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Biomedical subjects

G B Ferrara

Publications and source records attributed to G B Ferrara.

150 records · Page 9Linked to original sources
PubMedDate not supplied

Analysis of HLA specificity of human monoclonal antibodies by cytofluorimetry and cell ELISA.

In this study we report on the characterization of cytotoxic human monoclonal antibodies (HmAb) detecting polymorphic HLA class II specificities using cytofluorimetric analysis in combination with micro cell ELISA. In both techniques, five anti-HLA HmAb were tested against HLA-transfected murine L cells as target cells and the bound antibody was detected, either by cytofluorimetry or by cell ELISA reader, after addition of fluoresceinated or peroxidase-conjugated anti-human IgG+IgM antibodies, respectively. The results demonstrate that HmAb directed against HLA-DR, -DQ and -DP molecules can be efficiently discriminated by cytofluorimetry and cell ELISA, which appear to be highly sensitive and perfectly comparable to the standard cytotoxicity assay.

Animals↗
PubMedDate not supplied

RFLP analysis with cDNA probes for DQ/DP region in HLA identical sibling marrow transplants: lack of correlation with GvHD.

BACKGROUND: The role of HLA-DP as transplantation antigen in contributing to GvHD is a matter of current debate. HLA-DP, is encoded centromeric to DR-DQ and its alleles are in weak linkage disequilibrium with the rest of the MHC; thus a certain number of HLA matched pairs could be actually DP incompatible to study a possible correlation between HLA-DP matching and GvHD, 24 HLA identical BMT/donor-recipient sibling pairs (serologically tested for HLA Class I and DR antigens) were tested for DQ and DP genes using restriction fragment length polymorphism (RFLP) analysis. METHODS: DNA extracts were digested according to a standard procedure with two different restriction enzymes (HIND III and MSP I) and hybridised with DQ (alpha and beta) and DP (alpha and beta) specific probes. Highly stringent hybridization and washing conditions were used to prevent cross-hybridizations. RESULTS: Twenty four out of 24 pairs proved to be DQ and DP identical. GvHD developed in 16 out of 24 (66.6%) recipients. DISCUSSION: These data suggest that DNa analysis of DQ-DP regions, with the probes and enzymes used, does not give predictive informations for GvHD in HLA genotypically identical pairs.

Adolescent↗
PubMedDate not supplied

[Allelic variations of DPB1, DQA1, DQB1 and DRB1 and rheumatoid arthritis: further genetic and statistical considerations].

We used polymerase chain reaction amplification and hybridization with specific oligonucleotides to analyze the distribution of DPB1, DQA1, DQB1, and DRB1 allelic variants in 48 patients with rheumatoid arthritis (RA) and compared our results with those from 109 randomly chosen, healthy control subjects. Our work confirms a previously reported increase in DR4 specificity in RA: in particular, we found a statistically significant positive association of the DRB1*0401 and DRB1*0404 alleles with RA. When we compared the DR4 groups, however, none of the DRB1*04 alleles were increased in the RA group. Molecular analysis of the other DRB1 polymorphic variants disclosed the trend of a positive association of DRB1*0101 (DR1) in DR4 negative patients vs DR4 negative healthy control subjects, and an increase in DRw6 (DRB1*13,*14) in the DR4 and/or DR1 negative patient group. Moreover, analysis of the association between RA and a heptapeptide motif (positions 67-74) in the third hypervariable region confirmed that this epitope confers enhanced risk for the development of RA with respect to the allele DRB1*0404 (etiologic fraction = 0.53 vs 0.12). We also observed a statistically significant increase in DQA1*0301 and DQB1*0302 accompanied by a significant decrease in DQA1*0202, DQA1*0501 and DQB1*0201 in RA patients. Analysis of DPB1 alleles disclosed no significant differences between RA patients and healthy control subjects.

Adult↗
PubMedDate not supplied

Genomic PCR-SSCP analysis of the metastasis associated NM23-H1 (NME1) gene: a study on colorectal cancer.

To facilitate further mutational analysis of NM13-H1, a human metastasis suppressor gene, we have established its genomic organization. NM23-H1 is composed of five exons, spanning a genomic DNA fragment of 10 kb. Using oligonucleotide primers flanking each exon, PCR-SSCP analysis was performed on genomic DNAs of healthy individuals. A common polymorphism, a C to T transition, was detected 30 nucleotides upstream from the 5' splice site flanking exon 1. As NM23-H1 allele loss and altered expression have been reported in colorectal cancer, genomic DNAs of 20 colorectal tumors were analyzed for the presence of gene-specific mutations by PCR-SSCP: no abnormal sequences were detected within the coding and splice site regions of the NM23-H1 gene. This finding suggests that NM23-H1 mutations are rare events in human colorectal cancer.

Base Sequence↗