rbo-IFN-t occurred after accumulation of cytosolic UCRP and its conjugates. The extended time (24 h) that was required for release of UCRP might be related to a secondary secreted role. For example, the primary role for UCRP might be regulating cytosolic proteins. After UCRP and its […]

Pregnancy and Interferon: DISCUSSION(4)

In the present Western blot experiments, second antibodies used for detection cross-reacted with uterine proteins in the absence of primary anti-ubiquitin or anti-UCRP antibody (Fig. 4). This nonspecific staining occurred mostly with four bands of protein ^ 30 kDa, but it did not interfere with detection of ubiquitin or UCRP. Anti-UCRP antibody immunoreacts with UCRP, […]